Plant

Secondary
Metabolism
 ine of
Plant Secondar
y Metabolism

chlorophyll + C 0 + light
2

cyclitols, polyols
photosynthesis^
C C -compounds
6 1

t
glycosides
carbohydrates

pentose phosphate
shikimi c acid
erythrose 4-phosphate

I glycolysis

chorismic acid phosphoenol pyruvate

£so-chorismic cyanogenic glycosides

acid * glucosinolates
peptides • proteins
non-protein amino acids

f tricarboxyli c acid
amino acids cycle acids

alkaloids
t e
r^ °
alkaloids
n i d

s-
ternenes ^ roev . .

^ ^ / / waxes

I acetylenes

rotenoids / hydrocarbons
flavonoids polyketides
• phenols
phenylpropanoid \ naphthoquinones
coumarins compounds \ anthraquinones
Iigni n lignans condense
d tannins
 Plant
Secondary
Metabolism
]David S. Seigler
Department of Plant Biology
University of Illinois, Urbana

SPRINGER SCIENCE+BUSINESS M E D I A , L L C
Library of Congress Cataloging-in-Publication Data

Seigler, David S.
Plant secondary metabolism / David S. Seigler.
p. em.
Includes bibliographical references.
ISBN 978-1-4613-7228-8 ISBN 978-1-4615-4913-0 (eBook)
DOI 10.1007/978-1-4615-4913-0
1. Plants--Metabolism. 2. Metabolism, Secondary. 3. Botanical
chemistry. r. Title.
QK887.538 1995
581.1'33--DC20 89-70112
CIP

Copyright © 1998 by Springer Science+ Business Media New York

Originally published by Kluwer Academic Publishers in 1998
Softcm-er reprint of the hardcover 1st edition 1998
All rights reserved. No part of this publication may be reproduced, stored in a retrieval
system or transmitted in any form, or by any means, mechanical, photocopying, recording,
or otherwise, without the prior written permission of the publisher.

Printed on acid-free paper.

This printing is a digital duplication of the original edition.

Table of Contents

Preface vii Chapter 21 Sesquiterpenes 367

Acknowledgments ix Chapter 22 Diterpenes and Sesterterpenes 398
Chapter 1 Introduction 1 Chapter 23 Triterpenes and Steroids 427
Chapter 2 Fatty Acids 16 Chapter 24 Saponins and Cardenolides 456
Chapter 3 Acetylenic Compounds 42 Chapter 25 Limonoids, Quassinoid., and Related
Chapter 4 Plant Waxes 51 Compound. 473
Chapter 5 Polyketides 56 Chapter 26 Tetraterpenes or Carotenoids 486
Chapter 6 Benzoquinones, Naphthquinones, and Chapter 27 Introduction to Alkaloids 506
Anthraquinones 76 Chapter 28 Simple Amines, Simple Aromatic and
Chapter 7 Shikimic Acid Pathway 94 Pyridine Alkaloids 513
Chapter 8 Phenylpropanoids 106 Chapter 29 Pyrrolidine, Tropane, Piperidine, and
Chapter 9 Coumarins 130 Polyketide Alkaloids 531
Chapter 10 2-Pyrones, Stilbenes, Chapter 30 Pyrrolizidine, Quinolizidine, and
Dihydrophenanthrenes, and Xanthones 139 Indolizidine Alkaloids 546
Chapter 11 F1avonoids 151 Chapter 31 Alkaloids Derived from Anthranilic
Chapter 12 Tanuins 193 Acid 568
Chapter 13 Nonprotein Amino Acids 215 Chapter 32 Isoquinoline and Benzylisoquinoline
Chapter 14 Peptide. 234 Alkaloids 578
Chapter 15 Carbohydrate. 247 Chapter 33 Alkaloids Derived from Both Tyrosine
Chapter 16 Cyanogenic Glycosides and and Phenylalanine 617
Cyanolipids 273 Chapter 34 Indole Alkaloids 628
Chapter 17 Glucosinolates 300 Chapter 35 Ergot and Other Indole Alkaloids 655
Chapter 18 Introduction to Terpenes 312 Chapter 36 Alkaloids of Terpenoid Origin 668
Chapter 19 Monoterpenes 324 Chapter 37 Miscellaneous Types of Alkaloids 692
Chapter 20 Iridoid Monoterpenes 353 Index 713
Preface

Life has evolved as a unified system; no organism exists similar role also has been suggested for fatty acids from
alone, but each is in intimate contact with other organisms cyanolipids. Nonprotein amino acids, cyanogenic glyco-
and its environment. Historically, it was easier for workers sides, and the non-fatty-acid portion of cyanolipids also are
in various disciplines to delimit artificially their respective incorporated into primary metabolites during germination.
areas of research, rather than attempt to understand the entire Secondary metabolites of these structural types are accumu-
system of living organisms. This was a pragmatic and neces- lated in large quantities in the seeds of several plant groups
sary way to develop an understanding for the various parts. where they probably fulfill an additional function as deter-
We are now at a point, however, where we need to investi- rents to general predation.
gate those things common to the parts and, specifically, those The second type of relationship involves interaction of
things that unify the parts. The fundamental aspects of many plants with other organisms and with their environment. Bio-
of these interactions are chemical in nature. Plants constitute logical interactions must be viewed in the light of evolution-
an essential part of all life systems; phytochemistry provides ary change and the coadaptation, or perhaps coevolution, of
a medium for linking several fields of study. organisms. A plant that possesses the ability to synthesize
It is partially through teaching a course in phytochemistry a compound that disturbs the physiological functions of a
that I have come to realize more fully the potential of phyto- herbivore may have a selective advantage over one that does
chemical studies to unite and integrate a vast amount of not. The chemical abilities of the plant may be utilized to
material from different disciplines. The diversity of students
establish interlocking relationships within the contexts of
that have taken this course and their research interests also
pollination, seed dispersal, establishment of mycorrhizal
reflects the utility and value of this type of information.
fungi, or protection of the plant by another organism, such
Among the fields represented are Biology (Ecology, Ento-
as ants. The "advantage" gained from such interactions may
mology, Plant Biology, Botany, Physiology, Microbiology,
be offset by metabolic cost or self-toxicity, but the process
and Zoology), Geology (Organic Geochemistry), Chemistry
of natural selection optimizes these interacting factors. Thus,
(Organic Chemistry, Biochemistry, and Analytical Chemis-
assuming the system remains stable for a sufficient time,
try), Agriculture (Horticulture, Animal Science, Dairy Sci-
eqnilibrium will be reached. Finally, it must be remembered
ence, Forestry, Agronomy, and Plant Pathology), Veterinary
Medicine, and Food Science. Possibly because of the diffi- that all organisms in a given system are evolving. Interacting
culties involved in integrating such a diversity of interests, organisms have the ability to adapt to changes in plant chem-
no adequate text presently exists for this course. istry and morphology, and may eventually ntilize plant hosts
In this text, I propose to survey our present knowledge that were formerly unacceptable. Host selection, food prefer-
of the phytochemistry and the role that secondary metabo- ences, taste, toxicity, wounding responses, allelopathy, and a
lites play in biological relationships. These relationships bas- vast array of other topics are the results of these evolutionary
ically fall into two categories: the first concerns the function processes.
and value of the compounds within the plants themselves. An understanding of the biosynthetic pathways leading
Many compounds are catabolized for energy or used to fulfill to secondary metabolites and a recognition of the chemical
other requirements that involve nitrogen. For example, fatty structural types present and their distribution among plant
acids from triglycerides have long been recognized as energy groups have proven useful for the study of biosystematic
sources for the embryo of a germinating seed. Recently, a problems and for achieving an understanding of plant phylo-

vii
viii Preface

geny. The predictive value of chemotaxonomic information sects, marine organisms, bacteria, fungi, and other organisms
has been recognized by natural products chemists. has been incorporated where warranted.
Plant secondary compounds play an important role in in- No attempt has been made to reference exhaustively all
dustry and medicine. Many industries are based on flavoring information cited in the text. Many references given are to
agents and perfumes, rubber, and naval stores. Several sec- review articles and in many instances are not those of the
ondary compounds are physiologically active, which results workers who carried out the original work. Although an at-
in their use as insecticides, medicinal agents, or biological tempt has been made to give the structures of most com-
probes or "tools." Plant compounds that are toxic to man pounds given in the text, some are not included. Further, the
and domestic livestock are widespread and may be responsi- structures of some representative compounds are included
ble not only for accidental but also for chronic poisoning by in figures, but, as they are not specifically cited in the text,
common foods such as cassava, sago, lima, and fava beans. they are not given numbers.
Subjects are grouped on the basis of major chemical struc- I apologize to authors inadvertently misquoted and will
tural types. Each will be discussed in terms of biosynthesis attempt to correct errors in any future editions. As it is diffi-
and distribution, diversity of known structures, taxonomic cult to locate many references dealing with topics discussed
application, biological function, toxicity, and medicinal and in this text, I solicit comments and any pertinent information
industrial uses. I do not present an encyclopedic coverage that may be useful for correction of errors or for the prepara-
of all available literature. Rather, the goal is a readable, inte- tion of further editions and wish to thank those who have
grated text that will be suitable for instruction at the ad- already provided me with manuscripts, reprints, unpublished
vanced undergraduate as well as the graduate level. In gen- material, suggestions, and assistance in preparing this pres-
eral, only phytochemical (Le., plant chemical) data has been ent manuscript. ffitimately, of course, I stand responsible
included, although information about the chemistry of in- for the conclusions and statements made in this text.
Acknowledgments

I wish to thank Anita Brinker, H. David Clarke, Ute Ecken- David C. Breeden, Anita Brinker, Stacie E. Canaan, H.
bach, Richard Lindroth, Karin Readel, and Kurt Potgieter David Clarke, Paul R. Connelly, Katherine Dowd, Thomas
for reading earlier versions of portions or the entire manu- Dudman, Nicole Duffee, Candace Easter, Robert J. Eilers,
script and making suggestions for improvement. The techni- John Gerlits, Peter Gottschalk, Mark Hediger, Ellen Hei-
cal assistance of Susan Gibbons, Elizabeth Bartlett, Cheryl ninger, Tricia HooChung, Han-Young Kang, Frederick J.
Frankfater, Nathaniel Ohler, and Ulrike Nolte also is greatly Lang, Wei Jane Liao, Hengchen Lin, Vincent Ling, Randall
appreciated. The comments of J. Balsevich, K. Brown, S. A. Lovell, John Martini, Susan McCarthy, John Ng, James
A. Brown, E. E. Conn, K. R. Downum, G. Cordell, D. Gian- Nitao, Paul Ode, Mark S. Pavlin, Raymond Pedersen, Char-
nasi, J. Gershenzon, J. B. Harbome, E. Leistner, S. Mole, lotte Read, Karin Readel, Nelson T. Rotto, Claire Rutledge,
A. Nahrstedt, R. G. Powell, P. Reichard, G. A. Rosenthal, Michael J. Sophia, Andrew L. Staley, Larry J. Thompson,
T. J. Simpson, K. Schreiber, P. Waterman, and anonymous Martin B. Wolk, and Craig M. vanZyl.
reviewers have been extremely useful in revising the manu- This work would not be possible if it were not for some
script. Greg Payne of Chapman & Hall has provided many of my former teachers and mentors including Allie Marie
helpful suggestions. Hobbs, Donald Hamm, Jordan J. Bloomfield, Leon Cieres-
In a number of cases, topics were reviewed in term p'apers zko, Eric Conn, Dale M. Smith and Tom Mabry.
submitted by students of the Plant Secondary Metabolites I also wish to express my appreciation to my wife Janice
or the Chemical Ecology course. Information from these for her patience during the preparation of this book. She bas
papers was extremely helpful and I especially wish to thank assisted in many ways to further this work.
the following stndents: John Andersen, Du-Jong Baek,

Ix
1
Introduction

Developments in the Study of Plant Secondary niques used to separate complex mixtures of plant com-
Compounds pounds are paper (PC), thin-layer (TLC), column (CC), liq-
What Is a Secondary Metabolite? uid (HPLC and MPLC) and several types of countercurrent
The Origin of Plant Secondary Pathways and Compounds chromatography. Newer techniques for the identification and
The "Waste Product" Hypothesis characterization of compounds include lH and l3C-nuciear
Internal Functions of Plant Secondary Compounds magnetic resonance (NMR) (Derome, 1989; Sadler, 1988;
The "Overflow" or Excess Primary Metabolism Simpson, 1986), mass spectrometty (MS), ultraviolet (UV),
Hypothesis and infrared (IR) spectrophotometty, and x-ray crystallogra-
The "Increase in Fitness" Hypothesis phy. Development of computerized techniques has permitted
The "Cost" of Secondary Metabolite Synthesis utilization of Fourier transform methods for acquisition of
Variation of Secondary Compounds spectra and greatly increased the capabilities of these meth-
Site of Synthesis and Accumulation of Secondary ods. Today, it is possible to isolate, purify, and characterize
Compounds amounts as small as several nanograms of secondary com-
Interactive Roles of Plant Secondary Compounds pounds from complex mixtures.
Patterns of Simultaneous Evolution of Plants and Other At the same time that individual plants may be studied,
Organisms various plant organs and tissues can be examined. Immuno-
Evolution of Chemical Pathways logical methods and radioactive labeling techniques can be
The Value of Plant Secondary Chemistty for used to detect compounds at the cellular level. ELISA (en-
Understanding Evolutionary Relationships and zyme-linked immunosorbent assay) serves to localize the
Phylogeny of Plants site of accumulation of secondary metabolites by binding
Why Study Secondary Compounds? specific antibodies to a solid surface (Wilkinson et al., 1992).
References The development of plant cell culture techniques has fa-
cilitated major breakthroughs in the study of the biosynthesis
of plant secondary compounds (Barz and Ellis, 1981; Ellis,
DEVELOPMENTS 1l'I11fE SnIDV OF PLAl'IT 1988; Flores, 1987; Hutchinson, 1986; Wink, 1987). None-
SECOl'lDAKV COMPOUNDS theless, many cultures either do not produce secondary me-
tabolites or produce an array that differs from that found in
Formerly, relatively large amounts of plant materials were the plant. Thus, there are still many questions to be answered.
needed to isolate, purify, and characterize most secondary In instances where a culture fails to produce a particular
compounds. Only those compounds that crystallized readily, chemical, it is often assumed that the enzymes of the path-
were biologically active, colored, or were present in excep- way are not formed, but, sometimes, only one enzyme is
tionally large amounts tended to be studied. Recent advances repressed or there is no product formation because of poor
in chemical microtechniques permit studies of the chemical precursor supply, lack of storage capacity, or product degra-
composition of individuals of plants and other organisms. It dation (Wink, 1987). The addition of exogenous precursors
has become feasible to examine patterns of variation within to the culture medium may increase product yield. For exam-
and among populations of naturally occurring organisms that ple, many plants appear to contain an L-alanine:aldehyde
previously were not amenable to study. Among the tech- aminotransferase that results in amine formation; the supply
 Introduction

of substrate appears to be the controlling step in this reaction. In this case, tissue cultures contained 116 times more of the
Cultures of lupines rarely accumulate the levels of quinolizi- enzyme than the roots of the plant, clearly demonstrating
dine alkaloids found in the intact plants; in this example, an advantage to plant tissue culture methods. The partially
alkaloids are formed, but are rapidly degraded (Barz and purified enzyme was immobilized, which increased its tem-
Koster, 1981; Wink, 1987). Isolation and use of protoplasts perature stability and permitted the synthesis of strictosidine
(cells of bacteria, fungi, or plants from which the cell walls (4) (the first of the intermediates mentioned above) (Hutch-
have been removed) for biosynthetic studies sometimes inson, 1986). Two glucosidases were isolated which had
modifies restrictions on the rate or extent of the uptake of strict substrate specificity for strictosidine. Reduction of
precursors (Hutchinson, 1986). 4,21-dihydrogeissoschizine (7) gives geissoschizine (8),
In order to understand many of the biosynthetic pathways which had previously been thought to be an precursor to
described in this text fully, it will be necessary to isolate and ajmalicine. However, the study of the metabolism of this
purify many of the enzymes of the pathways to homogeneity. compound in vitro revealed that it is a shunt product of the
This has not been completed for any major pathway, al- pathway (Hutchinson, 1986). Study of the enzymes has clari-
though significant steps have been taken for a number of fied the early stages of events in this complex pathway (see
types of secondary metabolites (Hutchinson, 1986). Chapter 34).
Most of the recent progress in alkaloid enzymology has As another example of the value of enzymological stud-
been made possible by purified enzymes that catalyze the ies, the major enzymes of the benzylisoquinoline alkaloid
respective steps of alkaloid biosynthesis. This is illustrated pathway have also been isolated from plant tissue culture
by the application of these techniques to alkaloid biosyn- and studied by Zenk and co-workers (see Chapter 34) (Fig.
thesis. For example, formation of ajmalicine (1) from its 1.2). An enzyme, (S)-noriaudanosoline synthase, which cat-
precursors, tryptamine (2) and secologanin (3), involves four alyzes the conversion of dopamine and (3,4-dihydroxyphen-
key intermediates (Fig. 1.1). yl)acetaldehyde (9) into (S)-noriaudanosoline (10) has been
The first enzyme of the pathway, strictosidine synthase, purified about 40-fold from cell suspension cultures of
has been purified from Catharanthus roseus about 50-fold. Eschscholtzia tenuifolia (see also chapter 32 for up-dated in-

~H
~HN) ~
5SH~ 2 H"'"
.:'::::.""O-glllCOSYI
__
HN

+ ~ 0 strictosidine (4)
CH,02C
tryptamine (2) seco-Ioganin (3)

4,21-dihydro-
geissoschizine
(7)

HO

HO
geissoschizine (8) ajmalacine (1) cathenamine

Fig. 1.1. Biosynthesis of ajmalicine (Hutchinson, 1986; modified and used with pennission of the copyright owner, the Royal Society of Chemistry).
 Introduction

H0X)! H0)CJ(0 HO~

HO~I :H
I:
HO
CHO
HO
I: CO2- HO

HO
,;7

~
I

(3,4-dihydroxyphenyl)acetaldehyde (9) (3,4-dihydroxyphenyl)pyrovale (11) (S)-norlaudanosoline (10)

Fig. 1.2. Some intermediates in the biosynthesis of benzylisoquinoline alkaloids.

fonnation). The properties of this enzyme indicate that (3,4- and some proteins are identical in animals, bacteria, fungi,
dihydroxyphenyl)acetaldehyde is the actual precursor for plants, and other organisms. Nonetheless, differences in the
this series of alkaloids rather than (3,4-dihydroxyphenyl)py- pathways leading to these primary metabolites and the actual
ruvate (11), as previously suggested (Hutchinson, 1986) (see compounds involved in some fundamental processes differ
Chapter 32). in some instances (Haslam, 1986).
Because it is often difficult to obtain the quantity of pro- Secondary metabolites, produced by pathways derived
teins, especially enzymes, needed to study individual steps from primary metabolic routes, are numerous and wide-
of biosynthetic pathways, or those involved in transport or spread, especially in higher plants. More than 20,000 were
accumulation of plant secondary metabolites, DNA recombi- known in 1985 (Hartruann, 1985), and at least 1000 addi-
nant methods hold great promise for future studies. In in- tional compounds. are described each year. In practice, the
stances where the gene can be cloned and expressed in bacte- difference between the primary and secondary metabolites
rial cells, it frequently becomes possible to obtain relatively is fuzzy. Plant honnones such as gibberellic acid, indoleace-
large amounts of pure, biologically active protein (McPher- tic acid (auxin), ethylene, kinetin, and abscisic acid, as well
son and Parish, 1987). In order for this method to be more as compounds involved in plant cell wall structure such as
widely applicable, however, additional knowledge about the cinnamic acid and its polymeric derivative, lignin, are inter-
regulation of developmental genes and the control of gene mediate between primary and secondary metabolism (Birch,
expression is needed. 1973). In some instances, compounds nonnally considered
primary metabolites may accumulate in large amounts and
behave in a manner usually associated with secondary me-
WHAT IS A SECONDARY l'IETABOLlm? tabolites. Entities such as shikimic acid and squalene, which
initially were considered secondary metabolites, were subse-
The compounds in living organisms may be divided into two quently shown to be important intennediates in the fonnation
major groups: primary and secondary metabolites. Primary of primary metabolites (phenylalanine, tyrosine and trypto-
metabolites are those produced by and involved in primary phan, and steroids, respectively).
metabolic processes such as respiration and photosynthesis, The amount of any plant secondary compound found in
whereas others, including many pathways clearly derived an organism is the result of an equilibrium among synthesis,
from primary metabolic pathways, are considered "second- storage, and degradation. Regulation of secondary metabo-
ary." In practice, there is overlap between the two tenns lism is complex. The onset of secondary metabolism is
and, other than providing a pragmatic way to identify a large linked to the developmental stage of the organism and is
body of compounds, pathways and literature, too much sig- often closely linked to morphological and cytological
nificance should not be attached to the definitions. changes (Haslam, 1986). In general, the fonnation of prod-
Primary metabolites, as mentioned above, include the ucts in secondary metabolism appears to be enzyme limited,
components of processes such as glycolysis, the Krebs or but the level of substrates present influences the production
citric acid cycle, and, in photosynthetic organisms, photo- of other secondary metabolites, especially in artificial culture
synthesis and associated pathways. Primary metabolites, (Bu'Lock, 1980). The degree to which anyone prevails often
which are virtually identical in most organisms, include depends on the developmental stage of the plant and a variety
Ubiquitous small molecules such as sugars, amino acids, tri- of other factors. In the fungal genus Claviceps, biosynthesis
carboxylic acids, or Krebs cycle intennediates, the universal of the alkaloids begins shortly after fonnation of two key
building blocks and energy sources-as well as proteins, enzymes, DMAT synthetase (tryptophan dimethylallytransf-
nucleic acids, and polysaccharides, that differ in structural erase) and chanoclavine cyclase, at the beginning of the sta-
detail from one organism to another, but which appear to tionary phase of growth. The rate of synthesis is detennined
have universal functions as enzymes, structural, and heredi- by the first of these enzymes; L-tryptophan, a substrate of
tary materials. Many sugars (such as glucose), protein amino this enzyme, must be added early in the growth phase to
acids, low-molecular-weight organic acids, many fatty acids, serve as a precursor (Haslam, 1986).
 Introduction

In another example of the complex sequences of factors in cell cultures of Lupinus polyphyllus (a legume), Conium
controlling secondary metabolite biosynthesis, the enzyme maculatum (Apiaceae), Daucus carota (Apiaceae), Atropa
phenylalanine ammonia-lyase (PAL) in higher plants is often belladonna (Solanaceae), Chenopodium rubrum (Cheno-
associated with the rate of accumulation of flavonoids and podiaceae), Spinacia oleracea (Chenopodiaceae), and Sym-
has been considered by many to be the principal regulating phytum officinale (Boraginaceae) was induced by adding
factor in the synthesis of phenylpropanoids. However, in foreigu alkaloids and a number of other "disturbances"
parsley cell cultures, two groups of enzymes can be distin- (Wink and Witte, 1983) (see Chapter 30). Of these, only the
guished: enzymes of general phenylpropanoid metabolism legume would normally be expected to contain quinolizidine
(including PAL) and those specific to pathways leading to alkaloids. These and other studies suggest that we do not
flavones and flavonol glycosides. Both are induced by irra- know the full biochemical potential of any plant.
diation with UV light, but the first group reaches maximal
activity several hours before the second and then declines
more rapidly than the latter group. However, the level of THE ORIOIN OF SECOl'lDAKY PAntWAYS
free phenylalanine normally is very low. Hence, in this in- Al'ID CO!IfPOUl'lDS
stance, the amount of phenylalanine is more likely to be
limiting than the level of enzymes (Haslam, 1986). The ultimate origin of secondary compounds in all organ-
Several new approaches to the study of the variation of isms is based on accumulation of changes (or mutations) of
plant secondary compounds have been undertaken by Som- genetic materials originally associated with primary meta-
erville and Browse (1991). More than 10,000 individuals of bolic pathways. Although the process probably is not en-
Arabidopsis thaliana (Brassicaceae) have been screened for tirely random, changes in DNA-RNA responsible for the
mutants in the pathways of lipid metabolism, and mutants formation of particular types of secondary compound are
of at least 12 steps have been identified. None of these mu- more or less accidental. Most of the products of altered path-
tants can be distinguished from the wild type by visual in- ways are deleterious to the producing organism and, if ex-
spection under normal growth conditions. Most mutations pressed, the organisms (and the genes).that produce them
cause a loss or reduction in the amount of an unsaturated acid are quickly reduced in frequency or eliminated by natural
and affect the accumulation of a less saturated precursor. By selection. Changes in secondary metabolic pathways and the
this approach, it has been confirmed that, except for <l.9- resultant compounds generally are tolerated more readily
unsaturated C 18 fatty acids, double bonds are introduced into than changes in primary pathways and compounds. The ef-
fatty acids that are esterified to lipids rather than the acyl- fects of these changes may be witnessed as "modified en-
CoA or acyl-ACP derivatives of the acids. At least eight of zyme specificity," availability of substrates, production of
these genes now are known to control specific desaturases a modified structure, or a variety of other factors (Haslam,
in this species. The mutations have been genetically mapped 1986).
and this information forms the basis to isolate the corre- Most prokaryotic organisms have relatively simple path-
sponding genes (Somerville and Browse, 1991). Analysis of ways for respiration and synthesis of food materials. These
these mutations has provided insight into the regulation of organisms have a single copy of most genes, and any changes
membrane lipid desaturation. Other mutants had defects in that occur in the genetic information are expressed. Most of
genes involved in fatty acid elongation. The availability of these changes are probably lethal, but some have selective
specific alterations in membrane fatty acid composition pro- advantage and are maintained. The number of compounds
vides another method to examine the physiological conse- other than primary metabolites (those involved in respiration
quences in variation of lipid unsaturation. Certain types of and/or synthesis) produced by prokaryotic organisms is lim-
unsaturation are directly linked to cold tolerance. These stud- ited. Much of our knowledge of the origin of secondary
ies promise to shed light, not only on the biosynthesis but metabolites in prokaryotic organisms is based on bacteria
also control of synthesis and accumulation, as well as the and blue-green algae in culture, and few studies have been
influence of plant secondary compounds on the physiology made of metabolism in their natural habitats. There is evi-
and ecology of the plant (Somerville and Browse, 1991). dence, however, for antagonisms among soil organisms and
Our knowledge of secondary compounds is limited the antibiotics produced by some species appear to bind spe-
mostly to those that are accumulated, but, in some instances, cifically to receptors in other microbes (Williams et al.,
compounds normally not thought to be present have been 1989). Although microorganisms have been used exten-
found at very low levels. For example, azetidine-2-carbox- sively for the study of the biosynthesis and catabolism of
ylic acid, normally found in the Liliaceae and in some leg- secondary metabolites, the extent to which identical se-
umes, has been isolated from amino acid fractions of sugar quences occur in plants varies.
beets (Beta vulgaris, Chenopodiaceae) isolated as a by-prod- In contrast, the amount of genetic material in plants and
uct of sugar manufacture. A number of other unusual amino other eukaryotic organisms is greater than in prokaryotic
acids also were isolated from the mixture. About 500 kg organisms and is organized into chromosomes. Because
of these fractions is obtained from 106 tons of sugar beets there are two copies of each chromosome for diploid stages
(Fowden, 1972). The formation of quinolizidine alkaloids of organisms, the accumulation of recessive characters is
 Introduction

possible and many changes are not expressed immediately. compounds are leached from the above ground parts of
In addition. the existence of multiple copies of genetic mate- plants by precipitation. Others are exuded from roots of
rial produced by polyploidy or increase in the amount of plants (Tukey, 1970). Well-organized enzyme complexes in-
chromosomal material promotes the accumulation of genetic volving channeling of precursors are involved in the biosyn-
variation at particular alleles, because one of a group of thesis of many types of secondary compounds (Hrazdina and
multiple copies of a gene may change without a significant Jensen, 1992). The synthesis of these compounds requires
effect on other copies or on primary metabolic pathways. many kilobases of DNA. To suggest that such highly ordered
Many plant enzymes occur as mixtures of closely related processes arose from a series of neutral mutations that have
but distinct proteins (isozymes) that presumably originated accumulated contradicts the accepted basis of evolutionary
from genetic material altered in the manner described above. changes (Williams et aI., 1989). Further, it is now known
If a particular metabolite can be tolerated, or its synthesis that many secondary metabolites exist in a state of dynamic
is favored, the necessary mechanisms that pennit "pools" equilibrium Of "turnover" in which compounds are con-
of compounds to fonn also must exist, if the compound is stantly being synthesized and broken down by anabolic and
to be accumulated. catabolic enzymes (Barz and Koster, 1981). It has been pos-
Selection against deleterious modifications may lead to tulated that this process may involve further biosynthetic
an apparent increase in those changes with "positive" ad- transfonnation, polymerization, or catabolism to yield pri-
vantage for a particular organisms under a particular set of mary metabolic consituents (Haslam, 1986). Feeding of ra-
circumstances. Much confusion has been brought about by dioactively labeled precursors indicates that the half-lives of
"teleological" statements concerning the origin of second- many of these secondary metabolites are between 6 and 24
ary metabolites that have been suggested to play roles in h. Other compounds, particularly those of strnctural materi-
plants and their interactions with other organisms. The origin als and from storage tissues/organs such as seeds tum over
of secondary metabolites and the capability of an organism at much lower rates. The pathways by which "turnover"
occurs are known in few plants. Some interesting examples
to accumulate them is detennined largely by stochastic
are known from the work of Croteau and Gershenzon on
events influenced by preexisting starting materials (for in-
monoterpenes in mints (see Chapter 19) and from the work
stance, the initial pathways and precursors available for
of Lieberei and Selmar (Selmar, 1989; Selmar et aI., 1988)
modification, whereas accumulation of secondary com-
on cyanogenic glycosides in Hevea brasiliensis, and Gander
pounds with functions in living organisms is directed largely
(1962) on the cyanogenic glycoside dhurrin (see Chapter
by natural selection. The parameters involved in the process 16).
of natural selection which lead to this accumulation undoubt- The origin of certain types of compounds such as glyco-
edly are many and poorly understood. Attempts to account sides and methylated derivatives has been ascribed to cata-
for the mixtures of secondary metabolites found in many bolic changes and detoxication mechanisms (Hartmann,
organisms, without considering the complexity of the bio- 1985; Luckner, 1972). Indeed, there are marked differences
logical and physical environment in which these organisms in the biological activities of aglycones and the correspond-
arose and exist, in some simple "satisfactory" or "explicit" ing glycosides and methylated derivatives (e.g., see Chapter
manner seem naive. Several hypotheses about the origin and 11). However, detoxification seems inadequate to explain
accumulation of secondary compounds have been proposed. the variety of secondary metabolites found in plants (Wil-
liams et aI., 1989).
The line between catabolic and anabolic activity is as
TUB "WASTE PRODUCT" HYPOTHESIS
nebulous as that between primary and secondary metabo-
lism. The mycotoxins aflatoxin and patulin give every ap-
The relatively large number and amount of secondary metab-
pearance of being catabolic in nature (Haslam, 1986). Many
olites observed in nature and the notion that these com-
secondary metabolites possess elements of structure that di-
pounds arose from' 'errors" in primary metabolism, coupled
rectly parallel those observed in the catabolism of primary
with the apparent absence of excretory mechanisms in
metabolites such as a-amino acids (Haslam, 1986).
plants, led to the idea that secondary compounds arise and
If secondary metabolites are "waste products" and the
accumulate as "waste products" (Luckner, 1972, 1990;
"expense" involved in making, accumulating, and maintain-
Luckner et aI., 1976; Mothes, 1966c, 1976; Paech, 1950).
ing these compounds is significant (see below), plants that
Mothes originally felt that secondary plant substances are make them would be at a considerable selective disadvantage
not essential to the existence of plants. In support of this with regard to plants that do not. Less efficient organisms are
idea, he observed that they can be missing from individual often eliminated in the process of natural selection.
members of a related group (of plants) and that they are
not an "inherent taxonomic characteristic" (Mothes, 1966a,
INTERNAL FUNCTIONS OF PLANT SECONDARY
I 966b, 1966c).
COMPOUNDS
Several types of data suggest that the "waste product
hypothesis" is not a likely explanation for the origin of sec- Others have suggested that secondary metabolites once
ondary compounds. For example, the idea that plants cannot played a metabolic role in the organisms prOducing them or
excrete secondary compounds has been disproved. Many that they were primary metabolites in the past (Williams
 Introduction

et al., 1989). Many compounds do have roles intermediate phates (e.g., ATP). In this way, the synthesis of secondary
between primary and secondary metabolism. For instance, compounds is interpreted as an "overproduction" of metab-
some plant secondary compounds (e.g., the gibberellins, ab- olites (Haslam, 1986).
scisic acid, ethylene, indole acetic acid, kinetin, and a num- The addition of exogenous substrates of primary metabo-
ber of other plant hormones) are associated with plant growth lites to organisms in culture (or blocking of particular steps
and development. of biosynthetic pathways) can result in "overproduction"
Other secondary metabolites serve as reserves of energy of either primary or secondary compounds. Gross primary
and as precursors of important organic compormds or production by plants exceeds net primary production. Addi-
sources of nitrogen and may be recycled within the plant tionally as much as 38% of carbon fixed by photosynthesis
(Rosenthal, 1982). As nitrogen is often a limiting factor for may be lost through photorespiration. The process of cya-
the growth of plants, it is not surprising that plants use it nide-resistant respiration represents an obvious nonaccumu-
"economically"; for this reason, many nitrogenous com- lative mechanism by which plants can divide any overflow
pounds appear to be involved in multiple functions in plants, into carbon dioxide (Waterman and Mole, 1989).
which include storage of nitrogen, plant defense, and regula- Although "overflow" may occur in some instances,
tory roles (Jones, 1979; Seigler and Price, 1976; Swain, many plants that would not experience overflow possess a
1976). For example, nonprotein amino acids accumulated in variety of secondary metabolites and this theory does not
legume seeds disappear upon germination of the seeds, and explain adequately the vast number of secondary metabolites
the nitrogen appears to be reused (Rosenthal, 1982). Cyanol- known to occur. Neither does it address the functions many
ipids, which make up about 15% of the total biomass of the of these compounds are now known to possess (Williams et
seeds of Ungnadia speciosa, are mobilized and disappear in al., 1989).
about 2 days during germination (Selmar et al., 1990); the
nitrogen again appears to be reutilized, although in this in-
stance, it may be transferred to cyanogenic glycosides. Simi- THE "II'ICREASE II'I FITl'lESS" HYPOTHESIS
lar changes occur in instances when alkaloids are accumu-
lated in seeds. They undoubtedly play a defensive role but
Many, especially a number of ecologists, have come to ac-
are mobilized and recycled during germination (Waller and
cept the hypothesis that secondary metabolites increase the
Nowacki, 1978; Wink, 1987).
"fitness" of individuals that possess them and that those
Modified secondary metabolites have different solubility
individuals have been favored by the process of natural se-
and other physical properties than the parental compounds
lection (Harbone, 1986; Rosenthal and Jansen, 1979; Swain,
and may be involved in transport and accumulation mecha-
1977). Not all of the large number of compounds in any
nisms in plants (Wink, 1987). Nonetheless, there is little
particular plant will have obvious benefits. These benefits
evidence to support the notion that most or all secondary
simply may not be recognized at present, although it has
metabolites once had primary roles (Williams et al., 1989).
been suggested that "mechanisms must have evolved that
ensure generation and retention of chemical diversity." Re-
tention of inactive compounds is one way of increasing
THE "OVERFLOW" OR EXCESS PRIJIIARY chemical diversity and the probability of producing active
I'IETABOLISJllIIYFOTHESIS compormds in the face of consumer adaptation" (Jones and
Firn, 1991). The "increase in fitness" hypothesis is the ouly
In instances of unbalanced growth, secondary metabolites one that accounts for the fact that many natural products
have been envisioned by some as shunt metabolites produced trigger very specific physiological responses in other organ-
in order to reduce abnormal concentrations of normal cellu- isms and, in many cases, bind to receptors with a remarkable
lar constituents~ The synthesis of enzymes designed to carry complementarity (Williams et al., 1989). These workers sug-
out secondary metabolism permits primary metabolic en- gested that "natural products may ... aid an organism's sur-
zymes to continue to function until such time as circum- vival in the absence of an immune system."
stances are propitious for renewed metabolic activity and However, one of the key considerations in the hypothesis
growth. For example, this could be linked to the depletion is that there can be evolutionary selection against those indi-
of nutrients such as phosphorus or nitrogen (Bu'Lock, 1980; viduals in popUlations of organisms that lack certain second-
Haslam, 1986). ary metabolites and, in that manner, those organisms that
Phosphoenolpyruvate, pyruvate, acetyl coenzyme A, and have secondary metabolites with beneficial aspects are fa-
a number of amino acids involved in the biosynthesis of vored. Accordingly, those compounds which impart benefit
cyanogenic glycosides, alkaloids, glucosinolates, and antibi- to plants have become prominent. Metabolites with "neu-
otics are linked to the final portion of the glycolytic sequence tral" functions may be selected against because of the' 'met-
and immediately prior to entry into the Krebs cycle. A shift abolic cost" associated with their production, but if the cost
from normal sustained growth to that in which there is a of production is sufficiently low, many relatively inactive
decreased uptake of acetyl coenzyme A in the Krebs cycle compounds could be tolerated for a period of time (Jones
may reflect a diminished requirement for nucleotide triphos- and Fim, 1991).
 Introduction

THE "COST" OF SECOl'lDAKY METABOLITE In a genetically based series of experiments, seed produc-
SYNfHESIS tion of the tall morning glory, Ipomoea purpurea, was used
as a measure of fitness. No observable reduction in fitness
was observed in the presence of the herbivore sweet potato
The concept of "cost" in terms of metabolic energy, avail-
flea beetle, Chaetocnema confinis (Simms and Rausher,
ability of precursors and limiting factors, as well as ecologi-
1987). On the other hand, the magnitude of cost of produc-
cal considerations and evolutionary fitness has engendered
tion of furocoumarins in the wild parsnip, Pastinaca sativa,
a large body of literature. Many factors that must be consid-
in terms of umbel production, is significant (Berenbaum et
ered in making cost estimates have been discussed and evalu-
al., 1986).
ated (Chew and Rodman, 1979). Estimates of the energetic
Metabolic inefficiency, as implied by the waste product
outlay to produce and maintain plant secondary compounds
hypothesis, seems unlikely if a significant cost is associated
vary widely, from perhaps 8-10% of a plant's total meta-
with the production of secondary metabolites. Most plants
bolic efforts (Robinson, 1974, 1980) to nothing (Simms and
produce and accumulate relatively large amounts of mixtures
Rausher, 1987). Carbon usually is not limiting in plant bio-
of secondary compounds, despite any metabolic cost of syn-
synthesis, and formation of carbon-based compounds may
thesis and accumulation. The cost of maintenance of cata-
play a beneficial role to plants under high light conditions,
bolic enzymes must also be considered. Further, plants can
by helping to dissipate excess energy associated with photo-
excrete most of the compounds produced andlor limit their
synthesis. This may be a major function of photorespiration
production by other means. All these factors suggest that
(Selmar, 1992). In general, synthesis of secondary metabo-
plants with secondary compounds are at a selective advan-
lites containing limiting elements, such as nitrogen, incur
tage.
higher costs. Most approaches to estimate "cost" or "ex-
In summary, the cost of production of secondary metabo-
pense" attempt to assess the physiological cost to the plant
lites is difficult to estimate. The physiological cost of pro-
but fail to evaluate the effects of resistance on fitness or
duction varies for different secondary compounds; costs for
fecundity (Bazzaz et al., 1987), and, hence, the constraints
compounds containing limiting elements such as nitrogen
on the evolution of defense.
may be especially sensitive to enviromnental fluctuations in
Several theories related to the distribution of plant sec-
these nutrients. Physiological cost and evolutionary success
ondary compounds for defensive purposes have been pro-
are not linked in linear fashion; for evolutionary considera-
posed (Feeny, 1990). Many "qualitative" or "mobile" de-
tions, some estimate of cost linked to fitness is essential.
fensive compounds contain nitrogen, whereas most of the
Many aspects of the origin of secondary metabolites and
"quantitative" or "immobile" compounds do not (Coley et
their accumulation remain obscure. No one theory can ac-
al., 1985; Feeny, 1990; McKey, 1974, 1979). McKey (1974)
count for all the problems inherent to resolving this enigma.
proposed that the amount of defensive compounds in particu-
When all the possibilities are examined, some examples can
lar tissues should reflect the value of those tissues to the
be found that can be explained by each of the hypotheses.
plant. In practice, however, plants are subject to a variety
Further, some compounds may have arisen by different path-
of physiological constraints that vary with the availability
ways in an assortment of organisms in response to a variety
of resources and alter the relative costs of different types of
of factors.
defense (Feeny, 1990). Plants which are adapted to high light
intensities in nutrient-poor habitats, for example, are likely
to have a relative surplus of carbon and, hence, to deploy
carbon-based defenses (Bryant et al., 1983). Unless con- V ARIATIOl'l OF SECOl'lDAKY COMPOUl'lDS
strained by other factors, plants should maximize the bene-
fits conferred by secondary metabolites relative to the cost Genotypic variation gnarantees diversity in both murpholog-
of reclaiming or replacing them (McKey, 1979). In studies ical and chemical characters. In addition, more complex pat-
of light gap species in a lowland rain forest in Panama, Coley terns of variation or polymorphism occur in many plants;
concluded that "pioneers" or "light gap specialists" were individuals from natural populations of plants often differ in
grazed more than "persistents" that could tolerate only low the amounts and types of compound present. Roots, leaves,
light levels and grew in the shade. The former produced stems, seeds, fruit walls, flowers, and buds frequently differ
relatively fewer defensive compounds. She proposed that in chemical composition. Further, each of these parts may
plants adapted to favorable areas (such as light gaps) can vary at different stages of development and at various times
grow fast enough to escape the relatively high levels of dam- of the year (Luckner, 1990; Waterman and Mole, 1989;
age resulting from low levels of defense. Plants that typically Wink, 1987). Daily (diurnal) variation of many compounds
occupy unfavorable habitats are limited in their productivity, also occurs (Fliick, 1963; Luckner, 1990). The reasons that
cannot grow rapidly, and must rely on greater resistance to plants fail to produce or accumulate compounds are numer-
herbivores. She further stated that the availability of re- ous, but include regulatory processes, blockage of pathways
sources in the environment is the major determinant of both by mutations, and the absence of precursors at the site of
the amount and type of plant defense (Coley et al., 1985; synthesis.
Feeny, 1990). Plants are often quite "plastic." They vary in complex
 Introduction

ways, not only because of genetic differences as discussed great geographic distance. In contrast, examination of popu-
above but also in response to environmental factors. Both lations of other plant species often reveals a gradual variation
qualitative and quantitative variation of secondary metabo- in plant chemistry over a distance. These racial or elinal
lites is known to occur in response to various types of stress shifts in chemistry usually parallel changes in other charac-
(Waterman and Mole, 1989). Among these are biological teristics of the plant involved.
stresses such as attack by fungi, bacteria, nematodes, insects, When plants reproduce elonally or vegetatively, the
or by grazing by mammals, and abiotic stress such as ex- amount of variation in plant chemistry may be quite small.
tremes of temperature and moisture, shading, presence of
heavy metals, and injury (Fliick, 1963; Gershenzon, 1984).
Formation of many secondary compounds is influenced SITE OF SYl'ITIIESIS Al'ID ACClJlllULATlOI'l
by environmental stress in both general and specific manners OF SECOl'lDARY COJIIPOV1'lDS
(Waterman and Mole, 1989). Plant secondary metabolites
respond to changes or differences in soil nutrients, but not Although all cells of a plant harbor the genes for synthesis
all plants respond in similar ways. In general, production of of a particular secondary metabolite, ouly a limited number
non-nitrogenous shikimic-acid-derived metabolites is in- express them and produce the enzymes needed to make the
creased-when plants are grown under nutrient-deficient con- compound. This temporal and spatial expression of genes is
ditions (Waterman and Mole, 1989). Compounds from mev- a typical feature of eukaryotes (Kutchan, 1983; Wierman,
alonic acid pathways do not show consistent relationships. 1981; Wink, 1987). For example, highly colored flavonoids
An increase in light intensity tends to produce increases in and anthocyanins are typically formed in flowers or fruits.
phenolic compounds and terpenoids. Water stress has been Further, this compartmentation is observed within single
reported to lead to increases in cyanogenic glycosides, glu- cells (Luckner, 1990; Matile, 1978; Wink, 1987). However,
cosinolates, terpenoids, alkaloids, and condensed tannins even if a particular secondary metabolite is found in an organ
(Waterman and Mole, 1989). Grazing is known to increase or tissue, this does not necessarily mean that the compound
the amounts of many secondary compounds in plants; the was formed there. Long-distance transport must also be con-
changes observed are similar to general wound responses, sidered (Sehnar, 1989; Sehnar et al., 1988). This is particu-
although insect grazing caused more widespread and greater larly true in epidermal cells. Many secondary metabolites
response in birch than artificial damage (Waterman and are synthesized elsewhere and transported to these cells
Mole, 1989). which form an early line of defense for the plant (Luckner,
Other responses, such as formation of phytoalexins, are 1990; Wink, 1987).
more specific (Chappell and Hahlbrock, 1984; Luckner, Lupine alkaloids are formed in the green, aerial parts of
1980). It has been shown that production of secondary com- Lupinus polyphyllus that incorporate labeled cadaverine into
pounds in cell cultures also can be enhanced by environmen- the lupanine skeleton, consistent with the fact that the en-
tal stress. Treatment of cell cultures with fungal, bacterial, zymes of alkaloid biosynthesis, in this case, are located in
or plant cell wall materials often results in the formation of a the chloroplast stroma (Hartmann, 1985). Roots of the intact
number of flavonoids, stilbenes, terpenoids, anthraquinones, plants or in vitro cultured roots do not. A similar sitoation
rutacridone alkaloids, sanguinarine, and gossypol (Eilert, obtains for coniine in Conium maculalum, where the en-
1987; Heinstein 1985). In some cases, production of these zymes occur in both the chloroplasts and mitochondria.
metabolites is transient. Clearly, we need more information However, alkaloids are rarely formed in plastids (Hartmann,
on the basic principles of the defensive response, such as 1985), but are usually formed in the cytoplasm. Chloroplasts
structures and perception of defensive signals, transduction, are not only the site of photosynthesis, but also of lipid,
and translation of these signals into a response (Luckner, amino acid, and terpenoid biosynthesis (Schultz et al., 1985;
1980; Wink, 1987). Wink,1987).
Especially complex variation arises in situations in which All these factors are important for the development of
hybridization or the combination of two genetically different plant cell cultures. For example, cultures of Lupinus poly-
groups exists. In some plants, "additive" chemistry, appar- phyllus only synthesize alkaloids when kept in the light and
ently produced by the presence of two sets of enzyme sys- alkaloid content is correlated with chlorophyll content. This
tems, has been observed. In hybrids of Baptisia leucophaea may be explained by a better supply of lysine (formed in
and B. spherocarpa, the appearance of a hybrid compound the chloroplasts), the fact that the enzymes of alkaloid bio-
(a 7-0-glucoside-3-0-rutinoside) can be explained as such synthesis have a pH optimum of about pH 8, which is created
an additive combination (one parental type produced the 7- in the chloroplast stroma only in the light, and that the en-
O-glucoside, whereas the other produced the 3-0-gluco- zymes are subject to activation by reduced thioredoxin which
side). Other compounds, predicted on the basis of the chem- is generated only in the light (Wink, 1987). The alkaloids
istry of the putative parents, are mysteriously absent (Alston of Peganum harmala, which are formed in the roots of the
et al., 1965; Markham and Mabry, 1968). plant, act in an opposing manner. Cultures maintained in the
"Races" or chemically distinct populations of plants light fail to produce the alkaloids, whereas those kept in
often are observed when a plant species is studied over a the dark synthesize harman alkaloids (Barz and Hiisemann,
 Introduction

1982). Similar effects have been observed with antbraqui- a secondary metabolite that is subject to long-distance trans-
nones and lipoquinones in Morinda Lucida cell cultures port in the differentiated plant (Wink, 1987).
(Schultz et al., 1985). The presence of tissue-specific "pro-
moters" is linked to expression of secondary compounds in
many tissues. All these data indicate the need to understand IN'mRACTIVE ROLES OF PLANT SECOl'IDAKY
the basic biochemistry and physiology of the biosynthesis COMPOUl'lDS
of secondary compounds before it will be possible to initiate
synthesis in cell cultures in a controlled manner{Hutchinson, Plants exist in contact with a changing array of animals
1986; Rosahl et al., 1986). (mammals, insects, and nematodes), plants (algae, bry-
In addition to these factors, the mechanisms for product ophytes, ferns, and higher plants), fungi (mycorrhizal associ-
accumulation and storage are of prime importance, but little ates, pathogens, etc.), and bacteria. Secondary compounds
studied (James, 1950; Matile, 1978, 1984; Wink, 1987). In (allelochemics) are involved in interactions with and be-
general, there are few data concerning the site of storage of tween these organisms (Nordland et al., 1981; Swain, 1977;
secondary compounds within the plant. As observed above, Whittaker and Feeny, 1971). Many appear to confer protec-
storage at a particular site does not necessarily imply that tion to the plants that produce them (allomones); others are
the compound was synthesized there. For example, lupine involved in processes such as pollination and fruit/seed dis-
alkaloids are accumulated in epidermal cells (which lack persal (synomones); and yet others are used by the interact-
chloroplasts) but synthesized in mesophyll cells. The alka- ing organism to locate the plant host (kairomones). Similar
loids are transported to the epidermis via the phloem. Accu- considerations apply for insects which now are being recog-
mulation depends on the season and developmental stage of nized as rich sources of secondary metabolites. Although
the plant (James, 1950; Mothes, 1955). Sites of synthesis insects sequester plant-derived compounds, there is now
and accumulation often are separated in cells by compart- considerable evidence that many compounds can be synthe-
mentation. Lipophilic compounds tend to be accumulated sized de novo by the insects (Nahrstedt, 1989; Pasteels et
in membranes, vesicles, dead cells, or extracellular sites. al., 1988, 1989, 1990). For example, larvae of the burnet
Hydrophilic compounds tend to be stored in an aqueous envi- moth, Zygaena trifolii, feed on plants that produce the cyano-
ronment, typically the vacuole (Matile, 1978, 1984; Wink, genic glycosides linamarin and lotaustralin. Studies of incor-
1987). poration of labeled precursors indicate that this insect both
Improvements in techniques for isolation of protoplasts sequesters and synthesizes the compounds (Nahrstedt,
(plant cells without the cell wall) and vacuoles have facili- 1989). The origin of the pathways leading to these com-
tated studies of the uptake of plant secondary compounds. pounds is an important unanswered question. Some products
The concentrations of secondary metabolites in the vacuole and steps of the pathways are similar, suggesting a degree
may be higher than 500 mmollL. Compounds are usually of homology between insect and plant enzymes. Did these
stored in the vacuole against a concentration gradient; the pathways and compounds arise independently within the two
tonoplast membrane (the membrane surrounding the cell groups or have they been transferred from plants to insects?
vacuole) appears to contain a number of active proton-trans- Although either alternative would seem unlikely, recent
locating ATPases and pyrophosphatases. Import of com- work suggesting that mites have transferred transposable ele-
pounds into the vacuole is achieved by an H+-substrate anti- ments between insects makes interspecies transfer of genetic
port mechanism (Sze, 1984; Thom and Komor, 1984). These material a more acceptable possibility (Houck et al., 1991;
mechanisms for import of alkaloids into the vacuole are Robertson, 1993).
highly specific. In Fumaria capreolata vacuoles, (S)-scoule-
rine and reticuline are transported, whereas the R-forms are
not. This transport is activated by ATP, displays saturation PAtTERNS OF SIMULTANEOUS EVOLUTION
kinetics, depends on hydrogen ion concentration, and is sen- OF PLANTS Al'ID OTHER OROAl'llSIIIS
sitive to inhibitors (Deus-Neumann and Zenk, 1984, 1986).
In suspension cultures of cells of Lupinus polyphyllus, spar- As observed above, the chemistry and morphology of indi-
teine and lupanine are accumulated. The vacuoles take up viduals in most populations of any organism are variable.
these compounds (which are normally produced in the plant) In response to selection pressures such as herbivory, the
but discriminate against other alkaloids. The system is also abundance of certain phenotypes of plants within a given
pH and temperature dependent and can be activated by ATP population will be reduced. If selection pressure is sufficient
and KCI. In other plants, a variety of secondary compounds and is maintained for a sufficient period of time, plants of
such as coumaryl glycosides, flavonoids, and cardenolides this phenotype may become greatly reduced in number
appear to be taken up by similar mechanisms (Matern et al., within the population or, in extreme cases, disappear. In
1986; Werner and Matile, 1985; Wink, 1987). response, the numbers of the herbivore will decrease (assum-
Long-distance transport in plants is undoubtedly impor- ing that the herbivore is limited to this plant). If a plant
tant, but little studied. The mechanisms involved are not phenotype that is avoided by the herbivore exists within the
elucidated. Interestingly, few (if any) cell cultures produce population (or is formed by a mutation or by some new
10 Introduction

genetic combination), a gradual shift toward that phenotype bivore) further causes a change in gene frequency of the first
will occur. (the plant), the phenomenon is sometimes called coevolution
Variability occurs in both plant and herbivore popula- (Berenbaum, 1983; FutuymaandKeese, 1992; Futuymaand
tions. As a particular plant phenotype increases in numbers, Slatkin, 1983). Few, if any, proven examples exist in which
any phenotypes of interacting herbivores which exist or arise a one-on-one gene relationship of such changes has been
and are capable of utilizing the plants will be favored also. demonstrated.
This type of interaction probably is and has been very As the basic biochemical systems (including respiration
common in the course of evolution. As all organisms have and detoxication of various compounds) are similar in many
evolved in the presence of other organisms, it is difficult animals, it is not surprising that a large number of plant
to imagine circumstances in which this situation does not secondary compounds are deleterious to the growth and de-
prevail. velopment of many animals. The toxicity or poisonous prop-
One ofthe classic systems of this type involved butterflies erties of plants, in many cases, are determined by whether
and plants and, in particular, insect herbivores on Passiflora the animal has previously become coadapted to the plant
species (Ehrlich and Raven, 1964). From a systematic evalu- compounds. In some cases, medicinally useful compounds
ation of the kinds of plants fed upon by the larvae of certain (coincidentally?) modify the physiology of other organisms
groups of butterflies, these authors concluded that plant sec- in a beneficial way (Swain, 1972; Wagner and Hiirhammer,
ondary substances played the leading role in determining 1971). A new area of research involving examination of
patterns of utilization. They felt that this was true for other the role of the chemical "messages" involved in biological
groups of insects that were phytophagous or parasitic on interactions has emerged. A number of examples from this
plants as well. In general, whenever a plant produces chemi- new area of research, often called "chemical ecology," will
cal products that are deleterious or repellent to an insect (or be presented in this text.
other herbivore) in some way, the plant is at least partially
protected (Bemays and Chapman, 1987a,b; Jermy, 1976;
Jermy and Szentesi, 1978; Nahrstedt, 1989; Spencer, 1988). BVOLUI10N OF CHBlIUCAL PATHWAYS
Past evolutionary patterns have resulted in a great diversity
of plant secondary compounds and the herbivores on most As changes in secondary metabolism have occurred, new
plants represent only a small fraction of the total herbivores pathways have been elaborated. Modifications that involve
possible. These herbivores often appear to be "coadapted" relatively simple shifts of secondary metabolic pathways are
or "coevolved" although some herbivores are generalist encountered frequently. Precursors that are found in most
feeders and have the ability to consume a broad range of (if not all) plants, such as the compounds leading to gibberel-
plants. If a recombinant or mutation appears in an insect lins, carotenoids, or steroids, can serve as sources of many
population (or is selected from a variation that already exists) new secondary metabolites. Changes that parallel those pro-
that permits the insect to feed on some previously protected duced by widely distributed primary enzymes should be
plant group, selection will carry the line into a new adaptive more likely than those for which new enzymes would be
zone. Thus, the diversity of plants not only will tend to aug- required.
ment the diversity of insects, but the converse also appears In general, simply derived secondary compounds are
to be true. Changes in food source would be especially fa- more widely distributed than complex secondary compounds
vored in situations where the supply of the "preferred" food (Le., those involving numerous steps in their derivation from
plant is limited. In some instances, formerly repellent sub- primary compounds or readily available precursors, or in-
stances (allomones) might become cues that help the insects volving complex or chemically difficult steps) (Fig. 1.1). For
to locate the host plant (kairomones). Of course, the com- example, simple amines such as tryptamine (2) are widely
pounds may at the same time be allomones to noncoadapted distributed; alkaloids of greater complexity are less widely
insects. If the newly adapted insect herbivore is too success- distributed. Harmine (13), a J3-carboline alkaloid, is found
ful, selection for new resistant chemical variants in the plant in at least 20 plant families (Downum et al., 1991), several
will be favored. Thus, plant secondary chemistry appears to of which are not closely related. Even more complex com-
have played a strong role in the radiation of both plants pounds such as ajmalacine (1) are less widely distributed.
and herbivores. Clearly, other factors are involved and plant This alkaloid has been only isolated from three plant fami-
chemistry is not the dominant driving force for diversifica- lies. Quinine (14), an alkaloid derived from the same precur-
tion in many instances (Luckner et al., 1976). sor as ajmalacine and the product of a long pathway involv-
Why do we have so many secondary compounds? Radia- ing many complex steps, is known only from two or three
tion of chemically distinct plant popnlations in response to genera in the family Rubiaceae (Fig. 1.3) (Hansel, 1956;
herbivores and concurrent proliferation of herbivores with Waller and Nowacki, 1978).
different abilities to tolerate the presence of the resulting As a generality, evolutionarily "advanced" organisms
compounds illustrates at least one major way by which large have more "advanced" secondary compounds; that is, they
numbers of plant secondary metabolites could have arisen. are structurally and substitutionally more complex and more
In instances in which the second organism (here, the her- highly altered in comparison to the structure of the precursor.
 Introduction 11

0----0
~HN) ~ H2
CH,-O
CeQ
I~
~
I
HN
"""N

tryptamine (2) harmine (13)

found in at least 20 families, some not

widespread
closely related

CH3 0

ajmalacine (1) quinine (14)

found in 3 closely related found in 2 or 3 closely
families related genera of the Rubiaceae

Fig. 1.3. More complex members of series of plant secondary metabolites have restricted distribution.

There are many exceptions to these general observations. bers of the family (Gottlieb, 1980, 1989; Kubitzki and Got-
Small changes greatly affect the secondary compounds pro- tlieb, 1984, Kubitzki, 1987). Anthranilic acid is derived from
duced, if they occur early in the biosynthetic route. Further, a precursor of the shikimic acid pathway previous to those
it is easier to lose than to gain the ability to synthesize and leading to the phenylpropanoids which serve as lignin pre-
accumulate a particular compound. Thus, it is not surprising cursors. It is argued that because the pathway to lignins was
to find many instances of apparent reversals in biosynthetic blocked, the precursors were shunted into alkaloids and other
capacity. This should particularly be troe in instances where compounds. As the pathways to these alkaloids became
the initial selection pressures that favored the presence of the blocked, intermediates earlier in the pathway were shunted
compounds have disappeared. Species of plants that occur on into anthranilate-derived alkaloids. There was a concom-
islands (presumably with lower herbivore pressures) often mitant increase in the role of mevalonate (terpenoid) path-
have reduced secondary compound production and accumu- ways in more advanced groups; terpenoids (limonoids) and
lation in comparison to mainland species of the same genus coumarins replaced many compounds based on the shikimic
(Mabry, 1973; Seeligmann and Alston, 1967). acid pathway in the course of evolution (Gottlieb, 1980,
In instances where breakage of a biosynthetic route oc- 1989, 1990).
curs, precursors may be shunted into completely different
routes that may not have appeared in the particular species
previously or may be accumulated (Birch, 1973). THE VALUE OF PLAl'IT SECOI'IDAKY
Lignification, based on phenylpropanoid metabolites, is CHEJllISTRY FOR Ul'lDERSTAl'lDIl'IG
associated with the advent of land plants. It is thought by EVOLUfIONARY RELATIONSHIPS OR
many that early land plants were woody. In evolutionarily PHYLOGEl'IY OF PLAl'ITS
advanced plant groups, there is a tendency toward herba-
ceousness, or a decrease in the synthesis and accumulation Many simply derived secondary compounds are widespread
of lignin. In some of these plants, lignin precursors may and have limited usefulness for understanding the relation-
serve as the starting point for such secondary metabolites as ship of higher taxonomic groups of plants (Hegnauer, 1986).
phenylpropanoids, lignans, flavonoids, and alkaloids. Some Compounds derived from more complex pathways, how-
plants of the family Rutaceae accumulate alkaloids derived ever, are often restricted in distribution and can be valuable
from phenylpropanoid precursors, but these compounds ap- in this regard. Whereas simply derived compounds (espe-
pear to have been lost and replaced by alkaloids based on cially those involving common chemical steps from wide-
anthranilic acid in some more evolutionarily advanced mem- spread precursors) could have evolved several or even many
12 Introduction

times in the evolution or plants (or perhaps were present in of these compounds as "chemical messages" has proven
the ancestors of all modem plants), more complex com- important to our understanding of many ecological problems
pounds with restricted distribution are less likely to have and has led to the development of "chemical ecology."
arisen in this manner. Thus, plants that share these complex Plant secondary metabolites are not just randomly pro-
compounds are more likely to be related in an evolutionary duced compounds, but ones that have been shaped and opti-
sense than those which share simple compounds. mized during evolution. It is not surprising that many are
Certain groups of compounds characteristically are ob- used by man as pharmaceuticals, spices, fragrances, pesti-
served to occur in specific groups of plants. The patterns cides, poisons, hallucinogens, stimulants, or colors (Barz and
of distribution and information related to the biosynthetic Ellis, 1981; Luckner, 1990; Wink, 1988). Many of the com-
pathways responsible for secondary compounds have proven pounds also are important as medicines, pharmaceutical and
useful for understanding evolutionary relationships of plants. industrial precursors, fuels, pesticides, flavurings, perfumery
In some instances, the same molecular structure is known ingredients, plant hormones, adhesives, and drugs of abuse.
to arise by different biochemical pathways (convergence) Some still play exotic roles in other cultures as arrow poi-
and it is important to know not only the structure but also sons, piscicides (fish poisons), and hallucinogens.
the route of origin (Birch, 1973). Plants may appear to have The data provided by study of plant secondary metabo-
regressed as a result of mutations that block past evolution- lites have proven important in linking various aspects of
ary transformations (Birch, 1973). ecology, organic chemistry, biochemistry, both animal and
Secondary compounds are not, of course, the only fea- plant systematics, mycology, phycology, plant physiology,
tures of plants that have evolved. Many morphological and and evolutionary studies. There are important implications
anatomical characteristics also have been modified in the of these data for food science, range science, nutrition, an-
process of evolution. Studies in which a number of different thropology, archaeology, agronomy, horticulture, dairy sci-
aspects of evolutionary change are examined are especially ence, geology, and plant pathology.
desirable, as different aspects of plants are under different
selection pressures, and examination of several features will
be expected to give a more complete picture of the evolution- REFERENCES
ary history involved.
As one compares major groups of plants, it is clear that ALSTON, R. E., H. ROsLER, K. NAIFEH, and T. J. MABRY, Hybrid
no one line of secondary compounds has evolved indefi- compounds in natural interspecific hybrids. Proc. Nat. Acad.
nitely. Most major biosynthetic pathways are replaced even- Sci.,54, 1458-1465 (1965).
tually in more highly derived plant groups by other second- BARZ, W. and B. E. ELLIS, Potential of plant cell cultures for phar-
ary metabolic pathways. In some instances, these may have maceutical production. in Natural Products as Medicinal Agents
provided more effective systems of chemical defense. For (J. L. Beal and E. Reinhard, eds.), 471-507, Hippocrates Verlag,
example, in relatively primitive plants of the Rutaceae, ben- Stuttgart, 1981.
zylisoquinoline alkaloids are common. In more advanced BARZ, W. and W. HusEMANN, Aspects of photoautotrophic cell
groups within this family, acridone alkaloids almost com- suspension cultures, in Plant Tissue Culture 1982 (A. Fujiwara,
pletely replace the former type of alkaloids. In a similar ed), 245-247, Maruzen, Tokyo, 1982.
manner, condensed tannins are replaced by hydrolyzable tan- BARZ, W. and J. KOsTER, Tomover and degradation of secondary
nins among the more highly advanced groups of angio- (natural) products, in Secondary Plant Products, (E. E. Conn,
sperms (Gottlieb, 1980; Kubitzki and Gottlieb, 1984; Kubit- ed.), of Biochemistry of Plants (p. K. Stumpf and E. E. Conn,
zki, 1987). eds.), 35-84, Academic Press, New York, 1981.
Although the study of plant secondary metabolites has BAZZAZ, F. A., N. R. CHIARIELLO, P. D. COLBY, and L. F. i'rrELKA,
been of value for understanding the phylogeny of higher Allocating resources to reproduction and defense, BioScience,
plants, the reverse is true also. Knowledge of plant systemat- 37, 58-67 (1987).
ics and phylogeny has considerable predictive value for the BBRENBAUM, M. R.. Coumarins and caterpillars: A case for coevo-
study of secondary metabolites. lution, Evolution, 37, 163-179 (1983).
BERENBAUM, M. R., A. R. ZANGERL, and J. K. NrrAo, Constraints on
chemical coevolution: Wild parsnip and the parsnip webworm,
Evolution, 40, 1215-1228 (1986).
WHY STUDY PLAl'IT SECOl'IDAKY COl'lPOUl'IDS?
BERNAYS, E. A. and R. F. CHAPMAN, Chemical deterrence of plants,
in Molecular Entomology (J. Law, ed.), UCLA Symposium,
In the past, secondary compounds from plants have been Vol. 49, 108-116, Liss, New York, 1987a.
major targets for both structural and synthetic studies of or- BBRNAYs, E. A. and R. F. CHAPMAN, Evolution of deterrent re-
ganic chemists and have provided interesting challenges to sponses by phytophagous insects, in Perspectives in Chemore-
biochemists in the areas of biosynthesis and enzymology. A ception and Behavior (R. F. Chapman, E. A. Bemays, and J. O.
knowledge of the distribution of these compounds has Stoffolano, eds.), 159-174, Springer-Verlag, New York, 1987b.
proven useful to plant systematists and taxonomists and has BIRCH, A. 1., Biosynthetic pathways in chemical phylogeny, Pure
shed light on the phylogeny of plants and fungi. The role Appl. Chern., 33, 17-38 (1973).
 Introduction 13

BRYANT, 1. P., F. S. CHAPIN III, AND D. R. KLBIN, Carbon/nutrient lites under water and nutrient stress, in Phytochemical Adapta-
balance of boreal plants in relation to vertebrate herbivory. tion to Stress (B. N. Timmennann, C. Steelink, and F. A. Loe-
Oikos, 40, 357-368 (1983). WUS, eds.), Recent Advances in Phytochemistry, Vol. 18,
Bu'LocK, J. D., Mycotoxins as secondary metabolites, in The Bio- 273-320, Plenum, New York, 1984.
synthesis of Mycotoxins (p. S. Steyo, ed.), 1-16, Academic GOTILIEB, O. R., Micromolecular systematics: Principles and prac-
Press, New York, 1980. tice, in Chemosystematics: Principles and Practice (F. A. Bisby,
CHAPPELL, 1. and K. HAHLBROCK. Transcription of plant defense J. G. Vaughan, and C. A Wright, eds.), 329-352, Academic
genes in response to UV light or fungal elicitor. Nature (Lon- Press, London, 1980.
don), 3]], 76-78 (1984). GD'ITLIEB, O. R, The role of oxygen in phytochemical evolution
CHEW, F. S. and J. E. RODMAN, Plant resources for chemical de- towards diversity, Phytochemistry, 28, 2545-2558 (1989).
fense, in Herbivores (G. A Rosenthal and D. H. Janzen, eds.), GD'ITLIEB, O. R., Pbytochemicals: Differentiation and function,
271-307, Academic Press, New York, 1979. Phytochemistry,29, 1715-1724 (1990).
COLBY, P. D., J. P. BRYANT, and T. CHAPIN, Resource availability HANSEL, R., Pflanzenchemie und Pflanzenverwandschaft, Arch.
and plant antiherbivore defense, Science, 22, 895-899 (1985). Pharm., 259, 619-628 (1956).
DEROME, A E., The use of N.M.R. spectroscopy in the structure HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod.
detennination of natural products: Two-dimensional methods. Rep., 3, 323-344 (1986).
Nat. Prod. Rep., 6, 111-141 (1989). HARTMANN, T., Prinzipien des pflanzlichen Sekundlir Stoffwech-
DEUS-NEUMANN, B. and M. H. ZENK, A highly selective alkaloid sees, Plant Systematics and Evolution, 150, 15-34 (1985).
uptake system in vacuoles of higher plants, Planta, 162, HASLAM, E., Secondary metabolism-Fact or fiction, Nat. Prod.
250-260 (1984). Rep., 3, 217-249 (1986).
DeUS-NEUMANN, B. and M. H. ZENK, Accumulation of alkaloids HEGNAUBR, R., Phytochemistry and plant taxonomy-An essay
in plant vacuoles does not involve an ion-trap mechanism, on the chemotaxonomy of higher plants, Pbytochemistry, 25,
Planta, 167, 44-53 (1986). 1519-1535 (1986).
DoWNUM, K. R, L. A SWAIN, and L. J. FALBlRo, Influence of light HEINSTEIN, P. F., Stimulation of sesquiterpene aldehyde formation
on plant allelochemicals: A synergistic defense in higher plants, in Gossypium arboreum cell suspension cultures by conidia of
Arch. Insect Biochem. Pbysiol., 17, 201-211 (1991). Verticillium dahliae, J. Nat. Prod., 48, 905-915 (1985).
EHRUCH, P. R and P. H. RAVEN, Butterflies and plants: A study HOUCK, M. A, J. B. CLARK, K. R. 1'ETERs0N, and M. G. KIDWELL,
in coevolution, Evolution, 18, 856-608 (1964). Possible horizontal transfer of Drosophila proctolaelaps re-
En.ERT, U, Elicitation: Methodology and aspects of application, in galis, Science, 253,1125-1129 (1991).
Cell Culture and Somatic Genetics of Plants, Vol. 4 (F. HRAzoINA, G. and R. A JENSEN, Spatial organization of enzymes
Constabel and I. K. Vasil, eds.), 153-196, Academic Press, San in plant metabolic pathways, Annu. Rev. Plant Physiol. Plant
Diego, CA, 1987. Mol. BioI., 43,241-267 (1992).
ELLIS, B. E., Natural products from plant tissue CUlture, Nat. Prod. HUTCHINSON, C. R, Biological methods for studying the biosyn-
Rep., 5, 581-612 (1988). thesis of natural products, Nat. Prod. Rep., 3, 133-152 (1986).
FEENY, P., Theories of plant chemical defense: A brief historical JAMBS, J. 0., Alkaloids in plants, in The Alkaloids, Vol. I (R. H.
survey, Symp. Hung. ~iol., 39, 163-175 (1990). F. Manske and H. L. Holmes, eds.), 15-90, Academic Press,
FLORES, H. E., Use of plant cells and organ culture in the production New York, 1950.
of biological materials, in Biotechnology in Agricultural Chem- JERMY, T., Insect host-plant relationships: Coevolution or sequen-
istry (H. M. LeBaron, R. O. Mumma, R. C. Honeycutt, J. H. tial evolution, Symp. Bioi. Hung., 16, 109-113 (1976).
Duesing, J. F. Phillips, and M. J. Haas, cds.), ACS Symposium JERMY, T. and A SZBNTESl, The role of inhibitory stimuli in the
Ser. 334, 66-86, American Chemical Society, Washington, choice of oviposition site by phytophagous insects, Ent. Exp.
D.C., 1987. Appl., 24, 458-471 (1978).
FLUCK, H. t Intrinsic and extrinsic factors affecting the production JONES, D. A., Chemical defense: Primary or secondary function?
of secondary plant products, in Chemical Plant Taxonomy (T. Am. Nat., II3, 445-451 (1979).
Swain, ed.), 167-186, Academic Press, London, 1963. JONES, c. G. and R D. FIRN, On the evolution of plant secondary
FOWDBN, L., Amino acid complement of plants, Pbytochemistry, chemical diversity, Phil. Trans. Royal Soc. London, Ser. B, 333,
]],2271-2276 (1972). 273-280 (1991).
1'uTuYMA, D. J. and M. C. KeEsE, Evolution and coevolution of KUBITZKI, K., Pbenylpropanoid metabolism in relation to land plant
plants and phytophagous arthropods, in Herbivores: Their Inter- origin and diversification, J. Plant Pbysiol., 131, 17-24 (1987).
actions with Secondary Metabolites, Vol. 2 (G. A Rosenthal KUBITZKI, K. and O. R. GD'ITLIEB, Micromolecular patterns and
and M. R Berenbaum, eds.), 439-475, Academic Press, San the evolution and major classification of angiospenns, Taxon,
Diego, CA, 1992. 33,375-391 (1984).
FuroVMA, D. 1. and M. SLATKIN, Coevolution, Sinauer Associates, KUTCHAN, T. M., S. AVABE, R. J. KRUEGER, E. M. COSCIA, and
Sunderland, MA, 1983. C. J. COSCIA, Cytodifferentiation and alkaloid accumulation in
GANDER, J. E., Incorporation of C l4 into p-hydroxymandelonitrile- cultured cells of Papaver somniferum, Plant Cell Rep., 2,
j3-g1ucose and other phenolic substances in sorghum seedlings, 281-284 (1983).
J. BioI. Chern., 237, 3229-3232 (1962). LUCKNER, M., Was ist SekWldarstoffwechsel, Phannazie, 26,
GERSHBNZON, 1., Changes in the levels of plant secondary metabo- 717-724 (1971).
14 Introduction

LUCKNER, M., Expression and control of secondary metabolism, sect-Plant Interactions (P. Barbosa and D. Letourneau, eds.),
in Secondary Plant Products, Vol. 8 of Encyclopedia of Plant 235-272, Wiley, New York, 1988.
Physiology (E. A. Bell and B. V. Charlwood, eds.), 23-63, PASTIillLS, J. M., M. ROWBLL-RAHmR, J. C. BRAEKMAN, D. DALOZB,
Springer-Verlag, Berlin, 1980. and S. DUFFEY, Evolution of exocrine chemical defense in leaf-
LUCKNER, M., Secondary Metabolism in Plants and Animals, Aca- beetles (Coleoptera: Chrysomelidae), Experientia, 24,
demic Press, New York, 1972; third edition, Springer-Verlag, 3299-3302 (1989).
Berlin, 1990. ROBERTSON. H. M., The mariner transposable element is wide-
LUCKNER, M., K. MOTHES and L. NoVER (eds.), Secondary Metabo- spread in insects, Nature, 361,241-245 (1993).
lism and Coevolution, Nova Acta Leopoldina Suppl. No.7, ROBINSON, T., Metabolism and function of alkaloids in plants, Sci-
Deutsche Akademie der Naturforscher Leopoldina, 1976. ence, 184,430-435 (1974).
MABRY, T. J., The chemistry of geographical races, Pure Appl. ROBINSON, T., The Organic Constituents of Higher Plants, 4th ed.,
Chern., 34, 377-400 (1973). Cordus Press, North Amherst, MA, 1980.
MARKHAM, K. R. and T. J. MABRY, The identification of twenty- ROSAHL, S., P. EcKEs, J. SCHELL, and L. WlLLMITZER, Organ-spe-
three 5-deoxy- and ten 5-hydroxyflavonoids from Baptisia lec- cific gene expression in potato. Isolation and characterization
ontei (Leguminosae), Phytochemistry, 7, 791-801 (1968). of tuber-specific cDNA sequences, Mol. Gen. Genet., 202,
MATERN, u., C. REICHENBACH, and W. HEu.ER, Efficient uptake 368-373 (1986).
offlavonoids into parsley (Petroselinum hortense) vacuoles re- ROSENTHAL, G. A., Plant Nonprotein Amino and imimo Acids,
quires acylated glycosides, Planta, 167, 183-189 (1986). Academic Press, New York, 1982.
MATILE, P., Biochemistry and function of vacuoles in plants, Annu. ROSENIHAL, G. A. and D. H. JANZEN (eds.), Herbivores, Academic
Rev. Plant Physiol., 29, 193-213 (1978). Press, New York, 1979.
MATILE, P., Das toxische Kompartiment der Pflanzenzelle, Natur- SADLER, 1. H., The use of N.M.R. spectroscopy in the structure
wissenschaften, 71, 18-24 (1984). detennination of natural products: One-dimensional methods,
McKEY, D., Adaptive patterns in alkaloid physiology, Am. Nat., Nat. Prod. Rep., 5, 101-127 (1988).
J08, 305-320 (1974). SCHULTZ, G., J. SOLL, E. FIEDLER, and D. SCHULZE-DlEBBRT, Syn-
McKEY, D., The distribution of secondary compounds within thesis of prenylquinones in chloroplasts, Physiol. Plant., 64,
plants, in Herbivores (G. A. Rosenthal and D. H. Hanzen, eds.), 123-129 (1985).
56-13 Academic Press, New York, 1979. SBELIGMANN P. and R. E. ALsTON, Complex chemical variation
McPlmRSON, M. J. and J. H. PARISH, Applications of recombinant and the taxonomy of Hymenoxys scaposa (Compositae), Brit-
DNA in biotechnology, Nat. Prod. Rep., 4, 205-224 (1987). tonia, 19, 205-211 (1967).
MOTHES, K., Physiology of alkaloids, Annu. Rev. Plant Physiol., SEIGLER, D. S. and P. W. PRICE, Secondary compounds in plants:
6, 393--432 (1955). Primary functions, Am. Nat.,llO, 101-104 (1976).
MOTHES, K., Bemerkungen zur Biologie der Alkaloide, in Abhand- SELMAR, D., Cyanogenesis and metabolization of cyanogenic com-
lungen der Deutschen Akademie der Wissenschaften zu Berlin. pounds: An example for multifunctionality of secondary plant
Klasse ffir Chemie, Geologie, und Biologie, Akademie-Verlag. products, Planta Medica, 55, 592-593 (1989).
Berlin; 1965, 3rd Internationales Symposium Biochemie und SELMAR, D., Die Metabolisierung Cyanogener Verbindungen, Ha-
Physiologie der Alkaloide, Halle (Saale), Germany, 27-34 bilitationsschrift, Technischen Universitiit Carolo-Wilhehnina
(1966a). zu Braunschweig, Braunschweig, 1992.
MOTHEs, K., Biogenesis of alkaloids and the problem of chemotax- SELMAR, D., S. GROCHOLEWSKI, and D. S. SEIGLER, Cyanogenic
onomy, Lloydia, 29, 156-171 (1966b). lipids. Utilization during seedling development of Ungnadia
MOTHES, K., Zur Problematik der metabolischen Exkretion bei speciosa, Plant Physiol., 93, 631-636 (1990).
Pflanzen, Naturwissenschaften, 53, 317-323 (1966c). SELMAR, D., R. LIEBEREI, and B. BIEHL, Mobilization and utilization
MOTHES, K., Secondary plant substances as materials for chemical of cyanogenic glycosides, Plant Physiol., 86, 711-716
high quality breeding in higher plants, in Biochemical Interac- (1988).
tions between Plants and Insects (R. L. Mansell and J. Wallace, SIMMS, E. L. and M. D. RAuSHER, Costs and benefits of plant
eds.), Recent Advances in Phytochemistry, Vol. 10, 385-405, resistance to herbivory, Am. Nat., 130, 570-581 (1987).
Plenum, New York, 1976. SIMPSON, T. 1., I3C_NMR in metabolic studies, in Nuclear Magnetic
NAHRSTEDT, A., The significance of secondary metabolites for in- Resonance, in Modem Methods of Plant Analysis (H. F. Lin-
teractions between plants and insects, Planta Medica, 55, kens and J. F. Jackson, eds.), 1-42, Springer-Verlag, Berlin,
333-338 (1989). 1986.
NORDI.AND, D. A., R. L. JONES, and W. J. LEWIS, Semiochemicals, SOMERVILLE, C. and 1. BROWSE, Plant lipids: Metabolism mutants,
Wiley, New York, 1981. and membranes, Science, 252, 80-87 (1991).
PAECH, K., Biochemie und Physiologie der sekundaren Pflanzens- SPENCER, K. C.. Chemical Mediation of Coevolution, Academic
toffe, Springer-Verlag, Berlin, 1950. Press, San Diego, CA, 1988.
PASTEELS, J. M., S. DUFFEY, and M. ROWELL-RAHIER, Toxins in SWAIN, T. (ed.), Plants in the Development of Modem Medicine,
chrysomelid beetles, J. Chern. Ecol., 16, 211-222 (1990). Harvard University Press, Cambridge, MA, 1972.
PASTEELS, J. M., M. RoWBt.t.-RAHIER, and M. J. RAupp, Plant-de- SWAIN, T., Secondary compounds: primary products, in Secondary
rived defense in Chrysomelid beetles, in Novel Aspects of In- Metabolism and Coevolution (M. Luckner, K. Mothes, and L.
 Introduction 15

Nover, eds.), Nova Acta Leopoldina SuppL 7, 411-421, WHIITAKER, R. H. and P. P. FEENY, Allelochemics: Chemical inter·
Deutsche Akadernie def Naturforscher Leopoldina, Halle, Ger- actions between species, Science, 171, 757-770 (1971).
many, 1976. WIERMANN, R., Secondary products and cell and tissue differentia-
SWAIN, T., Secondary compounds as protective agents, Annu. Rev. tion, in The Biochemistry of Plants, Vol. 7 (E. E. Conn, ed.),
Plant PhysioL, 28, 479-501 (1977). 85-115, Academic Press, New York, 1981.
SZE. R., H+-translocating ATPases of the plasma membrane and WILKINSON, A. P., C. M. WARD, and M. R. A. MORGAN, Immuno-
tonoplast of plant cells, PhysioL Plant, 61,683-691 (1984). logical analysis of mycotoxins, in Plant Toxin Analysis (H. F.
THOM, M. and E. KOMOR, H+-sugar antiport as mechanism of sugar Linskens and J. F. Jackson, eds.), Modem Methods of Plant
uptake by sugarcane vacuoles, FEBS Lett., 173, 1-4 (1984). Analysis, VoL 13, 185-225, Springer-Verlag, Berlin, 1992.
TUKEY, H. B., JR., The leaching of substances from plants, Annu. WILLIAMS, D. H., M. 1. STONE, P. R. HAUCK, and S. K. RAHMAN,
Rev. Plant PhysioL, 21,305-324 (1970). Why are secondary metabolites (natural products) biosynthes-
WAGNER, H. and L. HORHAMMER (eds.), Pharmacognosy and Phyto- ized?, J. Nat. Prod., 52,1189-1208 (1989).
chemistry, Springer-Verlag, Berlin, 1971. WINK, M., Physiology of the accumulation of secondary metabo-
WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and Metabo- lites with special reference to alkaloids, in Cell Culture and
lism in Plants, Plenum, New York, 1978. Somatic Cen Genetics of Plants, Vol. 4 (F. Constabel and I. K.
WATERMAN, P. G. and S. MOLE, Extrinsic factors influencing pro- Vasil, eds.), 17-42, Academic Press, San Diego, CA, 1987.
duction of secondary metabolites in plants, in Insect-Plant In- WINK, M., Plant breeding: Importance of plant secondary metabo-
teractions, VoL 11 (E. A. Bemays, ed.), 107-134, CRC Press, lites for protection against pathogens and herbivores, Theoret.
Boca Raton, FL, 1989. AppL Genet., 75, 225-233 (1988).
WERNER, C. and P. MATILE, Accumulation of coumaroylglucosides WINK, M. and L. WIITE, Evidence for a wide-spread occurrence
in vacuoles of barley mesophyll protoplasts, 1. Plant Physio!., of the genes of quinoIizidine alkaloid biosynthesis, FEBS Lett.,
I18, 237-249 (1985). 159, 196-200 (1983).
2
Fatty Acids

Introduction INlKODUCTlOI'l
Primary Fatty Acids
Fatty Acids of Plant Vegetative Parts Most fatty acids occur in nature as esters of glycerol (com-
monly called triglycerides or triacylglycerols). These esters
Biosynthesis
Fatty Acid Biosynthesis are found in most living organisms and playa role in energy
storage. The molecular shapes of lipid molecules affect the
The Two-Pathway Model of Lipid Biosynthesis
biogenesis and function of various cell membranes (Browse
The Second 3-Ketoacyl ACP Synthase Isozyme
~d Somerville, 1991). Both phosphoinositides and methyl
Biosynthesis of Unsaturated Fatty Acids
Jasmonate are involved in signal transduction. Because of
The Prokaryotic Pathway
their economic and dietary impottance, triglycerides of seeds
The Eukaryotic Pathway
and the corresponding fatty acid derivatives have been inves-
Biosynthesis of Triacylglycerides
tigated extensively; there have been comparatively few stud-
Degradation of Fatty Acids
ies of glycerides and fatty acids from other plant palls. The
Unusual Fatty Acids in Plants
biochemistry of fatty acids, glycerides, and related com-
Fatty Acids from Unusual Starter Units
pounds has been reviewed (Browse and Somerville, 1991;
Fatty Acids with Unusual Patterns of Unsaturation
Goodwin and Mercer, 1983; Lie Ken Jie, 1989; Stumpf,
Hydroxy Fatty Acids
1976).
Epoxy Fatty Acids
Glycerides are major components of edible oils. Those
Methyl Branched, Cyclopropanoid, and Cyclopropenoid
which contain fatty acids with a relatively high degree of
Fatty Acids
unsaturation are thought to provide benefits in the diet. The
Cyclopentenoid Fatty Acids
ability of highly unsaturated oils to polymerize in the pres-
Fluoro Fatty Acids
ence of air has made them useful for paints. Glycerides that
Other Unusual Lipids
contain a predominance of saturated fatty acids can be con-
Metabolically Altered Fatty Acids
verted to salts by treatment with bases (saponification). So-
The Green Leaf Volatile Complex
dium and potassium salts are usefnI for soaps, whereas lith-
Functions of Fatty Acids and Their Derivatives in Plants
ium salts are components of lubricants. Triglycerides from
Jasmonic Acid and Related Compounds
seed oils possibly represent the most likely substitute for
The Requirement for Unsaturated Fatty Acids in the
petroleum in the future.
Diet of Animals Fatty acids also are stored as components of phospholip-
Effects of Fatty Acids and Their Derivatives on ids, glycolipids, and waxes, or converted to alcohols, alde-
Animals hydes, olefins, hydrocarbons, acetylenic compounds, and
Fatty Acids with Biological Activity other secondary metabolites.
Insect Pheromones Derived from Fatty Acids
Mammalian Pheromones Frimary Fatty Acids
Analysis of Fatty Acids ~any fatty acid.s are primary metabolites; that is, they
References are Involved In resprration and energy storage and are ubiqui-

16
 Fatty Acids 17

~CO,H capric acid

CO,H lauric acid (6)

CO,H myristic acid (7)

CO,H palmitic acid (1)

H H
CO,H palmitoleic
acid (13)
CO,H stearic acid (2)

H H
CO,H oleic acid (3)

CO,H eJaidic acid

H H H H

C01H linoleic acid

(4)
H H H H H H

CO,H linolenic acid

(5)

CO,H arachidic acid

(25)

H HH HH HH H
CO,H arichidonic acid
(61)

Fig. 2.1. Some naturally occurring primary fatty acids.

tous in plants. Among these are palmitic (1), stearic (2), oleic 1991). About 70% of the fatty acids ofleaves are linoleic and
(3), linoleic (4), and linolenic acids (5) (Fig. 2.1). Others, a-linolenic acids with smaller amounts of oleic, palmitic,
such as lauric (6) and myristic acid (7) also are widely dis- palmitoleic, and (E)- or trans-3-hexadecenoic acid (8) (Fig.
tributed in nature. 2.14) (found only in chloroplasts). Some plants contain siza-
Chain lengths of 16 and 18 are common for most animals, ble quantities of hexadecatrienoic acid. Lipids are responsi-
microbes, and plants. This may be because these chain ble for about 7% of the total weight of many plant leaves.
lengths provide optimal physical properties of bilayer mem- About 30% of these leaf lipids are comprised of phospholip-
branes (Ohlrogge et al., 1993). ids (including glycerophospholipids), glycolipids (Joyard
and Douce, 1987), and waxes (Harwood, 1980) (Fig. 2.2).
Fatty Adds of Plant Vegetative Parts Phosphatidylglycerol (9) makes up as much as 20% of the
Whereas plant seeds contain a wide variety of fatty acids, total phospholipids of thylakoids and envelope membranes
fatty acids from leaf tissue are closely linked with the pres- (Lawlor, 1993); monogalactosyl and digalactosyl diglycer-
ence and number of chloroplasts and are remarkably con- ides (10, 11) may represent 20-45% of the total leaf lipids
stant. Plant membranes differ in composition from those of (Elbein, 1980). Complex glycolipids appear to be wide-
animals and fungi. This is especially seen in the composition spread among plants (Kojima et al" 1990). No triacylglycer-
of chloroplast membranes which consist largely of glycolip- ols and only minor amounts of phospholipids occur in chlo-
ids. The lipid composition of plant chloroplasts is similar to roplasts (Stumpf, 1976), where as much as 30% of the total
that of cyanobacterial membranes (Somerville and Browse, lipids may be monogalactosyl diglycerides (Elbein, 1980).
18 Fatty Acids

o o

{ O~R
R = alkyl group II
O--1L. R OJLR

RTo {
0
R---,rO
II II
0
R---,ro{
II
0
II
CH CH
,~+ 3
o O~R O O-P-O'
I
0 O-p-O............. "CH
I 'V' 3
O·

triglyceride phosphatidic acid (19)0' pbospbatidylcholine (23)

o 0
~ {O~R O~R
R 0{0 0 R RIIO ~ RT O { ~ ~
IoI "
0-P·0~NH3
+0 O-P·O"'-"'" /"OH
,,(
0 O-P'O--v'O'P-O}
O' OH·' 0
6. o· OH 0 O..lLR
Rlr°
o
pbosphatidyletbanolamine (29) pbosphatidylglycerol (9) diphosphatidylglycerol

monogalactosyl dlgaJacrosyl sulfoquinovosyl

diacylglycerol (10) diacylglycerol (11) diglyccride (12)

Fig. 2.2. Glyceropho'phatid, and glycolipid, from plants. [Reprinted in modified form from Stumpf (1976).]

Glycolipids of this type are synthesized in the chloroplast tions and/or the fonnation of the oil storage organelle or
(Lawlor, 1993). Sulfoquinovosyl diglyceride (12) is wide· oil droplet (which contains only triacylglycerols) (Stumpf,
spread in leaves but usually occurs at low concentrations 1980, 1987). Some evidence suggests that lipid biosynthesis
(Stumpf, 1976). This lipid is linked to S04 - reduction in also may occur in mitochondria (Shintani and Ohlrogge,
the chloroplast (Lawlor, 1993). Sterol esters and acylated 1994).
steryl glycosides are also found in leaf lipids. Fatty acids and lipids usually are not transported between
Unusual fatty acid structures, often found in seed oils, the cells of higher plants, but are transported within cells
are rarely encountered in leaves. The presence of cyclopro· from chloroplasts to sites on the endoplasmic reticulum and
penoid fatty acids in leaves of plants of the Malvaceae and back into the chloroplasts (Somerville and Browse, 1991).
related families and conjugated unsaturated fatty acids in
leaves of plants of the Boraginaceae appear to be exceptions The Two·Pathway Model of Lipid
(Shoriand, 1963). Biosynthesis
In general, the linolenic acid of leaves is replaced by Research since about 1980 indicates that the leaves of
linoleic acid in roots. higher plants utilize two distinctive pathways for glyceroli·
pid biosynthesis. Palmitoleic acid (C I.) (13) and oleic acid
(CIS,I) (3) are synthesized de novo in the chloroplast and
BIOSYNI'HESIS may be used directly for the biosynthesis of chloroplast lipids
via the prokaryotic pathway (see below) or exported from
In tissues that nonnally do not store triacylglycerols, for the chloroplast as CoA esters that are used in the eukaryotic
example, leaf cells, the chloroplast is the primary source of pathway primarily at sites on the endoplasmic reticulum
fatty acids (Browse and Somerville, 1991). The endoplasmic (Browse and Somerville, 1991). However, in all higher
reticulum in the cell cytosol is involved in further modifica· plants, a proportion of the diglycerol moiety of phosphatidyl·
tions of the acyl moiety. In tissues high in oil content, the choline, synthesized by the eukaryotic pathway, again enters
proplastid is the source of the primary fatty acids. The endo· the chloroplast where it contributes to the production of thy-
plasmic reticulum is the site of further structural modifica· lakoid membranes (Browse and Somerville, 1991).
 Fatty Acids 19

In many families of angiospenns, phosphatidylglycerol NH,

(9) is the only major product of the prokaryotic pathway.
Other chloroplast lipids in these plants are synthesized only
by the eukaryotic pathway. In some primitive angiospenn
families, both pathways contribute to the synthesis of mono-
l):)
galactosyldiacylglycerol (10) and other lipids (Browse and
Somerville, 1991). HO ~ o-p-o-p-o~~
~ ~
I I
0

o OH OH CH NH)
Fatty Acid Biosynthesis

S
Ho-Lo OH O=\H
Fatty acids in plants are synthesized by a series of reac-
tions similar to those for fatty acid biosynthesis in animals,
yeasts, bacteria, and other organisms. The synthetases of
~H
prokaryotic cytoplasmic systems are freely soluble and read- acetyl-CoA (15) S-COCH,
ily separable as discrete proteins; those in mammalian sys-
tems are localized in the cytoplasm as large soluble synthe-
tase complexes. The exact nature of association of the
enzymes in plants is unknown (Ohlrogge et al., 1993). The
reactions of plant fatty acid biosynthesis are catalyzed by
individual polypeptides (Somerville and Browse, 1991);
'yO acetyl·CoA carboxylase ~"
S·CoA
once organelles are disrupted, the enzymes also are soluble S·CoA
and may be isolated (Somerville and Browse, 1991; Stumpf,
1976). In contrast to mammalian systems, plant fatty acid acetyl·CoA (15) malonyl·CoA (14)
synthetases are plastid localized.
Fig. 2.3. Acetyl-CoA and the origin of malonyl-CoA.
Synthesis of fatty acids occurs in the stroma but involves
mitochondria and the cytosol (Lawlor, 1993). Dihydroxyace-
tone phosphate (DHAP) moves from the stroma to the cyto- amino acid sequence surrounding the prosthetic group to
sol. Pyruvate is fonned from the triosephosphate DHAP and which acyl groups are attached is highly conserved. Further,
enters the mitochondria where acetyl-CoA and acetate are from other studies, it appears that precursors to ACP units
produced. This reaction also produces NADH, which pro- contain chloroplast transit peptides. Thus, ACPs are nuclear-
vides part of the reducing power needed for later reactions encoded proteins that must be imported into the plastid from
in the sequence. Acetate returns to the chloroplasts where their site of synthesis on cytoplasmic ribosomes (Ohlrogge
acetyl-CoA again is synthesized (Lawlor, 1993). Because et al., 1993).
acetyl-CoA does not cross membranes, this molecule must In subsequent steps of fatty acid synthesis, a fatty acyl
be resynthesized in plastids to provide the carbon precursors group that is linked by a thioester bond to the active site
for fatty acid biosynthesis (Ohlrogge et al., 1993). of 3-ketoacyl-ACP synthase condenses with malonyl·ACP.
Plastids of oil seeds appear to have a complete glycolytic One molecule of CO2 is liberated (Browse and Somerville,
pathway, which together with pyruvate dehydrogenase, can 1991 Conn and Stumpt, 1972, Lehninger, 1982) (Fig. 2.4).
result in the conversion of glucose to acetyl-CoA. Chloro- Although acetyl-ACP has been considered to condense ini-
plasts and plastids of oilseeds may use different pathways tially with malonyl-ACP, recent work indicates that the
to provide the necessary substrates and cofactors (Ohlrogge product of the first condensation, butyryl-ACP, is fonned
et al., 1993). by the condensation of acetyl-CoA and malonyl-ACP and
The synthesis of malonyl-CoA (14) from acetyl-CoA that acetyl-ACP is a minor participant in fatty acid biosyn-
(15), which also occurs in the stroma, with acetyl-CoA car- thesis (Jaworski et aI., 1993).
boxylase (ACCase) is the first committed step of fatty acid Three forms of 3-ketoacyl-ACP synthase have been dis-
biosynthesis (Fig. 2.3). Malonyl-CoA produced by plastids covered in plants. These fonns may be distinguished by their
primarily is used for fatty acid biosynthesis (Ohlrogge et al., substrate specificity; they are homodimers with molecular
1993). weights of 43,000 to 45,000 per subunit. One, KAS III, ap-
Malonyl-CoA condenses with an acyl carrier protein pears to be responsible for the first condensation of acetyl-
(ACP). This reaction is catalyzed by malonyl-CoA ACP CoA and malonyl·ACP (Browse and Somerville, 1991; Ohl·
trans acylase, an enzyme that has been purified from several rogge et al., 1993). The activity of this enzyme in plants
plants (Ohlrogge et al., 1993). ACP is a relatively small seems to bypass the need for acetyl-ACP, although that mol-
heat-stable protein of 9000 molecular weight that occurs in ecule is fonned and accumulated in some plants (Ohlrogge
several isofonns (Browse and Somerville, 1991; Lehninger, et al., 1993). KAS I or 3-ketoacyl-ACP synthase elongates
1982). Both ACP and coenzyme A have a 4'-phosphopan- the acyl chain to palmitoyl-ACP, whereas KAS II converts
tetheine prosthetic group (Ohlrogge et aI., 1993). Amino acid palmitoyl-ACP to stearoyl-ACP (Ohlrogge et al., 1993).
sequences for ACP molecules from more than 15 plants, The acetoacetate ester fonned in the initial condensation
including both monocots and dicots, are available. The step is reduced by 3-ketoacyl·ACP reductase in the presence
20 Fatty Acids

malonyl trausaeylase
malonyl-CoA + ACP-SH • malonyl-S-ACP + CoA

acetyl-CoA + malonyl-S-ACP - - - - - - - _ aeetoaeetyl-S-ACP + CO2

3-ketoaeyl-ACP reductase
acetoacetyl-S-ACP + NADPH + H+ • D(-)-3-hydroxybutyryl-S-ACP + NADP+
3-hydroxy-ACP dehydratase
D(-)-§-hydroxybutyryl-S-ACP • E-Ll.2-butenoyl-S-ACP + H 20
enoyl ACP reductase
E-Ll.'-butenoyl-S-ACP + NADPH + H+ • butyryl-S-ACP + NADP+

butyryl-S-ACP + e n z ( * ) - - - - - - - - - . butyryl-S-enz (0) + ACP

butyryl-S-enz(*) + malonyl-S-ACP - - - - - - . . 3-ketohexanoyl-S-ACP + enz(.) + CO2 etc_

enz(*) = 3-ketoacyl-ACP synthetase

Fig. 2.4. Summary of reactions leading to fatty acid biosynthesis.

ofNADPH to fonn 0-3-hydroxybutyryl-S-ACP. Isofonns of ACP dehydratase to yield E-t.. 2-butenoyl-S-ACP. This en-
the enzyme require either NADPH (the dominant fonn) or zyme has a molecular weight of 85,000 and appears to be
NADH. The native enzyme appears to be a tetramer with a a homotetramer (Ohlrogge et al., 1993). The olefin produced
molecular weight of 130,000 (Ohlrogge et al., 1993). 0-3- is reduced by NADPH in the presence of enoyl-ACP re-
Hydroxyburyryl-S-ACP is dehydrated by 3-hydroxyacyl- ductase to fonn buryryl-S-ACP (Fig. 2.5) (Lehninger, 1982).

~ ~CO~cc_ase
)Vl
0 0

ACP-SH~ SCoA ACP-SH

o COA~ AD~O 1 CO~-SHO
A
ATP SCoA 0

~YI-COA:ACP
transacylase malonyl-CoA:ACP
~ ~ /~
II
S-ACP HO S-ACP
malonyl-ACP

CO2 +
ACP-SHor
o o 0 CoA-SH
_ II KAS·I
~S-A-C-P-----=;::'::""'----
AAS-ACP
aeyl.ACP 3-keloaeyl-ACP
A enoyl-ACP reductase H20 3-ketoacyl-ACP reductase

NADP+" 0 ~ OH 0 ~NADPH
NADPH _ II - 1 I NADP·
/~S-ACP /~S-ACP
enoyl-ACP 3·hydroxyaeyl-ACP 3-bydroxy-ACP
dehydrase
Fig. 2.S. Reactions of fatty acid biosynthesis (Ohlrogge et aI., 1993; modified and used with pennission of the copyright owner, CRe Press, Boca
Raton, Florida. © 1993).
 Fatty Acids 21

biotin
O=< o 0

B: \ OH carboxylation
H~COA acylation

~ retention
H 'H ~SR inversion

~
COA
'H
(28) ~
'H
o

HQ~
o 0

./JlX JL. . . NADPH

'y·
3 H 2H
SCoA - - - - + /' ,)\! 'SCoA
elimination
3H2H

(lR,3R) (16)

H 0
o

~SCOA NADPH
~SCOA'H H
'H
(17)

Fig. 2.6. Stereochemistry of fatty acid biosynthesis (Torssell. 1983; modified and used with permission of the copyright owner, John Wiley and Sons,
Ltd., Chichester. © 1983).

This reductase has two isoforms, one of which is specific doplasmic reticulum which involves the second isozyme of
for NADPH and exists as a homotetramer with a molecular 3-ketoacyl ACP synthase (KAS TI) (Browse and Somerville,
weight of 115,000 to 140,000 (Ohlrogge et aI., 1993). 1991; Lehninger, 1982; Somerville and Browse, 1991). This
To begin the next series of reactions, a malonyl unit is system requires only NADPH and palmitoyl-ACP. Malonyl-
transferred from malonyl-CoA to the phosphopantotheine ACP is the specific C-2 substrate. Paimitoyl-CoA cannot
site of ACP. In a manner similar to the initial condensation, serve as a substrate in this system (Browse and Somerville,
the homolog containing two additional carbons (in this case, 1991; Stumpf, 1976).
butyryl-ACP) reacts with the malonyl unit, displaces CO2 , Both the de novo synthesis and the elongation step (i.e.,
and is reduced in the next three steps of the reaction sequence the systems that involve isozyme I and II of 3-ketoacyl ACP
(Fig. 2.5). Energy is required to provide the NADPH reduc- synthase) occur as ACP-derivatives (Fig. 2.7).
tant and (as ATP) to regenerate the thioester linkage of ace- These chain lengths may be maintained by the combined
tyl-CoA, and for formation of malonyl-CoA (Lehninger, action of the three keto-acyl synthase enzymes (KAS I-III),
1982). The cycle continues until palmitoyl-ACP is produced. and relatively specific thioesterases that convert acyl-ACP
A number of conclusions concerning the stereochemistry molecules to the corresponding CoA derivatives. Most
of the condensation have been established (Fig. 2.6). The hy- plants have an acyl-ACP thioesterase with the highest activ-
droxy acyl intermediate (16) has a (3R)-configuration. Upon ity for oleyl-ACP. Chain termination also may occur when
loss of water, the E- or trans-enoate (17) is preferentially fatty acids are transferred from ACP to glycerol-3-phosphate
formed. As the 'H derivative of (2R,3R)-3-hydroxy-2-'H-bu- by acyltransferases (Ohlrogge et al., 1993) (see below).
tenoate (16) gave trans-butenoate (17) with retention of 'H,
elimination must have occurred in a syn-fashion and the initial Biosynthesis of Unsaturated Fatty Acids
condensation must have proceeded with inversion of configu-
ration (Fig. 2.6) (Simpson, 1984; Torsseli, 1983). Desaturation of stearoyl-ACP (18), which requires the
soluble enzyme stearoyl desaturase, is probably the primary
Tbe Second 3·Ketoacyl ACP Synthase route for the synthesis of oleic acid (3) in .plants (Browse
Isozyme and Somerville, 1991; Ohlrogge et ai., 1993; Somerville and
Palmitoyl-ACP is subsequently elongated to stearoyl- Browse, 1991). Stearoyl desaturase is a homodimer with a
ACP in another biosynthetic system associated with the en- molecular weight of 68,000 (Ohlrogge et ai., 1993). The
22 Fatty Acids

--
KAS·II stearoyl-ACP
KAS·II1
elongation desaturase
C2 + 7C3 C16·ACP~ C1s-ACP - - - + C 18:r ACP
system

thioesterases
.~-j ~-j ~o~j
.wj
16:0 18:0 18:1

thiokinases CoA
.wj "'j CoA CoA

ff
16:0·CoA 18:O-CoA

j j 18:2 oA

metabolic pool: acylation of polar lipids, etc.

Fig. 2.7. The interrelationship between acyl-ACPs and acyl-eoAs in plants (modified from Stumpf, 1976; modified and used with permission of the
copyright owner, Academic Press, Orlando).

enzyme is highly specific for stearoyl·ACP. This aerobic stood, partially because the enzyme systems are membrane
mechanism for desaturation of fatty acids, found in all eukar- bound, which has made them difficult to study (Browse and
yotic cells, involves molecular oxygen and a reductant. Z- Somerville, 1991; Jaworski, 1987). Microsomal membrane
or cis-elimination of two hydrogens occurs to form a Z- preparations from the developing endosperm of castor bean
double-bond system. The oxidative desaturase is associated catalyzed the transfer of oleate from oleyl-CoA to phosphati-
with microsomal or membranous particles and requires dyl choline (Bafor et aI., 1991). There is strong evidence for
NADPH or NADH and O 2 • Cytochrome b5 is the electron the intermediacy of an a- or f3·oleyl phosphatidylcholine
carrier coupling the reductant with the desatorase (Fig. 2.8) intermediate in the formation of a-linoleyl phosphatidylcho·
(Browse and Somerville, 1991). line. Oleyl-CoA is a very effective and highly specific sub-
Lipids from the prokaryotic pathway of both leaves and strate for these same systems. Once linoleic acid (4) is syn-
seeds contain only oleic and palmitic acids when they are thesized, it is further desaturated to a-linolenic acid (5)
first synthesized in the chloroplast or proplastid (Browse and (Stumpf, 1976).
Somerville, 1991). In anaerobic bacteria, unsaturated fatly acids can be syn-
The mechanism for desaturation of oleic acid (3) to lino- thesized in the absence of oxygen (Fig. 2.9), but only monoe-
leic (4) and linolenic acids (5) in plants is not well under- noid fatty acids are produced. It seems likely that a branching

O,INADPH Z
CH,(CH')16COSX CH,(CH,J,CH=CH(CH,),cOSX
stearate (2) oleate (3)

O,INADPH z Z
CH,(CH,J.CH=CHCH,CH=CH(CH,J,COSX

linoleate (3)

O,INADPH z z z
CH,CH,CH=CHCH,CH=CHCH,CH=CH(CH,J,COSX

a-Iinolenate (4)

Fig. 2.8. Desaturation of stearoyl-ACP to oleyl-ACP and linoleyl-ACP.

 Fatty Acids 23

-
H D D
-H,O
SEnz ),6~
'1,) lr _
--,SEn.
K-
o H 0
H+

H>=(,[ _
If X "SEnz
CO,H

H D H D D D D

(3)

Fig. 2.9. Desaturation of stearic acid to oleic acid in a bacterial system (Simpson, 1984; modified and used with pennission of the copyright owner, the
Royal Society of Chemistry. London).

point in the synthesis occurs at C8 or C w and that the desatu- the chloroplast and are converted into palmitoyl and oleoyl-
ration occurs at this stage followed by chain elongation to CoA derivatives, respectively, by the action of thioesterases,
produce acids with 18 carbon atoms (Mann, 1987; Sedgwick, thiokinases, CoA, and ATP in the outer envelope (Browse
1973). Oleic acid (3) synthesized from [2-2H21 malonyl-CoA and Somerville, 1991). The resulting soluble CoA esters are
contained no deuterium at C w . As deuterium at that position used for the synthesis of phosphatidic acid (19) at the endo-
in the precursor is known to occupy the pro-S-position, that plasmic reticulum. However, in contrast to the prokaryotic
deuterium atom must have been abstracted in the fonnation pathway, the acyltransferases of the endoplasmic reticulum
of the olefm_ Further analysis showed that the deuterium at synthesize phosphatidic acid (19) with a C I8 fatty acid at
position C8 was in the pro-2R position and a proton must the sn-2 position.
have been added to the 2-si face (Fig. 2.9) (Gunstone, 1984;
Simpson, 1984). Biosynthesis of Triacylglycerides
The synthesis of triacylglycerides is located in the micro-
The Prokaryotic Pathway
somal fraction of the cell. The fonnation of sn-glycerol 3-
In most higher plants, the first enzyme on the prokaryotic phosphate (19) (Fig. 2.2) from free glycerol and ATP is
pathway, the stromal acyl-ACP:glycerol-3-phosphate acyl- essential for this synthesis. The glycerokinase that catalyzes
transferase is highly specific for oleyl-ACP, whereas the this reaction occurs as a soluble chloroplast enzyme in the
second acylation, catalyzed by a membrane-bound acyl- cell. The thiokinases and the acylating enzyme appear to be
ACP-l : lysophosphatidic acid acyltransferase specifically associated with microsomal particles (Stymne and Stobart,
uses palmityl-ACP to yield 1-0Ieyl-2-palmitylphosphatidic 1987). Chain termination also may occur when fatty acids
acid (Browse and Somerville, 1991). Much of the palmitic are transferted from ACP to glycerol-3-phosphate by acyl-
acid (30-70%) at the sn-2 position ofphosphatidylglycerol transferases (Ohlrogge et al., 1993). There are two enzymes
(9) is converted to the a 3 -isomer (8) (Fig. 2.14). Chloroplast in the plastid that carry out this reaction. One is a soluble
phosphatidylglycerol is synthesized from phosphatidic acid enzyme that prefers oleoyl-ACP as the substrate and trans-
(19) in all higher plants. In C 16,3 plants, a prokaryotic diacyl- fers the oleoyl group specifically to the sn-l-position of glyc-
glycerol pool is fonned by the action of phosphatidic acid erol-3-phosphate. The second enzyme is located on the inner
phosphatase_ In C 18 ,3 plants, only prokaryotic phosphatidyl- membrane of the chloroplast envelope and is specific for
glycerol is found, and a eukaryotic pool of diacylglycerol is palmitoyl-ACP. This enzyme transfers the palmitoyl unit to
used to synthesize chloroplast lipids. The most abundant the sn-2-position of 1-acyl-glycerol-3-phosphate (OhIrogge
chloroplast lipid, monogalactosyldiacylglycerol (10), is et ai., 1993).
fonned from diacylglycerol by action of UDP-galactose: dia- sn-G1ycerol 3-phosphate is acylated by acyl-CoA in two
cylglycerol galactosyltransferase (Browse and Somerville, steps to yield diacylglycerol3-phosphate (phosphatidic acid)
1991). .- (Fig. 2.10), which is dephosphorylated to give diacylglyc-
erols. The synthesis of phosphatidylcholine (23) (Fig. 2.2)
The Eukaryotic Pathway from diacylglycerols (20) by choline phosphotransferase is
reversible and phosphatidylcholine is a direct precursor of
The first committed step of the eukaryotic pathway is the highly unsaturated diacylglycerols in seeds (Browse and
hydrolysis of C I6 acid and C 18 ,I-ACP to the free fatty acids. Somerville, 1991). A specific a l2 fatty acid hydrolyase from
These free fatty acids move through the two membranes of castor bean (Ricinus communis) endospenn converts phos-
24 Fatty Acids

sn-! 0

~H,-O.--lL.R
o i
II i
/'-2
R ~O - C- H triacylglycerol (22)
1 0

!H,-O.--lL. R
~
sn-3

CH,--OH

I
CH,--OH
o ,
R.---lL.o_l_
I
H HO--C--H

0
CH,-o-_.JJII_- R

diacylglycerol (20) monoacylglycerol (21)

Fig. 2.10. Triacylglycerol structure (modified from Stymne and Stobart. 1987; used with pennission of the copyright owner, Academic Press. New
York).

phatidylcholine containing oleic acid at the sn-2 position quired for this process. The small amounts of odd-chain fatty
into ricinoleyl phosphatidylcholine. A specific phospholi- acids found in nature may be formed by this pathway, but
pase hydrolyzes ricinoleyl phosphatidylcholine into phos- a-oxidation is not a major route of metabolism of fatty acid
phatidylcholine and ricinoleic acid (24) (Fig. 2.15). The ri- in most plants.
cinoleic acid is then converted to ricinoleyl-CoA which is, The principal mechanism by which fatty acids are broken
in sequence, converted into diacylglycerols (Bafor et ai., down in plants is (3-oxidation (Fig. 2.11) (Kindl, 1987). On
1991). Diacylglycerols are further acylated in the presence germination, seeds with high oil content form glyoxysomes
of acyl-CoA and diacylglycerol acyltransferase to produce that contain the enzymes of (3-oxidation. In this process,
triacylglycerides (25) (Bafor et al., 1991; Stumpf, 1976; much of the energy stored in the lipids is converted to acetyl-
Stymne and Stobart, 1987). In the system from castor bean CoA and is trapped in the thioester bond. Acetyl-CoA then
endosperm, the enzymes exhibited selectivity for ricinoleyl- enters the tricarboxylic acid cycle (TCA). (3-0xidation in
containing diacylglycerols (Bafor et al., 1991). In instances plants appears to be identical to that of animals.
where the three acyl substituents differ, the product usually Palmitoleic (13) and similar acids may arise by (3-oxida-
is optically active, and specificity as to the position of attach- tion of oleic acid (3).
ment of particular fatty acids is exhibited (Fig. 2.10) (Good- The Z,z-methylene-interrupted system oflinoleic (4) and
win and Mercer, 1983; Styrnne and Stobart, 1987). linolenic acid (5) is readily susceptible to attack by lipoxy-
genase. This enzyme catalyzes the direct addition of oxygen
to the system with the formation of a Z,E-I,3-butadiene hy-
DEGRADATION OF FArrY ACIDS droperoxide. Lipoxygenase has been found widely in plant
seeds and leaf tissues. This enzyme serves as a starting point
for the breakdown of multiply unsaturated fatty acids and
When a seed germinates, lipids disappear and sugars appear
gives rise to a host of compounds (Goodwin and Mercer,
rapidly. Lipase activity rises sharply and participates in the
1983; Hamberg and Gardner, 1992; Stumpf, 1976).
stepwise hydrolysis of triacylglycerols to diacylglycerols
and monoacylglycerols, ultimately leading to glycerol and
free fatty acids. Once fatty acids are formed by lipase action
(Huang, 1987), they may be oxidized by several mecha- Ul'IUSVAL FArrY ACIDS Il'I PLA1"ITS
nisms, the principal being a- and (3-oxidation (Kindl, 1987;
Stumpf, 1976). Most plants have the standard complement of stearic, palmi-
Free fatty acids from C I2 to CIS may be attacked readily tic, oleic, linoleic, and linolenic acids, a certain number of
to yield either a fatty acid with one less carbon atom and lower-molecular-weight acids (lauric, myristic, palmi-
CO2, or a n-a-hydroxy fatty acid. Molecular oxygen is re- tooleic), and occasional higher-molecular-weight homologs
 Fatty Acids 25

~\o
R.CH,.CH,.CO,H ATP

\\OCOASH
II
t-
CO,
~ CH3·C·SCoA
II
R·CH,·C·SCoA
+0

II
several repetitions
(d;!n)lt
R·C·SCoA
sucrose COAS~ ,,~ 0
H
R·G=CH·C·SCoA
II
" "
R·C·CH,·C·SCoA H,OJ
/ )

~
·2H R.CHOH.CH,.C.SCoA
II
(pyridine nucleotide)
Fig. 2.11. The j)Moxidation cycle in plants (Stumpf, 1976; modified and used with pennission of the copyright owner, Academic Press, New York).

such as arachidic acid (25), (Fig. 2.1). These acids all are prise more than 50% of the total fatty acids of the oils. Coco·
primary metabolites. nut oil contains high percentages of hexanoic, octanoic, and
Some groups of plants accumulate unusual quantities of decanoic (capric) acids. Most other fatty acids have limited
primary fatty acids. For example, myristic acid (C I4 ) is distribution among plants (Harwood, 1980; Smith, 1970).
found in the seeds of Myristica fragrans (nutmeg) as a com· Unusual fatty acids typically are found in seed glycerides
ponent of the triacylglycerol, trimyristicin. Plant lipids often (often as major components) along with common fatty acids.
contain small amounts of fatty acids that differ from common
primary fatty acids only in chain length. Usually, these com· Fatty Acids from Unusual Starter Units
pounds represent a small percentage of the total fatty acid
content and are overlooked or ignored in analyses, but they In certain cases, starter molecules other than acetyl·CoA
may be major constituents. For example, C l2 (6) and C l4 are utilized. Small amounts of isa· and anteisa-fatty acids
(7) fatty acids occur in large quantities in the seeds of many which co-occur with normal fatty acids in most plants arise
groups of palms (Arecaceae), and these two acids may com· from precursors (26) and (27), respectively (Fig. 2.12). Fatty

JySCOA

(3) (4)

CO,H

an isofatty acid

CO,H
an anteisofatty acid

CO,H

a fatty acid based on a cyciohexylMCoA starter unit

Fig. 2.12. Fatty acids derived from iso, anteiso, and cyc10hexyl starter units.
26 Fatty Acids

acids with highly restricted distribution are found in the ther- likely, an intermediate in the lipoxygenase-type reaction
mophilic prokaryotic organism Bacillus acidocaldarius, in might stabilize in a number of ways, each leading to the
which the CoA derivative of cyclohexylcarboxylic acid is accumulation of a diene, dienol, or triene. Thus, 9Z, I 2Z-
an acceptable starting unit. In a group of closely related linoleic acid (4) gives either coriolic acid (l3-hydroxy, 9Z,
higher plants, cyclopentenoid fatty acids are synthesized liE) (38) or a-dirnorphecolic acid (9-hydroxy, JOE, 12Z)
from a cyclopentenecarboxylic acid starting unit (aleprolic (37), the 12Z-family (the 12Z-compounds of 31 and 35), and
acid) (see Fig. 2.18 and discussion below). the 9Z-derivatives of 33, 34, and 36. The 9Z, 12E-isomer of
linoleic acid yields the 12E-compounds of 31, 32 and 35
Fatty Acids with Unusual Patterns of and the 9Z, 13E-compounds 34 and 36, whereas 9E, 12E-
Unsaturation linoleate gives rise to 12E-compounds of 31, 32, and 35,
Chloroplasts from photosynthetic tissues of higher plants and the 9E-forms of 33,34, and 36. Significantly, the inter-
and algae contain E-3-hexadecenoate (8) (Fig. 2.13), an unu- mediates postolated occur in the same tissues in many cases.
sual palmitate-derived fatty acid. Desaturation of the 3-posi- The production of isomers with E-double-bonds remains
tion requires light and may occur after the palmitic acid unexplained (Hitchcock and Nichols, 1971).
precursor is attached to the acyl moiety. This acid is found
almost entirely at the 2-position of phosphatidyl glycerol (9) Hydroxy Fatty Acids
(Fig. 2.2).
Fatty acids with unsaturation in positions other than the Hydroxy fatty acids appear to derive from multiple routes
characteristic 9,1 O-position probably arise, in most in- (Butt and Lamb, 1981). Ricinoleic acid (24) (Fig. 2.15), for
stances, from modified A-9 desatorases. Petroselinic acid example, arises in the embryo (to a lesser extent in the endos-
(28), a C'8-monounsatorated acid with the double bond in the perm) of the castor bean, Ricinus communis. Synthesis be-
A6-position, from developing endosperm of carrot, Daucus gins about 12 days after pollination and ceases about 6 weeks
carota, and coriander, Coriandrum sativum (Apiaceae), after flowering. At that time, ricinoleic acid accounts for
comprised 70-75 mol% of the fatty acids of the triacylglyc- about 90% of the fatty acids in the triacylglycerols. Proplas-
erols, but only about JO mo1% of the phosphatidylcholine tids isolated from developing castor bean endosperm synthe-
(23) and phosphatidylethanolamine (29) (Fig. 2.2) fractions size only oleic acid (3), although the major product of fatty
(Cahoon and Ohlrogge, 1994). However, introduction of la- acid synthesis in these seeds is triricinoleoylglycerol. Oleic
beled acetate indicated that there was rapid incorporation acid is exported from the plastid, where it is synthesized to
into phosphatidylcholine and that most of this was in the the cytosol and activated to oleyl-CoA. An acyl-CoA: Iyso-
form of petroselenic acid. In time course stodies, the radiola- phosphotidylcholine acyltransferase transfers the oleate
bel initially entered phosphatidylcholine at the highest rates, from oleyl-CoA to Iyso-phosphatidylcholine to form oleyl
but later accumulated in triacylglycerols. Petroselenic acid, phosphatidylcholine. A specific A12-fatty acid hydroxylase
in this instance, was readily incorporated into both the sn- catalyzes the conversion of oleyl phosphatidylcholine into
1 and sn-2 positions of triacylglycerols (Cahoon and Ohl- ricinoleyl phosphatidyIcholine, mainly at position sn-2 (see
rogge, 1994). The coding of a cDNA for this desatorase has triglycerol synthesis above) (Baforetal., 1991). Synthesis of
about 65% sequence identity with the A9 -desaturase (Ohl- ricinoleic acid (24) involves microsomal action (presumably
rogge et aI., 1993). endoplasmic reticulum) that hydroxylates oleyl-CoA to ri-
Conjugated ethylenic acids (31-38) may arise from lino- cinoleyl-CoA; this product is then channeled into triacyl-
leic acid (18: 2,9Z,I2Z) and its Z,E- or E,E-isomers (Fig. glycerides and storage oil droplets (Stompf, 1980). The en-
2.14) (Hitchcock and Nichols, 1971). The two hydroxy zyme requires NADH and molecular oxygen. Oleyl-CoA
dienes (37 and 38) could arise directly from linoleic acid (or was the only reactive substrate (Stompf, 1976); notably pal-
its isomers) and give rise to the conjugated dienes, or more mitate, stearate, or linoleate do not serve as precursors. Re-
tention of label in the carboxyl group of ricinoleate demon-
strated that the compound was not made by breakdown and
resynthesis. In contrast, ricinoleic acid in ergot oil (produced
H H by the fungus Claviceps) apparently arises by hydration of
the double bond of linoleic acid.
12,13-Dihydroxyoleic acid (30) (Fig. 2.15) is formed
from vemolic acid (39) (which, in tom, is formed from lino-
petroselenicacld(l8}
leic acid) in crushed seeds of Xeranthemum annuum (As-
teraceae) and Euphorbia lagascae (Euphorbiaceae).
Laetisaric acid (9Z,I2Z,8-hydroxyoctadecadienoic acid)
(40) from the soil-dwelling fungus Laetisaria arvalis has
allelopathic activity and inhibits growth of the major root
{E).3.hexedeeeno!e acid (8)
pathogen Pythium ultimum (Bowers et aI., 1986). An anti-
Fig. 2.13. (E)·3-Hexadecenoic acid and petroselenic acid. fungal compound from taro, Colocasia antiquorum, infected
 Fatty Acids 27

?H
t
ZIE E
- CH= CH- CH= CH- CH- CH,- dlmorphecolic acid
• 12 1" • (9·hydroxy,10E,12Z) (37)

OOH
ZIE E I
-CH= CH- CH=CH-CH- CH,-

ZlE E Z
-CH= CH- CH=CH- CH=CH-
ZIE E E
-CH=CH- CH=CH- CH=CH-
"
(35)
\
--CH==CH--CH==CH--C--CH--
. / (31)

/ ZIE Z
-CH=CH-CH=CH-CH-CH-
\
ZIE ZIE (32)
-CH= CH-CH,-CH=CH-CH,-
Il 12 It 10 !II 8
linoleic acid (4)
E ZIE
(18:2,9Z,I2Z) -CH-CH--CH=CH-CH=CH-
:~ (33)

• E ZlE
--CH--C--CH==CH--CH==CH--

~~L~'~'~~\
-CH";'C-C~CH-CH~CH-
(36) ~.
OH
I -CH~C-C~CH-CH~ECH- •~ (34)

_ CH1-
I
C_
E ZIE
CH=CH- CH=CH-
I I'
cor 0 IC aCI
'd (38)
Il It !II (13·hydroxy,9Z,11E)

Fig. 2.14. Possible mechanism of biosynthesis of conjugated fatty acids.

with the pathogen Ceratocystis Jimbriata proved to be Methyl·BranchecL Cyc:lopropanoicL and

9,12, 13·trihydroxy-(E)-1 O-octadecenoic acid. Cyc:lopropenoid Fatty Acids

Epoxy Fatty Acids Several naturally occurring fatty acids contain additional
methyl groups andlor cyclopropyl rings. In some cases,
Epoxy fatty acids also are found as components of triglyc-
cyclopropyl fatty acids arise from starter units such as propi-
erides of the seed oils (Smith, 1970). This unusual fatty acid
onyl-CoA, but in others, the extra atoms are derived from
type is sporadically distributed in species of several taxo-
nomically unrelated plant families. 2-12, 13-Epoxyoleic acid S-adenosylmethionine (Buist and Findlay, 1985). DeRosa et
(vemolic acid) (39) (Fig. 2.15) is biosynthesized from lino- al. (1974) suggested that tuberculostearic acid (41) is fonned
leic acid in the seeds of Xeranthemum annuum and Euphor· from oleic acid and S-adenosylmethionine (Fig. 2.16). Cy-
bia lagascae (Hitchcock and Nichols, 1971). This compound clopropane containing fatty acids are probably fonned by a
and 2-9, lO-epoxystearate are synthesized by the introduction related process. Lactobacillic acid (42) and tuberculostearic
of oxygen from O2 across the double bonds of linoleic and acid are both known from bacteria; other cyclopropanoid
oleic acids, respectively (Butt and Lamb, 1981; Morris, and cyclopropenoid fatty acids also are known from higher
1970). The biosynthesis of epoxy fatty acids is related to plants. Sterculic acid (43) is fonned by oxidation of the
that of dihydroxy fatty acids, and epoxy fatty acids may cyclopropanoid fatty acid (44) derived from oleic acid
serve as precursors for the latter type of compound. (Mann, 1987). Cyclopropenoid fatty acids are found com-
28 Fatty Acids

H H

CO,H

ricinoleic acid (24) vaccenic acid (64)

H H
CO,H

lactobaclllic acid (42)

vernollc add (39)
R CO,H

sterculic acid (43)

OH
12,13 . cUbydroxyoielc acid (30)
CO,H
OH OH
dihydrosterculic acid

OR
laellsaric acid (40) CO,H
Fig. 2.1S. Selected hydroxy fatty acids.

monly in seed oils of the families Malvaceae, Stercuiiaceae,

and Bombacaceae (Fig. 2.17).

Cydopentenoid Fatty Adds

The nonprotein amino acid cyclopentenylglycine (45) is sterculynic acid
a constitnent of the free-amino-acid pool in seeds and leaves Fig. 2.17. Cyclopropanoid and cyclopropenoid fatty acids from plants.
of Hydnocarpus anthelminthica and Caloncoba echinata
(FJacourtiaceae), where it comprises up to 2% of the total
free amino acids. This unusnal compound is derived via the
pathway presented in Fig. 2.18. This amino acid is then con-
verted to cyclopentenylcarboxylic acid (aleprolic acid) (46)

CO,H
CO,H

sterculic acid (43)

tuberculostearic acid (41)

Fig. 2.16. Proposed biosynthesis of methyl, cyc1opropenyl, and cyclopropyl fatty acids in bacteria and in plants.
 Fatty Acids 29

a-ketoglutaric
acid
r~H3COSCOA
(

CO,"
CO,

""::::">-"","--,,.c!::...
f"
CO,"

T" --
CO,"

(45)

CO,H

aleprolic acid (46)

chaulmoogric acid (47)

CO,H

hydnocarpic acid H H

CO,H

Fig. 2.18. Biosynthesis of cyclopentenoid fatty acids (modified from Mangold and Spener, 1980; used with permission of Academic Press, Orlando,
FL).

which serves as a starter unit for the synthesis of cyclopen- chapetalum toxicarium (Dichapetalaceae) in west Africa
tenoid fatty acids such as chaulmoogric acid (47) and gorlic (Sierra Leone) (Meyer and O'Hagan, 1992; Smith, 1970).
acid (48), found in the family Flacourtiaceae. Cyc!openten- This shrub is known to be highly toxic to livestock; it contains
oid fatty acids also have been reported from red algae (Mira- w-fluorooctadec-Z-9-enoic acid, w-fluorooleic acid (49), and
lie et aI., 1990). The cyclopentenoid unit in higher plants w-fluoropa1mitic acids (Fig. 2.20). Although even-numbered
apparently is synthesized by the following pathway (Fig. compounds are highly toxic (they are metabolized to fluoroa"
2.18) (Mangold and Spener, 1980). cetic acid), odd-numbered chains are much less so.
Fatty acids with structures resembling those of prosta- Fluoroacetate is found in at least 34 species of Gastrolob-
glandins have been isolated from Lemna species (Fig. 2.19) ium and Oxylobium (Fabaceae) in Australia, and occurs to
(Monaco and Previtera, 1987; Previtera and Monaco, 1987). the extent of 0.25% of the fresh weight. Bush rats and rat
kangaroos in the areas where these plants grow are resistant
F1uorofatty Acids to poisoning, whereas those from other areas are not (Har-
A number of w-fluorofatty acids are known to occur in borne, 1986).
seeds of species of the genus Dichapetalum, especially in Di- The caterpillars of Sindris albimaculatus, which consume
30 Fatty Acids

U U CU,
U I
~
o ~
-J(CU,).

~
o
ou uo ouol
(12S)-hydroxyhexadeca-SZ,IOE,l4Z-trienoic acid
U H (CH,),

OH I
CO,H

E
co,U lipid from Ipomoea parasitlca seeds (51)

O_galaciOSYI

Fig. 2.19. Prostaglandin-like fatty acids from Lemna species.

O-galactosyl

the flowers, fruits, and young leaves of Dichapetalum cymo- O.C16,C18 fatty acids
sum, accumulate the toxin fluoroacetate as a defense against
predators (Meyer and O'Hagan, 1992).
(SO)

other Unusual Upids Fig. 2.21. Glycolipids from Briza spicata and Ipomoea parasitica
seeds.
Although glyceride structures with sugar substitutions,
especially galactoglycerols, are not uncommonly encoun-
and roots. To cite but one example, the flavor of Bartlett
tered as components of plant leaf lipids, a number of grasses
pears (44) (Fig. 2.22) is due in part to esters. Analysis of
accumulate digalactosylglycerols in their seeds (Joyard and
Douce, 1987). These compounds are partly responsible for these metabolically altered fatty acids is usually effected by
GC/MS (gas chromatography/mass spectruscopy) methods
the physical properties of flours and make them useful for
(Buttery et aI., 1987; Kameoka, 1986).
breadmaking. Compounds of this type have been isolated
from Briza spicata (50) (Fig. 2.21) (Smith and Wolff, 1996). Essential oils often are important as synomones (com-
In seeds of some species of the Convolvulaceae, normal pounds that are of benefit to both the sending and receiving
organism) in mutualistic relationships between plants and
glycerides are replaced by glycolipids (e.g., 51) (Fig. 2.21)
(Smith et aI., 1964). 6-D-Glucopyranosyl esters of palmitic, animals (especially insects). Synomones are especially im-
oleic, linoleic, and linolenic acids occur in rape, Brassica portant for pollination and fruit and seed dissemination.
napus, pollen (Grove et aI., 1978). The volatile resin exuded from the stem bark of Commi-
phora rostrata (Burseraceae) consists mainly of 2-decanone,
2-undecanone, 2-dodecanone, and hexadecanal. This sub-
stance seems to be involved in limiting herbivory on this
METABOLICALLY ALTERED FATTY ACIDS small tree. When branches or twigs are bent or cut, a resin
is released that rapidly covers the area around the injury
Metabolically altered fatty acids (i.e., fatty acids that have (McDowell et aI., 1988).
been reduced in chain length by (X- and/or f3-oxidation and The wild tomato, Lycopersicon hirsutum f. glabratum
have undergone various processes of oxidation or reduction) (Solanaceae), is covered with trichomes which contain 2-
occur in many essential oils. Many of these intermediary tridecanone (53) (Williams et aI., 1980). The level of this
metabolites are converted ultimately to esters, acids, alco- compound is much lower in the domesticated tomato, L.
hols, aldehydes, alkenes, and alkanes (Fig. 2.22). These fatty esculentum. The exudate is toxic to Manduca sexta (tobacco
acid derivatives are relatively volatile and occur widely as homworm) and to Helicoverpa zea (the cotton bud worm).
components of the essences of flowers, fruits, stems, leaves, The density of glandular trichomes was influenced by an
interaction between day length and light intensity; the toxic
U U
plant compound was significantly more abundant on foliage
of plants grown under long-day regimes. This fmding is of
considerable importance, as there is a possibility of intruduc-
F
ing this resistance into acceptable tomato cultivars (Kennedy
et aI., 1981; Williams et aI., 1980; Zamir et aI., 1984). The
sesquiterpene zingiberene was found in Lycopersicon hirsu-
m·Ouorooleic acid (49)
tum f. hirsutum and its presence coincided with resistance
Fig. 2.20. ro-Fluorooleic acid. to the Colorado potato beetle (Carter et aI., 1989).
 Fatty Acids 31

~ ~O~
o
3·octanone octyl acetate

~OH~CHO

I-octano) octanal

~~OH
2-undecanone I-dodecanol

2-tridecan.ne (53) IO-undecen·l·o1

H H

~COlCH3.rClH'-
(52)

Fig. 2.22. Metabolically altered fatty acid derivatives.

Plant-derived compounds such as (3Z)-hexenyl acetate, the strongest responses were obtained for the monoenoic C.
(3Z)-hexen-I-ol, (2E)-hexenal, (3E)-hexenal, 4-methyl-3- alcohols and aldehydes which make up much of the green
heptanol, (llZ)-hexadecenal, (9Z)-hexadecenal, and (7Z)- leaf volatile complex of many plants (Light et al., 1988).
hexadecenal are used by parasitoids to locate the host habitat Humans respond in a similar way; oct-l-ene-3-01 and (3Z)-
(Elzen et al., 1984; Kennedy, 1984; Turlings et al., 1991; hexenol contribute much of the characteristically pleasant
Wickremasighe and Emden, 1992). (2E)-Hexenal was "green odor" to the fruits of chayote (Sechium edule, Cu-
strongly inhibitory to tomato seed germination at concentra- curbitaceae) (MacLeod, 1990).
tions as low as 6.9 fJM (Bradow and Connick, 1990). Compounds such as E-2-hexen-I-ol, I-hexanol, Z-3-
hexen-I-ol, and E-2-hexenol and linalool (a monoterpene)
Tbe Green Leaf Volatile Complex are involved in the olfactory orientation of the Colorado
potato beetle, Leptinotarsa decimlineata, to the foliage of the
The leafy or grass-like smell of most green leaves is due potato, Solanum tuberosum (Solanaceae). However, none of
to the presence of a series of aldehydes and alcohols derived the individual components alone was attractive; the beetle
by oxidative degradation of fatty acids. (see Fig. 2.23.) The was attracted only to a mixture similar to that of the host
relative proportion of these compounds varies among plant plant (Visser and Ave, 1978; Visser et al., 1979). A similar
species and often seasonally within the same species. The complement of compounds seems to be involved in attrac-
composition of mixtures of these compounds also is affected tion of the alfalfa seed chalcid (Bruchophagus roddi) to al-
by aging and injury. falfa (Medicago sativa, Fabaceae) (Buttery and Karnm,
In electroantennogram responses of the Mediterranean 1980).
fruit fly, Ceratitis capitata, to a spectrum of plant volatiles, Whereas undamaged plants emit low levels of green leaf
32 Fatty Acids

acyl hydrolases
leaf lipids - - - - - -•., free fatty acids - - - - . linoleic acid (18:2)

.. oxidation
oxidatio,:--- linolenic acid (18:3) /
¥' ~CHO
~CHO isomerase

3Z·hexenal ---------... ~CHO hexanal

i 2E-hexenal ~CH,OH

3Z-hexenol
I-hexanol

,
~CH,OH

~CH,OH 2E-hexenol ' ...

~CH,OH
3E-hexenol 2-Z-hexenol

Fig. 2.23. Proposed biosynthesis of green leaf volatile complex.

volatiles, caterpillar-damaged plants emit higher levels. Fe- cell extension activity in the parenchymatous cells of the
male braconid parasitoids, Micropiitis croceipes and female bean pod mesocarp (Mandava, 1979).
ichneumonid parasitoids, Netelia heroica, oriented to the Modified fatty acids (such as 54 and 55) containing cyclo-
compounds and damaged plants in a wind tunnel, but seldom pentenoid rings have been isolated from the alga Eleocharis
to undamaged plants, suggesting that these parasitoids may mierocarpa and have been suggested to be involved in allelo-
orient toward plant damage to locate their caterpillar hosts pathic interactions with other aquatic organisms (Fig. 2.25)
(Whitman and Eller, 1990). (van Aller et a1., 1985). A series of structurally similar com-
pounds that have pheromonal properties is known from
brown algae (Fig. 2.24) (Jaenicke, 1974, 1991). In addition
to pheromonal properties, a mixture of two of these com-
f'Ul'ICTIONS OF FATTY ACIDS Al'ID THEIR pounds, dictyopterene A (50) and B (51), inhibits feeding
DERIVATIVES Il'I PLANTS on the brown alga, Dictyopteris de/ieulata, by reef fish (Hay
et a1., 1988). These substances also may indirectly protect
Many functions of fatty acids in plants have been reviewed small marine herbivores that fed on the algae.
(Fuller and Nes, 1987). The most important role of fatty
acids is as a component of energy storage systems in plant Jasmonic Acid and Related Compounds
seeds and other lipid-accumulating organs and as compo-
nents of plant membranes. Upon germination of the seed, Perhaps one of the greatest surprises in recent years is
fatty acids are usually broken down to smaller compounds the wide range of physiological activities in plants induced
and utilized by the plant. On a percent weight basis, glycer- by the presence of jasmonic acid (59) and structurally related
ides are the most efficient energy storage material commonly compounds (Fig. 2.25). Jasmonic acid and the corresponding
used by plants. Most plants store energy as either glycerides methyl ester are fragrant constituents of the essential oils of
or carbohydrates, but few produce large quantities of both. fasminum spp. (Oleaceae) as well as other perfumery plants
Seed lipids provide an excellent source of food not only for (Sembdner and Parthier, 1993). These compounds occur in a
plant embryos, but also for a variety of animals, fungi, and large variety of plant species. Jasmonic acid and its relatives
bacteria. Thus, there has been evolutionary selection to pro- possess structural similarities to prostaglandins from animals
tect seeds from predation by several mechanisms including (Fukui et aI., 1977a,b; Gross, 1980).
accumulation of toxic secondary metabolites in the seeds Jasmonic acid is derived from a-linolenic acid, 13(S)-
(Levin, 1974). In some instances, the poisonous materials hydroperoxylinolenic acid, 12-oxophytodienoic acid, and
are modified fatty acids stored in the seeds. These com- phytodienoic acid (Farmer and Ryan, 1990, 1992; Hamberg
pounds are later catabolized when needed as energy or nu- and Gardner, 1992). The biosynthesis of jasmonic acid and
trient reserves. related compounds has been reviewed (Sembdner and Par-
A wound hormone in plants, traumatin or l-decene-1, 10- thier, 1993).
dicarboxylic acid, was first isolated from bean pods. This (- )-Jasmonic acid (59) and its methyl ester are wide-
substance is capable of inducing renewed cell division and spread among plants and have demonstrated growth-inhibit-
 Fatty Adds 33

ing capabilities in plants (Dathe et aI., 1991) (Fig. 2.24). A seems to be involved with tuber formation in both yam (Di-
structurally similar group of compounds including cucurbic oscorea) and potato (Hamberg and Gardner, 1992). Jas-
acid has been isolated from pumpkin (Cucurbita pepo, Cu- monic acid may be active in the process of tendril coiling in
curbitaceae) and other plants (Dathe et aI., 1991). Cucurbic some lianas (Hamberg and Gardner, 1992; Staswick, 1992).
acid, (lS,2S,3S)-3-hydroxy-2-(2'-cis-pentenyl) cyclopen- When methyl jasmonate was applied to the surface of
tane-I-acetic acid (60), and the co-occuring 3-0-f3-D-gluco- tomato plants, the synthesis of a defensive proteinase inhibi-
side and methyl ester possess growth inhibiting ability in tor protein was induced, not only in the plant to which the
rice, but lack activity in Avena coleoptile bioassays. application was made but also in nearby plants. In subse-
The phytohormone effects of jasmonic acid are similar quent studies, this compound was shown to have a similar
to those of abscisic acid, an established phytohormone; it effect with plants of other families as well (Farmer and Ryan,
has been argued that jasmonic acid should be recognized as a 1990; Pearce et aI., 1991). In a series of experiments, it was
representative of a unique class of phytohormones (Hamberg demonstrated that methyl jasmonate can be a component of
and Gardner, 1992; Sembdner and Parthier, 1993; Staswick, interplant communication systems. Octadecanoid precursors
1992). Jasmonic acid is more effective than abscisic acid in of jasmonic acid were able to activate the synthesis of
promoting senescence, induces the expression of a specific wound-inducible proteinase inhibitors (Crombie and Mistry,
set of proteins (jips or jasmonate-induced proteins) in differ- 1991; Farmer and Ryan, 1992). Jasmonic acid is a signal
ent plants (Sembdner and Parthier, 1993; Staswick, 1992), transducer in elicitor-induced plant cultures of Rauvolfia
and, in some instances, may be involved with the induction canescens and Eschscholtzia californica. Tissue cultures of
of the formation of seed storage proteins. This compound 36 species of plants could be induced to accumulate second-

OH

linolenic acid
~-~
OH ~ctor fucoserraiene

O2 COz,HzO
OH
(2ATPlMgH)
~CO,H_""-",,,-_....

opP

(a) ~ • \.
V
~
~
/='- --:. . . . -
~~

Ov_~ __ ~,==

~~g: dictyopterene B (51)

dictyopterene A (SO)
(+)-ectocarpene

~Al
H~
(b) (+homosirene

Fig. 2.24(a & b). Patty acid-derived phennones from brown algae (Modified from Jaenicke, 1974; used with pennission of the copyright owner,
Springer-Verlag, Berlin),
34 Fatty Acids

Methyl jasmonate is a pheromone produced by the male

~CO'H H~~ oriental fruit moth (Staswick, 1992).

~
o
~C01H The Requirement for Unsaturated Fatty
Acids In the Diet of Animals
Although most plants appear to have the ability to make
jasmonic acid (59) cdcurbic acid (60)
linoleic and linoleic acids, animals cannot desaturate posi-
OH tions from C-9 to the methyl terminus of the fatty acid chain,

~CO'H
and must obtain unsaturated fatty acids such as linoleic acid
and linolenic acid from their diet. The requirement for these

~
acids involves formation of arachidonic acid (61) and, subse-
(55) quently, prostaglandins. Arachidonic acid, eicosatetra-
o 5,8,1l,14-enoic acid, is formed from Iinoleate by chain ex-
tension and by additional desaturation. Prostaglandins (such
as 62 and 63) are derived from eicosatrienoic and eicosate-
traenoic acids, respectively, although the actual path of bio-
synthesis is not completely elucidated (Fig. 2.26). These
compounds have pronounced hormonal and other effects on
o mammals and other organisms.
A range of prostaglandins have been reported from the
Fig. 2.25. Cucurbic, jasmonic acid. and algal cyclopentanoid fatty leaves of onions (Allium cepa, Liliaceae) and other plants
acids. (A1-Nagdy et aI., 1986; Levinet al., 1988). Compounds from
Bryonia (Cucurbitaceae) are not structurally similar but have
ary metabolites by the presence of methyl jasmonate (Gun- prostaglandin-like activity in animals (Fig. 2.27) (Panossian
dlach et aI., 1992). et al., 1983).
When cotton seedlings are infested by herbivorous mites,
volatile signals are released that attract additional mites. Effects of Fatty Acids and Their Derivatives
Downwind-uninfested plants are made more resistant to mite on Animals
attack by contact with these volatile compounds; this system The effect of many toxic secondary metabolites is proba-
also seems to involve methyljasmonate (Bruin et al., 1992). bly to prevent or limit feeding rather than to kill the herbivore

HO

HO
~ /
HO
I
arichidonic acid (61) PGF,a(62)
HO

~H ~COzli
l(~~
/ i
HO
I
HO H~
thromboxane A1
PGF",(63)
OH
HO I
\ !

~~ j ! HO 0 I
HO HA H~
thromboxane 8 1

Fig. 2.26. Representative prostaglandins.

 C
Fatty Acids 3S

OlH of lauric acid and glycerol) is the most potent compound in

~COlH
OH this regard (Kabara, 1987; Masui et aI., 1989).

s:x
Although similar in structure to common fatty acids, eru-
HO A _ HO ~ cic acid (Z-13-docosenoic acid) (66), when fed to rats at as
OH little as 15% of their caloric intake, produced myocarditis

C
OH (accumulation of lipids in the heart) (Fig. 2.28). Eventually,
myocardial fibrosis and other abnormalities were observed.
CO,H
COlH
Because rapeseed oil was widely used for margarine produc-
tion, a breeding program was initiated to produce new vari-
OH eties in which most or all of the erucic acid is replaced with
HO 6 OH
oleic acid. This new product is called canola oil. High erucic
HO
HO rapeseed oil is still used to prepare lubricants (Simpson and
OH
Conner-Ogorzaly, 1986). Both types are cultivated.
Fig. 2.27. Compounds from Bryonia alba (Curcubitaceae) with A number of mammalian and fungal enzyme systems,
prostaglandin activity.
whole cells, and entire animals have been shown to respond
differently to fatty acids that differ in number and position
outright, although many compounds could certainly accom- of double bonds, geometric isomers, or cyclopropyl ring po-
plish the latter, if enough were consumed. Seeds may be sitions (Seigler, 1983). Few olefinic or acetylenic fatty acids
made "distasteful" or "non-nutritious" by one mechanism are highly toxic, although there are large variations in the
or another. ease of utilization by different organisms.
Triglycerides from most plants are edible to most animals. Most unsaturated fatty acids contain Z- or (cis )-double-
In humans, they have mild laxative properties, especially bonds. When fatty acids with E- (or trans-) double-bonds
when consumed in large amounts. Glycerides have been are fed to rats along with adequate amounts of essential fatty
shown in at least two cases to serve as compounds that elicit acids, E- or trans-fatty acids have little effect on growth,
aggregation in insects. l-Palrnitoyl-2,3-diolein, 2-linoleoyl- longevity, or reproduction, but when fed as the sole source
1,3-dipalmitin, and I-palmitoyl-2-linoleoyl-3-0Iein were of lipids, E-fatty acids exaggerate the symptoms of fatty
identified in three active fractions from wheat germ that elicit acid deficiency. An effect on the metabolism of long-chain
aggregation of the confused flour beetle (Seigler, 1979; Ta- polyunsaturated acids was noted, however (Alfin-Slater and
mald et al., 1971). Similarly, 1,3-diolein was shown to "at- Aftergood, 1979; Seigler, 1983).
tract" houseflies (Musca domestica) to the fruiting bodies A pentane extract of tung (Aleurites fordii, Euphor-
of Amanita muscaria, and similar fractions were shown to biaceae) that contained a-eleostearic acid [(9Z,l1E,13E)-
"attract" houseflies to the fruiting bodies of other mush- octadecatrienoic acid] (67) and erythro-9,1O-dihydroxy-l-
rooms of the Tricholomataceae and Amanitaceae (Seigler, octadecanyl acetate (acetate of 68) was shown to be strongly
1979). 1,2-Dioleins are the major attractants for ants in elaio- repellent to Anthonomis grandis, the boll weevil, as well as
somes of a number of ant-dispersed seeds in the northeastern other insects (Jacobson et al., 1981).
United States. (Kusmenoglu et aI., 1989). Castor oil, which contains up to 90% ricinoleic acid (24)
Fatty acids are known to be feeding stimulants for certain (Fig. 2.15), possesses cathartic activity in humans, but not
insects. Linolenic acid (as the free fatty acid) in mulberry in rats.
leaves stimulates leaves potent feeding activity in Bombyx Several esters of hydroxy fatty acids (such as 3-acetoxy
mori, the silkworm. Linoleate and laurate also have activity, fatty acids) are involved in mutualistic relationships of some
whereas myristate, palmitate, stearate, elaidate, oleate, and plants (such as those of the genus Krameria, Kramericeae)
vaccenate (64) (Fig. 2.16) have none. Oleate promotes and their pollinating species. o-3-Hydroxydecanoic acid has
growth, but not feeding activity. A synergistic effect of (3- fungicidal properties (Seigler, 1983).
sitosterol has been observed with linolenate, linoleate, vac- Cyclopropenoid fatty acids (Fig. 2.17) inhibit fatty acid
cenate, and laurate. Linoleic acid and linolenic acids are desaturation in several species, including chicken, rat, pig,
phagostimulants for the fire ant, Solenopsis saevissima var. cow, and trout, causing a rise in the stearate to oleate ratio.
richter;' Palmitate and stearate produce a positive feeding This may have an effect on the composition and function of
response in the hide beetle (Dermestes maculata) and valeric membranes as variation in lipid composition is known to
acid (71) (Fig. 2.29) in the Khapra beetle (Trogoderma gra- alter permeability of membrane systems. Numerous types
narium). Fatty acids (CS-C ll ) are repellent to the red flour of deleterious effects from consumption of seed oils with
beetle (Tribolium castaneum). Honey bees are attracted to cyclopropenoid fatty acids have been observed (Seigler,
clovers by the action of the triene (65) (Mann, 1987). 1979). A cyclopropene fatty acid from Sterculia foetida oil
was lethal to the larvae of Callosobruchus maculatus at 0.1 %
Fatty Acids with Biological Activity in artificial diets (Janzen et al., 1977). The most important
The antifungal and antibiotic properties of fatty acids and edible oil with these compounds is cottonseed oil from Gos-
some derivatives have been studied; monolaurin (the triester sypium hirsutum (Malvaceae). Much of the cyclopropenoid
36 Fatty Acids

~ CO,H

(65)

(X-eleostearic acid (67)

OH

OH

erythro-9,lO-dihydroxy-l-octadecanol (68)

erucic acid (66)

echinacein (69)

neoherculin (70)

Fig. 2.28. Fatty acids and fatty acid derivatives with biological activity.

content is removed by processing of the oil, especially in INSECT PHEROMOrrnS DERIVED FROM
the process of deodorization, but the range of cyclopropenyl FAITY ACIDS
fatty acids in commercial cottonseed oil is from 0.1 % to
0.5%. A number of insect pheromones are derived from fatty acid
Oils containing cyclopentenoid fatty acids such as chaul- metabolism. These are liberated by the female insect to at-
moogric and hydnocarpic acids (Fig. 2.18) have long been tract the male from a distance and to induce copulation when
used for the treatment of leprosy. at close quarters. In other instances, the males liberate similar
Amide derivatives of unsaturated fatty acids (CID-C IS ) compounds to attract the females. In human terms, these
are especially common in members of the families Aristolo- compounds are equivalent to aphr0disiacs.
chiaceae. Asteraceae, Piperaceae, and Rutaceae; about 140 The first insect pheromone to be isolated and character-
compounds of this type are known (Greger, 1988). Many ized was bombykol (structure 72) from the silk moth, Bom-
contain only olefinic fatty-acid-derived portions, but those byx mori, by Butenandt and co-workers (Kelly, 1990). Pher-
of the tribes Anthemideae and Heliantheae of the Asteraceae omone glands (500,000) were extracted to yield 12 mg of
often contain acetylenic linkages as well. Among these, the the 4'-nitroazobenzene-4-carboxylic acid ester of the volatile
isobutyl amides [such as echinacein (69) and neoherculin alcohol. As few as 2500 molecules of the pheromone is
(70)] have pronounced insecticidal activity, others induce enough to elicit behavior in the male moth (Hecker and Bu-
local anesthesia (Greger et aI., 1985). Most of these com- tenandt, 1984; Kelly, 1990).
pounds have not proven practical for insecticides, as they Pheromones have now been studied and characterized
are highly irritating to mammals and are relatively unstable from many different species and at least 200 female sub-
(Jacobson, 1971). Leaves and inflorescences of Spilanthes stances are known from female lepidopterans and at least
olearacea (anamafana or anamalaho) that contain com- 60 from males (Fig. 2.29) (Harborne, 1982).
pounds of this type are widely used as an herb in Malagasy In terms of structure, the simplest is valeric acid (71), the
cooking. This plant contains both olefinic and acetylenic female pheromone of the sugar beet wire worm. The major-
isobutylamides (Greger, 1988), which may be responsible ity are long-chain unsaturated alcohols, acetates, or carbox-
for the tingling sensation in the mouth that is produced by ylic acids. In anyone insect, the s!mctures ofthe pheromones
this herb. are usually quite specific and small changes (position of
 Fatty Acids 37

double bonds, Z versus E double bonds, or stereochemistry} characterized as macrocyclic lactones, such as l8-octadeca-
usually diminish sharply or eliminate any activity. In some nolide (75). These compounds also are derived from fatty
cases, mixtures of compounds occur; the exact ratios must acid metabolism (Fig. 2.30).
often be present to maintain activity. Pheromones are usually The European cherry fruit fly, Rhagoletis cerasi, lays a
active at extremely low concentrations and have generally single egg into half-ripe cherries. An oviposition-deterring
proven difficult to study for this reason. pheromone, which prevents oviposition by a second female
Several aggregation pheromones are metabolically al- of the same species, was isolated from the feces of the insect.
tered fatty acids, or mixtures of these compounds. The most This compound was characterized as N[15(J3-glucosyl}oxy-
active attractants for the sawtoothed grain beetle, Orzyaephi- 8-hydroxypalmitoyIJ-taurine (76) (Fig. 2.30) (Hurter et aI.,
Ius surinamensis, from oats were (2E}-nonenal and (2E,4E)- 1987).
nonadienal (Mikolajczak et al., 1984). The most abundant
compound from a variety of food-related resources with
pheromonal activity for males of the driedfruit beetle, MAMMALIAN FHEKOMOrmS
Carpophilus hemipterus (Coleoptera: Nitidulidae), was
(2E,4E,6E,8E}-3,5,7-trimethyl-2,4,6,8-decatetraene (Bartelt A number of mammalian pheromones are combinations of
et aI., 1990). low-molecular-weight carboxylic acids. In other mamrnals,
In a few cases, pheromonal substances are taken from the steroids, sulfur-containing compounds, amines, and alcohols
plant and used directly (see Shorey and McKelvey, 1977), play pheromonal roles; some of these compounds also are
but more commonly, pheromones appear to be modified or derived from carboxylic acids (Albone, 1984; Harborne,
synthesized by the insect. The biosynthesis of pheromones 1982). Macrocyclic lactones such as civetone (77) and mus-
has been discussed (Tumlinson and Teal, 1987). cone (78) are the active odor compounds of the civet cat and
Brevicomin (73) and frontalin (74), probable products of the musk deer, respectively.
fatty acid metabolism, are part of the pheromone system of
bark beetles of the genus Dendroctonus. These compounds
are involved in interactions of these insects with their host ANALYSIS OF FATl'Y ACIDS
plants, many of which are important softwoods (gymno-
sperms) (Fig. 2.30). Historically, this was one of the most difficult groups of
The active constituents of the Dufour's glands of species compounds for chemical examination. Because the intact
of Colletes and Halictus bee species have been studied and glycerides are difficult to purify and to characterize, most

~CO,H valerie acid (71) sugar beet wireworm

Limonius californicus

o
~O"- z-7-dodecenyl acetate cabbage looper
Trichoplusia ni
o
~CO'H E-9-keto-Z-decenoic acid honey bee
Apia mellifern
o
~O~ Z-S-dodecenyl acetate oriental fruit fly
Dacus orienkllis

bombykol (72) silkworm moth

HO

o
/l-o~ red banded leaf roller

I"", I"", CO,H monarch butterfly

H02~

Douglas fir tussock moth

Z-6-heneicosen-ll-ooe Orygia pseudotsugata

Fig. 2.29. Fatty-aeid-derived insect pheromones (modified from Harhorne, 1982).

38 Fllf1.\' AcidJ

frontalin (73) brevicomin (74)

HO _ _<OH ____ 0, fH,

HO~CH(CH,J'<;H(CH,J.CONHCH1CH1SO,H
OH I
OH

N[15(P-glucosyl)oxy-8-hydroxypalmiloyll-taurine (76)

° °

Q
18-octadecanolide (75)

clvetone (77) muscone (78)

Fig. 2.30. Bioactive molecules derived from fatty acids.

studies have been carried out with the fatty acids or esters genated fals (in rats), in Geometrical and Positional Fatty Acid
derived from them. As is true for most esters, trlacylglycerols Isomers (E. A. Emkin and H. J. Dutton, eds.), 53-74, American
may be hydrolyzed with either acid or base tu yield an alco- Oil Chemical Society, Champaign, IL, 1979.
hol (glycerol) and free fatty acids. Glycerides may also be BAFOR, M., M. A. SMITII, L. JONSSON, K. STODART, and S. STYMNE,
transesterified to form fatty acid methyl esters. Ricinoleic acid biosynthesis and triacylglycerol assembly in mi-
With the advent of column chromatography, thin-layer crosomal preparations from developing castor-bean (Ricinus
communis) endosperm, Biochem. J., 280, 507-514 (1991).
chromatography, and gas liquid chromatography, the analy-
sis offatty acids and their methyl esters has become routine. BARTELT, R. J., P. F. DoWD, R. D. PLATTNER, and D. WBISLEOER,
Aggregation pheromone of the driedfruit beetie, Carpophilus
GCIMS makes the characterization of most fatty acids a
hemipterus: Wind-tunnel bioassay and identification of two
straightforward procedure.
novel tetraene hydrocarbons, J. Chern. Bcol., 16, 1015-1039
(1990).
BOWERS, W. S., H. C. HOCH, P. H. EVANS, and M. KATAYAMA,
REFERENCES Thallophytic a11elopathy: Isolation and identification of laeti-
sarie acid, Science, 232, 105-106 (1986).
ALBoNE, E. S.,Mammalian Semiochemistry: The Investigation of BRAoow, J. M. and W. J. CONNICK, JR., Volatile seed germination
Chemical Signals between Animals, John Wiley and Sons, inhibitors frum plant residues, J. Chern. Bcol., 16, 645-666
Chichester, 1984. (1990).
AL-NAGOY, S., M. O. ABOEL RAHMAN, and H. l. HBIBA, Extraction BROWSE, 1. and C. SOMERVll.LE, Glycerolipid synthesis: Biochemis-
and identification of different prostaglandins in Allium cepa, try and regulation, Annu. Rev. Plant Physiol. Plant Mol. BioI.,
Compo Biochem. Physiol., 85C(1), 163-166 (1986). 42,467-506 (1991).
ALFIN-SLATER, R. B. and L. AFrBIWOOO, Nutritional role of hydro- BRUIN, 1., M. DICKE, and M. W. SADELIS, Plants are better prolected
 Fatty Acids 39

against spider-mites after exposure to volatiles from infested GOODWIN, T. W. and E. I. MERCER, Plant Biochemistry, 2nd ed.,
conspecifics, Experientia, 48, 525-529 (1992). Pergamon Press, Oxford, 1983.
BUIST, P. H. and I. M. FINDLAY, The biosynthesis of cyclopropane GREGER, H., Comparative phytochemistry of the alkamides, in Con-
fatty acids. III. pH dependence of methyl hydrogen exchange: ference on Chemistry and Biology of Naturally-Occurring Acet-
gas chromatographic-mass spectral studies, Can. J. Chern., 63, ylenes and Related Compounds (NOARC) (J. Lam, H. Breteler,
971-974 (1985). T. Amason, and L. Hansen, eds.), 159-178, Elsevier, Amster-
BUTT, V. S. and C. J. LAMB, Oxygenases and the metabolism of dam,1988.
plant products, in Secondary Plant Products (E. E. Conn, ed.), GREGER, H., O. HOFER, and A. WERNER, New amides from
Vol. 7 of The Biochemistry of Plants (P. K. Stompf and E. E. Spilanthes oieraeea, Monats. Chern., 116, 273-277 (1985).
Conn, eds.), 627-665, Academic Press, New York, 1981. GROSS, D., Wachstomsregulatorisch wirksame PlIanzeninhaltss-
BUTTERY, R. G. and J. A. KAMM, Volatile components of alfalfa: toffe, Z. Chemic, 20, 397-406 (1980).
Possible insect host plant attractants, J. Agric. Food Chern., 28, GROVE, M. D., G. F. SPENCER, P. E. i'FEFFBR, N. MANDAVA, J. D.
978-981 (1980). WARTHEN, JR., and J. F. WORLEY, 6-D-Glucopyranosyl fatty
BUTTERY, R. G., R. TBRANlsm, and L. C. LING, Fresh tomato aroma acids esters from Brassica napus pollen, Phytochemistry, 17,
volatiles: A quantitative study, J. Agric. Food Chern., 35, 1187-1189 (1978).
540-544 (1987). GUNDLACH, H., M. J. MULLER, T. M. KUTCHAN, and M. H. ZENK,
CAHOON, E. B. and J. B. Om.RGGGE, Apparent role of phosphatidyl- Jasmonic acid is a signal transducer in elicitor-induced plant
choline in the metabolism of petroselinic acid in developing cell cultores, Proc. Nat. Acad. Sci. USA, 89, 2389-2393 (1992).
Umbelliferae endosperm, Plant Pbysiol., 104, 845-855 (1994). GUNSTONE, F. D., F.tty acids and glycerides, Nat. Prod. Rep., 1,
CARTER, C. D., J. N. SACALIS, and T. J. GIANFAGNA, Zingiberene 483-497 (1984).
and resistance to Colorado potato beetle in Lycopersicon hirsu- HAMBERG, M. and H. W. GARDNER, Oxylipin pathway to jasmo-
tum f. hirsutum, J. Agric. Food Chern., 37, 206-210 (1989). nates: Biochemistry and biological significance, Biochem. Bio-
CONN, E. E. and P. K. STUMPF, Outlines of Biochemistry, 3rd ed., phys. Acta, 1165, 1-18 (1992).
Wiley, New York, 1972. HAREORNB, J. B., Introduction to Ecological Biochemistry, 2nd ed.,
CROMBIE, L. and K. M. MISTRY, Synthesis of 12-oxophytodienoic Academic Press, London, 1982.
acid (12-oxoPDA) and the compounds of its enzymic degrada- HARBORNB, J. B., Recent advances in chemical ecology, Nat. Prod.
tion cascade in plants, OPC-8:0, -6:0, -4:0, and -2:0 (epi- Rep .. 4, 323-344 (1986).
jasmonic acid), as their methyl esters, J. Chem. Soc. Perkin HARWOOD, J. L., Plant acyl lipids: Structure, disrribution, and analy-
Trans. I, 1981-1991 (1991). sis, in Lipids (P. K. Stumpf, ed.), Vol. 4 of The Biochemistry
DATIIE, W., C. SClDNDLER, G. SCHNEIDER, J. SCHMIDT, A. PORZEL, of Plants (P. K. Stompf and E. E. Conn, eds.), 1-55, Academic
E. JENSEN, and I. YAMAOUCHI, Cucurbic acid and its 6,7-stereo- Press, New York, 1980.
isomers, Phytochemistry, 30, 1909-1914 (1991). HAy, M. E., J. E. DUFFY, W. FENICAL, and K. GUSTAVSON, Chemi-
DEROSA, M., A. GAMBACORTA, and J. D. Bu'LocK, Specificity cal defense in the seaweed Dictyopteris deliculata: Differential
effects in the biosynthesis of fatty acids in Bacillus acidocaldar- effects against reef fishes and amphipods, Marine EcoI.-Prog.
ius, Pbytochemistry, 13, 905-910 (1974). Ser., 48, 185-192 (1988).
ELBBIN, A. D., GLYCOLIPIDS, in Lipids (P. K. Stompf, ed.), Vol. 7 HECKER, E. and A. BUTENANDT, Bombykol revisited, in Techniques
of The Biochemistry of Plants (P. K. Stumpf and E. E. Conn, in Pberomone Research (H. E. Huntmel and T. A. Miller, eds.),
eds.), 571-587, Academic Press, New York, 1980. 1-44, Springer-Verlag, Berlin, 1984.
ELZBN, G. W., H. J. WILLIAMS, and S. B. VINSON, Isolation and HrrCHCOCK, C. and B. W. NICHOLS, Plant Lipid Biochemistry, Aca-
identification of cotton synomones mediating searching behav- demic Press, London, 1971.
ior by parasitoid Campoletis sonorensis, 1. Chern. Ecol., 10, HUANG, A. H. C., LIPASBS, in Lipids: Structure and Function (P.
1251-1264 (1984). K. Stumpf, ed.) Vol. 9 of The Biochemistry of Plants (P. K.
FARMBR, E. E. and C. A. RYAN, Interplant communication: Air- Stompf and E. E. Conn, eds.), 91-119, Academic Press, Or-
borne methyl jasmonate induces synthesis of proteinase inhibi- lando, FL, 1987.
tors in plantleaves. Proc. Nat. Acad. Sci. USA, 87, 7713-77116 HURTER, J., E. F. BOLLER, E. STADLER, B. BLATTMANN, H. R.
(1990). BUSER, N. U. BosSHARD, L. DAMM, M. W. KOZLOWSKI, R.
FARMER, E. E. and C. A. RYAN, Octadecanoid precursors of jas- SCHONI, F. RASCHDORF, R. DAHINDEN, E. SCHLUMPF, H. FRITZ,
monic acid activate the synthesis of wound-inducible proteinase W. J. RICHTER, and J. SCHREIBER, Oviposition-deterring phero-
inbibitors, Plant Cell, 4, 129-134 (1992). mone in Rhago/etis cerasi L.: Purification and determination
FUKUI, H., K. KOSIDMIZU, Y. YAMAZAKI, and S. USADA, Structures of the chemical constitotion, Experientia, 43, 157-164 (1987).
of plant growth inhibitors in seeds of Cucurbita pepo L., Agric. JACOBSON, M., The unsaturated isobutylamides, in Naturally Oc-
BioI. Chern., 41, 189-194 (1977.). curring Insecticides (M. Jacobson and D. G. Crosby, eds.),
FUKUI, H., K. KOSHIMIZU, S. USADA, and Y. YAMAZAKI. Isolation 137-176, Marcel Dekker, Inc., New Yori<, 1971.
of plant growth regulators from seeds of Cucurbila pepo L., JACOBSON, M., M. M. CRYSTAL, and J. D. WARTHEN, JR., Boll
Agric. BioI. Cbem., 41,175-180 (l977b). weevil feeding deterrents from tung oil, J. Agric. Food Chern.,
fuLLER, G. and W. D. NBS (eds.), Ecology and Metabolism of Plant 29,591-593 (1981).
Lipids, ACS Symposium 325, American Chemical Society, JAENICKE, L., Chemical signal transmission by gamete attractants
Washiugton, DC, 1987. in brown algae, in Biochemistry of Sensory Functions (L. Jae-
40 Fatty Acids

nicke, ed,} (25, Colloquium Gesellsch. BioI. Chern.), 307-309, MACLEOD, G. Volatile components of chayote. Phytochemistry,
Springer-Verlag, Berlin, 1974. 29, 1197-1200 (1990).
JAENICKE, L. Development: Signals in the development of crypto- MANDAVA, N. B., Natural products in plant growth regulation, in
grams, Prog. Bot., 52, 138-189 (1991). Plant Growth Substances (N. B. Mandava, ed.), ACS Sympo-
JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary sium Ser. III, 135-213, American Chemical Society, Washing-
compounds to the seed-eating larvae of the bruchid beetle Callo- ton, DC, 1979.
sobruchus maculatus, Phytochemistry, 16, 223-227 (l977). MANOOLD, H. K. and F. SPENER, Biosynthesis of cyclic fatty acids,
in Lipids (P. K. Stumpf, ed.), Vol. 4 of The Biochentistry of
JAWORSKI, 1. G., Biosynthesis of monoenoic and polyenoic fatty
Plants (P. K. Stumpf and E. E. Conn, eds.), 647-663, Academic
acids, in Lipids: Structure and Function, (P. K. Stumpf, ed.),
Press, Orlando, FL, 1980.
Vol. 9 of The Biochemistry of Plants (P. K. Stumpf and E. E.
Conn, eds.), 159-174, Academic Press, New York, 1987. MANN, J., Secondary Metabolism, Oxford University Press, Ox-
ford, 1978; 2nd edition, 1987.
JAWORSKI, J. G., D. POST-BElTENMILLER, and J. B. OHLROGGE, Ace-
tyl-acyl carrier protein is not a major intermediate in fatty acid MAsli, H., T. KONDO, and M. KOJIMA, An antifungal compound,
biosynthesis in spinach, Eur. J. Biochem.,213, 981-987 (l993). 9,12,13-trihydroxy-(E}-IO-octadecenoic acid from Colocasia
antiquorum inoculated with Ceratocystis jimbriata, Phytochem-
JOYARD, J. and R. DOUCE, Galactolipid synthesis, in Lipids: Struc-
istry, 28, 2613-2615 (1989).
ture and Function (P. K. Stumpf, ed.), Vol. 9 of The Biochentis-
McDoWELL, P. G., W. LWANDE, S. G. DEANS, and P. G. WATER-
try of Plants (P. K. Stumpf and E. E. Conn, eds.), 215-274,
MAN, Volatile resin exudate from stem bark of Commiphora
Academic Press, New York, 1987.
rostrata: Potential role in plant defence, Phytochemistry, 27,
KABARA, J. 1., Fatty acids and esters as antimicrobial/insecticidal 2519-2521 (1988).
agents, in Ecology and Metabolism of Plant Lipids (G. Fuller
MEYER, M. and D. O'HAGAN, Rare fluorinated natural products,
and W. D. Nes, eds.), ACS Symp. Ser. 325, 220-238, American
Chern. Br., 785-788 (1992).
Chemical Society, Washington, DC, 1987.
MIKOLAJCZAK, K. L., B. W. ZlLKOWSKI, C. R. SMITH, JR., and W.
KAMEoKA, GC-MS methods for volatile flavor components of E. BURKHOLDER, Volatile food attractants for Oryzaephilus suri-
foods, in Gas Chromatography-Mass Spectrometry (H. F. Lin- namensis (L.) from oats, J. Chern. Ecol., 10, 301-309 (1984).
skens and J. F. Jackson, eds.), 254-276, Springer-Verlag, Ber-
MIRAu.E, J., M. AKNrN, L. MICOUlN, E. GAYDOU, and J. KORN-
lin, 1986.
PROBST, Cyclopentyl and 00-5 monounsaturated fatty acids from
KELLy, D. R .. Insect semio-chemicals, Chern. Br. 124-127 (1990). red algae of the Solieriaceae, Phytochemistry, 29, 2161-2163
KENNEDY, B. H., Effect of multilure and its components on parasites (1990).
of Scolytus multistriatus, J. Chern. Ecol., 10, 373-385 (1984). MONACO, P. and L. PREvrrnRA, Oxygenated fatty acids from Lemna
KENNEDY, G. G., R. T. YAMAMOTO, M. B. DIMOCK, W. G. WlL- trisulca, Phytochentistry, 26, 745-747 (1987).
UAMS, and J. BORDNER, Effect of day length and light intensity MORRIS, L. 1., Mechanisms and stereochemistry in fatty acid metab-
on 2-tridecanone levels and resistance in Lycopersicon hirsutum olism, Biochem. J., II8, 681-693 (1970).
f. glabrum to Manduca sexta, J. Chern. Ecol., 7, 706-716 OHLROGGE, J. B., J. G. JAWORSKI, and D. PosT-BEITTENMlLLER, De
(l981). novo fatty acid biosynthesis, in Lipid Metabolism in Plants (T.
KINDL, H., ~-Oxidation of fatty acids by specific organelles, in S. Moore, Jr., ed.) 3-32, CRC Press, Boca Raton, FL, 1993.
Lipids: Structure and Function, (P. K. Stumpf, ed.), Vol. 9 of PANOSSIAN, A. G., G. M. UVETISSIAN, V. A. MMATSAKANIAN, S.
The Biochentistry of Plants (P. K. StumpfandE. E. Conn, eds.), G. BATROKOV, S. A. VARTANIAN, E. S. GABIDICAN, and E. A.
31-52, Acadentic Press, New York, 1987. AMROYAN, Unsaturated polyhydroxy acids having prostaglan-
KonMA, M., H. SHlRAKI, M. OHNlSHl, and S. ITO, Two diglycosyldi- din like activity from Bryonia Alba. n. Major Components
acylglycerol isomers in plant leaves, ferns, mosses and sea- Planta Medica; 47, 17-24 (1983).
weeds, Phytochentistry, 29, 1161-1163 (1990). PEARCE, G., D. STRYDOM, S. JOHNSON, and C. A. RYAN, A polypep-
KUSMENOGLU, S., L. L. ROCKWOOD, and M. R. GRETZ, Fatty acids tide from tomato leaves induces wound-inducible proteinase
and diacylglycerols from elaiosomes of some ant-dispersed inhibitor proteins, Science, 253, 895-898 (1991).
seeds, Phytochemistry, 28,2601-2602 (1989). PREVITERA, L. and P. MONACO, Further oxygenated fatty acids from
LAWLOR, D. W., Photosynthesis, 2nd ed., Longman, London. 1993. Lemna minor, J. Nat. Prod., 50, 807-810 (1987).
SEDGWICK, B., The control of fatty acid biosynthesis in plants, in
LEHNlNGER, A. L., Principles of Biochemistry, Worth, New York,
Biosynthesis and its Control in Plants (B. V. Milborrow, ed.),
1982.
179-217, Academic Press, London, 1973.
LEVIN, D. A., The oil content of seeds: An Ecological Perspective,
SEIGLER, D., Toxic seed lipids, in Herbivores, Their Interaction
Am. Nat., 108, 193-206 (1974).
with Secondary Plant Metabolites (G. A. Rosenthal and D. H.
LEVIN, E. D., V. E. CHEREPANOVA, I. A. ZIMOVSTSEVA. and T. O. Janzen, eds.), 449-470, Academic Press, New York, 1979.
SEDLOVA, Prostaglandins: Identification and content in higher SEIGLER, D. S., Role of lipids in plant resistance to insects, in Host
plants, Phytochemistry, 27,3241-3243 (1988). Plant Resistance to Insects (p. A. Hedin, ed.), ACS Symposium
Lm KEN JIE, M. S. F., Fatty acids and glycerides, Nat. Prod. Rep., Ser. 208, 303-327, American Chemical Society, Washington,
6,231-261 (1989). DC, 1983.
LIGHT, D. M., E. B. JANG, and J. C. DICKENS, Electroantennogram SEMBDNER, G. and B. PARTHmR, The biochentistry and the physio-
responses of the Mediterranean fruit fly, Ceratitis capitata, to a logical and molecular actions of jasmonates, Annu. Rev. Plant
spectrum of plant volatiles, J. Chern. Ecol., 14,159-180 (1988). Physiol. Plant Mol. BioI. 44, 569-589 (1993).
 Fatty Acids 41

SHINTANI, D. K. and J. B. OHLROGGE, The characterization of a in wheat genn as chemical stimuli eliciting aggregation of the
mitochondrial acyl carrier protein isofonn isolated from Arabi- confused flour beetle, Tribolium confusum (Coleoptera: Tene-
dopsis thaliana, Plant Physio!., 104, 1221-1229 (1994). brionidae),1. Agric. Food Chern., 19, 285-288 (1971).
SHOREY, H. H. and J. J. MCKELVEY, Chemical Control of Insect TORSSELL, K. B. G., Natural Product Chemistty, John Wiley and
Behavior, Wiley, New York, 1977. Sons, Chichester, 1983.
SHORLAND, F. B., The distribution of fatty acids in plant lipids, in TuMLINSON, J. H. and P. E. A TEAL, Relationship of structure
Chemical Plant Taxonomy (T. Swain, ed.), 253-311, Academic and function to biochemistry in insect pheromone systems, in
Press, London, 1963. Pheromone Biochemistry (G. D. Prestwich arid G. J. Blomquist,
SIMPSON, B. B. and M. CONNER-OOORZALY, Economic Botany, eds.), 3-26, Academic Press, Orlando, FL, 1987.
McGraw-Hill, New York, 1986. TuRLINGs, T. C. J., J. H. TuMLINSON, R. R. HEATH, A. T. PRovEAux,
SIMPSON, T. J., The biosynthesis of polyketides, Nat. Prod. Rep., and R. E. DOOLITrLE, Isolation and identification of a11elochem-
1,281-297 (1984). icals that attract the larval parasitoid Cotesia marginiventus
(Cresson) to the microhabitat of one of its hosts, J. Chern. Ecol.,
SMITH, C. R., JR., Occurrence of unusual fatty acids in plants, Prog.
17,2235-2251 (1991).
Chern. Fats Other Lipids, 11,139-171 (1970).
VAN ALLER, R. T., G. F. PEsSONBY, V. A ROGERS, E. J. WATKINS,
SMITH, C. R., JR. and I. A. WOLFF, Glycolipids of Briza spicata
and H. G. LEGGETT, Oxygenated fatty acids: A class of allelo-
seed, Lipids, 1, 123-127 (1966).
chemicals from aquatic plants, in The Chemistry of Allelopathy
SMITH, C. R., JR., L. H. NIECE, H. F. ZOBEL, and I. A WOLFF, (AC. Thompson, eds.), ACS Symposium Ser, 268, 387-400,
Glycosidic constituents of Ipomoea parasitica seed, Phyto- American Chemical Society, Washington, DC) 1985.
chemistty, 3, 289-299 (1964).
VISSER, J. H. and D. A AvE, General green leafivolatiles in the
SOMERVll.LE, C. and J. BROWSE, Plant lipids: Metabolism, mutants olfactory orientation of the Colorado beetle, L~ptinotarsa de-
and membranes, Science, 252, 80-87 (1991). cemlineata, Ent. Exp. Appl., 24, 538-549 (1978).
STASWICK, P. E., JASMONATE, genes and fragrant signals, Plant VISSER, J. H., S. VANSTRATEN, and H. MAARSE, Isolatkn and identi-
Physio!., 99, 804-807 (1992). fication of volatiles in the foliage of potato, Solanum tuberosum,
STUMPF, P. K., Lipid metabolism, in Plant Biochemistty, 3rd ed. a host plant of the Colorado beetle, Leptinotarsa decemlineata,
(1. Bonner and J. E. Varner, eds.), Academic Press, New York, J. Chern. Eco!., 5, 13-25 (1979).
1976. WHITMAN, D. W. and F. 1. ELLER, Parasitic wasps orient to green
STUMPF, P. K., Biosynthesis of saturated and unsaturated fatty leaf volatiles, Chemoecology, 1, 69-75 (1990).
acids, in Lipids (P. K. Stumpf, ed.), Vo!' 4 of The Biochemistty WICKREMASIGHE, M. G. V. and H. F. EMDEN, Reactions of adult
of Plants (P. K. Stumpf and E. E. Conn, ed,.), 177-204, Aca- female parasitoids, particularly Aphidius rhapalasiphi to vola-
demic Press, New York, 1980. tile chemical cues from host plants of their aphid prey, Physiol.
STUMPF, P. K., The biosynthesis of saturated fatty acids, in Lipids: Entom.,17, 297-304 (1992).
Structure and Function (p. K. Stumpf, ed.) Vo!' 9 of The Bio- Wll.LIAMS, W. G., G. G. KENNBnY, R. T. YAMAMOTO, J. D.
chemistty of Plants (P. K. Stumpf and E. E. Conn, eds.), THACKER, and J. BORDNER, 2-Tridecanon: a naturally occurring
121-136, Academic Press, New York, 1987. insecticide from the wild tomato Lycopersicon hirsutum f. gla-
STYMNE, S. and A K. STOBART, Triacylglycerol biosynthesis, in bratum, Science, 207, 888-889 (1980).
Lipids: Structure and Function, (P. K. Stumpf, ed.) Vol. 9 of ZAMIR, D., T. S. BEN-DAVID, J. RUDICH, and J. A. JUVIK, Frequency
The Biochemistty of Plants (P. K. Stumpf and E. E. Conn, eds.), distributions and linkage relationships of2-tridecanone in inter-
175-214, Academic Press, New York, 1987. specific segregating generations of tomato, Euphytica, 33.
TAMAKI, Y., S. R. LosclDAvo, and A J. McGINNIS, Triglyceride, 481-488 (1984).
3
Acetylenic Compounds

Introduction often occur as part of triacylglyceriiles and are restricted to

Biosynthesis seed oils in a few plant families. A second group of acetyle-
CIS Acetylenic Fatty Acids nic compounds consists of metabolically altered derivatives
AllyJic Oxidation and Rearrangement of crepenynic acid. These often are distributed throughout
Compounds Derived by Modification of CIS Acetylenic the plant, although, in some instances, the compounds may
Fatty Acids be found only in a single plant part, especially roots. This
Reduction of Chain Length by Ooe Carbon Atom second type is often found in the essential oils of plants of
Oxidative Cleavage of the Chain the Apiaceae, Ara1iaceae, and Asteraceae. Other acetylenic
Cyclization of Acetylenic Compounds compounds of this origin are found in seeds. A third group
Sulfur Derivatives of Acetylenes is formed by triple-bond introduction into compounds of
Distribution of Acetylenic Compounds in Plants and many different biosynthetic origins, such as terpenes. These
Fungi compounds are sporadically distributed in diverse plant
Physiological and Pharmacological Properties groups.
Biologically Active Acetylenic Compounds
Acetylenes as Phytoalexins
Plant-Insect and Plant-Nematode Relationships BIOSYl'lTHESIS
Phototoxic Effects of Acetylenic Compounds
Allenic Fatty Acids
CIS Acetylenic Fatty Acids
References
The facts that radioactively labeled acetate and malonate
label acetylenes in the same manner as fatty acids and that
INTRODUCTION few branched-chain acetylenic compounds are known
strongly suggest that naturally occurring acetylenes are de-
Although discovered in the late 1800s, there was little rived from fatty acids. Additionally, many acetylenic com-
progress in the study of acetylenic compounds from plants pounds with a Z double bond have the double bond in the
until the 1950s. These compounds are highly unstable to same position as the double bond of oleic acid. In a similar
light, oxygen, and heat. Plant acetylenes often occur in com- manner to fatty acids, introduction of double bonds into acet-
plex mixtures and the techniques needed to isolate, purify, ylenic compounds requires molecular oxygen. Although the
and characterize them were not widely available until that triple bond is usually introduced into a position where a
time; UV spectrophotometry permitted workers to assign double bond would be expected, in some instances no prior
structures and was a major breakthrough for study of this double-bond formation is observed. The enzymes responsi-
group. Acetylenic compounds are widespread in nature, but ble for the introduction of triple bonds have been little
they are concentrated in several groups of plants and fungi. studied.
At present, about 1000 naturally occurring acetylenes are Oleic and linoleic acids are precursors for acetylenes in
known. microorganisms and plants (Fig. 3.1), but crepenynic and
There are three main types of acetylenic compounds in dehydrocrepenynic acids also are incorporated into most
plants. Acetylenic acids, usually derived from oleic acid, acetylenic compounds. Crepenynic acid (1) is the first acety-

42
 AcetyJenic Compounds 43

z z
stearate --+ oleate __ CH3(CH,),CH=CHCH.CH=CH(CH,),COSX

IInoleate

z z
CH3(CH,).CH=CHC=CCH.CH=CH(CH.J,COSX _ CH3(CH,),C:CCH.CH=CH(CH,J,COSX

~ (2) crepenynate (1)

CH3(CH,).C:CC=CCH.CH=CH(CH,J,CO.R _ CH3(CH,).(C=C}zCH=CHCO(CH,),CO,R

1m !~
°II
CH3CH=CH(C:C),CH.CH=CH(CH,J,CO,R CH3CH=CH(C:C).CH=CHCO(CH,),CO.R?

""".00 ..' t.'. . .;--mt

. !
II z z
CH3CH=CH(C:C),CH=CHC(CH,),CO,R CH3CH=CH(C:C),CH=CHCO,CH3

~ Oz matricaria ester (4)

E
CH3(C:ChCH=CHCO.CH3

dehydromatricaria ester (5)

Fig. 3.1. Biogenesis of matricaria ester and dehydromatricaria ester.

lenic fatty acid in the reaction sequence. This compound is

then desaturated to yield dehydrocrepenynic acid (2) which
is, in turn, converted to the corresponding diacetylenic com- z
pound (3). A number of other acetylenic fatty acids are de- CH3(CH,),C:::CCHzCH=CH(CHz)7COzH
rived from both (1) and (2) by reactions involving allylic
oxidation and subsequent rearrangement (Bohlmann and crepenynie aeid (1)
Burkhardt, 1972; Bohlmann et al., 1973; Bu'Lock, 1966).

AllyUc Oxidation and ReillTangement ! [0]

Allylic oxidation is often followed by allylic rearrange-
ment (Fig. 3.2). Oxygen atoms are rarely introduced into
nonallylic positions in these molecules (Bohlmann et al.,
1973).

Compounds Derived by Modification of CIS

Acetylenic Fatty Acids allylie 1 rearrangement

A large series of compounds are derived by modification

of crepenynic acid and related C •• acetylenic fatty acids.
Whereas these acids may be accumulated in seed oils as
E
components of triacylglycerols, modified derivatives USUally CH3(CH,),C=CCH=CHCH(CHz),COzH
are found in vegetative portions of plants. The biosynthesis
of matricaria ester (4) and dehydromatricaria ester (5) (both
I
OH
with 10 carbon atoms in the acid portion of the molecule)
(Fig. 3.1) is typical for this series of compounds. In this from a Helichryaum species (Asterae..e)
instance, fatty acids with chain lengths of less than 18 carbon
atoms do not serve as precursors. Fig. 3.2. Allylic oxidation followed by allylic rearrangement.
44 Acetylenic Compounds

oleic acid

! Z
CH,(CH,J,C=CCH,CH=CH(CH,J,CO,H

/
crepenynic acid (1)

E
CH,(CH,J,C=CCH=CHiH(CHlhC01H

OH

Z
CH,(C=C),CH,CH=CH(CH,J,CO,H

Z / ! Z CH(CH-',CO,H
CH,(""'C_=C),CH1CH-_

1 1
CH,(C:C),CH,CH =CH(CH,),CO,H "

C lO polyacelylenes

C14 polyacetylenes C13 polyacetylenes

Fig. 3.3. Biosynthetic pathways from oleic acid to polyacetylenes (modified from Jente et aI .• 1988).

In the biosynthesis of metabolically altered acetylenic

compounds, various numbers of acetylenic bonds may be R,CHa·YHCOlR ..... R.CHrCHO
inserted and the chain shortened by a- or l3-oxidation or OH
both (Fig. 3.3). Triple bonds are usually introduced in a
conjugated manner, although they may be separated by a
methylene group from the initial double bond of oleic acid.
Other compounds maintain the Z double bond and do not
undergo an allylic rearrangement (Jente et al., 1988). " ~o
'0-
R·CH·CH,.C ~
In some plants, reduction of terminal carboxyl groups to I
the corresponding aldehyde occurs. Aldehydes may then be ( OH

reduced to alcohols, these dehydrated to olefins, and the H<

olefms saturated to hydrocarbons (Fig. 3.4). In other plants, Fig. 3.5. Fonnation of C17-tenninal olefins from Cl8 precursors.
the reverse (degradative metabolism) of this reaction series
occurs. These modifications occur frequently in the acetyle-
nic series and help to explain the large number of compounds
observed.

Reduction of Chain Length by One Carbon

Atom
Various combinations of a- and l3-oxidation lead to short- ----. HOCHl-e=C-R ---+ OCH-CEC·R
ened chain length as previously mentioned. In addition, in-
troduction of an oxygen alpha to the carboxyl group can
lead to the formation of an aldehyde with a chain length of
one carbon atom less (Fig. 3.5). The elimination of one car-
bon atom from the hydrocarbon end of the chain also is
observed (Fig. 3.6).
Introduction of an oxygen at the l3-position can lead to

Fig. 3.6. Possible sequence for fonnation of C17 acetylenes from CIS
precursors.
Fig. 3.4. Oxidation and reduction in biosynthetic sequences.
 ACt'tyfenit: Compounds 45

Cyclization of Acetylenlc Compounds

Extended conjugated acetylenic systems sometimes give
rise to aromatic rings in plants. These compounds usually
co-occur with other acetylenic compounds and have linear
side chains attached to the aromatic rings. (see Fig. 3.9)
Almost all aromatic compounds from acetylenic precursors
are found in the Asteraceae.
Acetate is incorporated in the manner predicted for fatty
acids (Weiss and Edwards, 1980). In some instances, aroma-
tization is known to occur under in vitro conditions and the
probable precursor and the corresponding aromatic com-
pound sometimes occur together in the same plant; for exam-
ple, compounds (6) and (7) often co-occur in Coreopsis spe-
cies (Fig. 3.10).

Sulfur Derivatives of Acetylenes

Conjugated dienes in some plants react with sulfur-con-
taining intennediates to yield thiophenes. In other instances,
methyl thioethers are fonned. Although not known with cer-
tainty, suggested sulfur donors are methionine, S-adenosyl
methionine (SAM), and cysteine.

Fig. 3.7. Baeyer-Villiger type oxidation possibly responsible for DISTRIBUTION OF ACETYLENIC COMPOUNDS
fonnation of CIO compounds.
IN PLANTS AND FUNGI

In contrast to the distribution of most fatty acids, acetylenic

decarboxylation and fonnation of an olefin with one less
compounds are found in a restricted number of plant taxa.
carbon atom. Compounds of both origins may then undergo
Acetylenic acids that are components of triacylglycerides
reduction or oxidation as indicated above (Bohlmann et aI.,
are occasionally found in the seed oils of members of the
1973).
Asteraceae (such as in species of Crepis and Helichrysum),
but are most commonly found in a group of closely related
Oxidative Cleavage of the Chain families: the Loranthaceae, Oiacaceae, Opiliaceae, and San-
talaceae (Order Santalales).
A reaction similar to Baeyer-Villiger oxidation (oxida-
Metabolically altered acetylenic compounds are common
tion adjacent to a carbonyl group) occasionally occurs and
constituents of many plants of the families Apiaceae, Aralia-
may be responsible for the fonnation of several C IO com-
ceae, and Asteraceae, but also occur in the Campanulaceae
pounds (Fig. 3.7).
and Pittosporaceae (Downum and Nemec, 1987). Among
A complex lipid from the seed oil of the Chinese tal-
the fungi, acetylenes of this type are quite common in basidi-
lowtree, Sapium sebiferum (Euphorbiaceae), appears to have
omycetes. Acetylenes usually occur free, are found in di-
arisen in this manner (Fig. 3.8). Although acetylenic com-
verse parts of the organism, and are found frequently as
pounds are not commonly found in the Euphorbiaceae, the
essential oil components. Other acetylenes only are obtained
presence of an aIIenic linkage is suggestive of their presence.
by extraction with nonpolar solvents.
Acetylenic compounds typical of the Apiaceae and Arali-

E
aceae characteristically contain C 17 acetylenes, falcarinone,
°II and related compounds. Those of the Asteraceae usually
O;(CH,J7CH=CHCH,CH :::CHCH,CH=CHCH,CH, have vinyl end groups (Fig. 3.11). Acetylenic compounds
of the Pittosporaceae resemble those of the Apiaceae and
Araliaceae in structure whereas acetylenic compounds of
the Campanulaceae resemble those of the Asteraceae. The
OC(CH,J7CH=CHCH,CH=CH(CH,J.CH,
occurrence of tetrahydropyran derivatives seems to be re-
stricted to the Campanulaceae.
The relationships of several of acetylene-containing fami-
OC(CH,J,CH==HCH,OCCH=CHCH=CH(CH,J.CH, lies (e.g., the Apiaceae, Araliaceae, Asteraceae, Campanu-
laceae, and Pittosporaceae) have long been enigmatic. Many
"o 0
"
Fig. 3.8. An unusual aOerne lipid from Sapium sebiferum seed oil.
of the groups with which the Apiaceae and Araliaceae have
been placed lack acetylenic compounds, but are known to
 0"
46 AcetyJenic Compounds

I (<ZC),CH,

""" CH'C=C~CH=CHCOOCH'

carlina oxide

O-O-CJ
S S S

a·lerlbienyl (15)

-0-
C-C

- C# ~C - -
S

-C:C-C::C- _ -C=CH--C=C--
I
S·CH,

Fig. 3.9. Aromatic acetylenic compounds, thiophenes, and methylthioethers.

possess iridoid monoterpenes. The Apiaceae and Araliaceae mon in the tribes Reliantheae and Anthemideae (Dahlgren
lack the latter group of compounds (Dahlgren et aI., 1981). et aI., 1981).
Some have felt that the presence of acetylenes (as well Acetylenic compounds are widely distributed in the basidi-
as other shared characters) indicates a relationship of the omycetes, mostly from families of the Agaricales and Pol-
Asteraceae, with the Apiaceae and Araliaceae (Dahlgren et yporales. Many of these compounds are polar, have extensive
aI., 1981). Similar arguments have been made for the rela- oxygenation, and chain lengths of 10 carbon atoms or less.
tionship of the Pittosporaceae with Apiaceae and Araliaceae The third group of acetylenic compounds is not related
(Regnauer, 1969) and the Campanulaceae and Asteraceae clearly to the first two. Acetylenic linkages are occasionally
(Mabry and Bohlmann, 1977). introduced into compounds of diverse biosynthetic origin
Within the family Asteraceae, acetylenic compounds are such as sesqniterpenes, tetraterpenes, and amino acids (Fig.
largely absent from the tribe Senecioneae and are most com- 3.12). These compounds are found in unrelated plant fami-
lies, among them the Annonaceae, Cupressaceae, Euphor-
CHz=CHCH=CHC=CC=CC=CCH=CHCH, biaceae, Fabaceae, Lauraceae, Myoporaceae, Sapindaceae,
Simaroubaceae, Sterculiaceae, Valerianaceae, and certain
(6) algae and mosses.

PHYSIOLOGICAL Al'ID PHARMACOLOGICAL

PROPERTIES

The quantity and type of acetylenic compounds present in

(7)
most plants vary considerably at different stages of develop-
Fig. 3.10. Compounds from Coreopsis species. ment and in different plant parts. For example, the roots of
 Acetylenic Compounds 47

Helianthus annuus contain a pentaynene that disappears NH2 NH,

when the fruits begin to ripen. Although Centaurea seeds I I
do not contain acetylenic compounds, I-day-old seedlings HC=CCHCH CHC0 H2 2 HC:CCHCH,CHCO,H
do. The turnover time of many acetylenic compounds in I I
CH,OH OH
plants is 6-48 h. The amount of polyacetylenes in plants
ranges from traces to about I %, but the amounts present
Euphorio longana, Sapindaceae
often vary widely within and between plants.
CH,
Callus cultures of Bidens pilosa which normally produce
no acetylenic compounds could be induced to do so by addi-
o
tion of culture filtrate from the fungus Pythium aphanider-
matum; cultures of this and other Bidens species that had
been transformed with the Ti plasmid of Agrobacterium tu-
mefaciens produced a root type profile of acetylenes (Ellis, Eremophila jreelingii, Myoporaceae
1988). Hairy root cultures of Ambrosia, Bidens, and Tagetes
have similar content of acetylenes as those of normal plant
roots (Flores et al., 1987, 1988).

E E
c 1" CH,(C=C),CH=CHCH=CHCH,CH,CH,OH
Chamaecyparis jormosensis, Cupressaceae

E E E
CH,CH=CH(C:C),CH=CHCH=CHCH,CH,CH,OH

taririe acid

E E Z
CH,CH=CHCH=CHC :CCH,CH=CH(CH,J,CH,OH

E Z
CH,CH=CH(C :C),CH,CH=CH(CH,J,CO,H

E E from a Dichranum species

C17' CH,(C=C),CH=cHCH=CH(CH,J,CH=CH,
Fig. 3.12. Compounds of diverse biosynthetic origin that possess
acetylenic linkages.
E E
CH,(C=C)'CH=CHCH=CHiH(CH,J,CH=cH,
Biologically Active Acetylenic Compounds
OAc
Acetylenic compounds that occur in essential oils proba-
E E E
CH,CH=CH(C :C),CH=CHCH=CH(CH,J,CH=CH, bly contribute to the overall taste-odor properties in several
spices and herbs derived from the Apiaceae. Compound (8)
(Fig. 3.13), in association with monoterpenes, contributes to
the flavor of caraway, Carum carvi.
Q-(C:C),CIf=CHCH,
In contrast, certain other acetylenes are highly toxic to
humans and domestic animals; for instance, the compounds
from Cicuta virosa (water hemlock) (10) and Oenanthe ero-
Some naturally occurring polyacetylenes of Dahlia species. cata (9). Relatively small variations in structure lead to
Fig. 3.11. Some naturally occurring acetylenic compounds of Dahlia
nearly nontoxic compounds. Other compounds such as falc-
(Asteraceae) (modified from Lam et al.. 1991; used with pennission of arinol (11) are less toxic to people, but still possess antifun-
the copyright owner, Pergamon Press, Oxford). gal properties. Compounds of this type occur widely in mem-
48 Acetylenic Compounds

bers of the Apiaceae and Araliaceae in plants such as carrots Acetylenes as Phytoalexins
(Daucus carota) and ginseng (Panax quinquefolium). There Although acetylenic compounds may served as pre-
do not appear to be enough acetylenic compounds present formed antifungal compounds, the amount of several of
in most cultivars of carrots, parsley, or celery roots to be these substances increases rapidly in response to attack by
really dangerous (Yates et al., 1983). pathogens and are phytoalexins (Downum and Nemec,
Many isolated acetylenic compounds have pronounced 1987). In safflower, Carthamus tinctorius, the amount of
antifungal activity (and often antibacterial activity), but their safynol, an ene-triyn-ene-diol (14) (Fig. 3.14), increases 20-
inherent instability limits their usefulness for this purpose. fold when the plant is attacked by the pathogen Phytophthora
One example is the antibiotic mycomycin (12) isolated from drechsleri. The best studied example of acetylenic phyto-
a basidiomycete, Nocardia acidophilus (Weiss and Edwards, alexins, however, is the broad bean, Vicia faba, which pro-
1980). duces a series of acetylenic furanoid ketones related to wyer-
Two African species of Aspilia (Asteraceae) which are one after infection with a Botrytis species. Wyerone and
used medicinally by man also are eaten by chimpanzees in related compounds also are known to occur in the lentil,
a similar manner. This plant contains a potent antibiotic, Lens culinaris. Although the relationships of the lentil have
thiarubrine A (13) (Rodriguez et al., 1985). An unusual anti- been debated, these data suggest an affmity with the broad
viral acetylene, chondriol (Fig. 3.13), and other related com- bean.
pounds have been isolated from a marine algae (Moore, Infection of the African marigold, Tagetes erecta, with
1978). highly pathogenic fungi results in a general decrease in thio-

z
o
II
C7HISCH=CHC(C=C),CCH=CH2
0
I HO)!?1
from Carum carvi (8) chondriol

E E,E E,E
HOCH2CH=CH(C=Clz(CH=CH),CH2CHC2H, HOCH2(CH2)2(C=Clz(CH=CH).CHCH2CH2CH.
I I
OH OH
oenantbetoxin (9) from Cicuta virosa (10)
o
/\
HCH=CHCH(C:C)2CH=CHCH2(CHz),CH. CH.CH2CH2C:CC:CCHCH -GH(CH2),CH=CH2
I
OH
I
OH
falcarinol (11) (SR,9R,IOS)-9,IO-epoxybepladec-I6-ene-4,6-diyne-8-o1 (16)

OH OH
I I z E
CH,(CH2).CH=CHCH(C=C),CHCH=CH2 HC=CC=CCH=C=CHC=CHCH=CHCH2C02H

falcarindiol mycomycin (12)

CH,C=C~C=CC:CCII=CH2
S-S

thiorubrlne A (13) laballenic acid (17)

Fig. 3.13. Some biologically active acetylenic compounds.
 Acefylenic Compounds 49

CH3CH2CH=CHC=cM-C}-CH=CHC02H wyerone acid

CH3CH2CH=CHC=C~-D-CH=CHC02CH3 wyerone

wyerone epoxide

OH
E E ' safyool (14)
CH3CH=CHC:::CC:::CC:::CCH=CHCHCH20H
saffiower

Fig. 3.14. Wyerone epoxide, wyerone, and safynol.

phene levels, whereas less pathogenic pathogens resulted weed bug Oneopeltus fasciatus. Upon purification and test-
in the accumulation of a-terthienyl (15) (Fig. 3.9) and two ing of individual compounds, these were shown to have anti-
bithiophene derivatives above the levels in noninfected juvenile hormone activity (see more discussion in Chapter
plants. This suggests that highly pathogenic fungi may sup- 18) (Bowers and Aregullin, 1987).
press the production of thiophenes (Downum and Nemec,
1987), or possibly that nonpathogenic fungi release sub- PHOTOTOXIC Bf'FECTS OF ACBTYLBl'IIC
stances that enhance production of thiophenes. COMPOUl'lDS

Plant-Insect and Plant-Nematode In addition to non-light-mediated toxic effects of acetylenic

Relationships compounds, many are potent photosensitizers that can enter
an excited state by absorption of light energy. This may
Many fatty-acid-derived acetylenic compounds in plants occur in the presence of sunlight or UV -A radiation (Dow-
are active in plant-insect relationships. 8-Z-Dihydromatrica- num and Rodriguez, 1986). In this excited state, such mole-
ria acid, found in the defensive secretion of the soldier beetle cules mediate a variety of phototoxic effects through two
(Chauliognathus lecontei), has been shown subsequently to major types of action. Straight-chain acetylenic compounds
have antifeedant properties against Phidippus spp. (jumping tend to interact directly with target molecules in the cell,
spiders) (Eisner et a!., 1981). Matricaria acid (5) also has whereas thiophenes mediate the oxidation of a variety of
antifeedant properties against the pink bollworm, bollworm, biomolecules (membrane lipids and proteins, in particular)
and tobacco hornworm (Binder et a!., 1979). presumably via singlet oxygen generation (Downum and
Several nematicidal acetylenic compounds have been iso- Nemec, 1987). In addition to killing or inactivating viruses,
lated; most possess thiophene structures and are from mem- bacteria, fungi, and nematodes, herbivorous insects are also
bers of the Asteraceae family. Another recently isolated affected (Downum and Rodriguez, 1986).
compound with this activity is (8R,9R,IOS)-9,IO-epoxyhep- Despite the fact that acetylenes are found in at least 15
tadec-16-ene-4,6-diyne-8-01 (16) from Cirsium japonieum families, phototoxic acetylenic compounds are only found
(Kawazu et a!., 1980) (Fig. 3.13).I-Phenylhepta-I,3,5-triyne in the Asteraceae (Downum et a!., 1991).
and 2-phenyl-5-(l'-propynyl)-thiophene from Coreopsis
laneeolata and Z-dehydromatricaria ester (5) from Solidago ALLBl'IIC FATTY ACIDS
altissima have been shown to be fly ovicidal substances
(Seigler, 1983). Several allenic acids are known. One of these, laballenic
Four of the major acetylenic compounds of Chrysanthe- acid (17), is from the seed oil of a mint, Leonotis nepetaefolia
mum coronarium (Asteraceae) were insecticidal to the milk (Fig. 3.13). An unusual allene containing lriacylglycerol is
SO Acetylenic Compounds

found in Sapium sebiferum (Euphorbiaceae) (see Fig. 3.8). ELus, B. E., Natural products from plant tissue culture, Nat. Prod.
In origin, these unusual compounds are probably related to Rep., 5, 581-612 (1988).
acetylenic compounds, although acetylenes are not known FLORES, H. E., M. W. HoY, and J. J. PICKARD, Soluble metabolites
from either the Larniaceae or Euphorbiaceae. from root cultures, Tibtech, 5, 64-69 (1987).
FLORES, H. E., J. J. PICKARD, and M. W. Hoy, Production of polya-
cetylenes and thiophenes in heterotrophic and photosynthetic
root cultures of Asteraceae, in Naturally Occurring Acetylenes
REFERENCES and Related Compounds (J. Lam, H. Breteler, T. Amason, and
L. Hansen, eds.), 233-254, Elsevier, Amsterdam, 1988.
HEoNAUER, R., Chemical evidence for the classification of some
BINDER, R. G., B. G. CHAN, and C. A. Eu.IGER, Antibiotic effect
plant taxa, in Perspectives in Phytochemistry (J. B. Harborne
of C IO-C 12 fatty acid esters on pink bnllwonn, bnllwonn, and
and T. Swain, eds.), Academic Press, London, 1969.
tobacco budwonn, Agric. Bio!. Chern., 43,2467-2471 (1979).
KAwAZU, K., Y. NISHR, and S. NAKAJIMA, Two nematicidal sub-
BOHLMANN, F. and T. BURKHARDT, Zur Biogenese des Matricari- stances from roots of Cirsium japonicum, Agric. BioI. Chern.,
aesters, Chern. Ber., 105, 521-528 (1972). 44,903-906 (1980).
BOHLMANN, F., T. BURKHARDT, and C. ZDERO, Naturally Occurring JENTB, R., E. RICHTER, F. BOSOLD, and G. A. OLANTuNn, Experi-
Acetylenes, Academic Press, New York, 1973. ments on biosynthesis and metabolism of acetylenes and thio-
BOWERS, W. S. and M. AREoUllIN, Discovery and identification phenes, in Chemistry and Biology of Naturally-Occurring Acet-
of an antijuvenile honnone from Chrysanthemum coronarium, ylenes and Related Compounds (NOARC) (J. Lam, H. Breteler,
Mem. Inst. Oswaldo Cruz, Riodegeneiro, 82, Supp!. 3, 51-54 T. Amason, and L. Hansen, eds.), 187-199, Elsevier, Amster-
(1987). dam, 1988.
Bu'LoCK, J. D., The biogenesis of natural acetylenes, in Compara- LAM, J., L. P. CHRlSTBNSEN, and T. THOMASEN, Polyacetylenes from
tive Phytochemistry (T. Swain, ed.), 79-95, Academic Press, Dahlia species, Phytochemistry, 30, 515-518 (1991).
London, 1966. MABRY, T. J. and F. BOHLMANN, Sununary of the chemistry of the
DAHLGREN, R. M. T., S. ROSENDAL-JENSEN, and B. J. NIELSEN, A Compositae, in The Biology and Chemistry of the Compositae,
revised classification of the angiospenns with comments on Vo!. 2 (V. H. Heywood, J. B. Harborne and B. L. Turner, eds.),
correlation between chemical and other characters, in Phyto- Academic Press, London, 1977.
chemistry and Plant Phylogeny (D. A. Young and D. S. Seigler, MOORE, R. E., Algal Nonisoprenoids, in Marine Natural Ptoducts,
eds.), 149-204, Praeger, New York, 1981. Vol. I (p. J. Scheuer, ed.), Academic PIess, New York, 1978.
RODIUGUEZ, E., M. AREoULUN, T. NISIDDA, S. UEHARA, R. WRAN-
DoWNUM, K. R and S. NEMEC, Light-activated antimicrobial chemi-
GHAM, Z. ABRAMowSKI, A. FINLAYSON, and G. H. N. TOWERS,
cals from plants: Their potential role in resistance to disease-
Thiarubrine A, a bioactive constituent of Aspilia (Asteraceae)
causing organisms, in Light-Activated Pesticides (J. R. Heitz
consumed by wild chimpanzees, Experientia, 41, 419-420
and K. R. Downum, eds.), ACS Symposium Ser. 339, 281-294,
(1985).
American Chemical Society, Washington, DC, 1987.
SBIGLER, D. S.• Role of lipids in plant resistance to insects, in Plant
DoWNUM, K. R. and E. RODRiGUEZ, Toxicological action and eco-
Resistance to Insects (p. A. Hedin, ed.), ACS Symposium Series
logical importance of plant photosensitizers, J. Chern. Bco!.,12, 208,303-327, American Chemical Society, Washington, DC,
823-834 (1986). 1983.
DOWNUM, K. R., L. A. SWAIN, and L. J. FALEIRo, Influence of light WEISS, U. andJ. M. EDWARDS, The Biosynthesis of Aromatic Com-
on plant allelochemicals: A synergistic defense in higher plants, pounds, Wiley, New York, 1980.
Arch. Insect Biochem. Physio!., 17, 2011-211 (1991). YATES, S. G., R. E. ENGLAND, W. F. KWOLEK, and P. W. SIMON,
EISNER, T., D. lIIll., M. GOETZ, S. JAIN, D. ALsOP, S. CAMIZINE, Analysis of carrot constituents: Myristicin, falcarinol, and falc-
and J. MElNwALO, Antifeedant action of Z-dihydromatricaria arindiol, in Xenobiotics in Foods and Feeds, ACS Symposium
acid from soldier beetles (Chlluliognathus spp.), J. Chern. Bco!., Series 234, (J. W. Finley and D. E. Schwass, eds.), 333-344,
7, 1149-1158 (1981). American Chemical Society, Washington, DC, 1983.
4
Plant Waxes

Introduction acid precursor seem to be formed in the same manner as

Biosynthesis those previously discussed. Chain elongation requires malo-
Taxonomic Usefulness nyl-CoA and NADPH as the elongating agent and reductant,
Function of Waxes respectively; exogenous stearic acid can be incorporated.
Commercial Value of Waxes Stearyl-CoA, but not stearyl-ACP, has been demonstrated
Biological Activity to be elongated by endoplasmic reticulum preparations from
Chemical Fossils and Petroleum the epidermis of leeks, Allium porrum (LiJiaceae). The
Cutin and Suberin length of acids produced depends on the plant species, but
Analysis of Waxes C2., C30 , and C32 acids are commonly encountered (Kolattu-
References kudy, 1980).
Long-chain alcohols are formed by reduction of long-
chain fatty acids. A fatty acyl-CoA reductase has been dem-
INTRODUCTION onstrated in cabbage, Brassica oleracea (Brassicaceae).
Fractionation of the protein fraction isolated in these studies
indicated that one protein fraction catalyzed acyl-CoA re-
Plant waxes are complex mixtures of hydrocarbons, alco-
duction to the aldehyde stage with NADH as the preferred
hols, aldehydes, ketones, esters, acids, and combinations of
reductant and another catalyzed reduction of the aldehyde
these that are deposited in a layer outside the epidermal cells.
to the alcohol with NADPH as the preferred reductant. The
This complex mixture of lipids probably is synthesized in
similarity of the naturally occurring fatty aldehydes and fatty
the epidermal cells of most plants and exuded onto the sur-
alcohols of a given plant is probably due to the fact that
face. Plants usually possess cutin, a layer of cross-esterified
aldehydes represent intermediates in the synthesis of fatty
hydroxy fatty acids which is also deposited on the surface
alcohols (Kolattukudy, 1980).
of the epidermal cells (Kolattukudy, 1980). This layer is
Wax esters in plants are formed by the combination of
impregnated with waxes. The combined lipid covering not
long-chain alcohols and long-chain acids. Although this can
only protects the plant from invasion by foreign organisms
occur in several ways, it is most probable that, in vivo, these
but also helps to regulate transpiration.
esters are formed by action of an acyl-CoA alcohol lransacyl-
Waxes are usually isolated by boiling, scraping, or wash-
ase that has been demonstrated to be present by Kolattukudy
ing the outside surface with a nonpolar solvent. Surface lip-
(1980).
ids differ markedly from both seed lipids and other leaf lip-
Wax esters from jojoba (Simmomisia chinensis, Simmonds-
ids. (See Chapter 2.)
iaceae) consist of molecules with mostly C20 acid (monoene)
esterified with about an equal mixture of Yo and C22 alco-
hols (monoene) (Yermanos, 1978; 1981; Miwa, 1971). In
BIOSYl'l'l1lESIS studies of the biosynthesis of the fatty acids and alcohols in
slices of fresh jojoba cotyledons, a radioactive label from
Most components of the wax layer are derived from long- glucose was incorporated into all carbons of both the C 20
chain fatty acids (C2.-C34 ) formed by chain extension of and C22 acids and alcohols. In contrast, exogenous acetate
shorter-chain fatty acids (Fig. 4.1). The shorter-chain fatty was used almost entirely for chain elongation from endoge-

51
S2 Plant Waxes

acelyl-CoA fatty acid syntbetase

palmiloyl-ACP
+ 7 malonyl-CoA

mnlonyl-CoA
-------I.~
NADPH
slearoyl-ACP stearic acid
-
stearoyl.CoA
malonyl-CoA

NADPH
CH,(CHzl,.COCoA
- NADH

-
' " acyl-CoA: alcohol
~ transacylase
NADPH
CH3(CH,),.CH,OH wax eslers

Fig. 4.1. Biosynthesis of long-chain fatty acids, aldehydes, alcohols. and wax esters. (Kolattukudy. 1980 modified and used with pennission of the
copyright owner. Academic Press. Orlando. FL).

nous oleate (pollard et aI., 1978). In this plant, fonnation of roles. For example, in species of Nepenthes, an insectivorous
oleic acid occurs on the ACP track. The acid is released and plant, wax particles inside the "trap" are easily detached
subsequent reactions leading to the fonnation of C 2G and Y2 and stick to the feet of insects, preventing them from getting
fatty acids and alcohols occur as CoA derivatives. a foothold inside the trap and escaping.
Hydrocarbons found in the wax layer of plants also are
synthesized from long-chain fatty acids. C ,2 , C ,4 , C ,6 , and
C ,S fatty acids are incorporated into Y9 alkanes. Com-
pounds that are efficient precursors for long-chain fatty acids COl'lllmRCIAL VALVE OF WAXES
are also efficient precursors of hydrocarbons. Both groups
of compounds are synthesized in epidennal cells. Exogenous Waxes are of considerable commercial importance, as syn-
C3G fatty acid is converted into C29 alkane in cabbage,
thetic materials have not been produced at a price that will
Brassica oleracea (Brassicaceae), and similar results have allow them tu compete with the natural substances. The most
now been obtained in a number of other plants. Oxygen
important natural waxes are camauba from palms of the
is required for this decarboxylation. Further, an <x-hydroxy
genus Copernicia, candelilla from Euphorbia antisyphlitica,
intennediate is involved (Kolattukudy, 1980).
and sugar cane wax-a by-product of the sugar industry
Secondary alcohols, ketones, and ketols also are synthe-
(Schery, 1972).
sized from long-chain fatty acids. Ketones generally appear
Camauba wax is comprised mostly of wax esters, cande-
to be synthesized from secondary alcohols by oxidation. The
Iilla contains a significant hydrocarbon fraction, and sugar
mechanistic details of these reactions remain obscure.
cane wax is a complex mixture of materials.
In an exceptional case, Pinus jeffreyi essential oils consist
For the last several years, there has been mnch interest
of about 98% n-heptane. Acetate can serve as a precursor
in jojoba, Simmondsia chinensis, a plant native to the south-
for this compound in the plant.
western United States and Mexico, because of the presence
of liquid wax esters in the seeds (Wisniak, 1988). Numerons
TAXOI'lOI'lIC VSEFULl'IESS schemes to convert desert land into orchards of these plants
have been proposed. The oil is an excellent lubricant, has
Virtually all plants have hydrocarbons in the same molecu- good emollient properties, and may replace spenn whale oil
lar-weight range; those of C 29 , C3l , and C33 are essentially for many uses. As the fatty acid combination of Limnanthes
ubiquitous. The same is true for wax esters and other long- douglasii (meadowfoam, Limnanthaceae) is quite similar to
chain lipid constituents of plants. Although a small number that of jojoba, the synthesis of a similar wax ester by hydroly-
of plants have been examined, there appears to be little varia- sis and reduction followed by esterification has been pro-
tion that would serve to differentiate plant groups (Eglinton posed (Miwa and Wolff, 1962).
and Hamilton, 1963). Hexadecanol and octadecanol have been used extensively
in the sonthwestern United States to produce a monolayer
I'Vl'ICDOI'l OF WAXES on the top of lakes and other nonfJowing bodies of water to
retard evaporation. These compounds also have potential
As previously mentioned, waxes function primarily to pro- agricultural applications (Taylor et aI., 1964). Gennination
tect the plant, by preventing entry of foreign organisms and of bluegrass seed was increased by application of these mate-
desiccation of the plant. However, waxes may play other rials to soil (Atsatt and Bliss, 1963).
 Plant Waxes 53

BIOLOGICAL ACTIVITY and the ants protect the wasps from predators as well. The
cuticular ratios of alkanes and alkenes of the ants and wasps
were found to be identical. The compounds from these two
It is generally assumed that long-chain hydrocarbons are
insect species proved to be identical to the cuticular hydro-
relatively inert and lack pronounced biological activity.
carbons from the spines of the tree (Espelie and Hermann,
However, octacosanol (C2S ) has androgenic activity in
1988).
chicks (measured by comb growth) similar to 11,000 times
Alkanes also play an important role in nest mate recogni-
the same amount of testosterone propionate.
tion of wasps (Polistes metricus). However, these same sig-
The application of similar compounds to crop plants has
nals may announce the wasp's presence to possible predators
been conducted (Taylor et aI., 1964). I-Triacontanol, a
(Espelie and Hermann, 1990; Espelie et aI., 1990).
straight-chain fatty alcohol, has been isolated from alfalfa
and is sometimes found in other leaf waxes as the palmitate
ester. This compound has been reported to be a powerful
plant growth stimulant. Application of amounts as low as 4 CHEMICAL FOSSILS AND PETROLEUM
mg per acre can lead to 10-40% increases in growth and
yield of food crops. Application of this compound to corn A major portion of the petroleum used today is derived from
and rice plants produces a response within minutes-even lipids of plants deposited in past eons. The structures of most
in the dark. Triacontanol increased hypocotyllength of ger- compounds isolated from petroleum suggest a derivation
minating cucumber seeds by 14% over controls, but it from fatty acids. Various chemical changes have occurred,
showed inconsistencies in field tests on agricultural crops. mostly reduction and decarboxylation, so that most petro-
I-Docosanol reportedly stimulates growth in auxin assays. leum is comprised of a mixture of odd- and even-chain-
Lower homologs with 16-28 c",bon atoms and structural length hydrocarbons. Some petroleums have a variety of
isomers with other hydroxyl substitutions are ineffective other compounds that appear to be derived from chemical
(Gross, 1980). modification of other types of secondary metabolites.
Surface waxes of plants are important in contact chemore- These compounds may be viewed as chemical fossils. In
ception by insects (Stadler, 1984, 1986). After being at- some instances, the mixture of compounds may be correlated
tracted to the plant by visual and olfactory cues, the undam- with a particular plant or plant groups, although such correla-
aged plant surface is normally the first point of contact. The tions are usually difficult in coal and in petroleum.
physical nature of the surface also is important in determin-
ing whether the insect will accept the plant. Long-chain fatty
acids are involved in the biting behavior of Locusta migra- CUTIN AND SUBERIN
toria (Chapman, 1977). Undoubtedly, many more volatile
compounds are dissolved in the surface waxes, but the waxes Cutin, the lipid that covers the outside of epidermal cells,
themselves also are important for sutface recognition. and suberin, which is associated with cork cells in plants,
Some nest parasites in social insects are protected because are largely comprised of cross-linked hydroxy fatty acids.
their cuticular chemistry is very similar to that of the hosts. The composition of each type of compound varies from plant
The complex cuticle hydrocarbon blends of parasite and host to plant. Generally, cutin is distinct from suberin in the type
are virtually identical and the hosts cannot distinguish the of monomeric units present (Harwood, 1980).
parasites from conspecifics (Stowe, 1988). Nest parasites Cutin is composed chiefly of a C I6 andlor a CIS family
of the genus Trichopsenius biosynthesize their hydrocarbon of monomers (Fig. 4.2). The usual components of the former
mixture; the pattern matches the species-specific mixture of are palmitic acid, 16-hydroxypalmitic acid, and 1O,16-dihy-
their termite hosts. In contrast, the scarabaeid beetle, Myr- droxypalmitic andlor its positional isomers in which the mid-
mecaphodius excavatacollis, which associates with ants, ac- chain hydroxyl group is at C 9 , C g , or C7 • Monomers that
quires its hydrocarbons by association with the host and can could be derived from the above acids by further oxidation
invade the nests of several different species of ants with or reduction also have been found in cutin from some plants.
different hydrocarbon blends (Stowe, 1988). In another bi- The C I8 family consists of stearic acid, oleic acid, linoleic
zarre example, larvae of the neuropteran, Chrysopa slosso- acid, 18-hydroxyoleic acid, 18-hydroxy-9,IO-epoxystearic
nae, cover themselves with wax plucked from their wax- acid, and 9,1O,18-trihydroxystearic acid together with their
coated aphid prey. The ants that tend these aphids fail to 11.-12 unsaturated analogs. Members of the C I6 family are
recognize the neuropterans and do not attack them (Stowe, the dominant components of the cutin offast-growing plants,
1988). whereas the cutin of slower-growing plants with a thicker
The social wasp. Parachartergus aztecus, an ant Pseu- cuticle consists of a mixture of C16 and CIS monomers (Har-
domyrmex ferrugineus, and the tree they inhabit, Acacia col- wood, 1980).
lins;;, exemplify a complex biological interaction (Espelie The major aliphatic components of suberin are w-hydroxy
and Hermann, 1988). The ants feed on and protect the tree acids, the corresponding dicarboxylic acids, very long acids
from herbivory and infringement by other plants and fungi, (greater than Czo), and similarly long alcohols. Among the
the trees provide a place for the ants and wasps to nest, w-hydroxy and dicarboxylic acids, monounsaturated CI8 and
54 Plant Waxes

CH,(CH')'4COZH iHZ(CH,),CH=CH(CH,),COZH
I
OH OH

CHz(CH,),GH-CH(CHz),COzH
I
OH
"i
y = 8, 7, 6, or 5 CHz(CH,),CH-CH(CHz),COzH

x+y=13
I I I
OH OHOH

OH
I
HOzC(CH,).CH(CH,).COzH
i H
OHC(CHz).CH(CHz),COzH

OHOH
I I ~HIH ~H~H
HO(CH,).CH-CH(CH,),COzH HO(CH,).CH-CHCHzCH-CH(CHz),COzH

o
II
HO(CH,).C(CHz).COzH

Fig. 4.2. The major components of cutin (Harwood, 1980; Kolattukudy and Espelie, 1985; modified and used with pennission of the copyright owners,
Academic Press, Orlando and Springer-Verlag, Berlin).

saturated C,. acids are often the dominant components (Har- BAGNERES, A. G. and E. D. MORGAN, A simple method for analysis
wood, 1980). of insect cuticular hydrocarbons, J. Chem. Ecol., 16. 3263-3276
Both cutin and suberin contain phenolic components that (1990).
are presumably esterified to the polymeric matrix. p-Coum- CHAPMAN, R. F., The role of the leaf surface in food selection by
arlc acid and lesser amounts of ferulic acid are found (Kolat- Acridids and other insects, Comportement des insectes et milieu
trophique, Colloques Intern.tionaux CNRS, 265, 133-149
tukudy and Espelie, 1985). However, a recent reevaluation
(1977).
of the composition of suberin indicates that there is no solid
DAVIN, L. B. and N. G. LEWIS, Pbenylpropanoid metabolism: Bio-
evidence for aliphatic-aromatic linkages, nor is it under-
synthesis of monolignols, lignans and neolignans, lignins and
stood how the phenolic units are joined, nor are their identies
suberins, in Phenolic Metabolism in Plants (H. A. Stafford and
clear (Davin and Lewis, 1992). R. K. Ibrahim, eds.), 325-375, Plenum, New York, 1992.
EGl.INTON, G. and R. J. HAMn.TON, The distribution of alkanes, in
Al'IALYSIS OF WAXES CbenticalPlantTaxonomy (T. Swain, ed.), 187-217, Acadentic
Press, London, 1963.
Hydrocarbons from waxes are usually removed by dissolu- ESPEUE, K. E. and H. R. lIERMANN, Congroent cuticular hydrocar-
tion in a nonpolar solvent such as purified hexane and ana- bons: Biochemical convergence of a social wasp, an ant and a
lyzed by capillary gas chromatography. Most common hy- host plant, Biochem. Sys!. Beol., 16, 505-508 (1988).
drocarbons can be identified readily by mass spectrometry EsPEUE, K. E. and H. R. lIERMANN, Surface lipids of the social
(Bagneres and Morgan, 1990). Analysis of wax esters is wasp Polistes annularis (L.) and its nest and nest pedicel, I.
Chern. Ecol.,16. 1841-1852 (1990).
more difficult, as several different compounds in the rather
ESPEUE, K. E., J. W. WENZEL, and G. CHANG, Surface lipids of
complex mixtures obtained possess the same molecular
social wasp Polisites metricus Say and its nest and nest pedicel
weight. Analysis of these mixtures has been effected by tan-
and their relation to nesbnate recognition, I. Chern. Ecol., 16.
dem mass spectrometry (MS-MS) methods (Spencer and 2229-2241 (1990).
Plattner, 1984).
GROSS, D., Wachstumsregulatorisch wirksame Pflanzeninhalts-
stoffe, Z. Chern., 20. 397-406 (1980).
REFERENCES HARWOOD, J. L., Plant .cyllipids: Structore, distribution, and analy-
sis, in Lipids: Structore and Function (p. K. Stumpf, ed.), Vol.
ATSATI, P. R. and L. C. Buss, Some effects of emulsified hexa- 4 of The Biochemistry of Plants (P. K. Stumpf and E. E. Conn,
octadecanol on gennination, establishment, and growth of Ken- eds.), I-55, Acadentic Press, New York, 1980.
tocky bluegrass, Agron. J., 55, 533-537 (1963). KOLA'ITUKUDY, P. E., Cutin, suberin, and waxes, in Lipids: Struc-
 Plant Waxe,f 55

ture and Function (P. K. Stumpf, ed.), Vol. 4 of The Biochemis- STADLER, E., Contact chemoreception, in Chemical Ecology of In-
try of Plants (P. K. Stumpf and E. E. Conn, eds.), 571-645, sects (W. J. Bell and R. T. Carde, eds.), 3-35, Chapman &
Academic Press, New York, 1980. Hall, London, 1984.
KOLATIUKUDY, P. E. and K. E. EsPELIE. Biosynthesis of cutin, STADLER, E., Oviposition and feeding stimuli in leaf surface waxes,
suberin, and associated waxes, in Biosynthesis and Biodegrada- in Insects and the Plant Surface (B. Juniper and R. Southwood,
tion of Wood Components (T. Higuchi, ed.), 161-207, Aca- eds.), 105-121, Arnold, London, 1986.
demic Press, Orlando, FL, 1985.
STOWE, M. K., Chemical mimicry, in Chemical Mediation of Coe-
MrwA, T. K., Jojoba oil wax esters and derived fatty acids and volution (K. C. Spencer, ed.), 513-580, Academic Press, San
alcohols: Gas chromatographic analysis, J. Am. Oil Chem. Soc., Diego, CA, 1988.
48, 259-264 (1971).
TAYLOR, S. A., S. R. OLsEN, J. P. VAVRA, W. J. ROBERTS, G. M.
MrwA, T. K. and I. A. WOLFF, Fatty acids, fatty alcohols, and wax
AUBERTlN, G. W. GoRSLINE, J. P. LAw, JR., and F. M. RHOADBS,
esters from Limnanthes douglasii (meadowfoam) seed oil, J.
Can fatty alcohols reduce water losses?, Crops Soils, 16(8), 7-9
Am. Oil Chern. Soc., 39, 320-322 (1962).
(1964).
POLLARD, M. R., J. B. OHLROGGE, and P. K. STUMPF, Wax ester
formation in the developing jojoba seed, in Proceedings of the WISNIAK, J., The Chemistry of Jojoba, American Oil Chemical
3rd International Conference on Joboba, (D. M. Yermanos, ed.), Society, Champaign, JL, 1988.
71-81, International Committee on Jojoba and the Dept. of YERMANOS, D. M. (ed.), Proceedings of the 3rd International Con-
Botany and Plant Science, University of California Press, River- ference on Joboba, International Committee on Jojoba and the
side, 1978. Dept. of Botany and Plant Science, University of California
SCHERY, R. W., Plants for Man, 2nd ed., Prentice-Hall, Englewood Press, Riverside, 1978.
Cliffs, NJ, 1972. YERMANOS, D. M., Jojoba, in Resource Materials (T. A. McClure
SPENCER, G. F. and R. D. PLATTNER, Compositional analysis of and E. S. Lipinsky, eds.), Vol. 2 of Handbook of Biosolar Re-
natural wax ester mixtures by tandem mass spectrometry. 1. sources (0. R. Zaborsky, ed.), 245-257, CRC Press, Boca
Am. Oil Chern. Soc., 61, 90-94 (1984). Raton, FL, 1981.
5
Polyketides

Introduction plants, for example, as flavonoids, naphthoquinones, and an-

Biosynthesis of Polyketides in Plants and Fungi thraquinones.
Tetraketides Many polyketides have pronounced physiological effects
Pentaketides and several are important antibiotics.
Hexaketides
Heptaketides
Octaketides BIOSYl'fl'HESIS OF POLYKETIDES Il'I PLAl'ITS
Tetracyclines Al'ID FUl'IQI
Decaketides
Macrolide Antibiotics Few areas of natural products chemistry have seen as many
Ionophores major advances in the study of biosynthetic pathways as
Acetogenins have occurred in polyketide compounds. Birch and Donovan
Polyketide Compounds from Unusual Starter Units (1953) demonstrated that a wide range of structural types
Methylenebisphloroglucinols and Related Compounds are derived from acetate (later shown to be acetate and malo-
Polyketides from Lichens nate). In experiments with deuterated precursors, acetate
Mycotoxins serves preferentially as a starter unit for the formation of 6-
Functions of Polyketide Compounds methylsalicylic acid in Penicillium griseofulvum (Simpson,
Phytotoxins 1983). Thus, polyketides are derived from the same precur-
Phytoalexins sors as fatty acids and the initial step seems to be similar (Fig.
References 5.1). Extensive purification of 6-methylsalicylate synthetase
from Penicillium patulum has been performed. This enzyme
system is distinct and separable from the co-occurring fatty
Il'ITKODUCTlOI'l acid synthetase and has a molecular weight approximately
half that of the former enzyme. NADPH is required as a
Polyketide or polyacetate compounds are derived from ace- coenzyme for methylsalicylate synthetase from this source
tate-malonate pathways and, in terms of biosynthesis, are (O'Hagan, 1990; Packter, 1980).
related to fatty acids. Polyketides are assembled by conden- In contrast to the reaction sequence observed with fatty
sation of acetate and malonate units; however, in contrast acid synthases, the sequential reduction, elimination, and
to fatty acid biosynthesis, the carbonyl groups may not be reduction of fatty acid biosynthesis does not occur. The re-
reduced and intermediate compounds typically condense to sulting intermediates usually are cyclized into phenolic sys-
produce aromatic ring systems, usually with phenolic substi- tems; although in practice these compounds show varying
tutions (Packter, 1980). degrees of reduction andlor oxygenation (Simpson, 1987).
Many polyketides have restricted patterns of distribution, It has been suggested that as each malonate unit is added,
but members of this group are common in algae, bacteria, the newly added portion may undergo any or none of the
fungi, and lichens. In general, polyketides from these organ- steps above, and in this way, "polyketide precursors,"
isms are water soluble and formed in high yield, late in the which show varying degrees of reduction and deoxygena-
growth cycle. However, polyketides also are found in higher tion, can be assembled in a stepwise manner on the synthase

56
 o
Polyketides 57

o 0
II~ II lie-
CH3C-CoA CH3CCH,C-CoA
on polyketide synthetase
H'cl""",O enzyme complex
H,~
c;:iCOSACP
H/I ......... 0.
u/1n

/r
ACPSCO '-.)'

-
000
II II II
CH3CCH,CCH,CSACP

bound on enzyme surface

from acetyl·CoA
)JL( from the latest
malonyl.ACP added

ACP =acyl carrier protein

Fig. 5.1. Fonnation of polyketide intennediates.

before being released from the enzyme by a stabilizing-ring lecular Claisen condensation produces another (phloroglu-
condensation (Simpson, 1987). While the intennediate is cinol structures) (Fig. 5.3).
still bound, alkylation of some activated methylene groups In order for these condensations to occur, the reactive
may occur. Acyl-CoA species are involved, but the exact sites must approach more closely than is possible in a linear
nature of enzyme-bound intennediates has yet to be estab· precursor molecule. One proposed mode of action of polyke-
lished. Although there is a pronounced "starter effect" in· tide synthetase enzymes is to facilitate fonnation of a Z dou-
volving acetyl CoA, other starter units such as methylmalo· ble bond into the molecule (Fig. 5.4). In this manner, the
nate and ethylmalonate occur (Simpson, 1987). reactive groups for the above types of condensations can
The "chain length" of the polyketide intennediate in- become proximal. This could be accomplished either by eno-
creases by units of two carbon atoms to various lengths. lization of a carbonyl function or by reduction and dehydra-
Cyclization of intennediates occurs by one of two pathways. tion. The fonner process is reversible and should result in
An intramolecular aldol condensation produces aromatic loss of label (by exchange) at position a. Reduction and
rings with one substitution pattern (Fig. 5.2) and an intramo- dehydration should result in relative loss of label at position

rZ?
oUo
CO,R

000
R
~COzR ---=:::.

HOVOH
R

(yCOZR

HOVOH
tv-
orsellenic acid (2)
Fig. 5.2. Orsellenic acid and aldol~type condensation.
 v
58 Polyketides

_:
0\JR
OO~
_
o~R:~_ ('OR
0
R
0
0
-----"'-
'-. - «::""""

o 0
o

HOXOH

Y OH engenone (8)
Fig. 5.3. Eugenone and Claisen-type condensation.

b. Examples of both mechanisms have been discovered (Fig. odds that a carbon adjacent to a pair of acetate-derived car-
5.4) (Simpson, 1984). bon atoms in which I3C_I3C is enriched also will be 13C
As is true in other groups of secondary metabolites, sev- labeled are low. Because of this, only the I3C_I3C of acetate-
eral types of reactions follow the initial cyclizations: addition derived groups will be seen. By means of these and a variety
and removal of hydroxyl and methoxyl groups, oxidation of of other isotopic labeling and NMR techniques, details of
methyl groups to alcohols, aldehydes, and acids, the reverse many proposed pathways for polyketide compounds have
of this series, and decarboxylation. Decarboxylation of 13- been investigated (Simpson, 1984; 1987; Torssell, 1983).
keto esters is especially common.
Many recent studies have used precursors labeled with
13C, I·C, 2H, 3H, and 18 0 (Simpson, 1986, .1987). Use of TETRAIrnTIDES
2H and 180 precursors permits detection of specific hydrogen
and oxygen atoms by means of the isotope shifts that are Several examples are known in which four C2 units are incor-
induced in I3C-NMR (nuclear magnetic resonance) absorp- porated and cyclized to produce phenolic molecules; 6-meth-
tions. Further, much of the information that formerly re- ylsalicylic (I) and orsellenic acid (2) are formed in this man-
quired degradation of compounds can now be obtained by ner. In the presence of malonyl-CoA and NADPH, 6-
these nondestructive methods. 3H-NMR spectroscopy has methylsalicylic acid, triacetic acid, lactone (3), and fatty
proven useful as this isotope has a nuclear spin of 112, a acids are produced by an enzyme preparation from Penicil-
slightly higher sensitivity than IH and gives sharp peaks and lium patu/um. In the absence of NADPH, only the lactone
chemical shifts and coupling constants similar to those of (3) is formed (Fig. 5.5).
1H. Deuterium is inexpensive, has a nuclear spin of 1, is Formation of 6-methylsalicylic acid involves condensa-
stable and nonradioactive, and can be accurately integrated, tion of an acetyl-CoA unit with three units of malonyl-CoA.
but sensitivity is low. 13C also has a spin of 112 and a large The entire sequence is enzyme bound and intermediates are
chemical shift range (200 ppm). Sensitivity is also lower not released. Furthermore, attempts to introduce exogenous
than that for IH, but this is partially compensated by determi- intermediates in feeding studies failed. As mentioned above,
nation of spectra with proton decoupling (Torssell, 1983). extensive purification of 6-methylsalicylate synthetase from
I3C-NMR spectroscopy is especially useful when it is possi- Penicillium patu/um yielded an enzyme complex distinct and
ble to introduce precursors enriched in 13C. Determination separable from fatty acid synthetase and approximately half
of I3C_ 13C spin-spin couplings often provides information in molecular weight. NADPH is required for production of
about which intact two-carbon units are present (when 6-methylsalicylic acid (packter, 1980). The application of
[1,2- 13C2]-acetate is used as a precursor) (Torssell, 1983). NMR spectroscopy to study of the biosynthetic steps in the
From casual observation of structures in which I3C_I3C formation of 6-methylsalicylic acid and other polyketides
couplings of intact acetate units are marked, it might appear has been reviewed (Simpson, 1987).
that all adjacent carbons would be coupled (if all the carbon Some more complex compounds are formed by union of
atoms in the molecule come from acetate). However, as the tetraketide-derived rings. For example, usnic acid (4) is
overall percentage oflabeling is relatively low, the statistical derived by oxidative coupling of two molecules of methyl-
 Polyketides S9

phloracetophenone (5), each produced from four C2 units Another complex polyketide compound, stipitatonic acid
(Fig. 5.6). The enzymes responsible for most in vivo coup- (7), is derived from four C2 units (Fig. 5.9) (Weiss and Ed-
lings are peroxidases which function in the presence of a wards, 1980).
porphyrin cofactor. A series of lactones from Melodorum fruticosum (Annon-
Patulin (6), also derived from a tetraketide precursor, is aceae) may be of tetraketide derivation (Fig. 5.10). These
a potent carcinogen produced by certain Penicillium and As- compounds have cytotoxic activity in the KB and P-338 cell
pergillus species, sometimes found in moldy apples and a lines (Tuchinda et aI., 1991).
variety of other fruits, as well as barley (Beier and Nigg,
1992). Patulin inhibits respiration of higher plant cells and
has a neurotoxic effect on animals. 6-Methylsalicyclic acid
(1) (see above) is an intermediate in the biosynthetic pathway PENI'AKEnDES
leading to patulin (6) (Fig. 5.7) (Gaucher, 1979; Simpson,
1983; Zamir, 1980). The oxidation steps are carried out by A series of pentaketides derived from five C 2 units is also
monooxygenases. The pattern of label introduced from intro- known. Eugenone (8) (Fig. 5.3) occurs in the essential oil
duction of 14C-labeled acetic acid was of value in the deter- of cloves, Syzygium aromaticum (Myrtaceae). Citrinin (9),
mination of the reaction path (Fig. 5.8) (Gaucher, 1979). a carcinogenic antimicrobial metabolite produced by several

:yY~
R~SEnz
:W· ~SEnz
o 0 ~hYdration !.nolizati:n in D2~
/<:088

u9(oo- R((_---: <xl

~ of protons at position a)

,,_:,.. 0

CR30~Oy
D'8~
OR OR 0
/ 0 0 ~,,:,.,.:.

RO;
Wi
0.3 ~ I
~
0
OR

OR 0
by enolization

o OR 0 OR

CR30:@1 CR30~1
I I 0

0.4 :::,...
I I OR 0.4 ,..
..........
0
OR 0
OR 0 OR ,(6oR OR 0 OR
o ,:y I by enolization
RO :::,.. OR
1

Fig, 5.4. Preliminary steps of polyketide biosynthesis (Simpson, 1984; modified and used with permission of the copyright owner, the Royal Society of
Chemistry. London).
60 Polyketides

.~- -
0 0 HO 0

~-
acetyl·CoA NADPH NADPH

2 malonyl.CoA methylsalicylate
synthetase

/ 1
D V - ~" -
OH

o 0
~
C02H
OH D~
rr2COCOA
(3) SEm
6.methylsalicylic
acid (1) CO..

Fig. 5.5. Proposed biosynthesis of 6-methylsalicylic acid.

oy rs:w
V
o
Y
"(1" .
SCoA

C,' '-r 0

o
. o
---

rY.
COMe OH

.A( ----..~
HOyYOH radical formation

oOXOH

(5)
OH
o
OH HO

.~ovrrOH
"-.J0 ~
0 •

HO~ 0 ~ -
HO 0
o
0
OH HO

- ·H2O

HO 0

usnic acid (4)

Fig. 5.6. Biogenesis of usnic acid (Herbert, 1981; modified and used with pennission of the copyright owner, Chapman & Hall, London).
 Polyketides 61

JyCO'H
acetyl-CoA
3 malonyl-CoA
-- UOH

6-methylsalicylic
acid

V - (r0H
OH OH
oxygenase
¢roo OH

m-cresol OH
m-hydroxybenzyl
alcohol
OH o

N CHO
O~I
Y
O"NADPH
dehydrogenase OH
oxygenase ,f - -
H I
HO' H
OH

genistaldehyde isoepoxydon (49)

(a)

--
phyllostine (55)

HO
D' 0
..

HO
It -:?' h

OH
21"
,r

o
h

OH

isopatulin E-ascladiol patulin (6)

(b)

Fig. 5.7. (a & b) Biogenesis of patulin in Penicillium urticae (modified from Stoessl, 1981; used with pennission of the copyright owner, Academic
Press, New York).

:1'(X
3~"

#7
Ii'

OH
COH--
8 2
8
.7'"
3

5 4 0
2'
.d

1
.,0

OR
7

6-methylsalicylic acid (1) patulin (6)

OH

Fig.5.S. Incorporation of 14C label from carboxyl labeled acetic acid into
6-methylsalicylic acid and patulin (Gaucher, 1979; modified and used with stipitatonic acid (7)
pennission of the copyright owner, the International Association of Milk,
Food, and Environmental Sanitarians, Des Moines, IA). Fig. 5.9. Stipitatonic acid.
62 Polykelides

o ~ O~ 0 -
H
O~
)l
CH,

o O~ "0/ \ "" O~
O~CH' O~

Fig. S.10. Lactones from Meladorumfruticosum.

[1-1"c]-a<etate
OW""·
, ..
.
.
HOC:::"" :::,... 0
OH
C1

0:0:):/'0.. HOW
CH,-CO,Na --+
• elK
0 0
SR
- I 0
""" SR
o 0 HO 0

! C-I reduction

HO

WWW ,p-

"""
HO
I
'-'I ")

(11)
0;-'
\
0

H'
L --
---".
HO 17

"""
HO
I
CHO
OH C-3_
_
reduction
HO 17

"""
HO
(14)
I
CliO
0

O~ O~
~b
OH
(10)
- HO'C~~')
OH citrinin (9)

··.W
R R
HO~
AyJyb
OH 0 OH 0

(IS) (12)R=CH, (13)R=H

Fig. 5.11. Biogenesis of citrinin (Simpson. 1983; modified and used with pennission of the copyright owner. the Royal Society of Chemistry. London).
 Polykerides 63

Aspergillus species and by Penicillium citrinum, also is a (12) was incorporated into citrinin, whereas the correspond-
representative of this series, This pentaketide was at least ing compound (13), without methyl groups, was not. This
partly responsible for fatalities caused by "yellow rice dis- suggests that methylation occurs early in the biosynthetic
ease" in Japan during the early part of this century and to pathway, probably before aromatization. Compound (12)
a lesser extent following World War II (Ciegler, 1975), was shown by other methods not to be an obligate precursor
Several of the proposed steps of the biosynthetic pathway of citrinin, however. Compound (14), but not compound
for this compound (Fig, 5.11) have been reexamined. The (15), was incorporated into citrinin.
fungus that produces citrinin has been grown in media made
with 0 20. By adding normal protium eH)
containing pre-
cursors to the cultures, it is possible to measure incorporation
of that isotope. By study of the extent and sites of incorpora- IrnXAKBTJDES
tion of protium ('H) with combined I3C_, 2H_ and 'H-NMR
techniques, the origin of various hydrogen atoms in the ci-
trinin molecule could be determined. For example, the hy- The biosynthesis of ascochitine (16), a phytotoxic hexake-
drogen on C-4 of citrinin (9) is acetate derived. The marked tide from the fungus Ascochyta labae, has been investigated
difference in the amount of protium at C-I and C-3 suggests by means of I3C-Iabeled acetates and methionine and 14C_
that reductions at these two sites occur at markedly different labeled precursors (Ballio, 1981; Stoessl, 1981) (Fig. 5.12).
stages in the biosynthesis (Simpson, 1983). Ascochitine is derived from a single hexaketide precursor
Further, in the I3C-NMR spectrum of [1_I3C, "Oz]-ace- with introduction of three methyl groups from methionine
tate-enriched citrinin, isotopically shifted signals are ob- via a quinone-methide structure (17) similar to that involved
served for the resonances due to C-3, C-6, and C-8. This in the biosynthesis of citrinin. The aldehyde (18) and qui-
indicates that oxygen atoms attached to each of these carbons none-methide (17) were specifically incorporated.
come from acetate. The quinone-methide structure (as in The naphthoquinones plumbagin (19) from Plumbago
compounds 9 and 10) must be formed by the elimination of spp. (Plumbaginaceae) and 7-methyljuglone (R = CH3 ) (20)
hydroxyl (as water ?) from the hemiacetal (11). Compound from Drosera spp. (Droseraceae) are derived from six Cz

0X)):f0 -- H0)O¢D
C,
1\
--+
• ,If
I °
0 0
SR "'" SR
C,
o 0 HO 0

~
O~ HO~ HoxX0
~6 I ~6 I
I °
""
HO
CHO
HO OH
OH
(17) (18)

i
":~
Ho,c~6
OH
I
ascochitine (16)

Fig. 5.12. Biogenesis of ascochitine (Simpson, 1983; modified and used with permission of the copyright owner, the Royal Society of Chemistry,
London).
64 Polyketides

[0] [0] HEPTAKETIDES

! ! Griseofulvin (21), an antifungal antibiotic representative of

W-
the heptaketide series, is obtained from Penicillium griseo-
fulvum and probably is biosynthesized as indicated in (Fig.
o 0
COA ~
y y
5.14). Formation of this molecule involves both a Claisen
and an aldol condensation. The intermediate compound
o
OU OU
griseophenone B (22) is then cyclized via a free-radical
coupling to normethyldidehydrogriseofulvin, as discussed in
o 0
Chapter 6.

W
o

OCTAKETIDES

Although many anthraquinones in plants are derived from

OU 0 shikimic acid pathways (see Chapter 6), others, such as an im-
portant plant and fungal anthraquinone, emodin (25), are syn-
plumbagiD (19) '-methyljuglone (20)
thesized from octaketide units. This compound is widely dis-
Fig. 5.13. Biogenesis of plwnbagin and 7·methyljuglone. tributed in fungi, especially in the genera Penicillium and
Aspergillus, and is an important intermediate in the biosyn-
units (Fig. 5.13) (Durand and Zenk, 1971; Packler, 1980). thesis of many other fungal metabolites (Sanakawa, 1980).
In this unusual instance, the acetate arises from L-tyrosine The biosynthesis of helminthosporin (26) from Helmin-
(Haslam, 1986). Most naphthoquinones, however, are de- thosporium gramineum is similar to that of emodin (Fig.
rived from shikimate pathways and will be discussed in the 5.15).
following chapter. Because of secondary reactions (such as loss and/or sub-

OU 0 OU

~O -~
CU,COCoA
r
l
6Cj,COCOA

co,u UO~~ ~OU

O~coAJ-I ..
OU

OU 0 OH OH 0 OCH,

--{()c}1
I ~ - (xJi1
CH,O
I"'"
:::"... OH
.& OH CH,O
~.&
OH OH

griseofulvin C
griseophenone B (22)
(24)

~¢:Y
coupling
CH,o~
OH

CI
I
0

0-
OCH,

0
~
¢0:r
CH,O:::"'"
0CH'

I
0

O. -
OCH,

0
_ _ ~OCH'
0 OCH,

CH,O:::"'"
I
0
0

CI· CI

normethyldidehydrogriseofulvin 5 ,6 -didehydrogriseofulvin griseofulvin (21)

(23)

• label from acetate preferentially incorporated here

Fig. 5.14. Biogenesis of griseofulvin (modified from Vleggaar and Steyn. 1980; used with pennission of the copyright owner, Academic Press. New
York).
 Polyketides 6S

.
-- ~
o 0 O. OH 0 OH

• • •
OSEDZ ~COSEn.
? ~
CH,CO,H • 000 --- I I
o ••• HO% ~

HO~ -
OH OH

~~ HO

1 emodin anlbrone
o
emodin (25)

OH 0 OH o OH
OH

o
helminthosporin (26) alizarin (27)

Fig. 5.15. Probable biosynthesis of emodin and helminthisporin.

sequent addition of hydroxyl and methyl groups), it is some- toxic and carcinogenic natural products known. Various
times difficult to predict the biosynthetic origin of com- strains of Aspergillus (and other fungi) produce these potent
pounds of this series. Generally, those anthraquinones with carcinogens, which cause lesions in the mammalian liver
substituents on both A and C rings [such as emodin (25)1 (Beier and Nigg, 1992; Coulombe, 1991; Simpson, 1984;
are derived via the acetate-malonate pathway, whereas those Steyn, 1980). Aflatoxins are found in many food products,
with substituents in only one ring derive from shikimate but are most commonly encountered in corn (Zea mays) and
and malonate (Fig. 5.15). The structurally similar compound peanuts (Arachis hypogaea).
alizarin (27) is synthesized from iso-chorismic acid, a-keto-
glutarate, and mevalonate. Anthraquinones derived from
shikimate pathways will be discussed in Chapter 6. MACROLIDE Al'lTlBIOTICS

A number of polyoxygenated fungal metabolites are assem-

bled from c" C 3 , and C. units. Starter units of propionyl-
TETRACYCLIl'IES
CoA and successive additions of methylmalonyl-CoA are
especially important in the formation of many of these com-
Tetracyclines, formed from the condensation of eight C 2
pounds (O'Hagan, 1990). Most of these metabolites possess
units and a malonamide starter, are produced by a number
large rings and are potent antibiotics; they are known collec-
of Streptomyces species from an unusual group of bacteria,
tively as macrocyclic or macrolide antibiotics. One well-
the actinomycetes (Fig. 5.16). Many tetracyclines, such as
known example of this group of compounds is erythromycin
terramycin (28), aureomycin (29), and tetracyline (30), are
A (34) from Streptomyces erythreus (Fig. 5.18) (Omura,
broad-spectrum antibiotics (Thomas and Williams, 1984,
1984; Simpson, 1984). An extensive literature deals with
1985; Weiss and Edwards, 1980).
these antibiotic compounds but is beyond the scope of this
text.

DECAKETIDES
IOI'lOPHORES
The fungal metabolite aflatoxin B1 (31) is derived from a co-
occurring xanthone sterigmatocystin (32) which is, in turn, A group of natural products, ionophores, which disrupt nor-
produced by oxidative cleavage of the anthraquinone versi- mal ion gradients, function as antibiotics by binding to metal
colorin A (28) (Fig. 5.17). Aflatoxins are among the most ions and transporting them across cell membranes. One of
66 Polyketides

CONH, 8 CH,COCoA O~SEnz

tH1COCO; bOi ---- ~NH'--

o 000 0

-+
NH,

OH OH HO HO o OH OH HO HO o

OH ~HO
,:? H
,
O
OH

NH, "I
- . . ..........
" :
1 NH
1

OH OH o o OH 0 OHO 0 0

/
anhydrotetraeycline dehydrotetracycllne

~
~
I
P
I
OH H R H N(CH,),
I! ~ OH «m(
PI'
""-
CI OH
H

""- •
H
-
H N(CH,),
OH
I
NH,
""- ""- • NH, HO
HO OH 0 HO 0 0
OH 0 HO 0 0

R =OH, terramycin (oxytetracycline) (28) aureomycin (29)

R =H, tetracycline (30) (7·chlorotetracycline)

Fig. 5.16. Biogenesis of terramycin, tetracycline, and aureomycin (Thomas and Williams, 1984, modified and used with pennission of the copyright
owner, the Royal Society of C.hemistry, Cambridge),

CH,CO,H - -
O~
0 0 --
HO~""OH
I I
o ""- h
OOCoAOOO OOOHOH

averantin

averufin
--~
HO~I
o
0

0
",,0

OH
0yO
OHH

versiconal hemiacetal acetate

~O~
-
--+

versicolorin A (33) sterigmatocystin (32) aflatoxin B1 (31)

Fig. 5.17. Biogenesis of aflatoxin (Coulombe, 1991; modified and used with pennission of the copyright owner. the CRC Press. Boca Raton, FL. ©
1991).
 Polyketides 67

CH,CH,COCoA

CH,
I
6CHCOCoA
I
co·

dadlno.se
erythromycin A (34)

Fig. 5.18. Biogenesis of erythromycin.

these compounds, lasalocid A (35) (Fig. 5.19) has complex POLYKETlDE COlllPOUI'IDS FROM UNUSUAL
stereochemistry and oxygen substitution that serve to coordi- STARTER Vl'IITS
nate metal ions (Williams et al., 1989).
Many polyphenols are derived from malonate units and a
starter other than acetate. The starter unit for tetracyclines
(previously discussed) is malonamido-CoA (Fig. 5.16).
ACETOGEl'IIl'IS Members of the Anacardiaceae characteristically produce a
series of compounds based on unsaturated fatty acid precur-
sors. The surface of a number of plants of this family is
A series of acetate-derived compounds have been reported
covered with a mixture of phenols well known for their irri-
from plants of the family Aunonaceae (Annona cherimolia,
tating properties to humans. Poison ivy, Toxicodendron radi-
A. muricata, A. squamosa, and other Annona species; Asim-
cans, commonly is involved in human poisoning in eastern
ina triloba, pawpaw; Goniothalamus giganteus; and several
North America. The family of phenols known as urushiols
Rollinia and Uvaria species) (Fig. 5.20) (Alkofahi et al., are the major irritant constituents (Fig. 5.21), but the CoA
1989; Lieb et al., 1990; Nahrstedt, 1985; Rupprecht et al., ester of palmitoleic acid usually serves as a precursor of
1986). Most of these compounds exhibit potent antimicro- this group of compounds. The side chain structure of active
bial, antiparasitic, cytotoxic, and antitumor activity (Bories compounds is mono-, di-, or triunsaturated, suggesting that
et al., 1991). For example, asimicin (36), from the bark of a number of unsaturated acyl-CoA starter units can be em-
Asimina triloba, is extremely cytotoxic (EDso <10- 7 tJiml ployed. The pure compounds are very active allergens in
with several cell cultures); fractionation of the plant material man; those compounds with a catechol structure are the most
was monitored by the brine shrimp bioassay (AIkofahi et aI., potent in this regard. Related compounds from Mangifera
1989). This compound also is toxic to the striped cucumber indica, the mango, are thought to be responsible for resis-
beetle, Mexican bean beetle, mosquito larvae, blowfly lar- tance to the fungal pathogen black spot disease, Alternaria
vae, melon aphid, two spotted spider mite, and the free- alternata (Harborne, 1989). Similar substances have been
living nematode, Caenorabditis elegans (Rupprecht et al., isolated from Amphypterygium atistringens, a plant placed
1986). in the family Julianaceae (Navarette et al., 1989). These and

Iasalocid A (35)

Fig. 5.19. The ionophore antibiotic lasalocid A.

68 Polyketides

R' other data confinn that this family should be merged with
the Anacardiaceae.
7
Pest-resistant geraniums (Pelargonium species, Gerania-
R" 0
0 ceae) contain primarily C22 and C24 unsaturated anacardic
acids (Walters et aI., 1990). For example, geranicardic acid
(37) and similar compounds from the cultivated geranium
(Pelargonium X hortorum) are largely responsible for resis-
R R R' tance of the plants to the two-spotted spider mite, Tetrany-
R"
asirnjcin (36) OH OH OH chus urticae (Craig et aI., 1986; Gerhold et aI., 1984). In
asimicintriacetate OAc OAc OAc Pelargonium X hortorum. anacardic acids were shown to
uvaridn OAc OH H be biosynthesized from saturated and w-saturated C 16 and
OH CIS fatty acids by chain elongation with three acetate groups
(Walters et aI., 1990).
These and similar compounds are found in the Anacardia-
ceae, Geraniaceae, Ginkgoaceae, Myristicaceae, Myrsina-
ceae, Poaceae (Gramineae), and Proteaceae.

METHYLBNBBISPHLOKOGWCINOLS
OH
colliRODe
AND ReLATBD COMPOUI'IDS

Fig. 5.20. Acetogenins from the Annonaceae. Several ferns, mostly those of the genus Dryopteris, elabo-
rate compounds in which methylated phloroglucinol units
(such as 38) are coupled through methylene groups to yield

Z
CH,(CH,),CH=CH(CH,hCOSCoA

palmitoleic acid

7'0
/CH,-C"-.,
CH,(CH,),CH=CH(CH,),C CH
II / '
0 CH~
/ I 0
COSCoA \ OH

CH'(CH,),CH=CH(CH'h-Q'
- CH,(CH,),CH=CH(CH,),
-Q~
anacardic acid CO2" OH -
\ CO,H OH

j
CH'(CHzl'CH=CH(CHzl,-Q \

urushiol no on
OH

CH,<CHzl,CH=CH(CHzlr-Q CH'(CHzl'CH=CH(CH'),Q

cardanol OR cardol OH

OC
HO
CO'H

'>--
I Z
(CH,),CH=CH(CHzl,CH,

geranicardic acid (37)

Fig. 5.21. Compounds from poison ivy and geranicardic acid (modified from Geissman and Crout, 1969).
 Polyketides 69

compounds such as 39-43 (Fig. 5.22). Each of the phloro- A series of compounds which share many structural fea-
glucinol molecules is based on a C4 starter unit and subse- tures occur in dicotyledonous plants. Drummondin A, B, and
quent addition of three units of malonate. The extensive C- C [44,45, and 46, respectively, (Fig. 5.23)] from Hypericum
methylation is derived from S-adenosylmethionine and ap- drummondii (Clusiaceae) have significant activity against
pears to occur while the polyketide chain is bound to a pro- the gram-positive bacteria Staphylococcus aureus, Bacillus
tein surface (Geissman and Crout, 1969). subtilis, and the acid-fast bacterium Mycobacterium smeg-
Even more complex molecules are formed by coupling matis. Japonicin A (47), from the same plant, has antimalar-
of these units. Most of these couplings are through methyl ial activity. Robustaol A (48), from Eucalyptus robusta
groups that are ultimately derived from S-adenosylmethio- (Myrtaceae), has activity against Plasmodium berghei (Boris
nine. Filixic acid (39) is one of the most common of these and Schaeffer, 1992). Similar compounds, isolated from
compounds. These compounds have been used medicinally Mallotusjaponicus (Euphorbiaceae), have cytotoxic activity
as anthelminthics and purgatives. (Jayasuriya et aI., 1989).

CH,CH,CH,COSCoA
. _ CH,CH=CHCOSCoA
. biotin,COz

ATP

CO,H

.~
(
I
CH,CH=CHCOSCoA
•
--
.
CH,CH,CH,COCH,CH=CHCOSCoA
.
CH CH CH CO-SCoA
3 1 2

oxidation .
CH,CH,CH,COCH,COCH,CH,COSCoA
. C-methylation

hydration malonyl-eoA

.
CH,CH,CH,COCH,COCHCOCH,COSCoA _
. CH,CH,CH,CO - -
•
HOO·OH
~
I
1

:o:56r
CH, OH
(al

H
HO OH 'r OH
1 1 1

¢CO'
CH3CH2CH2CO :::-.. -COCH zCHzCH3
o OH CH
p·aspidin(39)
HO "'" OH "'" OH
H0X)c:(XIHOOH I I
1 1 1 1 CH,CH,CH,CO"'- ~ ~ "cOCH,CH,CH,
CH,CH,CH,CO COCH CH CH OH OH
o 0 2 2 3
margaspidin (41)
albaspidin (40)
CH, CH, :Q:5O:CH'

~ ~I
\/ HO OH"",OH
HO?, 0 0 r OH I I
CH,CH,CH,CO COCH,CH,CH,
CH,CH,CH,CO OH OH COCH,CH,CH, 0 OH
desaspidin (43)
methylenebisaspidinol
(42) COCH,CH,CH,

H000HO~OHH000H filixic acid (38)

CH,CH,CH,C~COCH,CH,CH'
(b)
o OH 0

Fig. 5.22. (3 & b) MetbyJenebispbloroglucinols from ferns (modified from Geissman and Crout, 1969).
70 Polyketides

drummondiD A (44)

drummondln C (46) japonicln (47)

robustaol (48)

Fig. 5.23. Antimicrobial compounds of mixed polyketide origin.

POLYKETlDI!S FROM LICHEI'IS ously discussed, among them are the aflatoxins patulin (6)
and citrinin (9). A few other common and important myco-
Lichens, organisms in which fungi and algae live in associa- toxins are discussed below.
tion, produce numerous polyketide-derived compounds, Animals that consume moldy feeds excrete the estrogenic
often in very large amounts in proportion to the dry weight polyketide mycotoxins zearalenone (51) and zearalenol (52)
of the organism (Culberson, 1969). In general, it appears in milk and milk products (Fig. 5.24). The same compound
that the fungal member of the pair synthesizes these com- sometimes is found in beer. Zearalenol and zearalenone are
pounds from carbohydrates provided by the alga. Many of synthesized by Fusarium species; these compounds produce
the compounds also are found in free-living fungi, but only sterility and other reproductive problems in livestock (Beier
rarely in algae. A substantial number of these substances are and Nigg, 1992).
unique to lichens. These polyketides are used frequently to Fumonisins are from Fusarium moniliforme, a fungus that
assist in the identification of the approximately 18,000 occurs worldwide on a variety of plant hosts, including
known species of lichens (Culberson, 1969, 1970, 1977). wheat, barley, soybeans, rice, maize, and oats (Beier and
The chemistry of lichen substances has been reviewed (Elix Nigg, 1992). Although man and domestic livestock are af-
et aI., 1984; Huneck and Yoshimura, 1996). fected, horses seem to be particularly sensitive. They suffer
Depsides, in which esterification to links two polyketide- a neurotoxic syndrome in which the white matter of one
derived units [such as lecanoric acid (50») (Fig. 5.24), are or both cerebral hemispheres liquifies. Fumonisins also are
quite common among lichens (Weiss and Edwards, 1980). highly carcinogenic. Of this group of compounds, fumonisin
Many of these compounds are strongly allelopathic and are B, (53) is the most common, often representing about 70%
responsible for the death of trees inhabited by the dense of the total fumonisins detected (Plattner et aI., 1992).
growth of lichens (Cutler, 1992). Methods for detection and analytical determination of
mycotoxins at low levels have been reviewed (Wilkinson et
aI., 1992).

MYCOTOXJI'IS
fUNCTIONS OFPOLYKETlDE COMPOVNDS
Many fungi produce compounds that are highly toxic to man Polyketides frequently possess potent physiological activity
and his domestic animals. A large number of these are de- which is expressed in a variety of ways. Some compounds
rived from polyketide pathways. Several have been previ- of this group, such as morphogens, appear to be involved in
 Polyketides 71

~y
O~O
OH
(+)-isoepoxydon (49) patulin (6)

1-(3,5-dichloro-2,6-dihydroxy- lecanoric acid (50)

4-melhoxyphenyl}-1 hexanone (53)

HO HO

OH o
zearalenone (51)
zearalenol (52)
OH
~
o
o 0

OH OH o OH

O~OH
fumonisin 8 1 (53)
8 o~A~OH.. 8
Fig. 5.24. Some biologically active polyketides.

fundamental developmental levels, whereas others are in- dom exert any apparent function in the physiology of the
volved in biological interactions with other organisms. producing organism. However, in nature, interactions of
Morphogens are signal molecules in embryos that are, as fungi and other organisms are complex and may involve
one example, involved in establishing the spatial pattern of plants, animals, and other fungi (Wicklow, 1984, 1988). For
cells during development. 1-(3,5-Dichloro-2,6-dihydroxy-4- example, (+ )-isoepoxydon (49) has been established as the
methoxyphenyl)-I-hexanone (54), apparently derived from major causative agent of interference between Poronia punc-
polyketide pathways, is required for producing the tata, a late fungal colonist of cattle dung, and two earlier
prestalkJprespore pattern in the aggregate formed by cells colonists, Ascobolus furfuraceaus and Sordaria fimicola
of the slime mold Dictyostelium in response to starvation (Gloer and Truckenbrod, 1988).
(Morris et al., 1987). Several homologs and analogs have
since been isolated (Jaenicke, 1991). Phytotoxins
Many of the polyketides produced by filamentous fungi
are water soluble, formed in high yield, and removed from Phytotoxins, compounds produced by pathogenic organ-
the intracellular pool by excretion into the culture medium. isms that contribute to the death and decomposition of the
Although these compounds frequently possess antibiotic ac- host plant (Ballio, 1981; Harborne, 1986, 1989; Stoessl,
tivity and a number have commercial importance, they sel- 1981), are well represented among polyketides.
72 Polyketides

0
OH

0
HO~
o
Fig. 5.25. Phytotoxins from the Dutch elm disease fungus, Ceratocystis ulmi.

cytochalasin B (56) cytochalasin D

Fig. 5.26. Some representative cytochalasins.

Dutch elm disease, caused by the fungus Ceratoeystis ical tools. Cytochalasin B (56), for example, disrupts micro-
ulmi, is disseminated by a bark beetIe of the genus Seolytus. filament assembly and contractile processes within cells
The fungus produces toxins that cause necrotic lesions and (Stoessl, 1981). These compounds are both phytotoxins and
leaf wilting. These toxins consist of a mixture of glycopro- mammalian toxins.
teins and three low-molecular-weight phenolic compounds Other powerful phytotoxins, T-Toxins from the fungus
(Fig. 5.25) (Claydon et aI., 1974; Harborne, 1982, 1986; Helminthosporium maydis race T, are derived from acetate
Wood et aI., 1972). (Fig. 5.27) (Kono et aI., 1981).
A compound that serves as an intermediate in the biosyn-
thesis of patulin (6), phyllostine (also known as epoxydon) Phytoalexins
(55), (Fig. 5.7) has been identified as a phytotoxin from a
Phyllostieta species that is pathogenic to red clover (Ballio, Another group of biologically active compounds, phyto-
1981). Patulin also has been reported to possess phytotoxic alexins, are produced by plants in response to invasion by
activity (Stoessl, 1981). Further, several structurally related foreign organisms, typically fungi and bacteria.
compounds from a number of different fungal species seem One of the earliest reported phytoalexins was 6-me-
to be involved in phytotoxicity (Stoessl, 1981). thoxymellein (57) (Fig. 5.28) produced by carrot root slices
Cytochalasins, a complex group of polyketide-derived after inoculation with Ceratoeystis fimbriata. This fungus,
microbial metabolites, exert profound effects on a variety of which causes black rot disease in sweet potatoes (Ipomoea
cellular processes (Fig. 5.26) and are used widely as cytolog- batatas, Convolvulaceae), is not pathogenic in carrots (Dau-

o 0

Fig. 5.27. T~Toxins from Helminthosporium maydis.

 Polyketides 73

--
ow
CO,H CH3-C0,H
co,
4CHj'"'C0,H I
enzyme 4CHf"'COEnz enzyme

o 0 0 0

~Enz--
o 0

HOW
-- 0-- 1.,9
OH 0

'·methoxymellein (57)

Fig. 5.28. 6-Methoxymellein biosynthesis (modified from StoessI. 1981; used with pennission of the copyright owner Academic Press).

cus carota, Apiaceae). Treatment of carrot tissues with ethyl- N. Nigg and D. S. Seigler, eds.), 117-158, Plenum, New York,
ene and a number of other chemicals also has been shown to 1992.
induce fOimation of 6-methoxymellein, as do several carrot CIEGLER, A" Mycotoxins: Occurrence, chemistry, biological activ-
storage fungi (Bailey and Mansfield, 1982; Yates, 1987). ity, Lloydia, 38, 21-35 (1975).
CLAYDON, N., J. G. GROVE, and M. HOSKEN, Phenolic metabolites
of Ceratoeystis ulmi, Phytochemistry, 13, 2567-2571 (1974).
COULOMBE, R. A., Oflatoxins. in, Mycotoxins and Phytoalexins (R.
REFERENCES P. Sharma and D. K. Salunkhe, eds.), CRC Press, Boca Raton,
Florida, 1991.
ALKOFAHI, A, 1. K. RupPREClIT, 1. E. ANDERSON, J. L. McLAUG- CRAIG, R., R. O. MUMMA, D. L. GERHOLD, B. L. WINNER, and R.
HLIN, K. L. MIKOLAJCZAK, and B. A. SCOIT, Search for new SNETSINOER, Genetic control of a biochemical mechanism for
pesticides from higher plants, in Insecticides of Plant Origin (J. mite resistance in geraniums, in Natural Resistance of Plants to
T. Amason, B. J. R. Philogene, and P. Morand, eds.), AC.S. Pests (M. B. Green and P. A Hedin, eds.), ACS Symposium
Symposium Series 387, 25-43, American Chemical Society, Series 296, 168-176, American Chemical Society, Washington,
Washington, DC, 1989. DC, 1986.
BAILEY, J. A and J. W. MANSFIELD, Phytoalexins, Wiley, New CULBERSON, C. F., Chemical and Botanical Guide to Lichen Prod-
York,1982. ucts, University of North Carolina. Press, Chapel Hill, NC,
BALLIo, A., Structure-activity relationships, in Toxins in Plant Dis- 1969.
ease (R. D. Durbin, ed.), 395-441, Academic Press, New York, CULBBRSON, C. F., Supplement to Chemical and Botanical Guide
1981. to Lichen Products, Bryologist, 73, 177-377 (1970).
BEIER, R. C. and H. N. NIGG, Natural toxicants in foods, in Phyto- CULBERSON, c.
F., CULBERSON W. L., and A. JOHNSON, Second
chemical Resources for Medicine and Agriculture (H. N. Nigg Supplement to Chemical and Botanical Guide to Lichen Prod-
and D. S. Seigler, eds.), 247-367, Plenum, New York, 1992. ucts, American Bryological Lichenological Society, S!. Louis,
BIRCH, A J. and F. W. DONOVAN, Studies in relation to biosyn- MO,1977.
thesis. I. Some possible routes to derivatives of orcinol and CUTLER, H. G., Herbicidal compounds from higher plants, in Phyto-
phloroglucinol, Aus!. 1. Chern., 6, 360-368 (1953). chemical Resources for medicine and agriCUlture (H. G. Nigg
BoRIES, C., P. LoISEAU, D. CORTES, S. H. MYlNT, R. HOCQUE- and D. S. Seigler, eds.), 205-226, Plenum Press, New York,
MILLER, P. GAYRAL, A CAYE, and A LAURENS, Antiparasitic 1992.
activity of Annona muricata and Annona cherimolia seeds, DuRAND, R. and M. H. ZBNK, Biosynthesis of plumbagin (1,5-
Planta Medica, 57, 434-436 (1991). hydroxy-2-methyl·1,4·naphthoquinone) via the acetate pathway
BORIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from plants, in higher plants, Tetrabedron Lett., 3009-3012 (1971).
in Phytochemical Resources for Medicine and Agriculture (H. Eux, J. A, A A WHl'ITON, and M. V. SARGBNT, Recent progress
74 Polyketides

in the chemistry of lichen substances, Fortschr. Chern. Org. Vol. 4 of The Biochemistry of Plaots, (P. K. Stumpf and E. E.
Naturst., 45, 103-234 (1984). Conn, eds.), 535-570, Academic Press, New York, 1980.
GAUCHER, G. M., Mycotoxins-Their biosynthesis in fungi: Pa- PLA1TNER, R. D., D. WEISLBDER, D. D. SHACKLEFORD, R. PETER-
tulin and related carcinogenic lactones, 1. Food Protect., 42, SON, and R. G. POWELL, A new fumonisin from solid cultures
810-814 (1979). of Fusarium moniliforme, Mycopathologia, 117, 23-28 (1992).
GEISSMAN, To' A. and D. H. G. CROUT, Organic Chemistry of Sec- RUPPRECHT, J. K., C. CHANG, J. M. CASSADY, 1. L. McLAUGHLIN,
ondary Plant Metabolism, Freeman Cooper, San Francisco, K. L. MIKOLAJCZAK, and D. WEISLEDER, Asimicin, a new cyto-
1969. toxic and pesticidal acetogenin from the pawpaw, Asimina tri-
GERHOLD, D. L., R. CRAIG, and R. O. MUMMA, Analysis of trichome loba (Annonaceae), Heterocycles, 24, 1197-1201 (1986).
exudate from mite-resistant geraniums, I. Chern. Ecol., 10, SANAKAWA, U., The biosynthesis of anthraquinonoid mycotoxins
713-722 (1984). from Penicillium isandicum Sopp and related fungi, in The Bio-
GLOER, J. B. and S. M. TRUCKENBROD, Interference competition syothesis of Mycotoxins (P. Steyo, ed.), 357-394, Academic
among coprophilous fungi: Production of ( + )-isoepoxydon by Press, New York, 1980.
Poronia punctata, Appl. Environ. Microbiol., 54, 861-864 SIMPSON, T. 1., The biosynthesis of polyketides, in Specialist Peri-
(1988). odical Reports, Biosynthesis, Vol. 7 (R. B. Herbert and T. J.
HARBORNE, J. B., Introduction to Ecological Biochemistry, Aca- Simpson, eds.), 1-44, Royal Society of Chemistry, London,
. demic Press, London, 1977; 2nd edition, 1982. 1983 .
HARBORNB, J. B., Recent advances in chemical ecology, Nat. Prod. SIMPSON, T. J .. The biosyothesis of polyketides, Nat. Prod. Rep.,
Rep., 4, 323-344 (1986). 1,281-297 (1984).
SIMPSON, T. 1., 13C_NMR in metabolic studies, in Nuclear Magnetic
HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod.
Resonance (H. F. Linskens and J. F. Jackson, eds.), 1-42,
Rep., 7, 85-109 (1989).
Springer-Verlag, Berlin, 1986.
HASLAM, E., Secondary metabolism-Fact or fiction, Nat. Prod.
SIMPSON, T. J., Applications of multinuclear NMR to structural and
Rep., 4, 217-249 (1986).
biosynthetic studies of polyketide microbial metabolites, Chern.
HERBERT, R. B., The Biosynthesis of Secondary Metabolites, Chap- Soc. Rev., 16, 123-160 (1987).
man & Hall, London, 1981.
STEYN, P., (ed.), The Biosyothesis of Mycotoxins, Academic Press,
HuNECK S. and I. YOSHIMURA, Identification of Lichen Substances, New York, 1980.
Springer-Verlag, Berlin, 1996.
STOESSL, A., Structure and biogenetic relations: Fungal non-host-
JAENICKE, L., Development: Signals in the development of cryp- specific, in Toxins in Plant Disease (R. D. Durbin, ed.),
togams, Prog. Bot., 52, 138-189 (1991). 109-219, Academic Press, New York, 1981.
JAYASURIYA, H., J. D. MCCHESNEY, S. M. SWANSON, and 1. M. STOESSL, A., Biosynthesis of phytoalexins, in Phytoalexins (1. A.
PEzzUTO, Antimicrobial and cytotoxic activity of rottlerin-type Bailey and J. W. Maosfield, eds.), 133-180, Wiley, New York,
compounds from Hypericum drummondii, J. Nat. Prod., 52, 1982.
325-331 (1989).
THOMAS, R. and D. J. WILUAMS, Oxytetracycline biosyothesis:
KONO, Y., H. W. KNOCHE, and J. M. DALY, Structure: Fungal host- Mode of incorporation of [l- 13 C,2H3]acetate, J. Chern. Soc.,
specific, in Toxins in Plaot Disease (R. D. Durbin, ed.)~ Chern. Comm., 443-444 (1984).
221-257, Academic Press, New York, 1981.
THOMAS, R. and D. J. WILLIAMS, Oxytetracycline biosynthesis:
LIBR .. F., M .. NONFON, V. WACHBNDORFF-NEUMANN, and D. WEN- Mode of incorporation of [1- 13C, 180 2 ]acetate, J. Chern. Soc.,
DISCH, Annonacine und Annostatin aus Annona squamosa, Chern. Comm., 802-803 (1985).
P1anta Medica., 56,317-319 (1990).
TORSSELL, K. B. G., Natural Product Chemistry, John Wiley and
MANN, J" Secondary Metabolism, Oxford University Press, Ox- Sons, Chichester, 1983.
ford, 1978; 2nd edition, 1987.
TuCHlNDA, P., J. VDCHACHON, V. REUTRAKUL, T. SANTISUK, W. C.
MORRIS, H. R., G. W. TAYLOR, M. S. MASENTO, K. A. JERMYN, and TAYLOR, N. R. FARNSWORTH, 1. M.I'Ezzuro, and A. D. KINGH-
R. R. KAy, Chemical structure of the morphogen differentiation ORN, Bioactive butenolides from Melodorumfruticosum, Phyto-
inducing factor from Dictyostelium discoideum, Nature, 328, chemistry, 30, 2685-2689 (1991).
811-814 (1987). VLEGGAAR, R. and P. STEYN, The biosynthesis of some miscella-
NAHRSTEOT, A., Neue pharmakologisch interessant Naturstoffe neous mycotoxins, in The Biosynthesis of Mycotoxins (P.
(1983-1984), Oesterreichische Apotheker-Zeitung, 39, Steyn, ed.), 395-422, Academic Press, New York, 1980.
733-741 (1985). WALTERS, D. S., R. CRAIG, and R. O. MUMMA, Fatty acid incorpora-
NAVARETIE, A., R. MATA, and G. DELGADO, Alky1anacardic acids tion in the biosynthesis of anacardic acids of geraniums, Phyto-
from Amphypterygium adstringens, Planta Medica, 55, 579 chemistry,29, 1815-1822 (1990).
(1989). WEISS, U. and J. M. EDWARDS, The Biosynthesis of Aromatic Com-
O'HAGAN, D., Evolution of the polyketide metabolites, Chern. Br., pounds, Wiley, New York, 1980.
246-250 (1990). WICKLOW, D. T., Ecological approaches to the study of mycotoxi-
OMURA, S., Macrolide Antibiotics, Academic Press, Orlaodo, FL, genic fungi, in Toxigenic Fungi-Their Toxins and Health Haz-
1984. ard (H. Kurala and Y. Veno, eds.), 33-43, Kodaosha, Tokyo,
PACKTER, N. M., Biosyothesis of acetate-derived phenols (polyke- 1984.
tides), in Lipids: Structure and Function (P. K. Stumpf ed.), WICKLOW, D. T., Metabolites in the coevolution of fungal chemical
 Polyketides 7S

defense systems, in Coevolution of Fungi with Plants and Ani· WOOD, R. K. S., A. BALLlO, and G. GRANITI (eds.), Phytotoxins in
rnals (K. Pirozinski and D. Hawksworth, eds.), 173-201, Aca- Plant Diseases, Academic Press, London, 1972.
demic Press, London, 1988. YATES, S. G., Increased concentration of myristicin and 6-me·
WILKINSON, A. P., C. M. WARD, and M. R. A. MORGAN, Immuno· thoxymellein in carrot root upon irradiation with UV light, in
logical analysis of mycotoxins, in Plant Toxin Analysis (R. F. Al1elochemicals: Role in Agriculture and Forestry (G. R. Wal-
Linskens and J. F. Jackson, eds.), 185-225, Springer-Verlag, ler, ed.), ACS Symposium Series 330, 295-299, American
Berlin, 1992. Chemical Society, Washington, DC, (1987).
WILLIAMS, D. H., M. 1. STONE, P. R. HAUCK, and S. K. RAHMAN, ZAMIR, L. 0., The biosynthesis of patulin and penicillic acid, in The
Why are secondary metabolites (natural products) biosynthes· Biosynthesis of Mycotox ins (P. Steyn, ed.), 223-268, Academic
ized? J. Nat. Prod., 52, 1189-1208 (1989). Press, New York, 1980.
6
Benzoquinones, Naphthoquinones, and
Anthraquinones

Introduction properties of quinones result from their tendency to form the

Benzoquinones aromatic hydroquinone system (Morrison and Boyd, 1973).
Biosynthesis Both para- (3 and 6) and ortho-quinones (4) (Fig. 6.3) are
Distribution of Benzoquinones Among Higher Plants known. Quinonoid compounds in which a second aromatic
Biological Functions of Quinones in Plant-Plant ring is fused to the benzoquinone ring are called naphthoqui-
Interactions nones (6 and 7); those in which an aromatic ring is fused to
Biological Functions of Quinones in Plant-Animal both sides of the benzoquinone ring are called anthraqui-
Interactions nones (9-11). Anthrones (12) are related structurally tu an-
Naphthoquinones thraquinones. Anthrones are readily oxidized to anthraqui-
Biosynthesis of Naphthoquinones nones (12 is converted to 11).
Distribution of Naphthoquinones Among Higher Plants Many quinones are derived from acetate-malonate path-
Biological Functions of Naphthoquinones ways (discussed in Chapter 5), some from shikimate path-
Anthraquinones ways (discussed in Chapter 7), and others are derived by
Biosynthesis of Anthraquinones oxidative modification of secondary metabolites from a vari-
Distribution of Anthraquinones ety of other pathways. Quinones of this last type will be
Biological Functions of Anthraquinones discussed with the compounds to which they are biosyntheti-
Economic hnportance and Uses cally related.
Anthrones and Anthrone Dimers Most quinonoid compounds probably occur in plants as
Hypericin and Anthrone Dimers glycosides and many of the benzoquinones, naphthoqui-
References nones, and anthraquinones that have been studied are arti-
facts of isolation.
The IH_NMR spectra of anthraquinones and anthraqui-
INl'RODVCTIOI'l none glycosides have been discussed (Kalidhar, 1989).

Quinones, naphthoquinones, and anthraquinones are found

in many types of higher plants and fungi. About 600 natu- BEl'IZOQVIl'IOl'lES
rally occurring quinones have been described. More than
half of these are from one plant family, the Rubiaceae (Has- Simple quinones or benzoquinones are found commonly in
lam, 1974; Harhorne, 1982). Quinones are cyclic Ol,J3-dike- nature. Some quinonoid compounds such as the electron car-
tones of such a structure that they are converted by reduction rier ubiquinone (13), sometimes known as coenzyme Q, play
into hydroquinones, that is, phenols contaiuing two hydroxyl important primary roles in plants. Structurally related plasto-
groups such as (2) (Fig. 6.1) (Morrison and Boyd, 1973). quinones are important as electron carriers in photosynthesis.
The reduced and oxidized forms are closely balanced ener- Secondary compounds with similar structures, for exam-
getically and are easily interconverted. ple, polygonaquinone (5) (Figs. 6.1 and 6.2), are encountered
The highly conjugated oxidized forms of quinones are in species of Fagopyrum (buckwheat), Polygonum (smart-
colored; for example,p-benzoquinone (1), is yellow. In con- weed), Rumex (dock), and Rheum (rhubarb) in the Poly-
trast, the reduced forms usually lack color. Many chemical gonaceae (Mathis, 1966).

76
 Ben::oquinones. Napluhoquinones, and Anthraquinones 77

o
o

o
OK
reduction ....
"'" oxidation
o OK
p-benzoquin.ne (1) hydroquinone

*?
yellow (1)

W
I
I I
KO
o OK 0

arbutin (14) pyrolatin perez.ne (3) plumbagin (6)

o o

QQ OK 0

jugl.ne (7)
~
~ OK 0
belminth08porin (9)
® OK 0

(8)

~
~
o
emodin (11) emodin anthrone (12) 2-methylanthraquinone
(10)

Fig. 6.1. Quinones from higher plants.

Biosynthesis oxidation of many different types of primary and secondary

As mentioned above, quinones commonly arise by po- compounds. For example, hydroquinone and its J3-n-gluco-
lyketide pathways, by the shikimic acid pathway, and by side, arbutin (14) (Fig. 6.1), are known to occur in many
oxidation of a number of secondary metabolites of varied plants. This compound appears to arise by oxidative decar-
biosynthetic origin. In some instances, the same compound boxylation of p-hydroxybenzoic acid. Although arbutin is
is known to arise by distinct pathways in different organisms. widespread, it is especially common in the family Ericaceae.
In general, polyketide pathways are favored in bacteria, A number of naphthoquinones are also found in this family.
fungi, and lichens, whereas shikimic acid pathways are fa- The Myrsinaceae are known to produce and accumulate
vored in higher plants (Kubitzki and Gottlieb, 1984a, many benzoquinones. Among these are embelin (15), rapa-
1984b), although there are many exceptions to this generali- none (16), and maesaquinone (17) (Fig. 6.2). Maesanin (18),
zation. It is not always possible to determine the pathway from the fruit of Maesa lanceolata, is a powerful antibiotic
of origin for a particular quinone by structure alone, but a agent. A hot-water extract of the fruits is used locally in
knowledge of plant families and their relationships permits Africa to prevent cholera (Harbome, 1986).
an educated guess. In order to be certain, the pathway must Many ortho- and para-quinones based on terpenoid struc-
be established by biosynthetic studies in each case. tures are found in the Lamiaceae (Labiatae, the mint family)
especially in the genus Salvia (4, 19,20) (Fig. 6.3), the Mal-
Distribution of Benzoquinones Among vaceae (Gossypium and Hibiscus), the Bombacaceae (Bom-
HlgberFlants bax), Boraginaceae (Cordia, 21-25), the Asteraceae (Cac-
Benzoquinones are widely distributed among higher alia) (26, 27), as well as other families. The wood of the
plants. Compounds with quinonoid structure may arise by blue mahoe, Hibiscus elatus (Malvaceae), has a dark bluish
78 Benzoquinones, Naphthoquinones, and Anthraquinones

pene from cotton (see Chapter 21), although cotton is not a

~
O.& H host for this parasite. A compound which serves as a germi-
;....- vitamin Kl (37) nation stimulant for seeds of Striga also has been isolated
"" I I 3 from Sorghum bicolor, a normal host for the parasite. This
compound, 2-hydroxy-5-methoxy-3-[ (8'Z, 11 'Z)-8', 11',14'-
o pentadecatrieneJ-p-hydroquinone (28) (Fig. 6.4) and the cor-
° responding p-benzoquinone (29) co-occurred. Only the hy-

::~,
droquinone was biologically active (down to 10-7 M), but
ubiquinone (13) was readily converted to the benzoquinone (Chang et al.,
1986). Haustorium formation is tightly regnlated in Striga.
If the seed germinates and the parasite seedling root does

»N
not approach the root of the host closely within about 5-7
° days, the parasite seedling will die. Haustorial induction
must occur within about 50 f.LIll of the host root for successful
I plastoquinone attachment (Chang and Lynn, 1986, 1987). In experiments
"" H with the roots of Sorghum bicolor and germinating Striga
o ... asiatica seeds, a haustorial inducing substance, 2,6-dimeth-
oxy-p-benzoquinone (30) (Fig. 6.4) was isolated (Chang and
Yr0H {POH {POH Lynn, 1986, 1987). This compound is widespread among
higher plants, has been shown tn have allelopathic proper-
HO~n-Cl1Ra3 Hoyn.CllH23Ho~n.C13H27 ties, and may be involved in host defense. Further, several
0 0 0 fungi release this compound by the action of laccase en-
polygonaqulnone embellin (IS) rapanone (16)
(5)
zymes. It was shown that the meristematic tips of seedlings
of AgaUnis and Striga produce similar laccases (Chang and
o Lynn, 1987). It is proposed that compounds such as syringic
~OH maesaquinone(17)
acid (31) from Sorghum bicolor roots are oxidized by lac-
cases from Striga asiatica roots to produce 2,6-dinlethoxy-
HOY(CH2)13CH=CH(CHV3CH3 p-benzoquinone (30) (Fig. 6.5). Radicle development termi-
o nates with the induction of haustorial development (Chang
o Z and Lynn, 1986, 1987).
CH,o«(CH,)'CH~H(CH')3CH3 The haustorial inducer from Lespedeza sericea for the
I I maesanln (18)
parasite AgaUnis purpurea, however, proved to be a
OH triterpene, soyasapogenol B (Lynn, 1985).
o
Fig. 6.2. Benzoquinones occurring in flowering plants. Biological Functions of Quinones
in Plant-Animal Interactions

gray color that changes to yellow in the presence of light, Although most benzoquinones do not appear to be highly
which is caused by quinonoid sesquiterpenes present in the toxic to other orgauisms, a few, such as arbutin (14) found
wood (Leistner, 1985). in the nectar of Arbutus unedo (Ericaceae), are known to be
involved in biological interactions. Apparently harmless to
man, this compound is toxic to honeybees (Harborne, 1982).
Biological Functions of Quinones
In most plants, arbutin co-occurs with a l3-glucosidase that
in P1ant-FJant Interactions
converts the glucoside into hydroquinone. Hydroquinone
Parasitic plants are known from about 10 families of also is one of a complement of phenolic compounds leached
higher plants; the Scrophulariaceae and Orobanchaceae con- from the leaves of chaparral plants (in particular from Arc-
tain many parasitic species. Some, such as AgaUnis purpurea tostaphylos and Arbutus species, Ericaceae) in the western
(Scropulariaceae), a hemiparasite, have wide host ranges, United States. This compound inhibits the growth of compet-
whereas others, such as Striga asiatica (Scropulariaceae), a ing species and is important in plant-plant interactions
holoparasite, are restricted to a single host family, in this known as "allelopathy," a phenomenon which will be dis-
case, grasses. Because Striga attacks cereal grains and sugar cussed more fully (see Chapters 7, 8, and 19).
cane (grasses), this parasitic plant has major economic im- The wood of Dalbergia species (a tropicallegnme) con-
portance. Two levels of recognition of the host plant exist: tains quinones with microbiocidal and algicidal activity, is
one associated with seed germination and the other with resistant to marine borers, and causes allergic reactions. The
haustorial formation (a haustorium is a specialized attach- intensely orange heartwood of Dalbergia retusa contains
ment organ found in parasitic plants). Germination of Striga two major pigments: obtnsaquinone (32) (3% of the weight
seeds can be induced by compounds such as strigol, a diter- of the undried wood) and a black pigment, 4-methoxydalber-
 8en::.vquinones, Naphr/lOquinones. and Anrhraquinones 79

()I~
Q$d
o

~:O-<
yu
'" 0

C~';
yqy
(19) (4) (20)
o

cordiachrome A (21) (22) (23)

~
Ytb=
il-Mil
cordiachrome 6 (24) maturinone (26) decompositin (27)

M
R OH

~ ~ ~ ~

OH OH

(25)

Fig. 6.3. Some terpenoid quinones.

gione (33) (Fig. 6.6). Obtusaquinone is highly toxic to fish

(Leistner, 1985).
OH
Other quinones from Australian blackwood, Acacia mela-
OH noxylon (Fabaceae), are also capable of inducing allergenic
reactions. Among these are 2,6-dimethoxybenzoquinone
CH30 (see section above on plant-plant interactions) and 2-
OH methyl-6-methoxyfuranobenzoquinone (34) (Fig. 6.6). 2,6-
Dimethoxybenzoquinone (30) also is found in the wood of
(28)
Bowdichia nitida (Fabaceae) (Leistner, 1985) and in Ama-
ranthus palmeri (Amaranthaceae), where it is responsible
for allelopathic effects (Fischer and Quijano, 1985).
Simple phenols that can be converted readily to quinones
0 by enzymatic oxidation are used by arthropods as defensive
OH secretions. The biosynthetic origin of the phenolic substrates
is not known. Probably the most remarkable of these is the
CH3
defense of bombardier beetles, of the genus Brachynus. A
secretion as hot as IOO'C is produced by a reaction among
0
(29) hydroquinone, a phenolic substrate, H20 2 and the enzyme
Fig. 6.4.A gennination stimulant for Striga asiatica seeds from catalase. A highly exothermic reaction occurs as hydroqui-
Sorghum hie%r and a possible precursor. none is oxidized to benzoquinone, the major product of de-
fense. The beetle, when endangered, discharges a hot explo-
80 Benzoquinones, Naphthoquinones. and Anthraqu;nones

2) -- ¢
CO,H CO,H
I

000. -- OH
I

shikimic acid
:'
LaphenylalBRine
OH
E.4.hydroxycoumaric acid

/
\
HO
4 ~ OH
/"

4
OH

/ galJicacid
0

CH3 "'"'

OH
OCH,
- rnp~_ 0

syringlc acid (31) 2,6-dimethoxy-p-benzoquinone

(30)

Fig. 6.S. Proposed biosynthesis of 2,6-dimethoxy-p-benzophenone.

sive cloud of toxin in the direction of the attacker (Harborne, anthraquinones often have similar structures, the biosyn-
1982). thetic paths leading to individual compounds are sometimes
The spider, Vonones sayi, produces a mixture of quinones different (Inouye and Leistner, 1988).
that presumably serve as defensive compounds. These com- As is true for benzoquinones and anthraquinones, naph-
pounds are both crystalline at ambient temperatures but exist thoquinones are derived both by acetate-malonate and by
as a liquid in the proportions found in the spider. This ap- shikimic acid pathways.
pears to be necessary to permit secretion of the compounds Plumbagin (2-methyljuglone) (6) (and probably 7-meth-
(Mann, 1987). yljuglone) in Plumbago (Plumbaginaceae) and Drosera
(Droseraceae) species is derived from acetate. However, at
least in the family Droseraceae, the acetate is derived from
l'IAPIITIIOQUIl'IOI'IES L-tyrosine (Haslam, 1986).
The biosynthesis of compounds derived from shikimic
Phylloquinones, such as vitamin K J (37) (Fig. 6.2) are found acid is closely linked to that of isomers of vitamin K (35)
in all green plant tissues; menaquinones and other naphtho- (Fig. 6.7). In plants and in microorganisms, the aromatic
quinones are found in bacteria, fungi, and a number of plant ring is formed via the shikirnate pathway, which does not
families. Naphthoquinones occur free or as glycosides within exist in animals. Only recently has it been established that
the plants. They are usually colored and may serve as a vitamin K synthesis branches from iso-chorismic acid (36)
source of pigmentation in some plants. and not from chorismic acid (37). iso-Chorismic acid (36) is
derived from shikimic acid (see Chapter 7) (Leistner, 1986).
Biosynthesis of l'Iapbthoquinones Both of the cyclization steps leading to naphthoquinones and
Notably, many types of naphthoquinones and anthraqui- vitamin K are unusual in plants.
nones are synthesized and accumulated in plant tissue cul- The key intermediate in the formation of these com-
tures. This fortuitous situation has facilitated much recent pounds is o-succinylbenzoic acid (OSB) (38) which is
work on the biosynthesis of naphthoquinones and anthraqui- formed from iso-chorismic acid and a-ketoglutararic acid
nones (Ellis, 1988; Leistoer, 1981, 1986; Simpson, 1986). (39), a tricarboxylic acid cycle intermediate (Fig. 6.7) (Si-
Plant tissue cultures often do not accumulate other types mantiras and Leistoer, 1989). a-Ketoglutaric acid can be
of secondary metabolites. Although naphthoquinones and converted to glutamic acid, which previously was proposed
 Benzoquinones, Naphthoquinones, and Anthraquinones 81

as a precursor to OSB by transamination. In a cell-free sys- 1988; Kolkmann and Leistner, 1987; Leistner, 1986). The
tem prepared from Escherichia coli, iso-chorismic acid and final steps of formation of vitamin K2 (35) involve the addi-
a-ketoglutarate were converted to o-succinylbenzoic acid in tion of a mevalonate-derived unit (Fig. 6.7) (see Chapter
an almost 90% yield (Leistner, 1986). A subunit of the en- 18), and methylation.
zyme system, OSB synthase, is responsible for decarboxyl- Juglone (7) (from Juglans regia) and lawsone (41) (from
ation of a-ketoglutaric acid, and a thiamine pyrophosphate Impatiens balsamina) both come from shikimic acid (Fig.
(TPP) adduct of succinic semialdehyde, the direct product 6.8) (packter, 1980). The initial steps of biosynthesis are
of decarboxylation, has been isolated (Leistner, 1986). identical to those of vitamin K2 (Inouye and Leistner, 1988;
Formation of the coenzyme A ester of the aliphatic car- Kolkmann and Leistner, 1987). Either 1,4-dihydroxy-2-
boxyl group of o-succinylbenzoic acid (38) appears to acti- naphthoic acid (40) or 2-carboxy-4-oxotetralone (COT) (43)
vate the methylene proton sufficiently to permit attack at the is incorporated efficiently into both juglone and lawsone
aromatic carboxyl group (Fig. 6.7). 2-Carboxy-4-oxotetra- (Leistner, 1981). Biosynthesis of juglone proceeds through
lone (COT) (43) andlor 1,4-dihydroxy-2-naphthoic acid a symmetrical intermediate; 1,4-naphthoquinone (42) is an
(DHNA) (40) are likely intermediates for later stages of syn- efficient precursor (Leistner, 1981). In contrast, the biosyn-
thesis. The enzymatic conversion to DHNA with naphthoate thesis oflawsone (41) does not appear to involve a symmetri-
synthase (which consists ofOSB CoA synthetase and DHNA cal intermediate (Haslam, 1974; Inouye and Leistner, 1988).
synthetase) requires ATP, Mg2+, and coenzyme A. 1,4-Dihy- Phylloquinone (37), vitamin K J (Fig. 6.2), closely related
droxy-2-naphthoic acid (40) is a precursor of vitamin K2 to vitamin K2 (35) (Fig. 6.7) in biosynthesis, has a "menadi-
(35) and menaquinones (such as 44) (Inouye and Leistner, one" nucleus and a phytyl side chain (Fig. 6.9) and occurs

Dalbergia retusa
4-methoxydalbergione (33)
OCR,

o
CR3 o A , 0 , - - CR,-O
oR I I%
: O 0
~. RO
o I I
o OCR,
acmelin (34)
o
Acacia melanoxylon
Dalbergia Ian/oUa bowdichione
OR 0 latinone

0 OCR,
diospyrin diosindigo A (53)

CR,O OR 0 OR 0 OR 0
OR

Ro~R ~ '" OR
0 0
OR OCR,
alternin 2-methoxystypandron
diosindigo B

Fig. 6.6. Quinones from Dalbergia and Acacia species (modified from Leistner, 1985; used with pennission of the copyright owner, Academic Press,
Orlando. FL),
82 Benzoquinones, Naphthoquinones, and Anthraquinones

-- 0/"0
coo

w-
chorismie acid (37) iso-ehorismie acid (36)
a-ketoglutarate (39)

TPP = thiamine pyrophosphate

OC1
;:?'

""- I
oa °
SCoA

-
°

°
!° ortho-succlnyl-
benzoic acid (38)
2-carboxy-4-oxotetralone
(COT) (43)

~~~~ ~COOH
~COd_~ -

° OH

1,4-dihydroxy-2-naphthoie acid vitamin K2 (menaquinone)

(40) (35)

Fig. 6.7. Biosynthesis of the naphthoquinone skeleton from o-succinylbenzoate (modified from Inouye and Leistner, 1988; used with pennission of the
copyright owner, John Wiley & Sons, Ltd., Chichester).

in the chloroplasts of all green plants. Under certain environ- homogentisic acid (52) and mevalonate (Fig. 6.10) (Haslam,
mental, hormonal, and cultural conditions, photosyntheti- 1974).
cally active cell cultures of Morinda lucida cease synthesis Naphthoquinones such as alkannin (52) seem to be de-
of phylloquinone and accumulate anthraquinone pigments rived from p-hydroxybenzoic acid by the addition of geranyl
(see Section 6.4.1). pyrophosphate (Fig. 6.3). These naphthoquinones are espe-
When [1-carboxy-14C]o-succinyl benzoate is adminis- cially common in the family Boraginaceae (Leistner, 1981).
tered to plants of Catalpa ovata (Bignoniaceae), it is incor- Dimers of naphthoquinones [such as diosindigo A (53),
porated into a-Iapachones (such as 45), catalponol (46), and (Fig. 6.6)] are formed by free-radical coupling of monomeric
catalpalactone (47). Examination of the 3H/14C ratio in catal- units. These compounds are widespread in plants and usually
ponol (46) after administration of [1-carboxy- 14C,2'-3H2 ]o- co-occur with the corresponding monomeric compounds
succinylbenzoate reveals that the two protons at the 2'-posi- (Scott, 1967).
tion are both retained in the 3-position of catalponol (Fig.
6.9). Thus, prenylation occurs at the 2-position and does Disbibution of l'Iaphthoquinones Among
not involve an aromatic compound such as 1,4-dihydroxy- Higher Plants
2-naphthoic acid (DHNA) (40) (Inouye and Leistner, 1988).
2-Carboxy-4-oxotetralone (COT) (43) or 2-carboxy-4-hy- The distribution of naphthoquinones and anthraquinones
droxy-l-tetralone (48) are possible acceptors for the prenyl among higher plants is closely linked. In some instances,
unit. When 2-carboxy-4-hydroxy-l-tetraione (48), or its however, naphthoquinones are accumulated in families that
methyl ester, was introduced into the plant, the prenyl deriva- normally do not accumulate anthraquinones. Among these
tives of 2-carboxy-4-oxotetralone (COT) (43) and 2-car- are the Balsaminaceae, Boraginaceae (Mathis, 1966), Dros-
boxy-4-hydroxy-l-tetralone (50) were isolated as intermedi- eraceae, Ebenaceae, Ericaceae, Iridaceae, Juglandaceae,
ates (Inouye and Leistner, 1988). Plumbaginaceae, and Proteaceae (Zenk and Leistner, 1968).
Chimaphilin (51), a naphthoquinone found in members Juglone and related compounds occur in the family Juglan-
of the Ericaceae, is derived from shikimic acid, but by a daceae (Carva and Juglans) and the Proteaceae (Conosper-
quite different route. In this case, the precursors appear to be mum, Lomatia, and Stenocarpus) (Leistner, 1985).
 Benzoquinones, Naphthoquinones. and Anthraquinones 83

Biological Functions of l'Iaphthoquinones toxin, juglone. The bound form has been identified as the 4-
glucoside of 1,4,5-trihydroxynaphthalene (54) (Harborne,
A number of naphthoquinones are known to have pro-
1982). (Fig. 6.11). In practice, both root exudation and leaf
nounced physiological effects. One well-known example in-
leachates may be involved in the resulting allelopathic effects
volves the compounds juglone (7) and the [3-g1ucoside of the
corresponding hydroquinone (54) found in walnut, Juglans (Harborne, 1982). Juglone strongly inhibits growth and nitro-
regia (Juglandaceae) and in related species such as Jug/ans gen fixation by Frankia species in European black alder trees
nigra (Leistner, 1985). Juglone is a water-soluble yellow (Alnus g/utinosa) (Neave and Dawson, 1989).
pigment which causes the characteristic brown staining of In order to determine if juglone is effective in "chemical
the hands caused by handling walnuts. It has long been rec- warfare" (or allelopathy) among plants, a number of other
ognized that many garden plants such as tomatoes and lettuce effects, such as shading and mineral and water depletion
do not grow well when planted near walnut trees. Juglone should be considered. Further, if juglone is to be effective,
is toxic and an effective inhibitor of seed germination for it must persist in the soil. Bacteria that degrade juglone have
many plants. At a concentration of 0.002%, it completely been isolated from soil and from beneath walnut trees (In-
prevents germination of lettuce seed. ouye and Leistner, 1988). Some Pseudomonas species are
When tomato and alfalfa plants were planted different dis- able to use juglone as their sale carbon source (Inouye and
tances from walnut trees, plants close to the tree did not sur- Leistner, 1988; Schmidt, 1988). In the field, a number of
vive. The position at which the tomato plants were unaffected presumably coadapted plants grow well near walnut trees
by the "allelopathy" coincided with the extent of the root and appear able to tolerate the effects of juglone.
growth of the tree and it was assumed that the plants were Juglone also has been shown to be inhibitory to many
killed by the exudation of toxin from the roots. Later work insects. Differences in sensitivity exist, and, often, related
suggested that the effect might actually be caused by leaching insect species are not all equally sensitive to a particular
of a bound form ofjuglone from the leaves and stems followed allomone. For example, juglone is a feeding deterrent to the
by hydrolysis and oxidation in the soil to release the actual smaller European elm bark beetle (Scolytus multistriatus)

«r :?' . . o)oH
o o
ricOOrCOOH
0y- o
:,..,
I
o
Impatiens
C02H
balsamina
o

/
(38) (43) lawsone (41)

W - oQ.
OH OH
r
:,..,
I I luglans regia
C02H
:,..,
l';
h
~
vy
OH OH o
(40) ~ (42) ~

~ ~ ~
~- yy
~ ~ OH 0
"" H ?I O-glucose ? / jugJone (7)

"" I I n 't /

yQ:?' y:):?'
OH OH

o (44)
menaquinone ~ ~! --. : :. . . ~ I
(vitamin K 2)
OH OR OH O-glucose

Fig. 6.8. Biosynthesis of juglone and lawsone (modified from Inouye and Leistner, 1988; used with permission of the copyright owner, John Wiley &
Sons, Ltd., Chichester).
84 Benzoquinones. Naphlhoquinones, and Anlhraquinones

(YCOO(COOH ~COOH ~IC~.:~ ~ 0

0y-Vy o 0
-~-Vy
0 0

:
(38) (43) (49)

wro~
" ~
I • I

c¢Y
''''\': OR ? '0 0

I I (48) ~ r (2R,4S).::'ponol (46) ::,... 0

~

¢¢X
....... // catalpalactone (47)

~
?I
R,
00

0
IVY·
R, ~

OH
(2R,4S). (50)
#
Wok-
0

a·lapochones (45)
R,

Fig. 6.9. Biosynthesis of modified naphthoquinones in Catalpa ovata (modified from Inouye and Leistner, 1988; used with permission of the copyright
owner, John Wiley & Sons, Ltd., Chichester),

but not to the closely related hickory bark beetle (Scolytus A number of fungal naphthoquinones, including juglone,
quadrispinosus) (Seigler, 1983). It is probable that the host have been shown to act as phytotoxins (Stoessl, 1981). Jug·
(Carya spp., Juglandaceae) of the hickory bark beetle con· lone is highly toxic to the fungus Fusicladium effusum, a
tains juglone or related compounds and that the insect is pathogen of pecans (Carya illinoensis, Juglandaceae) in the
probably coadapted to the presence of this naphthoquinone. southeastern United States (Leistner, 1985).
The larvae of the chrysomeJid beetle GastroUna depressa Plumbagin (6) is as inhibitory to seed germination as jug·
accumulate juglone, presumably from the leaves of the host lone (7); the effect of a number of naphthoquinones on seed
plant Juglans, on which they feed (Pasteels et aI., 1988). germination of Abutilon theophrastii have been evaluated

6 CO~::, HX
OH

yIy,
I
¢ r o H COOH
y opp _ _

::?
I
--- """ -. ~ COOH CH,OH

"'" OH OH ""- /

(52) ~\

W-W-'C¢r
o 0 OH

o 0 OH
chimaphilin (51)

Fig. 6.10. Biosynthesis of chimiphilin (Haslam, 1974),

 Benzoquinones, Naphthoquinones, and Anthraquinones 85

«>
OR o AI'lTHRAQUIl'IONES

~ Anthraquinones are among the most common quinones

W
1) hydrolysis
found in natnre. Over 200 of these compounds are known
2) oxidation from flowering plants, and many others are produced by
OR O.glucose OR 0 lichens and fungi (see also Chapter 5). Many anthraquinones
occur as glycosides within the plant, but are converted into
4·glucoside of
l,4.5-trihydro"ynapbtbalene JUglone(')
the aglycones by (3-glucosidases or oxidative processes. An-
thraquinones are especially common in the families Faba-
(54)
ceae (Cassia), Liliaceae (Aloe), Polygonaceae (Rheum,
Fig. 6.11. Hydrolysis of the 4-glucoside of 1.4,5-trihydroxynaphthalene Rumex), Rhamnaceae (Rhamnus), Rubiaceae (Asperula,
to produce juglone (modified from Harbome, 1982). Coelospermum, Coprosma, Galium, Morinda, and Rubia),
andScrophulariaceae (Digitalis). Members of the Rubiaceae
(e.g., madder, Rubia tinctorial are characterized by brightly
(Powell and Spencer, 1988). To date, only juglone has been colored anthraquinone pigments that have been used in the
demonstrated to produce allelopathic effects in the field. past for dyeing. These compounds are partly responsible for
Prenylated naphthoquinones occur in the roots of several the colors of several tropical woods. In basic solution, most
genera of the Boraginaceae (Leistner, 1985). The shikonins
anthraquinones produce a deep red or blue color.
(55), an important group of prenylated naphthoquinones
Anthraquinones derived from shikimic acid are usually
used in Japan for their anti-inflammatory properties, antibac-
accompanied by substitnted naphthoquinones. A knowledge
terial activity, and. as dyestnffs, have become one of the of the structures of these compounds has been useful in es-
most important groups of secondary compounds produced
tablishing several portions of the biosynthetic pathways for
in tissue cultnre (Ellis, 1988; Flores et al., 1987). These com-
anthraquinones.
pounds come from the roots of Lithospermum erythrorhizon
(Boraginaceae). Although the roots accumulate up to 2%
Biosynthesis of Antbraquinones
naphthoquinones, several years are required for the plant to
reach commercial size. In a 23-day fementation period, cells As is true for the naphthoquinones, anthraquinones are
in a 750-L tank accumulated 23% of their dry weight as synthesized by a variety of routes in plants and fungi (Pack-
shikonins (Flores et al., 1987). ter, 1980). Most are derived from either acetate-malonate
A number of bisnaphthoquinones from the genera Euclea or isochorismate-cx-ketoglutaric acid-mevalonate path-
and Diospyros (Ebenaceae) (Fig. 6.6) have antifungal, anti- ways. Anthraquinones also are produced frequently in plant
bacterial, and temite-resistant properties. Scrapings of wood tissue cultnres (Ellis, 1988). Tissue cultures of Cinchona
are used in African folk medicine for the treatment of lep- species (Rubiaceae) produce at least 30 different anthraqui-
rosy, and the ichthyotoxic properties of the bark of some nones.
species are used for captnre of fish. One of these compounds, Emodin (11) and helminthosporin (9) (Fig. 6.1) (see also
diosindigo A (53), is blue and responsible for the blue color Chapter 5) are derived from acetate-malonate precursors.
of the heartwood of some Diospyros species (Fig. 6.6) (Leis- An enzyme that provides strict substrate contrul, emodin 1-
tner, 1985). O-methyltransferase, was purified 89-fold (Hutchinson,
Several woods that contain naphthoquinones, anthraqui- 1986). Chrysophanol (57), found in Rumex (polygonaceae)
nones, and related compounds are highly resistant to attack and Rhamnus (Rhamnaceae) species, appears to result from
by marine borers (Southwell and Bultman, 1971). The wood folding of the polyketide chain in one arrangement (58),
of Tabebuia guayacan (Bignoniacae) was shown to contain whereas aloesaponarin (59) seems to involve a second form
lapachol (56) and several related compounds (Manners et (60) (Fig. 6.13). Both chrysophanol and aloesaponarin occur
al., 1975) and it is thought that these compounds are respon- in the roots of Aloe saponaria (Liliaceae) (Leistner, 1981).
sible for the observed resistance (Fig. 6.12). Under special conditions, callus cultures of Rhamnus species

«roo ~ @
: I
0

0
lawsone (41)
I
0

0
::,..,

OR 0
,&

lapachol (56) shikonin (55)

Fig. 6.12. Shikonin from Lithospermum erythrorhizon.

86 Ben:oqu;nones. Naphrlwqu;IIolles. ({lid Amlwaquillones

O~
o ~
~
0 0 _

CO,H
000 OH 0 OH

""
(58)
chrysophanol (57)

°fuC o
o

0
0

o COOH
_
c;.¢yc;
""" I
OH
°

0
I ~
OH

0
OCH,

(60)
aloesaponarin (59)

Fig. 6.13. Different folding mechanisms involved in the synthesis of chrysophanol and aloesaponarin (Leistner, 1981; modified and used with
pennission of the copyright owner, Academic Press, New York).

produce labile anthrone and dianthrone derivatives. Bark of intermediate compounds such as 2-carboxy-4-oxotetralone
these same plants contains similar, but distinct, compounds (COT) (43) and/or 1,4-dihydroxy-2-naphthoic acid (DHNA)
(Ellis, 1988). (40) (Fig. 6.8) or 2-carboxy-4-hydroxy-l-tetralone (48) (Fig.
Certain other anthraquinones of this pathway begin with 6.9). Attack at both positions is observed in the biosynthesis
more complex starter units. The CoA derivative of hexanoic of anthraquinones. Clues as to the site of prenylation in bio-
acid (n-C,HllCO-SCoA) is utilized to produce the some- synthesis of anthraquinones can be found not only by label-
what unusual anthraquinone solorinic acid (61) (Fig. 6.14). ing studies or by examination of prenylated but uncyclized
Anthraquinones derived from acetate-malonate path- naphthoquinones that co-occur with anthraquinones in sev-
ways are particularly common in fungi and lichens, but are eral plants and in plant tissue cultures.
often found in higher plants as well. Acetate-malonate-de- Phylloquinone (37), plastoquinone, a-tocopherol, and
rived anthraquinones usually can be distinguished by their ubiquinone (13) (Fig. 6.2) are produced by a cell suspension
structures because they possess substituents in both benze- culture from Morinda lucida (Rubiaceae). Changes in envi-
noid rings of the anthraquinone nucleus (also see Chapter ronmental, hormonal, and other cultural conditions result
5), although there are some exceptions to this generalization. in accumulation of anthraquinones in cultures of Galium,
Anthraquinones in which only one ring is substituted are Morinda, and Rubia (all Rubiaceae). The addition of o-succi-
particularly common in the Biguoniaceae, Rubiaceae, and nylbenzoic acid (38) causes an increase in the amount of
Verbenaceae, but do not appear to occur in fungi, the Poly- anthraquinones produced. 2-Carboxy-4-oxotetralone (43) or
gonaceae, Rhamnaceae, Fabaceae, CaesaIpinioideae, or the 1,4-dihydroxy-2-naphthoic acid (40) appear to be intermedi-
Liliaceae (Aloe). These compounds are derived from a path- ates in the synthesis of both anthraquinones and phylloqui-
way similar to that previously discussed for naphthoqui- nones.
nones, but with subsequent addition of mevalonate-derived In robiaceous plants, anthraquinones are related closely to
units (Fig. 6.8). prenylnaphthoquinones with which they co-occur. Specific
As indicated under the biosynthesis of naphthoquinones incorporation of 14C from [7-14C]shikimic acid into the 9-
above, prenylation can occur at either the 2- or 3-position in position of anthraquinone provides evidence that prenylation

OHO OHO

-
o 000 0
~n'C5HIl
o~
n'C5HIl __
"
"C5H11 CSCoA
+ malonyl.eoA
o 0
CO,H HO~OH
o
solorinic acid (61)

Fig. 6.14. Biogenesis of solorinic acid (Geissman and Crout. 1969).

 Benzoquinones, Naphthoquinones, and Anthraquinones 87

07
OH o OH
'CO'H CO,H CO,H OH

I --
""
1 /
. "'
o OH
(38) (40) o alizarin (62)

«tr
OH o OH

- -HO I I .&-

oq
~
(65) OH OH 0

1
morindone (63)
OH 0 OH o OH

CO'CH'c¢Cc OH
"'" I "'" "'" "'" OH

~ .&- I ~ I I.&- O-primereverosyl OH

o 0 o
lucidin
lucidin primeveroside
mollugin (64)

Fig. 6.15. Proposed biosynthesis of antbraquinones in species of Rubiaceae (modified from Inouye and Leistner, 1988; used with pennission of the
copyright owner, John Wiley & Sons, Ltd., Chichester).

occurs at the position corresponding to C(3') of o-succinyl- with madder (Haslam, 1974). 1,4-Dihydroxy-2-naphthoate
benzoate (38) in the biosynthesis ofthe anthraquinones aliza- (DHNA) (40) and 2-carboxy-4-tetralone (COT) (43), formed
rin (62) and morindone (63) in Rubia tinctoria and Morinda from o-succinylbenzoic acid (OSH) (38), undergo prenyla-
citrifolia. This and the co-occurrence of mollugin (64) and tion, decarboxylation, and hydroxylation. Labeled meva-
2-methoxycarbonyl-3-prenyl-I,4-naphthoquinone (65) in lonic acid labels the C ring of Rubia tinctoria anthraquinones
Galium mollugo support the idea that prenylation in these (Fig. 6.17) (Leistner, 1981).
plants occurs at the 3-position of 1,4-dihydroxy-2-naphthoic Administration of [5_ l4C]_ and [2-14C]-MVA (mevalonic
acid (DHNA) (40) (Fig. 6.15) (Inouye and Leistner, 1988). acid) to the two plants resulted in incorporation of C-5 of
Intact plants and cell cultures of Streptocarpus dunnii MVA into C-4 of alizarin (62) and C-2 of MVA into C-lI
(Gesneriaceae) contain several 1,2-naphthoquinones with a of pseudopurpurin (70) (Inoue et aI., 1984).
reversed prenyl side chain, such as that in dunnione (66) In order to establish unambiguously the biosynthetic ori-
(Fig. 6.16). I-Hydroxy-2-methylanthraquinone (67) and 1- gin of a particular anthraquinone, feeding experiments with
hydroxy-2-(hydroxymethyl)anthraquinone (68) were also labeled precursors must be done, although other evidence is
isolated from this culture. Administration of [I-carboxy- helpful in the determination of the biogenetic origin. For
l3C]o-succinylbenzoate revealed that l3C was incorporated example, a number of species of the Rubiaceae and Verbena-
into the I-position of dunnione and the lO-position of anthra- ceae that synthesize anthraquinones also contain a series of
quinones. These results, together with those of feeding ex- nonquinonoid compounds that are structurally related to in-
periments in which lawsone (41) and its 2-prenyl ether (69) termediates in the biosynthesis of the anthraquinones (Fig.
were applied, suggested that dunnione (66) was formed by 6.18) (Geissman and Crout, 1969).
a Claisen-type rearrangement at the 2-position of 2-carboxy-
4-oxotetralone (COT) (43), whereas anthraquinones were Distribution of Antbraquinones
formed by prenylation at the 2-position of 2-carboxy-4-oxo-
tetralone (COT) (43) or 1,4-dihydroxy-2-naphthoic acid (40) Anthraquinones derived from acetate-malonate (polyke-
(Inouye and Leistner, 1988). tide) pathways are widespread in fungi and lichens, but less
The anthraquinones of madder, Rubia tinctoria (Rubia- common in higher plants. Emodin (11) is among the com-
ceae), are well known. Most of the anthraquinones occur as pounds frequently found in all of these groups.
glycosides within the plant. One of these, ruberythric acid Polyketide-derived anthraquinones have a wide and scat-
(67), a diglucoside, can be converted to alizarin (62) by hy- tered occurrence in the following families: Monocotyle-
drolysis. Many biosynthetic studies have been carried out dons-Iridaceae, Liliaceae (Asphodelaceae) (Aloe), Zingib-
88 B('II;/J'Iuill(Jf1I'.\". Nat'hrIIlJCffliIlOIlf'S. dlld Anrhra'l"illOIl('.\·

. o 0

r-yCO',(CO,H

~~
~oo~
l
1 0

W
(40) (68) 0 HO '" 0 HO
(38)

~ .
o
eo'H o ~$~:
I I
I -- .Q

l
~ ~

(43) 0 o

C\f
(40) ? 0 (57)

!~ I
OHeo,H ~~
I
~- - w·~·:
I
~

«r\-of)-
~ ::::,.. ::::,.. .Q

!
o 0 OH

0 0 0

c¢'~ o
dunnione (66)
Iawsone (41) (69)

Fig. 6.16. Proposed biosynthesis of anthraquinones and prenylated naphthoquinones in species of the Gesneriaceae (modified from Inouye and Leistner,
1988; used with permission of the copyright owner, Jolm Wiley & Sons, Ltd., Chichester).

eraceae; Dicotyledons-Anacardiaceae, Asteraceae, Biological Functions of Antbraquinones

CaryophylJaceae, Chenopodiaceae, Clusiaceae, Combreta-
ceae, Ericaceae, Euphorbiaceae, Fabaceae, Lythraceae, Po- Anthraquinones are involved frequently in plant-insect
lygonaceae, Rhamnaceae, Rhizophoraceae, Saxifragaceae, interactions. For example, two anthraquinones (72) and (73)
Scrophulariaceae (?), Simaroubaceae, Sonneratiaceae, Sola- (Fig. 6.19) and the related anthrones have been found in
naceae, Urticaceae, and Xanthorrheaceae. In the Fabaceae, the larvae of the elm leaf chrysomelid, Pyrrhallta luteola.
anthraquinones are almost exclusively found in the genus Because these compounds do not occur in elm leaves, it is
Cassia (Senna) (Dahlgren et al., 1981; Zenk and Leistner, probable that they are synthesized by the insect or its sym-
1968). biotic microorganisms (Harborne, 1986).
The fresh bark of Rhamnus frangula and most Cassia The chrysomelid beetle, Galeruca tanaceti, feeds on Ta-
species contains anthrones and not anthraquinones (Leistner, necetum vulgaris or Achillea millefolium (Asteraceae), nei-
1985). After storage and extraction, anthraquinones and di- ther of which produce anthraquinones. However, the beetles
anthrones are isolated but are probably artifacts. Except for or their microbial associates produce two anthraquinones,
.a dubious record of acetate-derived anthraquinones in the chrysophanol (57) and chrysazin (74). The eggs of this insect
Scrophulariaceae, these compounds are not yet recorded contain these two compounds and are protected against the
from orders of plants that produce iridoid monoterpenes (see ant, Myrmeca ruginodus. Both compounds are potent micro-
Chapter 20). bial growth inhibitors (Cudlin et al., 1976; Hilker and
Anthraquinones derived from shikimic and isochorismic Schulz, 1991).
acid are known to occur only in certain dicotyledonous fami- Several types of woods that contain anthraquinones and
lies: Bignoniaceae, Gesneriaceae, Rubiaceae, Scrophularia- related compounds are highly resistant to attack by marine
ceae, and Verbenaceae (Gentianales, Larniales, and Scro- borers (Southwell and Bultman, 1971). 2-Methyl- (57), 2-
phulariales). These families also characteristically hydroxymethyl- (75) and 2-formylanthraquinones in the
accumulate iridoid monoterpenes (Dahlgren et al., 1981). heartwood of teak (Tectono grandis, Verbenaceae) are effec-
 X
Benzoquinones, Naphthoquinones, and Anthraquinones 89

HO
cq~" '(~ +--
COOH CH,OH
•

0,,/
$0
(38)

$0
""'" $.0
0: OH
I I I -- I IC I I
.
-+--
~ ~.d' ~ .d'

o 0(57) 0

! alizarin (62)

#-'
o O.p.primeverosyl

purpurin (71) pseudopurpurin (70) ruberythric acid (67)

Fig. 6.17. Localization of label from shikimic acid in anthraquinones and naphthoquinones of Rubia tinctoria (modified from Haslam, 1974).

tive in inhibiting tennite activity (Rudman and Gay, 1961; drolysis ofruberythric acid (67) (Fig. 6.17), is the most im-
Seigler, 1983). portant compound. An aluminum lake prepared from alizarin
is utilized to effect binding of the dye to the fabric. Both
Economic Importance and Uses the lake and various mordants are used to adjust the colors
Dyestuffs from Rubia tinctoria (madder, Rubiaceae), obtained.
sometimes known as turkey red, have been used extensively A red carbon-carbon-bonded anthraquinone glycoside,
in the past. Although the importance of vegetable dyestuffs carminic acid (76), from the scale insect Coccus cacti (Ho-
has diminished greatly, madder is still used in certain Near moptera, cochineal), has also been used as a dyestuff. This
Eastern countries. Alizarin (62), derived by enzymatic hy- compound was first isolated and utilized by the Indians of

«»-
o OH
~
o
~OCH,

Fig. 6.18. Compounds that accompany anthraquinones in the Rubiaceae and Verbenaceae (Geissman and Crout. 1969).
90 Benzoqltinones, Naphthoquinones, and Anthraquinones

OH 0 OH OH 0 OH

6¢6 o ~ o
(72) (73)

t¢5 o
chrysophanol
OH chrysazin (74)

~
"
c¢u HO~
I
o

I
~
OH HO o~HOH
0 COH
r
~ I I
",'
h
HO OH
o OH 0
2-hydroxymethylanthraquinone (75) carminic acid (76)

Fig. 6.19. Biologically active anthraquinones.

0 OH OH 0 OH

OH 0 OH

~$
CO,H CH,OH

CH,OH HO

H H

O-glc 0 OH OH 0 OH
emodin anthrone (12)
sennoside B palmidinA

OH 0 OH
OH 0 OH

HO

HO

hypericin (77) fagopyrin (78)

Fig. 6.20. Anthones and anthrone dimers.

 Benzoquinones. Naphthoquinones. and Anthraquinones 91

08 0 08 08 0 08
08 0 08

~
.0$-00
80 "'" 8 h
80 80
-+
80
<7 '<:::

i.
::,.. I I h

08 0 08 08 0 08 OH 0 OH
bv
(79) byperico-dehydroanthrone

08 0 OH 08 0 08 08 0 OH

bv 80
80
-80
HO

08 0 08
08 0 08 08 0 OH

penilliopsin protohypericin (80) hypericin (17)

Fig. 6.:Z1. Proposed biosynthesis of hypericin (Scott. 1967; modified and used with pennission of the copyright owner, Marcel Dekker, Inc., New
York).

Mexico (Fig. 6.19). When prepared as an aluminum-cal- HypericUm (Clusiaceae) (the names Hypericaceae and Gut-
cium lake, the compound is known as "carmine." tiferae which have formerly been used for this family are
Anthraquinone glycosides have long been used medici- incorrect). Hypericin and related compounds serve as pig-
nally as cathartics and laxatives. Plant-derived drugs of this ments in certain fungi, for example, Polys/ictus versicolor
type include aloes (Aloe species), cascara sagrada (Rhamnus and Dermocype austrovena/a (Gill and Gimenez, 1991). In
purshianus), frangula (Rhamnusfrangula), rhubarb (Rheum Hypericum species, the compolJ!ld is found in small, opaque
officinale), rumex or yellow dock (Rumex crisp us) and glands on the leaves, sepals, and petals. This substance co-
senna (Cassia spp.). Many of the commercial preparations occurs with a related compound, pseudohypericin. Both
(patent medicines) based on these plants are readily avail- compounds are a brilliant red color. In alkaline solution,
able. hypericin appears to be dark green and can be distinguished
Antbrones and Antbrone Dimers from anthraquinones, which are red (Knox et al., 1987).
Structurally related compounds occur in other plants such
Anthrones e.g., emodin anthrone (12) are probable inter- as Cassia (Fabaceae) (Scott, 1967).
mediates in the biosynthesis of anthraquinones such as em- Emodin (11) also has been found in most of the plants
odin from polyketide pathways (Fig. 6.22) and occur widely
and fungi that produce hypericin. The first intermediate (79)
in nature. The position para to the carbonyl oxygen of the
in the biogenesis of hypericin probably is joined by a single
center (B) ring is sensitive to oxidation and, in many in-
bond between the two anthrone molecules. This compound
stances, oxidation products may be the major or only com-
is then converted to the corresponding dehydru compound
pounds isolated from plants which contain anthraquinones
after storage (Leistner, 1985). and by light into protohypericin (80) (Fig. 6.21). Protohyper-
In addition to conversion to anthraquinones, compounds icin is converted by light to hypericin.
such as emodin anthrone can be coupled by free-radical pro- Hypericin and pseudohypericin have been shown to in-
cesses to produce a series of complex dimers such as hyperi- hibit the retroviral infection and replication cycle of mV-I
cin (77) and fagopyrin (78) (Fig. 6.20) (Brockman and Lack- at two or more points (Kinghorn, 1992).
ner, 1979; Scott, 1967). st. John's wort (or Klamath weed as it is called in the
western United States), Hypericum perjoratum, was intro-
Hypericin and Anthrone Dimers duced quite early into the United States from Europe and
Hypericin (77), a complex anthrone dimer, is found in became a major weed problem. By the early 1950s, this plant
approximately half of the nearly 200 species of the genus caused considerable mortality among livestock in the west
92 Benzoqu;nones, Napluhoqu;nones, and Anthraqu;nones

and was a major weed on about two million acres of range chemistry and Angiosperm Phylogeny (D. A. Young and D. S.
land. When animals, chiefly cattie, graze on Klamath weed Seigler, eds.), 149-204, Praeger, New York, 1981.
and are subsequently exposed to sunlight, they become sus- DOWNUM, K. R., Light-activated phmt defence, New Phytol., 122,
ceptible to sunburn and other tissue damage. This photody- 401-420 (1992).
namic activity is caused by the presence of hypericin which ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod.
is absorbed by the animals and enters peripheral circulation. Rep., 5, 581-612 (1988).
Serious necrosis of the skin and infection often occurs (Dow- FiSCHER, N. H. and L. QUUANO, AI1elopathic agents from common
num, 1992; Knox et aI., 1987). weeds, in The Chemistry of Allelopathy (A. C. Thompson, ed.),
In Europe, where the plant is native, several species of ACS Symposium Series 268, 133-147, American Chemical
the genus Chrysolina feed on species of the genus Hyperi- Society, Washington, DC, 1985.
cum. One of these beetles, Chrysolina brunsvicensis, appar- FLORES, H. E., M. W. Hoy, and J. J. PICKARD, Secondary metabo-
ently requires the presence of hypericin as a feeding stimu- lites from root cultures, Tibtech, 5, 64-69 (1987).
lant and will eat only species of Hypericum that contain GEISSMAN, T. A. and D. H. G. CROUT, Organic Chemistry of Sec-
the compound. Plants of H. androsaemum (which lack the ondary Plant Metabolism, Freeman Cooper, San Francisco,
1969.
compound) are untouched even though plants of H. perfora-
tum growing near them are eaten. A similar species of beetle, Gn.r., M. and A. GIMENEz, Austrovenetin, the principal pigment
of the toadstool Dermocybe austroveneta. Phytochemistry,30,
Chrysolina quadrigemina, from Europe was introduced into
951-955 (1991).
the United States in an effort to control Klamath weed. By
HAREORNE, J. B., Introduction to Ecological Biochemistry, 2nd ed.,
the late 1950s, this noxious weed had been reduced to about
Academic Press, London, 1982.
I % of its former level. Interestingly, the plant was untouched
HAREORNE, J. B., Recent advances in chemical ecology, Nat. Prod.
when it grew in shady areas; apparently, the insect was not
Rep., 3, 323-344 (1986).
able to locate the plant host under those conditions. Although
HASLAM, E., The Shikimate Pathway, Wiley, New York, 1974.
it has been reported that some Chrysolina species accumu-
late hypericin in their bodies, this has not been confirmed HASLAM, E., Secondary metabolism-Fact or fiction, Nat. Prod.
Rep., 4, 217-249 (1986).
(Pasteels et al., 1988).
Hypericin is phototoxic to generalist feeding lepidopteran lIn.KER, M. and S. SCHUlZ, Anthraquinones in different develop-
mental stages of Galeruca tanaceti (Coleoptera: Chrysomel-
larvae such as Helicoverpa zea and Planynotaflavedana on
idae), J. Chern. Ecol., 17, 2323-2332 (1991).
artificial diets in the presence of light. In nature, Planynota
HUTCHINSON, C. R., Biological methods for studying the biosyn-
flavedana survives by tying together leaves and feeding in-
thesis of natural products, Nat. Prod. Rep., 4,133-152 (1986).
side the ties where the larvae are protected from light (Sand-
iNOUE, K., Y. SmoBARA, H. NAYESHIRO, H. iNOUYB, G. Wn.sON,
berg and Berenbaum, 1989).
and M. H. ZENK, Biosynthesis of anthraquinones and related
compounds in Galium mollugo cell suspension cultures, Phyto-
chemistry, 23, 307-3II (1984).
REFERENCES INOUYE, H. and E. LEISTNER, Biochemistry of quinones, in The
Chemistry of Quinonoid Compounds, Vol. 2 (S. Patai and Z.
Rappoport, eds.), 1293-1349, John Wiley & Sons, Ltd., Chich-
BROCKMAN, H. and H. LACKNER, Zur Konstitution des Fagopyrins, ester, 1988.
Tetrahedron Lett., 1575-1578 (1979). KALIDHAR, S. B.o Location of glycosylation and alkylation sites in
CHANG, M. and D. G. LYNN, The haustorium and the chemistry of anthraquinones by IH NMR, Phytochemistry, 28, 2455-2458
host recognition in parasiUc angiosperms, J. Chern. Eco1., 12, (1989).
561-579 (1986). KINGHORN, A. D., Plants as sources of medicinally and pharmaceut-
CHANG, M. and D. G. LYNN, Phmt-plant recognition: Chemistry- ically important compounds, in Phytochemical Resources for
mediating host identification in the Scrophulariaceae root para- Medicine and Agriculture (H. N. Nigg and D. S. Seigler, eds.),
sites, in Allelochemicals: Role in Agriculture and Forestry (G. 75-95, Plenum, New York, 1992.
R Waller, ed.), ACS Symposium Series 330, 551-561, Ameri- KNox, J. P., RI. SAMUELS, and A. D. DODGE, Photodynamic action
can Chemical Society, Washington, DC, 1987. of hypericin, in Light-Activated Pesticides 0. R Heitz and K.
CHANG, M., D. H. NETZLY, L. G. BUTLER, and D. G. LYNN, Chemi- R. Downum, eds.), ACS Symposium Series 339, 265-270,
cal regulation of distance: Characterization of the rrrst natural American Chemical Society, Washington, DC, 1987.
host germination stimulant for Striga asiatica, J. Am. Chern. KOLKMANN, R and E. LEISTNER, 4-(2'-Carboxyphenyl)4-oxobu-
Soc., J08, 7858-7860 (1986). tyryl coenzyme A ester, an intermediate in vitamin K2 (mena-
CUDLIN, J., M. BLUMAUEROVA, N. STEINBROVA, J. MATEJU, and quinone) biosynthesis, Z. Naturforsch., 42c, 1207-1214 (1987).
V. ZALABAK, Biological activity ofhydroxyanthraquinones and KUBflZKI, K. and O. R. GoTTLIEB, Micromolecular patterns and
their glucosides toward microorganisms, Folia Microbiol., 21. the evolution and major classification of organisms, Taxon, 33,
54-57 (1976). 375-391 (1984a).
DAHLGREN, R. M. T., S. ROSENDAL-JENSBN, and B. J. NIELSEN, A KUBITZKI, K. and O. R. GOTILIEB, Phytochemical aspects of angio-
revised classification of the angiosperms with comments on sperm origin and evolution, Acta Bot. Neer., 33, 457-468
correlation between chemical and other characters, in Phyto- (1984b).
 Bell:oqll;nolles, Napll,lwqll;mmes, and AIl,hraquinones 93

LEISTNER, E.. Biosynthesis ot' plant quinones. in Secondary Plant POWELL, R. G. and G. F. SPENCER. Phytochemical inhibitors of
Products (E. E. Conn. ed.). Vol. 7 of The Biochemistry of Plants velvetleaf (Abutilon theophrasti) germination as models for
(P. K. Stumpf and E. E. Conn, eds.), Academic Press, New new biorational herbicides. in Biologically Active Natural Prod-
York, 1981. ucts: Potential Use in Agriculture (H. O. Cutler, ed.), ACS Sym-
LEISTNER. E., Occurrence and biosynthesis of quinones in woody posium Series 380, 211-232, American Chemical Society,
plants, in Biosynthesis and Biodegradation of Wood Compo- Washington, DC, 1988.
nents (T. Higuchi, ed.), 273-290, Academic Press, Orlando, RUDMAN, R. and F. J. GAY, The causes of natural durability in
fL, 1985. timber, Holzforschung, 15, 117-120 (1961); 17, 21-25 (1963).
LEISTNER, E., Biosynthesis of iso-chorismate-derived quinones. in SANDBERG, S. L. and M. R. BERENBAUM, Leaf-tying by tortricid
The Shikimic Acid Pathway (E. E. Conn. ed.), (Recent Ad- larvae as an adaptation for feeding on phototoxic Hypericum
vances Phytochemistry Vol. 20, 243-262, Plenum, New York, peiforatum, J. Chern. Ecnl.,15, 875-885 (1989).
1986.
SCHIMlDT, S. K., Degradation of juglone by soil bacteria, 1. Chern.
LYNN. D. G., The involvement of allelochemicals in the host selec- Ecnl., 14, 1561-1571 (1988).
tion of parasitic angiosperms, in The Chemistry of Allelopatby
SCOIT, A. 1., Some natural products derived by phenol oxidation,
(A. C. Thompson, ed.), ACS Symposium Series 268, 55-81,
in Oxidative Coupling of Phenols (W. I. Taylor and A. R. Bat-
American Chemical Society, Washington, DC, 1985.
tersby, eds.), Marcel Dekker, Inc., New York, 1967.
MANN, I., Secondary Metabolism, Oxford University Press, Ox-
ford, 1978; 2nd edition, 1987. SEIGLER. D. S., Role of lipids in plant resistance to insects, in Plant
Resistance to Insects (P. A. Hedin, ed.), ACS Symposium Series
MANNERS, O. D., L. IURD, R. WONG, and K. PALMER, Constituents
208, 303-327, American Chemical Society, Washington, DC,
of Tabebuia guayaean, Tetrahedron, 31,3019-3024 (1975).
1983.
MATHIS, C., Comparative biochemistry of hydroxyquinones. in
StMANTIRAS, M. and E. LEISTNER, Formation of o-succinylbenzoic
Comparative Phytochemistry (T. Swain, ed.), 245-270, Aca-
acid from iso-chorismic acid in protein extracts from anthraqui-
demic Press, New York, 1966.
none-producing plant cell suspension cultures, Phytochemistry,
MORRISON, R. T. and R. N. BoYD, Organic Chemistry, 3rd edition, 28, 1381-1382 (1989).
Allyn and Bacon, Boston, 1973.
SIMPSON, T. J., 13C_NMR in metabolic studies. in Nuclear Magnetic
NEAVB, I. A. and 1. O. DAWSON, Juglone reduces growth, nitroge-
Resonance, (H. f. Linskens and J. F. Jackson, eds.), 1-42,
nase activity, and root respiration of actinorhizal black alder
Springer-Verlag, Berlin, 1986.
seedlings, 1. Chern. Ecol.,15, 1823-1836 (1989).
PACKTER, N. M., Biosynthesis of acetate-derived phenols {polyke- STOESSL. A .• Structure and biogenetic relations: Fungal nonhost-
specific, in Toxins in Plant Disease (R. D. Durbin, ed.),
tides}, in Lipids: Structure and function (P. K. Stumpf, ed.),
109-219, Academic Press, New York, 1981.
Vol. 4 of The Biochemistry of Plants (P. K. Stumpf and E. E.
Conn, eds.), 535-570, Academic Press, New York, 1980. SOUTIlWElL, C. R. and J. D. BULlMAN. Marine borer resistance of
PASTEELS, J. M., M. ROWELL-RAHIER, T. RANooux, J. C. BRAEK- untreated woods over long periods of immersion in tropical
MAN, and D. DALOZE, Pyrrolozidine alkaloids of probable host- waters, Biotropica, 3, 81-107 (1971).
plant origin in the pronotal and elytral secretion of the leaf ZENK, M. H. and E. LEISTNER, Biosynthesis of quinones, Lloydia,
beetle Oreina eaealiae, Ent. Exp. Appl., 49, 55-58 (1988). 31,275-292 (1968).
7
Shikimic Acid Pathway

Introduction pound an oddity of plant metabolism and failed to recognize

Biosynthesis its importance in primary metabolism. Several of the inter-
Use of Mutants in Biosynthetic Studies mediates of the shikimate pathway [e.g., (- )-shikimic and
Fonnation of Chorismic Acid ( - )-quinic acids1 are now known to occur free in plants.
Derivatives of Chorismic Acid Most of these compounds are actively metabolized during
Biosynthesis of Tryptophan growth.
Indole 3-Acetic Acid Plants and microorganisms are capable of making aro-
Avenalumins from Oats matic amino acids, but most animals cannot. In particular,
DIMBOA and Related Compounds animals cannot synthesize L-phenylalanine and L-tryptophan
Biosynthesis of Phenylalanine and Tyrosine and are dependent on bacterial or plant sources of these es-
Compounds Derived from Shikimic Pathway sential compounds. Many animals can make L-tyrosine from
Intermediates L-phenylalanine.
Derivatives of iso-Chorismic Acid
Arbutin and Hydroquinones
References
BIOSl'1'ITHESIS

Il'fI'KODVCTlOI'l
The shikimic acid (or shikimate) pathway can be divided
Most aromatic compounds in plants are derived from shiki- into three parts: condensation of erythrose-4-phosphate and
mic acid metabolism; many of these substances are phenols. phosphoenolpyruvate and the subsequent cyclization and
Compounds derived from this pathway are extensively mod- production of shikimic acid (Fig. 7.1), alteration of shiki-
ified and considered under other classes of plant secondary mate-3-phosphate to chorismic acid (Fig. 7.1), and the con-
metabolites. Although many types of secondary compounds version of charismate into other products (Fig. 7.2). The
are produced from intermediates of the shikimic acid path- shikimic acid pathway might more appropriately be called
way (e.g., certain naphthoquinones and anthraquinones dis- the chorismic acid pathway, as that compound is the key
cussed in Chapter 6), most are derived from four aromatic intermediate and branching point for most plant secondary
amino acids: phenylalanine, tyrosine, anthranilic acid, and compounds produced.
tryptophan. Aromatic compounds that arise from the shiki- Most of the enzymes of this pathway have now been
mic acid pathway usually can be distinguished from those isolated and studied (Floss, 1986; Jensen, 1986). The shiki-
of other origins by their substitution patterns and by a knowl- mic acid pathway is predominately found in plastids in
edge of the compounds with which they co-occur. higher plants; isolated spinach chloroplasts can assimilate
Shikimic acid, for which the pathway is named, was fIrst CO2 or shikimate and produce aromatic amino acids. The
discovered in the plant Illicium religiosum in 1885 and was controls for this pathway have been reviewed (Jensen, 1986).
named after the Japanese name for the plant, shikimi-no-ki. The following discussion will consider the reactions re-
The compound makes up 20% of the dry weight of the fruits sponsible for the production of chorismic acid and then reac-
of this plant. For many years, chemists considered this com- tions leading to other metabolites.

94
 Shikimic Acid Pathway 9S

HO

HO,OH
xS pp:u °
H

O--+HO!
OH
H

;
po~ ·4°o
_D_A_H_P_S_YD_th_e_tase
__

(1M
erythrose phospboenolpyruvate OH

D
4·phosphate (1) (2) DAHP(3)

3·dehydroquinate
synthetase

NAD+,Co++ o
D
BO

i
COr 3-dehydroquinate
debydratase

o~~======' 0
~
eOl

i
-

OH

OH OH
3-debydroquinate 3·dehydroshikimate

I
(4)
NADPHf?;': (5)
V/
3-dehydroshikimate
reductase

ooD~ ~H
ooh~ ~ oo~~ ;
OH OH
(a)
quinate shikimate (6) gallic acid

HO I
OH
OH ATP PO
h i
OH
)1,",
OH EPSP synthase
hol~
e OH
shikimate (6) shikimBte 3-phosphate
(7)

PO = phosphate
glyphosate
inhibits here

~
co,. o 0

II II
~)l,""
chorismate
~oJlCO" __8yn=-th_e_IB_se_.
/
C.CH,.NH·CHrP.OH
I
, HO OH
OH
tm glyphosate (10)
5-enolpyruvylshikimate-3-phosphate N-(phosphonomethyl)glycine
chorismate (9)
(b) (8)

Fig. 7.1. (a & b) The biosynthesis of chorismate in microorganisms 1995; (modified and reprinted from Phytochemistry, Vol. 39, J. Schmid and N.
Amrhein. The pathway from erythose 4-phosphate to chlorismate. pp. 737-749. Copyright 1995 with kind pennission from Elsevier Science Ltd.• The
Boulevard, Langford Lane, Kidiingto. OXS 1GB, UK.)
96 Shikimic Acid Pathway

U5 OO~H
HO"" , PPo
u HO~
~CO,-
NU,+ --

HO , ~ HO"''', 0 CO,- / L-tyrosine L.phenylalanine

NH,
D-glu::e erythrose 4-Ph~s:hate ~ Jl --------- UN)
(; ~CO'H
..
~ )l.-/ fm 0 CO,- ¢H'N L·tryptophan

PO COr chorismate r
PhOSPhoenOIPyruvate~
/ "" I

6:. Jl -
/ CO,H
COzH $~aminObenzoate
,OH I I ""
""
?'

"'- 0 COH
~
, 0
isochorismate isoprenoid quinones

Fig. 7.2. An outline of the products derived from chorismate (modified from Haslam, 1974).

Use of Mutants in Biosynthetic Studies is not used by the organism into a compound capable of
sustaining growth. Although not normally an intermediate
Most steps of the shikimic acid pathway have been stud-
in the shikimic acid pathway, quinic acid may be used in-
ied by means of mutants, isotopically labeled precursors,
stead of 3-dehydroquinic acid when present. It is sometimes
and studies of the specific enzymes from cell-free systems.
difficult to distinguish between "precursors" and "interme-
The use of mutants for examination of various steps of the
diates" (Weiss and Edwards, 1981).
shikimic acid pathway has been of great value for the study
In some instances, an established intennediate may not
of the enzymes and intermediates involved. Use of mutants
be utilized by intact cells of mutants. Several obligatory in-
permits interruption of the pathway at a stage that normally
termediates of the shikimic acid pathway are not utilized
would not be seen. Mutants that involve one step of the
when introduced exogenously. This phenomenon usually has
pathway are especially usefuL In some situations, intermedi-
been attributed to failure of the intermediate to reach the site
ates are accumulated in large quantities, facilitating analysis
of synthesis or utilization, a problem which sometimes can
of the sequence of steps in the pathways. Sometimes, how-
be averted with cell-free systems. The amount of a particular
ever, the accumulation of intennediates exerts control over
precursor introduced is also important. The presence of
earlier stages of the biosynthetic process and inhibits the
larger than normal amounts of an intermediate may also
activity or represses fonnation of enzymes at a previous
cause deviation from the usual sequence of events (Weiss
point in the pathway. In these circumstances, the biosyn-
and Edwards, 1981).
thetic intermediate before the break in the pathway often
In early examinations of the shikimate pathway leading
will be unable to serve as a precursor. Mutants of this type
to aromatic compounds, a number of possible precursor
normally do not survive unless provided with an outside
compounds were evaluated. Surprisingly, out of about 50
source of an intermediate or product that occurs after the
compounds tested in mutant systems, only shikimic acid was
point at which the pathway is blocked.
found to be utilized. This compound could replace all three
Employment of mutants for the study of biosynthetic
common aromatic amino acids and p-aminobenzoic acid
pathways is often complicated and is usually feasible only
(Weiss and Edwards, 1981).
for microorganisms or plants in tissue culture. The mutant
must be unable to grow in the absence of the products of
Formation of Chorismic Acid
the pathway of interest (Weiss and Edwards, 1981).
The products accumulated and isolated from the cultures The starting materials for the shikimic acid pathway,
of mutants are sometimes not those of the actual pathway, erythrose-4-phosphate (1) and phosphoenolpyruvate (2) are
but modified products derived from intermediates involved. both involved in the primary metabolism of sugars and have
For example, in Neurospora mutants blocked after the for- key roles in the carbon assimilation cycle of photosynthesis,
mation of dehydroshikimic acid, only protocatechuic and a process principally found in higher plants and algae
vanillic acids (derived from dehydroshikimic acid) accumu- (Bonner and Varner, 1976).
late. In other instances, an intennediate or a derivative of The first step of the pathway is a condensation reaction
an intermediate may be converted by a pathway that usually involving phosphoenolpyruvate (1) and D-erythrose-4-phos-
 Shikimic Acid Pathway 97

phate (2) (Fig. 7.1). This initial condensation is catalyzed

by 3-deoxY-D-arahino-heptulosonate 7-phosphate (DAHP)
synthase, an enzyme system specific for these precursors.

--
Recent evidence indicates that this is actually a complex of
several related enzymes that are sensitive to end-product
control by tyrosine and tryptophan (in E. coli). The isozymes anti

responsible for the fonnation of tryptophan and tyrosine B:

H
seem to differ somewhat from those that are involved in the B:H
fonnation of phenylalanine (Dewick, 1989; Jensen, 1986;
Floss, 1986). '"yXCO'H P~CO'H
The first acyclic intennediate, 3-deoxY-D-arabino-heptu-
losonic acid 7 -phosphate (DAHP) (3), is then cyclized by p~3H '~3H
3-dehydroquinate synthase to 3-dehydroquinate (4), the first
cyclic intennediate in the pathway (Dewick, 1989; Knowles, HO·R OR
1989). ~ anti
In the next step of the sequence, 3-dehydroquinate (4) is
dehydrated via a Z-elimination to produce 3-dehydroshiki-
B,
mate (5). This reaction is catalyzed by 3-dehydroquinate de-
hydratase. 3-Dehydroshikimate (5) is, in tum, reduced to
shikimate (6) by 3-dehydroshikimate reductase, an enzyme
that requires NADPH as a cofactor. Shikimate (6) is then
POSfCO'H 3H

converted to the 3-phosphate (7) by the action of shikimate

kinase and ATP.
The conversion of shikimate-3-phosphate (7) to 5-enolp- 2 OR
yruvylshikimate (EPSP) (8) by 5-enolpyruvylshikimate-3-
phosphate synthase represents a rare type of reaction in
which the enolpyruvate fragment of phosphoenolpyruvate
l·PO
(2) is transferred to a molecule of (- )-shikimate-3-phos- H,B

phate (7) (Figure 7.1). The herbicide glyphosate (10) blocks

the enzyme that catalyzed this reaction (Fig. 7.3). In 1984,
sales of this herbicide totalled $480 million (Amrhein, 1986;
Floss, 1986).
By means of phosphoenolpyruvate labeled with both tri-
tium and deuterium (E-[3-2H,,3Hdphosphoenolpyruvate), it
5-enolpyruvylskikimate-3-phosphate (8)
is possible to detennine the stereochemistry of both the addi-
tion and the elimination step involved in the fonnation of 5- Fig. 7.3. Stereochemical course of the 5-enolpyruvylshikimate 3-
enolpyruvylshikimate-3-phosphate (EPSP) (8) (Floss, 1986) phosphate synthase reaction (adapted with permission from Asano et a1.,
1985, copyright 1985, American Chemical Society).
(Fig. 7.3). The EPSP synthase reaction involves addition and
elimination at the 2,3 double bond with opposite stereochem-
istry (i.e., either syn addition and anti elimination or anti Although the fonnation ofp-aminobenzoic acid (36) (Fig.
addition followed by syn elimination) (Floss, 1986). 7.12) can be explained by amination and loss of pyruvate
A 1,4-conjugate elimination of phosphoric acid then from iso-chorismic acid, enzyme extracts from Enterobacter
transfonns 5-enolpyruvylshikimate-3-phosphate (8) into aerogenes and two Streptomyces species contain p-amino-
chorismate (9) (Fig. 7.1). The 6-pro-R hydrogen atom is benzoate synthase and iso-chorismate synthase activity. Ki-
labile in the chorismate synthetase reaction; hence, the reac- netic data suggest that synthesis of p-aminobenzoic acid oc·
tion occurs with overall trans-geometry. curs from chorismic acid (Johanni et aI., 1989). p-
Many aspects of the shikimic acid pathway have been Aminobenzoic acid is important in the fonnation of folic
reviewed in detail (Conn, 1986; Weiss and Edwards, 1981). acid in fungi and bacteria (Haslam, 1974).

Derivatives of Chorismic Acid

BIOSYl'ITIIESIS OF TRYPTOPHAN
From chorismic acid, four major pathways lead to essen-
tial metabolites: tryptophan, phenylalanine and tyrosine, p- Tryptophan is an essential amino acid for most organisms. In
aminobenzoic acid and the folate group of coenzymes, and plants and bacteria, this compound is derived from chorismic
the isoprenoid quinones (Fig. 7.2). Numerous secondary acid. Many groups of secondary compounds are fonned from
compounds in plants and other organisms are fonned from tryptophan; among these are several simple amine deriva-
products and intennediates of these pathways. tives and a number of alkaloids.
98 Shikimic Acid Pathway

l:l Hx'H
) : 2
+ glutamate

3nthron;}at.
synthase

anthranilic acid
chormismate (9) (11)

Fig. 7.4. Steric course of the anthranilic synthase reaction (adapted with pennission from Asano et al., copyright 1985 American Chemical Society).

The fIrst step in the formation of tryptophan involves then to indoleacetic acid (17) (Fig. 7.6). Two enzymes are
conversion of chorismate (9) to anthranilate (11) (Fig. 7.4). involved: tryptophan 2-monooxygenase and indoleacetam-
Although the reaction is not well understood, it is catalyzed ide hydrolase (Kosuge and Sanger, 1986).
by the enzyme anthranilate synthase and utilizes L-gluta-
mine. By means of specifically labeled chorismic acid, it was
determined that the protonation involved in the formation of AVBI'IALUJIIINS FROM OATS
anthranilic acid had occurred from the re face (Figure 4)
(Floss, 1986). Anthranilic acid (11) also serves as an inter- Three compounds, avenalumin I, II, and III (19-21), are
mediate for the synthesis of a number of secondary com- formed in resistant lines of oats (Avena sativa, Poaceae) in
pounds and occurs free and as various derivatives in many response to attack by crown rust, Puccinia coronata f. sp.
plants and other organisms (Dewick, 1989). avenae (Fig. 7.7) (Mayama, 1983). The structures of these
The second enzyme of the pathway leading to tryptophan, compounds suggest that they are derived from anthranilic
5-phosphoribosy1-pyrophosphate transferase (PR transfer- acid.
ase), causes addition of a phosphoribosyl unit (12) to anthra-
nilic acid. An additional series of enzymes then brings about
a rearrangement and the ultimate formation of indoleglycerol
DllImOA AND RELATED COJIfPOUNDS
3-phosphate (13).
Indoleglycerol 3-phosphate (13) is converted into trypto-
phan (14) by the action ofL-tryptophan synthase. The mech- A series of hydroxamic acid derivatives with 4-hydroxy-
anism of this enzymatic reaction involves formation of a 1,4-benzoxazin-3-one structures occur in a number of cereal
Schiff base with an enzyme-bound pyridoxal phosphate. The grain species (Poaceae or Gramineae) (Fig. 7.8) (Niemeyer,
a-aminoacrylate Schiff base formed undergoes the addition 1988). These compounds usually occur in plants as the
of a j3-substituent to produce tryptophan (Floss, 1986) (Fig. 2-j3-0-o-glucosides; enzymes that hydrolyze the com-
7.5). pounds usually co-occur. DIBOA [2,4-dihydroxy-I,4(2H)-
benzoxazin-3-onel (22) is the major compound in rye and
DIMBOA [2,4-dihydroxy-7 -methoxy-I ,4(2H)-benzoaxa-
Indole 3·Acetic Acid
zin-3-one1 (23) in wheat and maize. Although these com-
Indoleacetic acid (IAA) (17) is involved in many aspects pounds are found in most plant patts, hydroxamic acids typi-
of plant growth and development (Bonner and Varner, 1976; cally occur in highest concentration in young seedlings and
Kosuge and Sanger, 1986). This hormonal substance is de- roots. Young leaves usually contain higher levels than old
rived in most plants by conversion of tryptophan to indole leaves. Higher concentrations of hydroxamic acids are found
3-pyruvic acid (15) (tryptophan amino transferase), decar- in the vascular bundles (Niemeyer, 1988), DIMBOA gluco-
boxylation to the indole 3-acetaldehyde (16) (indole pyru- side is found in the honeydew of the aphid Rhopalosiphum
vate decarboxylase), and oxidation to indole 3-acetic acid padi feeding on wheat (Givovich et aI., 1992). DIMBOA
(17) (indole acetaldehyde oxidase) (Fig. 7.6) (Goodwin and tastes sweet to humans (Hamilton et aI., 1962).
Mercer, 1983). Benzoxazinones share a major patt of the biosynthetic
Indole 3-acetic acid (IAA) (17) also is found in certain pathway of tryptophan. Label from quinic acid and anthra-
bacteria. IAA is a virulence factor in the relationships of nilic acid is incorporated into the aromatic ring of benzoxazi-
one bacterium, Pseudomonas syringae pv savastanoi, and nones. The carbon atoms of the heterocyclic ring stem from
its plant hosts, Nerium oleander (oleander, Apocynaceae) ribose. C-2 comes from C-2 of ribose, whereas C-3 comes
and Olea europaea (olive, Oleaceae). Virulence is assessed from C-I of ribose (Niemeyer, 1988). A number of questions
by the production of tumor-like outgrowths by the plants concerning the nature of the biosynthetic reaction sequence
in response to secretion of phytohormones by the pathogen remain to be answered. Heating these compounds produces
(Kosuge and Sanger, 1986). The bacterium produces IAA benzoxazolin-2-ones and fonnic acid. Benzoxazinones de-
by conversion of L-tryptophan to indoleacetamide (18) and compose readily at higher pH (Niemeyer, 1988).
 Shikimic Acid Pathway 99

Both OIMBOA and OIMBOA glucoside inhibit photo· (Campos et aI., 1989). Other reports have shed doubt on the
phosphorylation by spinach chloroplasts (Queirolo et al., role of these compounds in disease resistance (Lyons and
1981). OIMBOA is a potent inhibitor of cyanide-insensitive Nicholson, 1989; NyhUS et al., 1989).
respiration in potato mitochondtia (Hediger and Seigler, un- Compounds of these classes also are thought to be respon-
published data). sible for the allelopathic effects of some cereal grains. In
Both hydroxamic acids and benzoxazolinones have been most instances, the hydroxamic acids rather than the corre-
linked to resistance to insect pests and fungal and bacterial sponding benzoxazolinones are thought to be active. OIBOA
diseases (Niemeyer, 1988). Although many of these com- glucoside inhibits germination and seedling growth of Abuti-
pounds are toxic or deterrent to a variety of organisms, hy- Ion theophrastii, a common field weed in many parts of the
droxamic acids of this type are not stable in culture media world (Niemeyer, 1988). OIBOA has been shown to be an
at pH 7; the results of many previous feeding studies should allelopathic agent from winter rye, Secale cerale. This com-
be reexamined. In other instances, concentrations of the pound is released from the corresponding glycoside, and
compounds thought to be active are not sufficiently high in although it does not inhibit seed germination, it inhibits root
the appropriate tissues to explain the levels of resistance growth at concentrations of 0.37 mmoVthn (Harbome,
observed. More recent studies, however, demonstrate that 1989). The synthetic herbicide Bentazon (25) (Fig. 7.8) has
OIMBOA in the diet of the European com ear worm, Os· a structure reminiscent of OIBOA.
trinia nubilalis, increases the mean time to pupation and 6·Methoxybenzoxazolinone (MBOA) (24), the decompo-
adult emergence. Both pupal and adult weights decreased sition product of OIMBOA, stimulates reproduction in the

O1
E: ...... 0
CO'_HN~CO,.

I~ X
~
J H H NH,+

anthranilate
CO
'. ~'Hy'y'
Jf"~-
c!H 0
",
CO,·
synthase
~ 0 CO,.

chorlsmate (9)

(XI CO,H
phosphoribosyl
(XI ~H
:
CO,.

H PRA isomerase
'-~--+-. NH CH,OP •
~ NH, transferase OH o.R
anthranilic acid H H H O~~
(11) ~,A----h CH,OP

~-~
OPP O/H

(X
(12)
cor
: I IGP .yntbetase
NHHO H
~CH'OP
o H OH
1·(o·carboxyphenyJamlno)-
I-deoxyribulose pbosphate Indoleglycerol phosphate
(CRDP) (lGP) (13)
NH,

tryptophan ~CO'H
l0l ) ••
synthetase N

L·tryptophan (14)

Fig. 7.5. Biosynthesis of tryptophso (modified from Luckner, 1990).

100 Shikimic Add Pathway

cc"'
NH,

--+
h
CO'H_
NH,
~
I I
P"
CO'H
:::,... NH
H

IRPlaY
chorismate (9) anthranilic acid (11) IAryptophnn (14)

1 in bacteria

°
~CO'H
° ~NH'
UN) UJ
indole-3-acetamide (IS)
(15)

°
~OH
UJ
indole-3-acetic add

~ ~ .
Fig. 7.6. . (modi'fled from Smith. 1980; used with pemnSSlon
.aCId
Biosynthesis of indole aceUc . . 0f the cop yright owner, Springer-Verlag, BerlIn),

avenalumin I (19) avenalulDin II (20) avenalumin m (21)

Fig. 7.7. Avenalumin It II, and m.

CH,-0yY° X O-glucosyl
CH'-°yY°XOH
~NI ° ~NI °
OH OH

a:x:H
DIMBOA glucoside DIMBOA(23) BentazoD (25)

CH,'°yY>=O
I
OH ~NH
DmOA(22) MBOA(24)

Fig. 7.8. l,4-Benzoxazin-3-ones and benzoxazolin-2-ones from the Poaceae.

 Shikimic Acid Pathway 101

montaine meadow vole (Microtus montanus) and may serve 1986; Jensen, 1986). These essential amino acids serve as
as an environmental cue affecting reproductive cycles in precursors to a large number of other types of secondary
many mammals (Berger et aI., 1981; Sanders et aI., 1981). compounds.
Although benzoxazinones mostly have been reported In bacteria, two soluble, multiactivity enzymes or enzyme
from grasses (Aegilops, Arundo, Chusquea, Coix, Elymus, complexes function in the utilization of chorismate for L-
Secale, Sorghum, Tripsacum, Triticale, Triticum, and Zeal, phenylalanine and L-tyrosine synthesis. A complex contain-
DIBOA glucoside has been reported from Acanthus moWs ing chorismate mutase and prephenate dehydratase activities
(Acanthaceae) (Powell and Spencer, 1988; Wolf et aI., 1985) has been observed. Although the reaction proceeds by what
and MBOA bas been found in Scoparia dulcis (Scrophularia- appears to be a Claisen rearrangement, the rate-limiting tran-
ceae) (Chen and Chen, 1976). sition state appears to be different in the enzymatically con-
Methods for the analysis of DIMBOA and related com- trolled process. It is not known if fonnation of the higher-
pounds have been reviewed (Lyons et aI., 1988; Niemeyer, energy diaxial confonner (27) occurs after binding to the
1988). enzyme or is from an equilibrium mixture (Dewick, 1984;
Floss, 1986; Jensen, 1986).
Stereochemical studies indicate that the branch for the
BIOSYl'lTHESIS 01' PHENYLALANINE pathway leading to tyrosine and that leading to phenylala-
ANDTYKOSIl'IIl nine of the chorismate mutase enzyme systems function with
the same stereochemistry (Floss, 1986) (Fig. 7.10).
In both bacteria and plants, two additional amino acids, phe- Prephenate (26) is converted to phenylpyruvate (28) in
nylalanine and tyrosine, are fonned from chorismic acid. many bacterial systems. In some organisms, such as Esche-
From chorismate, two separate routes diverge and lead to richia coli, prephenic acid also is oxidatively aromatized to
the amino acids L-phenylalanine and L-tyrosine. However, p-hydroxypbenylpyruvic acid (29) by a soluble, NAD+-de-
the pathways in bacteria and plants are distinct and involve pendent enzyme, prephenate dehydrogenase. p-Hydroxy-
different intennediates. Both of these pathways pass through phenylpyruvic acid is then transaminated by the addition of
the same intennediate, prephenic acid (26) (Fig. 7.9) (Floss, L-glutamate and pyridoxal phosphate to yield L-tyrosine (30).

***phenyIpyruvate aminotransferase

chorismate mutase-
6~
CH'COCO" PLP

I~
***
L-phenylalanine
prephenate dehy~~;ta/
(28) / (31)
(bifunction/ (enzyme bound)
(27) /arogenate dehydratase
CO2-

OoJlco,_ -- .. ·0 ( J " C O ' . _o,~.. CO,·

2 "~. 0 PLP ~'" NH3+

II ~II
****

arog~:te ~at~:;drogenase
i
OH
prePhe:: (26) (32)
chorismate (9)

** chorismate mutase
prePhenat~
dehydrogenase ;;COCO,.~ ~C02"
PLP = pyridoxal 5 -phosphate
**** prephenate aminotransferase
V HO~-NH3+
OH L-tyrosine (30)
"* p-hydroxyphenylpyruvate aminotransferase
p-hydroxyphenylpyruvate
(29)

Fig. 7.9. Formation of prephenic acid from chorismic acid (modified from Dewick, 1984; used with permission of the copyright owner, the Royal
Society of Chemistry, London),
102 Shikimic Add Pathway

&I
C:. 'HR'IEH r HOC @j-
2
o,:O'Hl
R
28 E.coU
'gHCO~:,
'H R

l
o
E.•oU.
CO. cborismate 1,1
P'I
j
OH
2 mutase ~
OH OH
I

HOD-CO"
~1b-
H j CO,.
~.
~

H OH
diequatoriaJ dimal

m-
OH
°it~(
enzyme-bound
diaxial chorismate
oo
'
- d:
=
HO
prepbenate (26)
co:.

Fig. 7.10. Stereocbemistry of the formation of prephenic acid from chorismic acid (modified from Baldwin and Davidson, 1983; used with pennission
of the copyright owner, Elsevier Science, Amsterdam),

Phenylalanine (31) may be fonned by conversion of choris· distinct series of enzymes and reactions to that discussed for
mate (9) to prephenate (as an enzyme·bound intennediate) phenylalanine above. Although both prephenic and arogenic
and subsequent fonnation of phenylpyruvate (28). Transami· acids are intennediates, they are part of a different' 'branch"
nation of this intennediate produces phenylalanine (31). leading to tyrosine. In higher plants, arogenic acid (32) is a
In higher plants, phenylalanine seems to be fonned in direct precursor to tyrosine. L·Phenylalanine does not appear
an alternative manner by fonnation of prephenate (26) and to be a precursor of L·tyrosine in either plants or bacteria;
conversion of this intennediate into arogenic acid (32). they lack phenylalanine hydroxylase, which is an important
Chorismate mutase occurs as two isozymes which have been enzyme in animals.
purified to homogeneity from mung bean and sorghum. All In plants, cinnamic and p-coumaric acids and their deriva-
chorismate mutase isozymes show allosteric activation by tives are ubiqnitous. These compounds, in torn, give rise to
chorismate (Poulsen and Verpoorte, 1991). One chorismate phenylpropanoids, lignans, lignin, flavonoids, and tannins.
mutase isozyme is inhibited by phenylalanine or tyrosine Both phenylalanine and tyrosine are precursors to numerous
and activated by tryprophan, whereas the second is not af· alkaloids.
fected by any of the aromatic amino acids.
Arngenic acid is converted into phenylalanine by the ac·
tion of arogenate dehydratase. Although plants nonnaIIy do COJllPOlJI'IDS DERIVED fROM SHIKIJIIlC
not appear to make phenylpyruvic acid (28) and p·hydroxy· PAntWAY IN1ERJIIEDlATES
phenylpyruvic acid (29), there is some evidence that these
compounds can serve as precursors for phenylalanine and Several secondary metabolites derived from intennediates
tyrosine, respectively (Jensen, 1986; Widholrn, 1974). in the shikirnate pathway are widely distributed in higher
Fonnation of tyrosine in plants involves a similar hut plants. As previously mentioned, shikimic acid and quinic
 Shikimic Acid Pathway 103

6
OH

HO COzH 0 ~OH
~ ~
o H
HO i OH H o z c f fOH
OH OH OH

sbikimic acid (6) quinic acid (33) chlorogenic acid (34)

Fig. 7.11. Chlorogenic acid. shikimic acid. and quinic acid.

COZHr;:-
~H
0:)00:-
..
RNHz
z
~:XCOZH
+
•
isochorismic acid (35)

(yCOzH

V1--0H
!

salicylic acid
¢ NHz
p-aminobenzoic acid
(36)

FIg. 7.12. Origin of salicylic acid from iso-chorismic acid (modified from Luckner. 1990).

troy
0 0 0

t:c
O)lPh o O)lPh OApb

troy
o 0 ::,.. 0 0 ::,.. 0

o O~ O~ O~Ph
crotepoxide (36) senepoxide (37) pipoxide (38)

Croton macrostachys Uvarla catocarpa Piper hoo"ri

Euphorbiaceae Annonaceae Piperaceae

Fig. 7.13. Crotepoxide, senepoxide, and pipoxide.

104 Shikimic Acid Pathway

¢ -- ¢ -- ¢ --
COO COO COO

~CO'-
::7 I H Fe"
02 or
""- NH,+- HOO·
HO

0- O· 0
L~tyrosine

H'

"-~O
6~Q -- ¢
OH
---.... OOH

I~ 1--
0") OH Oglucosyl
0
H' hydroquinone (39) arbutin (40)

Fig. 7.14. Biosynthesis of hydroquinone and arbutin (modified from Torssell, 1983; used with pennission of the copyright owner, John Wiley & Sons.
Ltd" Cbichester).

acid are found free in many plants. They also occur as esters In some plants, these compounds appear to arise by oxida-
and in other forms. For example, quinic acid (33) occurs tive decarboxylation of p-hydroxybenzoic acid. p-Hydrox-
frequently as the caffeoyl acid ester, chlorogenic acid (34) ybenzoic acid (36) has vitamin-like activity in some organ-
(Fig. 7.11). Up to 10% of the dry weight of a cup of coffee isms_ This growth factor requirement is associated with the
is comprised of this compound. Chlorogenic acid is pro- formation of ubiqninone (coenzyme Q) and serves as an
duced as a response to wounding in many plants_ Other iso- effective precursor for the benzoquinone ring of ubiquinone
mers, such as isochlorogenic acid, also occur. The physio- in mammals, bacteria, fungi, and plants (Haslam, 1974; also
logical importance of chlorogenic acid will be discussed later see Chapter 6).
(see Chapter 8).

Derivatives of iso-Chorismic Acid

REFERENCES
iso-Chorismic acid (35) is derived from chorismic acid
in plants. This compound gives rise to a number of primary
and secondary metabolites. In some organisms, salicylic acid AMRHEIN, N., Specific inhibitors as probes into the biosynthesis
is known to arise from iso-chorismic acid (Fig. 7.12). and metabolism of aromatic amino acids. in The Shikimic Acid
Pathway (E. E. Conn, ed.), Recent Advances io Phytochemistry
However, salicylic acid may be synthesized directly from
Vol. 20, 83-117, Plenum Press, New York, 1986.
benzoic acid by means of a benzoate hydroxylase enzyme
system (LeOn et al., 1993; Yalpani et al., 1993) (see Chapter ASANO, Y., J. J. LEE, T. L. SHIEH, F. SPRBAFlCO, C. KOWAL, and
H. G. FLOSS, Stereochemical course of the enol pyruvylsbimi-
8). The relative importance of these systems in higher plants
mate-3-phosphate synthase, chorismate mutase. and anthrinilate
is not known. synthase, J. Am. Chern. Soc., 107. 4314-4320 (1985).
As discussed in Chapter 6, iso-chorismic acid is involved
BALDWIN, G. S. and B. E. DAVIDSON, Kinetic studies on the mecha-
in the biosynthesis of naphthoquinones and anthraquinones
nism of chorismated mutaselprephenate dehydratase from Esch-
in a number of plants and possibly other organisms (Leistner, erichia coli, Biochim. Biophys. Acta, 742, 374-383 (1983).
1986; see Chapter 6).
BERGER. P. J., N. C. NEGUS, E. H. SANOERS, and P. D. GARDNER,
Another series of compounds that appear to be derived
Chemical triggering of reproduction in Microtus montanus, Sci-
from iso-chorismic acid are cyclohexene oxides such as ero- ence, 214, 69-70 (1981).
tepoxide (36), senepoxide (37), and pipoxide (38) from Cro-
BONNER, J. and J. E. VARNER, Plant Biochemistry, 3rd ed., Aca-
ton macrostackys (Euphorbiacee), Uvaria zeylanica (An- demic Press, New York, 1976.
nonaceae), and Piper kookeri (Piperaceae), respectively
CAMPOS, F., J. ATKlNSON, J. T. ARNASON, B. J. R. PHlLOOENE, P.
(Fig. 7_13). These compounds have demonstrated antitumor
MORAND, N. H. WERSTIUK, and G. 1'IMM:INs, Toxicokinetics of
activity (Jolad et al., 1981). 2,4-dihydroxy-7-methoxy-1 ,4-benwxazio-3-one (DIMBOA) in
the European com borer, Ostrinia nubilalis (Hiibner), 1. Chern.
Arbutin and Hydroquinone Bcol.,IS. 1989-2001 (1989).
Hydroquinone (39) and the glucoside arbutin (40) are CHEN, C. and M. CHEN, Methoxybenzoxazolinone and triterpenoids
derived variously from acetate-malonate, shikimic acid, or from roots of Scoparia dulcis, Phytochemistry,IS, 1997-1999
from tyrosine (Fig. 7.14). (1976).
 Shikimic Acid Pathway lOS

CONN, E. E. (ed.), The Shikimic Acid Pathway, Recent Advances LYONS, P. C., 1. D. HIPSKIND, K. V. WOOD, and R. L. NICHOLSON,
in Phytochemistry, Vol. 20, Plenum Press, New York, 1986. Separation and quantification of cyclic hydroxamic acids and
DRWICK, P. M., The biosynthesis of shikimate metabolites, Nat. related compounds by high-pressure liquid chromatography, 1.
Prod. Rep., I, 451-469 (1984). Agric. Food Chern., 36,57-60 (1988).
DEWICK, P. M., The biosynthesis of shikimate metabolites, Nat. MANN, 1., Secondary Metabolism, Oxford University Press, Ox-
Prod. Rep., 7, 263-290 (1989). ford, 1978; 2nd edition 1987.
FLOSS, H. G.. The shikimate pathway-An overview, in The Shiki- MAYAMA, S., The role of avenalumin in the resistance of oats to
mic Acid Pathway (E. E. Corm, ed.), Recent Advances in Phyto- crown rust, Mem. Fac. Agric. Kagawa Univ., No. 42., 1-62
chemistry Vol. 20 Plenum Press, New York, 1986. (1983).
GIVOVICH, A., S. MORSE, H. CERnA, H. M. NIEMEYER, S. D. WRAT- NIEMEYER, H. M., Hydroxamic acids (4-hydroxy-I,4-benzoxazin-
TEN, and P. 1. EDWARDS, Hydroxamic acid glucosides in honey- 3-ones), defence chemicals in the Gramineae, Phytochemistry,
dew of aphids feeding on wheat, J. Chern. Ecol.,18, 841-846 27, 3349-3358 (1988).
(1992). NyHUS, K. A., W. A. RUSSELL, W. D. GUTHRIE, and C. A. MAR-
GOODWIN, T. W. and E. 1. MERCER, Plant Biochemistry, 2nd ed., TINSON, Reaction of two maize synthetics to anthracnose stalk:
Pergamon Press, Oxford, 1983. rot and northern com leaf blight following recurrent selection
lIAMn.TON, R. H., R. S. BANDURSKI, and W. H. REUSCH, Isolation for resistance to Diplodia stalk rot and European com borer,
and characterization of a cyclic hydroxamate from Zea mays, Phytopathology, 79, 166-169 (1989).
J. Cereal Chern., 39, 107-113 (1962). POULSEN. C. and R. VERPOORTE, Roles of chorismate mutase iso-
HARBORNE, 1. B., Recent advances in phytochemistry, Nat. Prod. chorismate synthase and anthranilate synthase in plants, Phyto-
Rep., 7, 85-109 (I989). chemistry,30, 377-386 (1991).
HASLAM, E., The Shikimate Pathway, Wiley, New York, 1974. POWELL, R. G. and G. F. SPENCER, Phytochemical inhibitors of
JENSEN, R. A., Tyrosine and phenylalanine biosynthesis: Relation- velvelleaf (Abutilon theophrasti) germination as models for
ship between alternative pathways, regulation, and subcellular new biorational herbicides, in Biologically Active Natural Prod-
location, in The Shikimic Acid Pathway, (E. E. Corm, ed.), ucts: Potential Use in Agriculture (H. G. Cutler, ed.), ACS Sym-
Recent Advances in Phytochemistry, Vol. 20, 57-81, Plenum posium Series 380, 211-232, American Chemical Society,
PTess, New York, 1986. Washington, DC, 1988.
JOHANNI, M., P. HOFMANN, and E. LEISTNER, Origin ofp-aminoben- QUEIRoLO, C. B., C. S. ANDREa, R. H. VALLBJOS, H. M. NIEMEYER,
zoic acid from chorismic rather than iso-chorismic acid in En- and L. J. CoRCUERA, Effects ofhydroxamic acids isolated from
terobacter aerogenes. and Streptomyces species, Arch. Bio- Gramineae on adenosine 5'-triphosphate synthesis in chloro-
chern. Biophys., 271, 495-501 (1989). plasts, Plant Physiol., 68, 941-943 (1981).
loLAD, S. D., 1. 1. HOFFMANN, K. H. SCHRAM, 1. R. COLE, M. S. SANDERS, E. H., P. D. GARDNER, P. 1. BERGBR, and N. C. NEGUS,
1'BMPEsTA, and R. B. BATES, Structures of zeylenol and zeylena.
6-Methoxybenzoazolinone: A plant derivative that stimulates
constituents of Uvaria zeylanica (Annonaceae). J. Org. Chern.• reproduction in Microtus montanus, Science, 214, 67-69
46,4267-4272 (1981). (1981).
KNOWLES, J. R., Mechanistic ingenuity in enzyme catalysis: Dehy-
SMITH, T. A., PLANT AMINES, in Secondary Plant Products (E. A.
droquinate synthase, Aldrichim. Acta, 22, 59-66 (1989).
Bell and B. V. Charlwood, eds.), 433-456, Springer-Verlag,
KOSUGE, T. and M. SANGER, Indoleacetic acid, its synthesis and Berlin, 1980.
regulation: A basis for tumorigenicity in plant disease, in The
Shikimic Acid Pathway (E. E. Corm, ed.), Recent Advances in TORSSELL, K. G. B., Natural Product Chemistry, 10lan Wiley &
Sons, Ltd., Chichester, 1983.
Phytochemistry Vol. 20 147-161, Plenum Press, New York,
1986. WEISS, U. and J. M. EDWARDS, The Biosynthesis of Aromatic Com-
LEISTNER, E., Biosynthesis of iso-chorismate-derived quinones. in pounds, Wiley, New York, 1981.
The Shikimic Acid Pathway (E. E. Corm, ed.), Recent Advances WIDHOLM, 1. M., Control of aromatic amino biosynthesis in cul-
in Phytochemistry, Vol. 20, 243-262, Plenum, New York, 1986. tured plant tissues: Effect of intermediates and aromatic amino
LEON, J., N. YALPANI, I. RASKIN, and M. A.l.AWTDN, Induction of acids on free levels, Physiol. Plant, 30, 13-18 (1974).
benzoic acid 2-hydroxylase in virus-inoculated tobacco. Plant WOLF, R. B., G. F. SPENCBR, and R. D. PLATTNER, Benzoxalinone,
Physiol., 103, 323-328 (1993). 2,4-dihydroxy-l,4-benzoxazin-3-one, and its glucoside from
LUCKNBR, M., Secondary Metabolism in Microorganisms, Plants, Acanthus mollis seeds inhibit velvetleaf gennination and
and Animals, Springer-Verlag, Berlin, 1990. growth, 1. Nat. Prod., 48, 59-63 (1985).
LYONS, P. C. and R. L. NICHOLSON, Evidence that cyclic hydroxa- YALPANI, N., J. LEON, M. A. LAWTON, and I. RASKIN, Pathway
mate concentrations are not related to resistance of com leaves of salicylic acid biosynthesis in healthy and virus-inoculated
to anthracnose, Can. 1. Plant Pathol., II, 215-220 (1989). tobacco, Plant Physiol., 103, 315-321 (1993).
8
Phenylpropanoids

Introduction In this chapter, the origin of cinnamic and p-coumaric acids

Cinnamic Acid, p-Coumaric acid, and Related and their simple derivatives will be discussed initially, as
Compounds these intermediate substances are precursors to most other
Biosynthesis groups of related compounds. Various other modified C6 -C,
Physiological Roles compounds, such as simple phenylpropanoids, lignans, neo-
Volatile Phenylpropanoid (C6 -C,) Compounds lignans, lignin, and hydroxybenzoic acids, will then be cov-
Biosynthesis ered.
Some Important Phenylpropanoids Two separate routes of biosynthesis diverge from choris-
Biological Activity mic acid (3); one leads to L-phenylalanine (4) and another
Lignin to L-tyrosine (5) (see Fig. 7.9) compounds 3, 6, 7, and 10
Composition of Lignin are illustrated therein; Jensen, 1986). Both pathways involve
Origin of the Monomeric Units the common intermediate prephenate (6) that is derived from
Polymerization of Lignin chorismate by a Claisen rearrangement. There are two
Lignans chorismate mutases: one is plastidic and extensively regu-
Biosynthesis lated by phenylalanine and tyrosine, and the other is a cyto-
Biological Activity solic form that is unregulated. Prephenate is then converted
Neolignans into arogenate (7) via the action of prephenate aminotrans-
Hydroxybenzoic Acids (C6-C I ) Compounds ferase. Subsequently, arogenate is converted into phenylala-
Biosynthesis
nine by the enzyme arogenate dehydratase and into tyrosine
Distribution of Hydroxybenzoic Acids and Their
by arogenate dehydrogenase (Dewick, 1984; Jensen, 1986).
Derivatives
Although plantS and bacteria do not have the ability to
Biological Activity
do so, most animals can convert phenylalanine to tyrosine
Allelopathy
(Haslam, 1974). C6 -C2 compounds are probably formed by
References
a-oxidation of ketoacids such as phenylpyruvic (10) and p-
hydroxypyruvic acid (11). One well-known example, phen-
INTRODUCTION ylacetic acid (8), is common in plants (Gross, 1981). Mam-
Compounds derived from phenylalanine and/or tyrosine are mals and some microorganisms convert phenylalanine and
among the most common of all secondary metabolites in tyrosine to another C6-C2 compound, homogentisic acid (9)
plants, bacteria, and fungi. These include phenylpropanoid (Fig. 8.3).
compounds as well as many C6 -C I compounds (Fig. 8.1) Although both phenylpyruvic acid (10) and p-hydroxy-
(Gross, 1981). Relatively simple C6 -C3 compounds, such phenylpyruvic acid (11) can serve as precursors for phenylal-
as cinnamic (1) and p-coumaric acids (2), are modified to anine and tyrosine, respectively, it does not appear that plants
produce more complex derivatives (Fig. 8.2) (Conn, 1981, synthesize either compound. In E. coli, however, these two
1986). The term "phenylpropanoid" is sometimes used to intermediates are involved in the synthesis of phenylalanine
refer to any compound bearing a 3-carbon chain attached to and tyrosine. Both phenylpyruvic acid and p-hydroxyphen-
6-carbon aromatic ring (C6 -C, compounds). Most phenyl- ylpyruvic acid have been postulated to serve as precursors
propanoids are formed from cinnamic or p-coumaric acids. for a number of other compounds in plants-in particular,

106
 Phenylpropanoids 107

HO

HO~O HO-P-'Ll

L-phenylalanine (4) J' HO caffeic acid (12) OH CH3-0 OH

I via quinate Intermediate /' S·hydroxyferulic acid

~ Ho~:aq/:einterm~~~_o
t
OH CH3 -r ~H HO ~_ ~ ~ 0

! !
E·cinnamlc acid (1) ferullc acid (13) CH]" 0 OK
slnapic acid (14)

HO~O HO~O CH3~OH

OHCH3-O H HO~O
E-4-hydroxycinnamic acid (2) coniferaldehyde CH3" 0 H
sinapaldebyde

~
(p-coumaric acid)

+ HO~~
HO~~
- ~
0)=/
CH -O
~OH 3
H coniferyl alcohol

-
t lignans
sinapyl alcohol (43)

HO~ OH
neolignans

"=T'L.I lignin

p-coumaryl alcohol (44)

Fig. 8.1. Phenylpropanoid compounds and their derivatives.

alkaloids_ The role of these compounds will be discussed in (E2) and remains attached to the enzyme until the elimina-
more detail in Chapters 32, 33 and 37_ tion step. At that point, the "amino enzyme complex" either
releases ammonia and cinnamate or reacts again with cinna-
mate to regenerate L-phenylalanine (Fig. 8.4). PAL operates
CII'II'IAMIC ACID. p·COlJMARlC ACID. in a clearly defined stereochemical manner. Exclusive loss
Al'ID RELATED COlllPOUl'IDS of the pro-3S-hydrogen of phenylalanine is observed (Fig.
8.4) (Davin et al., 1992).
Some grasses (poaceae or Gramineae) have the corre-
Cinnamic acid and p-coumaric acid, derived from L-phenyl-
sponding enzyme, L-tyrosine ammonia lyase (TAL), capable
alanine, probably occur in all plants and fungi_ Their biosyn-
of converting L-tyrosine (5) into p-coumarate (2), but this
thesis was discussed in Chapter 7_
enzyme is not widely distributed. TAL also acts in a highly
stereospecific manner. The reaction also proceeds with re-
Biosynthesis moval of the pro-3S-hydrogen from the j3-methylene atom
Phenylalanine is converted to cinnamic acid by the action of L-tyrosine. It is not completely resolved that this enzyme
of the Ubiquitous enzyme L-phenylalanine ammonia lyase is distinct from PAL (Hanson and Havir, 1981).
(PAL) (EC 4.3_11.5) (Davin et al_, 1992; Hanson and Havir, E-Cinnamic acid (1) is the precursor of E-p-coumaric
1981)_ This enzyme, which catalyzes loss of ammonia and acid (2) in plants, and the presence of enzymes that catalyze
production of E-cinnamic acid (I), has been detected in a the hydroxylation of cinnamic acid to p-coumaric acid has
large number of vascular plants and several genera of basidi- been demonstrated (Butt and Lamb, 1981). These mixed-
omycetes_ PAL occurs mainly in the cytoplasm, but some function oxidases (in peas) are associated with the microso-
plants contain low levels of PAL in plastids, mitochondria, mal fraction and require molecular oxygen, NADPH, and
and microbodies (Hanson and Havir, 1981)_ Cinnamate is mercaptoethanol for the production of E-p-coumaric acid.
produced by the action of PAL in an elimination reaction p-Coumaric acid has a strong regulatory effect on the en-
108 Phenylpropanoids

go
~
o OH SCoA ~OOH ~ umbelliferone
Q QQ.... ~nA a coumarin
,:? __ ...... ~,:? O~··· HO % 0 0

I ,:? I I { alkaloids
~ ~ ~ cyanogenic glycosides

OH OR OH .. ". ~;:;:;~~::smino adds

p·coumaric acid (2) OH

....
HO~OO'Q"O HO~O
OH
OH
OH"
~ I I
OH I o
gallic acid ~ OH: I a flavonoid
OH
hydroxyacetophenones neoflavonoids stilbenes OH

gallotannins
eUagitannios

Fig. 8.2. Other major groups of secondary metabolites derived from phenylpropanoid metabolism (modified from Haslam, 1974).

zyme at 3 X 10-7 to 10-6 M concentration. When cinnamic the second OH group is introduced (Grand, 1984; Kiihnl
acid is fed to plants, it is recovered as p-coumaric acid (2) et al., 1987). The reaction requires NADPH and molecular
(usually) glucosylated at the carboxyl group. Cinnamic acid oxygen (Heller and Kiihul, 1985). Although shikimate and
is a precursor of caffeic (12), ferulic (13), and sinapic (14) other esters may be involved, quinates appear to represent
acids in most plants; quinate or shikimate esters appear to be the major route of synthesis of hydroxybenzoic acids (Kiihul
involved as intermediates (see below) (Gross, 1981; Haslam, et al., 1987).
1974) (Fig. 8.5). Cinnamic and related acids often occur in Once formed, these phenolic acids playa central role in
plants as esters. A number of methyltransferases (EC 2.1.1.6) the synthesis of lignin, flavonoids, and a wide range of other
can convert the hydroxycinnamates to methyl ethers (Davin phenolic secondary constituents.
et al., 1992; Poulton, 1981). Cinnamic acid and its derivative In plants, phenylpropanoic acids generally are bound to
acids, their ethers, and their esters are the principal constitu- various alcohols, especially sugars; mono-, di-, and trisac-
ents of the phenylpropanoid pool in plants (Haslam, I 974). charide esters are common (Mf/llgaard and Ravn, 1988). Mo-
Some relatively nonspecific enzymatic formation of caf- noglycosides are common in all dicotyledonous plants,
feic (12), ferulic (13), and synapic (14) acids has been noted whereas di- and triglycosides are most common in sympetal-
(Davin et w..,
1992). Monooxygenases of microsomal frac- ous plants. Quinic acid esters, such as chlorogenic acid (15),
tions appear to be involved. For example, a specific p-coum- are widespread in the Asteraceae, Solanaceae, and Rubia-
arate-3-hydroxylase has been isolated from mung beans. ceae, whereas rosmarinic acid (16) and related compounds
However, other work suggests that the carboxyl group of p- are restricted to the Lamiaceae and Boraginaceae. Disaccha-
coumaric acid must be esterified as aquinic acid ester before ride esters are found primarily in the Scrophulariaceae and
Brassicaceae (Mf/llgaard and Ravn, 1988). Complex glyco-
sides are known (Calis et al., 1990; Klick and Herrmann,
OH
1988); for example, 3,4-dihYdroxY-13-phenethyl-O-13-D-glu-

~-
copyranosyl- (1- 3}-4-0-caffeoyl- 13- D-glucopyranoside,
plantamajoside (17), has been isolated from callus of Reh-
mannia glutinosa (Scrophulariaceae) (Fig. 8.6) (Ravn and
Brimer, 1988).
OH
phenylacetic acid (8) Physiological Roles
homogentisic acid (9)
Cinnamic acid, p-coumaric acid (2), and several related
Fig. 8.3. Phenylacetic and homogentisic acid. compounds modify growth in plants (also see Section 8.8).
 PhenyJpropanoids 109

!1=
•
. ' \.
q-NH
°
CH~ .' \.
UN .;
° B:,~~
~ c::.J~~+Jl _
~
H,N ''''''H
CO,-
"..
HO '.
k-NH
....

H H
H~~
>-;0'H~+ · "·"H - CO,.

° HO'.l L-phenylalanine(4) ...• ~ 'H

'.~O"'\
HO
....

/"\,;-NH
NH
-
P<~~
H
-O,C I ---

d
E-cinnamic acid (1)

H1CO'-

0-:~~+
HoN ~~ HO
17

"""-
I HB

L-tyrosine (5) p.coumaric acid(l1)

Fig. 8.4. Stereochemistry of PAL action (modified from Havir and Hanson, 1970; published with pennission of copyright owner, Academic Press,
Orlando, FL.)

For example, E-cinnamic acid (1) was shown by Bonner nounced biological properties. These compounds are fre-
and Galston (1944) to be strongly inhibitory to seedlings of quently found in essential oils from plants and some are
Partheniurn argentaturn, guayule (Asteraceae). In attempts quite important as flavorings and perfumery ingredients.
to cultivate this plant, plants in the center of fields were Pltenylpropanoids are especially common in plants of the
stonted in appearance. Presumably, those at the periphery Magnoliales and the Apiaceae.
were exposed to lower concentrations of the active com-
pound. In later work, it was shown that E-cinnamic acid is Biosynthesis
liberated from the roots of adult plants; the leachate from Cinnamic (I), p-coumaric (2), and related acids may be
20,000 roots yielded 1.6 g of E-cinnamic acid. In the labora- activated by conversion to CoA esters by CoA ligases [e.g.,
tory, this compound proved to be inhibitory to Partheniurn 4-coumarate:CoA ligase (BC 6.2.1.12)] in much the same
growth at a concentration of 0.0001 %, whereas tomato seed- way that fatty acids are activated. The reduction of the CoA
lings were affected only if treated with a solution containing esters of cinnamic acids to cinnamyl alcohols involves two
100 times this concentration (Harborne, 1982). enzymes: cinnamoyl-CoA oxidoreductase (which forms the
A series of phenylpropanoid compounds from Pirnpinella aldehydes) and cinnamyl alcohol dehydrogenase (Grisebach,
spp. (Apiaceae) that contained epoxide rings were powerful 1981). Phenylpropanoids appear to be syuthesized from the
seed germination inhibitors (Cutler, 1992). CoA esters of this series of acids by conversion to the corre-
sponding aldehydes, then to the alcohols, and finally, by
elimination of a phosphate group, to allyl and propenyl com-
VOLATILE PHBNYLPROPAl'IOm (c,,-C3 ) pounds. In many plants. mixtures of all types co-occur (Fig.
COlllPOlll'IDS 8.7) (Gross. 1981; Mann, 1987). Reduction of the side chain
to produce dihydrocinnamic acids and related compounds is
The term phenylpropanoid also is used in a more restricted also known to occur in nature.
sense to indicate a number of relatively volatile compounds
in which the carbonyl function has either been removed, Some Important Phenylpropanoids
reduced, or otherwise modified. A number of volatile phen- Many phenylpropanoids are of ecological and economic
ylpropanoid compounds found in essential oils have pro- importance. These compounds are the principal volatile con-
110 Pllenylpropanoids

~O-
cionamic acid (1) p-coumarate (2)
L-phenylalanioe (4)
* clnnamic 4-hydroxylase
o

HO~"", CH'-O~""'- CH'-O~,<::::

""'- 0 0- 0 0- ""'- 0-
~ -- I __ HO~
HO HO ~
OCH,
caffeic acid (12) ferulic acid (13) sinapic acid (14)

E-S-O-(4-coumaroyI)-D-qulnate E-5-0-(4-caff.oyl)-D-quinale
•• These oxidations probably occur as quinic and shikimic esters, not as the free acids
Fig. 8.5. Biosynthesic steps of the hydroxycinnamic acids.

stituents of several of the most important spices and herbs. Biological Activity
Others are involved in plant-animal interactions as allo- An epimeric pair of dehydrodiconiferyl alcohol ~-D-gly­
mones, kairomones, and synomones. cosides (23, 24) from crown gall (Agrobacterium tumefa-
Among those from spices are eugenol (18), the major ciens) tumors in transformed Catharanthus roseus cultures
component (85%) of oil of cloves (Fig. 8.8), and cinnamalde- has been shown to promote cell division and replace the
hyde (19), the principal flavor constituent of oil of cinnamon. cytokinin requirement in tobacco cells (Fig. 8.10) (Binns et
However, the latter compound occurs in a bound form as al., 1987; Lynn et al., 1987).
cinnamyl acetate in the plant and fresh bark. During the Methyl Z-ferulate is a powerful inhibitor of uredospore
fermentative processing of the bark, this ester almost com- germination in a range of rust fungi (Friend, 1979). This and
pletely disappears and is not found in any quantity in com- related- compounds occur in the fungi themselves. They are
mercial cinnamon. Studies with radioactively labeled materi- gradually leached from the spores on the host surface. This
als confirm that the ester is converted into cinnamaldehyde. process must take place before germination of the spores
A number of phenylpropanoids are perceived as pungent occurs.
by humans and presumably by other organisms. Among Safrole (25) and isosafrole were once important as the
these are capsaicin (20), the pungent principle of peppers flavoring for root beer. These compounds are present in the
plant extracts used to make this beverage. By the 1950s,
(Capsicum species, Solanaceae) and piperine (21), the pun-
however, most root beer was flavored with synthetically de-
gent principle of black pepper (Piper nigrum, Piperaceae)
rived safrole. Both safrole and isosafrole were shown to be
(Fig. 8.9) (Harborne, 1982).
weakly carcinogenic and now have been banned for this
E-3'-Methylsulfonylallyl E-cinnamate (22) has been iso- purpose (Fishbein et aI., 1967). These compounds have a
lated from the bark of Phoebe brenesii (Lauraceae) (Castro methylenedioxy strocture and are known to inhibit mixed-
et al., 1985). function oxidase enzymes. Two compounds of similar stroc-
The precursors of some phenylpropanoid odorlflavor ture, piperonal (26) (naturally occurring) and piperonyl bu-
components of raspberry fruit occur as the 4'-O-~-D-gluco­ toxide (27) (synthetic) are mixed function oxidase inhibitors
pyranoside of 4-(4'-hydroxyphenyl)butan-2-one (raspberry and are used as synergists for insecticides such as pyrethrins,
ketone) and the 3'-O-~-D-glucopyranoside of 4-(3',4'-dihy- rotenoids, and Sevin (a carbamate insecticide) (Fishbein et
droxyphenyl)butan-2-one (Pabst et aI., 1990). al., 1967).
 Plienylpropanoids 111

Myristicin (28), which occurs in oil of nutmeg as well as the next instars, fifth instar larvae molted normally to adults
in essential oils of many members of the Apiaceae (Umbelli- but the ovaries were irreversibly affected. In such adults,
ferae), is a hallucinogenic compound. This, and a related the ovaries remained permanently immature (Saxena et ai.,
compound, elemicin (29) (Fig. 8.8), are largely responsible 1977).
for the hallucinogenic effects of nutmeg. The synthesis of One of the most interesting examples of the biological
myristicin in carrots is increased by exposure of carrot roots activity of phenylpropanoids in plant-insect interactions is
to light (Yates, 1987). Myristicin has also been shown to that of the oriental fruit fly and methyleugenol (31). This
have antifeedant properties for certain insects (Seigler, insect, Dacus dorsalis, uses methyleugenol as an aggrega-
1983), to be insecticidal, and to serve as a synergist for a tion pheromone. Pheromonal activity is exhibited both in
number of natoral and synthetic pesticides. Such synergists feeding and male aggregation. The same compound is pro-
in leaf tissue can significantly increase the toxicity of an duced by the flowers of several plant species, including Cas-
insecticide (Berenbaum and Neal, 1985, 1987; Lichtenstein sia flstulosa (Fabaceae: Caesalpinoideae). As a result, male
and Casida, 1963; Neal, 1989). fruit flies congregate on Cassia trees and perceive the pres-
The essential oil of Acorus calamus (Araceae) contains ence of methyleugenol as a sign to eat the leaves. If there
f3-asarone (30) that causes depression of development of is enough methyleugenol in the air, the flies will continue
the gonads and ovaries of the insect Dysdercus koenigii. to eat until they die. As little as 0.01 fLg of methyleugenol
Although third and fourth instar larvae molted normally to will produce a response in a single fly in a cage. Other ana-

o
o
HO~0-60
fiOH
~ \d OH
o H HO~ HO........... _

HO'c#0H o ~ b OH

OR OR cblorogenic acid (15) rosmarinic acid (16)

0.1:
fO,H OH

:~OH
HCOH

0' -
I HO

HC~
I ~ o
CO,H ~ h OH 0
g1u<osyJ-O HO monardein
H OH

t;
OH
p-coumaroyl m-tartrate
0H OHOH

~/7 OH

~ ~OLQ OH
o
benzyl clnnamate
~O
HO~ .
HO
4-p--coumaroyl myoinositol

HO~
~~ ~ O~OH
~
HOHO
OH
H
00
0

OH
H
0
Y"--Q-OH
_
OH

plantamajoside (17)

Fig. 8.6. Naturally occurring phenylpropanoid acid esters.

III Phenylpropanoids

o o o

HO
~ "":::'"
,& --
OH CH'.O~:::'"
I"
HO
OH
..& coenzyme A t
CH'.O~:::'"
1 "
HO'&
SCoA
NADPjf
~umaric acid (2) ferulic acid (13)

CH'.O~O
1 "" ~
CH'.O~:::'"
I" ~
CH,.o~:::,..?H
1 O-~=O
HO ,& NADPH HO b HO'& OH

coniferYI~/

eugenol (18) isoeugenol (83)

Fig. 8.7. Biosynthetic pathways for volatile phenylpropanoids in plants.

CH'.O~"
" ~ CH'.O~ CH'.O~:::'"
1 OH
HO'& HO~ HO ,&

eugenol (18) isoeugenol (83) coniferyl alcohol (42)

0
~H
1,,9 <0O~
I .# <0 ° v 1,,9
OCH,
cinnamaldehyde (19) safrole (25) myristicin (28)

CH'O~
1 .
~
CH'O~
CH,·O
Ib CH,·O ,& CH,.O 1 ,&

methylisoeugenol (32) metbyleugenol (31) anethole (36)

0""""0

~~
°OCHO
<0 1 .& CH,·O ~
1,&

methylchavicol
piperonal (26)
O(CH,),O(CH),OC,H, or estragole (35)

(a) piperonyl butoxide (27)

Fig. 8.8. (a & b) Some common phenylpropanoids.

 Phenylpropanoids 113

isoasarone (37) p-asarone (30) elemicin (29)

OCH,
CH,O

~
HOV

chavicol (34) veratrole (33)

3' d:C
r"

4' "'"
,pOl'
I l' "'"fl 0 ~2 S02CH3

E-3'-methylsulfonyl-E-cinnamate (22) piperenone (38)

~ OH

~ \~O O~O-CH'-CH'{}-OH
O~ H
H~~~\.-
IU~ 'ci\(
0 OH

(b) H mussatioside I (41)

Fig.8.8 (continued)

logs generally have reduced or no activity (Metcalf et ai., and chavicol (34) were active in early and middle August
1975), however. Western corn rootworrn (Diabrotica virgifera vir-
E-Methyl isoeugenol (32), E-asarone (2,4,5-trimethoxy- gifera) adults were attracted to a different set of compounds:
I-propenylbenzene), and, to a lesser extent, the leaf alde- estragole (4-methoxy-l-allylbenzene) (35), E-anethole (36),
hydes hexanal, E-2-hexenal, and heptanal (see Chapter 2) and indole. Estragole (35) was an effective attractant from
gave strong electroantennogram responses for the female early August (corn tasseling) until early October. 4-Me-
carrot rust fly Psila rosae (Guerin et ai., 1983). A mixture thoxycinnamaldehyde and 4-methoxy-1-propylbenzene also
of E-asarone and hexanal is more attractive than either com- are potent attractants (Metcalf and Lampman, 1989). Euge-
pound alone and probably serves as an oviposition stimulant nol (18), isoeugenol, and cinnamyl alcohol significantly at-
for the female fly. A synthetic mixture of E-methylisoeu- tracted northern corn root worm (Diabrotica barberi) adults
genol, E-asarone, osthol (a coumarin), bergapten (a furano- (Lampman et ai., 1987; Metcalf and Lampman, 1989).
coumarin), xanthotoxin (a furanocoumarin), and falcarindiol The effects of a series of phenolic compounds in artificial
(an acetylenic compound) was equally attractive as raw car- diets on biotype B greenbugs (Schizaphis graminum) has
rot extract and the components exhibited synergistic action been evaluated (Levin, 1976, Todd et ai., 1971). Z-Caffeic
(Stadler and Buser, 1984). acid (but not E-caffeic acid) caused a drastic reduction in
Southern corn rootworm (Diabrotica undecimpunctata growth and inhibited reproduction. Less than 20% of the
howardii) adults are attracted late in the season to many greenhugs survived on diets that contained vanillic, sinapic,
aromatic compounds: phenylacetaldehyde, benzyl acetone, gentisic, or ferulic acids (Todd et ai., 1971).
phenethyl alcohol, phenyl acetate, indole, veratrole (33), Isoasarone (37) (from Piper Jutokadzura, Piperaceae) and
methyleugenol (31), methylisoeugenol (32), eugenol (18), piperenone (38) are highly active as antifeedants against
and isoeugenol. Only veratrole (33), phenylacetaldehyde, Spodoptera litura (Seigler. 1983).
114 Phenylpropanoids

zingerone gingerol (40) paradol

Zingiber officinak

CH,NHCO(CH,),CII=CHCH(CH,), CH=CHCII=CHCQ---NQ

¢loc~
OH
~, 0---.1
ar-turmerine capsaicin (20) piperine (21)
Curcuma /onga Capsicum annuum Piper nigrum

Fig. 8.9. Pungent principles from plants (modified from Harhorne. 1982).

A series of structurally similar compounds derived by LlGl'IIl'I

acetate extension of phenylpropanoid precursors is also
found in the Piperaceae. Pipercide (39) from Piper nigrum
Lignin is the most abundant and most widely distributed
was insecticidal, but the mixture of amides from this plant
phenolic natural product in higher plants. This material,
was significantly more toxic and it appears that co-occurring
compounds exert synergistic effects (Fig. 8.11) (Berenbaum, which is associated with the advent of land plants, is not
1985). found in algae, nor is true lignin found in bryophytes (Lewis
Gingerol (40) (Fig. 8.9), a pungent principle of Zingiber and Yamamoto, 1990; Zinsmeister and Mues, 1987). Only
officinale (Zingiberaceae), ginger, has cardiotonic activity with lignified walls was it possible to build the rigid stems
(Shoji et aI., 1982). of woody plants and trees and the conducting elements for
Phenylpropanoid glycosides such as mussatioside I (41) water transport (Grisebach, 1981).
(Fig. 8.9) from a Mussatia species (Bignoniaceae) are re- It is not possible to designate a definitive structure for
ported to have medicinal properties. Some species are this amorphous polymeric material. Lignin is difficult to iso-
chewed with coca or alone in Peru and Bolivia (Jimenez et late in its natural state and is difficult to degrade chemically.
al.,1987). Nonetheless, there is general agreement concerning the prin-

OH OH
12

OCH, OCH,

OH

~
"OH "
HO~< _''''''0, 0
HO
HO
.0 ,• 0 ,. HO~
OH OCH,
OHI" OCH,

A(+)(23) 8(.)(24)

Fig. 8.10. Dehydrodiconiferyl alcoholj3-D-glycosides.

 Phenylpropanoids 115

o
<o NH~
o
plpercide (39)
o
<o NH~
dihydroplpercide

o
<o
o
guineensine
o

~ NH~
o
piperstachine

piperine (11) pellitorlne

Fig. 8.11. Extended phenylpropanoids from Piper species.

cipal Structural features and the manner in which it is formed of monomers from the cytoplasm into the cell wall and the
in the plant. subsequent polymerization are not well understood (Davin
Analytical methods for the detection and study of lignin et aI., 1992; Lewis and Yamamoto, 1990). Only p-coumaric,
have been reviewed (Lewis and Yamamotu, 1990). ferulic, and sinapic acids are viewed as bona fide lignin
precursors; they are converted into the corresponding mono-
Composition of Lignin mers without modification of the aromatic ring (Lewis and
Plant lignins can be divided into three broad classes that Yamamoto, 1990). The principal monomeric units (monolig-
are usually referred to as softwood (gymnosperm), hardwood nols) are p-coumaryl alcohol (44), coniferyl alcohol (42),
(dicotyledonous angiosperms), and grass or annual plant and sinapyl alcohol (43), although other compounds also
(monocotyledonous angiosperm) lignins. All are polymers may be involved. Most of the enzymes required for monomer
formed largely, ifnot entirely, from C.-C3 monomeric units. formation have heen purified and reasonably well character-
Coniferous or softwood lignins often are derived from coni- ized (Davin et aI., 1992; Lewis and Yamamoto, 1990). An
feryl alcohol (42) (Fig. 8.8), but more variety exists than has oxidoreductase, cinoamyl alcohol dehydrogenase (EC
heen recognized in the past (Lewis and Yamamoto, 1990). 1.1.1.195), catalyzes the fmal reductive step of monolignol
Syringyl-hased lignins also are found in many gymno- synthesis. This enzyme has been detected in various gymno-
sperms. Hardwood lignins are formed mostly from coniferyl sperm and angiosperm species (Davin et aI., 1992). Ferulic
and sinapyl (43) alcohols [e.g., in beech lignin the ratio is acid is incorporated into coniferyl alcohol and the lignin of
about I: I (Fig. 8.12)], but, again, many exceptions exist. wheat without degradation.
Finally, lignins from grasses and from such plants as bamboo In general, the levels of monomeric compounds in plants
and palms involve p-coumaryl alcohol (44) in addition to is small. Coniferyl alcohol occurs in the sap of gymnosperms
the coniferyl and sinapyl monomers. as the glucoside, coniferin (45) and sinapyl alcohol as the
glycoside, syringin (46) (Fig. 8.13). These compounds are
Origin of tbe Monomeric Units accumulated only rarely in angiosperms. In has heen sug-
The formation of the monomeric units has been described gested by some that the corresponding f3-glucosides are in-
in Section 8.3. The mechanism and regulation of transport volved in transport of the precursors through the plasma
116 Phenylpropanoids

9'
~H,OH CH,OH
CH I 'fL
A
OHCCH, CH

0
- , HOCH, CH· CH CJr"" - ---..CH, C
co *CH_~H ~:,OH

0 :(
I : CH,O"f' I ..., CH- CH,o O---CH
~H, CH,·O

*
OCH, 0-
"OCH,
CH~~CH' CH----O CH,OYOCH,
CH· Q
0 - CH OH CH9
1-<

°---CH - OCH ~ CH~HOCH, CH,OH

CH,O~ I h ~' 'I ~ o----~H / 0
I ~ OCH, I - IH"
CHP ~CH~:::-O *C~OH OCH, ~c'"
• ______CH,OH CH,OH I I ~
~H,OH C~H,O-y

M" I
CH OCH OCH,
~~O--CH co ~ ,
- CH "=(OCH, I I HOCH,·CH·C~
~
~
'" I
CH 0
'0
'" "" CH,OH
O-CH
CH,OH
~H OCH,
0
·CH,
OCH, II~

*
I , I

~ HOCH,.CH.CHO ;OCO ~CH~O~:_

~H-CHO '" I I '" OCH',¢-
- CH CH,O CH,O.& I
~H*OH
~ O~H OCH, CH,.O ~
CH",I c~ «;I

*
C"--
.. - - 0 OCH, I I
HOCH, CH

~
'"
I
OCH3
CH,OHcH.O
"
CHOH
CII
""

0
OCH
'0
OCH~
3 ~
I
CH

?
CH-CHO
,
CH,OH CH,o '"
I
OCH,
CH,·O I .&
OH
0-

Fig. 8.12. Structure of beechwood lignin (modified from Grisebach, 1981; used with permission of the copyright owner, Academic Press, Orlando, FL).

o o
~O.gIUCOSYI
SCoA ~SCOA
HO
via quinate
intermediate• HOY ----....~ RO~
OCH,
OH
coniferin (p·D-glueosyl) (45)

via quinate intermediate

+
+ + 0
~OI~HrO~S~:~·O~O.g1UCOSYI
HO
~ HOY -- OCH,
RO~
OCH,
p.coumaryl alcohol (44) syringin (R = ~·D·g1ucosyl) (46)

Fig. 8.13. Fonnation of coniferin and syringin.

 Phenylpropanoids 117

membrane and to the cell wall where lignification occurs; viewed (Ayres and Loike, 1990). Lignans are related closely
the corresponding l3-glucosidases are present (Davin et aI., to intermediates in the biosynthesis of lignin and probably
1992). Alternatively, these glucosides may serve only as are formed from analogous pathways. It should be noted,
storage products. however, that the majority of Iignans occur in distinctive
Coniferin can be incorporated into lignin in a variety of chiral forms, whereas most Iignins do not; no optically active
species (Grisebach, 1981) and, because a l3-glucosidase has degradation product of lignin has yet been isolated. Calli of
been located in the xylem of some plants, it is possible that Podophyllum peltatum produce the same array of Iignans
this enzyme controls the rate and location of lignification. [podophyllotoxin (51) (Fig. 8.15), a-peltatin (52), and 13-
Although coniferin is incorporated into lignin, there is little peltatin (53) (Fig. 8.17) as the plant rhizome. Lignans also
evidence that it is an obligatory precursor. Compounds of have been found in cell cultures of plants for which no previ-
this type may be involved in transport of the monomers ous record of Iignans occurs. For example, root organ cul-
(Lewis and Yamamoto, 1990). tures of Unum flavum yield very toxic extracts. This toxicity
Although Z-monolignols occur in many plants (e.g., in is caused by the presence of 5-methoxypodophyllotoxin, ap-
the bark of Fagus grandifolia), these appear to arise from parently as the 4-0-glucoside in the culture (Ellis, 1988).
E-monolignols by enzymatic reactions (Davin et a1., 1992).
Biosynthesis
Polymerization of Lignin
Many Iignans occur in optically active forms, suggesting
The phenylpropanoid monomeric units responsible for that the coupling process is stereoselective. In other in-
the formation of lignin are polymerized by the action of stances, however, enantiomeric mixtures are encountered
peroxidase enzymes (Grisebach, 1981; Lewis and Yama- (Davin et a1., 1992). This may require the presence of two
moto, 1990). Coupling of phenoxy radicals leads to dilignols stereoselective enzymes with different activities. Alterna-
and, afler reoxidation and radical coupling, to oligomeric tively, optically active compounds may be racemized in later
units, and, finally, to the lignin macromolecule (Grisebach, steps in some instances.
1981). The half-life of the typical flee-radical intermediate Coniferyl alcohol (42) is the specific precursor of (-)-
is about 45 s. secoisolariciresinol (54) in the presence of NADH, H2 0 2 ,
Lignin is deposited initially at the outer layers of the cell and a crude cell extract from Forsythia intermedia (Ume-
wall, and the enzymes which control the oxidative process zawa et a1., 1990a). A specific dehydrogenation then occurs
must be located near these sites. A carbohydrate matrix may to afford (- )-matairesinol (55). In Forsythia suspensa, coni-
be required for polymerization of Iignins (Haslam, 1974). feryl alcohol is the precursor of ( + )-pinoresinol (56). For-
The molecular architecture of lignin is complex but not mation of these compounds involves a direct stereochemi-
completely random, with a methodical distribution of certain cally controlled coupling of couiferyl-alcohol-derived
types of linkage and certain building units throughout the moieties. This conversion has been demonstrated both in
structure (Haslam, 1974; Lewis and Yamamoto, 1990). vivo and in vitro with a crude enzyme preparation (Davin
There is much need for additional study of lignin (United et a1., 1992; Umezawa et a1., 1990b). Nonspecific coupling
States Department of Energy, 1988). However, new ap- occurs in the presence of H 20 2 and horseradish peroxidase.
proaches such as the use of enzymes from Iiguin-degrading Subsequent processes are dependent on the regiochemistry
fungi and solid-state 13C-NMR spectroscopy promise to pro- and stereochemistry of the initial linkage (Fig. 8.16) (Davin
vide resolution of problems long considered intractable et a1., 1992; Weinges and Spiinig, 1967).
(Lewis and Yamamoto, 1990). The results of these studies have been explained tenta-
tively in the following manner (Davin et a1., 1992). IriFor-
sythia suspensa stems, couiferyl alcohol undergoes stereose-
L101'1A1'1S lective condensation to form the putative (-) (8R,8R,)-
quinone methide (Fig. 8.16), and not the corresponding ( + )-
Dimeric compounds derived from precursors related to those enantiomer. Subsequent intramolecular cyclization of the
involved in the formation of lignin are distributed widely ( - )-form yields ( + )-pinoresinol (56). In contrast, coniferyl
among higher plants (Davin et a1., 1992; MacRae and Tow- alcohol in cell-free extracts of Forsythia suspensa is not
ers, 1984; Pelter, 1986; Weinges and Spiinig, 1967). About properly compartmentalized and is in contact with nonspe-
450 Iignans from at least 55 families are known (Ayres and cific peroxidases as well as the stereospecific coupling sys-
Loike, 1990), especially from the Asteraceae, Pinaceae, and tem producing both ( + )- and ( - )-pinoresinol. When F or-
Rutaceae. Four major subtypes occur: dibenzylbutane deriv- sythia . intermedia cell'free extracts are incubated with
atives (47), dibenzylbutryolactones (lignanolides or deriva- coniferyl alcohol, both ( + )- and ( - )-quinone methides are
tives of butanolide) (49), monoepoxy Iignans ·or derivatives produced when H 20 2 is present as a cofactor. The (-)-
of tetrahydrofuran (49), and bisepoxylignans or derivatives antipode is reduced by a stereoselective NAD(P)H requiring
of 3,7-dioxabicyclo(3.3.0)-octane (50) (Fig. 8.14) (MacRae reductase to form (- )-secoisolariciresinol (54); the (+)-
and Towers, 1984). The use of spectroscopic methods for form is not reduced. Lastly, in a stereoselective enzymatic
characterization of this group of compounds has been re- NAD(P)-requiring dehydrogenation, only (- )-secoisolar·
118 Phenylpropanoids

x ~x
~1'1 - -
OCH,
I
I
I
OCH,
o 0

l o HO
H

X HO
CH,O H".. H
x
o

~
\

{~ )
Ho?cfCJ H""'"

h, , , , 0
"" H
::::::...
\

(. )-dihydrosesamin
OCH, OH
dibenzylbulane type (47)
OH nordihydroguaiuretic acid

CH,- 0 "" monoepoKy or tetrahydrofuran type (49)

OCOC~,CH' ~ 0

CH,O
o
CH,'O
o
asarinin
justicidin-A
o
' - - 0 3,7-dioxabicyclo(3.3.0)·octane type (50)

lignanolide or butenolide type (48)

Fig. 8.14. Types of lignans and their (probable) biosynthetic relationships (modified from Whiting, 1985; used with permission of the copyright owner,
the Royal Society of Chemistry).

iresinol is converted to ( - )-matairesinol (Davin et aI., 1992; stances. They may also be active in plants. A lignan (34)
Umezawa et aI., 1990a,b), isolated from the seed hulls of Aegi/ops ovata (Poaceae) is
Phenylalanine and p-coumaric acid, but not 3,4-dihydrox- a potent germination inhibitor for lettuce seeds (Lavie et aI.,
ycinnamic (caffeic) acid or acetate, were incorporated into 1974).
podophyllotoxin (51). In Podophyllum hexandrum (Berberi- Lignans are known to be active in plant-insect relation-
daceae), both cinnamic and ferulic acids are incorporated ships. Sesarnin (58) (Fig. 8.17) occurs in Sesamum indicum
into both halves of the molecule. The carbon of the O-methyl (Pedaliaceae) seed oil, as well as in a number of other plants.
group of ferulic acid is distributed into both rings A and C. This compound was isolated from Magnolia kobus (Magno-
Sinapic and 3,4,5-trimethoxycinnamic acids are not incorpo- liaceae) and demonstrated to be a growth-inhibiting sub-
rated. This suggests that other oxidation and methylation stance for Bombyx mori (Karnikado et aI., 1975). Lignans,
occurs after the initial dimerization reaction (Jackson and for example, those of Piper Jutokadzura and Libocedrus va-
Dewick, 1984; Pelter, 1986; Poulton, 1981). teensis, have antifeedant propereties as well (Davin et aI.,
1992).
Biological Activity Sesarnin (58) and asirinin (59) are effective in synergizing
Many of the biological activities of lignans have been the activity of insecticides (MacRae and Towers, 1984). The
reviewed (Davin et aI., 1992). These compounds may be methylenedioxy groups deactivate mixed-function oxidases
antimicrobial, antifungal, or antifeedant in a number of in- needed to detoxify the insecticides.
 Phenylpropanoids 119

~ ~:~o 1');0.:
~
<O~o
o~....{
() ._ oo~, ~
~OCH' / HO
I
00

¥ i,~,
OH
LOH

CH,-O""'-
OCH,
OCH, CH,-0 ~O
2''':::'''''- OH

podophylloloxln (51) HO """ HO

ferulicadd OCH, pinoresinol (56)

Fig. 8.15. Proposed precursors of some lignans.

ooq -
OH HO

OCH,
CH,O OCH,
coniferyl alcohol
o
NADPH!
H 0
2 2

HO
! ome:O:oH
(-)-(8R,8 R)-quinone

::?" H 8H """ OCH,

O
HOV'
CH,o OCH, OCH, (+)'pinoresinol (56)

CH,O

HO

OH
OH

(.)-.e<o·ISOIBrlclresinol
.. (54)
ynlheSls of (_)-mataIresinol
. and related r (.)-mataireSlnol
. (55)
Fig. 8.16. Bios . Ignans (based on data fro m Umezawa et aI., 1990 a,b).
120 Phenylpropanoids

OH

sesamio (58)
OR
OCH,

o
OH
OCH,

OCH,
guaiaretic acid
justicidin A (R=CH,) (85)
justicidin B (R=H) (86)

CH,.O OCH,

HO OH

(57)

a-(-)-peltatin (R=OH) (52)

~-peltatin (R=OCH,J (53)
(a)

Fig. 8.17. (a & b) Some biologically active lignans.

Justicidin A and B (85, 86) (Fig_ 8_17), from Justicia modified derivatives with reduced toxicity may be useful in
hayatai, have piscicidal activity as powerful as that of rote- this regatd (Kinghorn, 1992). Podophyllum resin has long
none (Davin et al_, 1992). been used for treating venereal watts; podophyllotoxin, 13-
Nordihydtoguaiaretic acid (NOGA) (60) is found in latge peltatin (53), deoxypodophyllotuxin, picropodophyllotoxin,
quantities in Larrea divaricata (creosote bush, Zygophylla- and a-peltatin (52) all have activity against measles and
ceae), a plant that is extremely abundant in the Chihuahuan, herpes simplex type I viruses. A variety of lignans inhibit
Sonoran, and Mojave deserts, as well as in southern South activity of certain enzymes. Among these ate the enzymes
America. This compound was widely used in the food indus- of respiration and cAMP phosphodiesterase (MacRae and
try for several yeats as an antioxidant, but its use has recently Towers, 1984).
been restricted. NOGA possesses light-activated toxicity Compounds structurally related to podophyllotoxin often
(Downum, 1992). have antimitotic activity. They arrest cultured cells at meta-
At least 40 lignans ate known to have antitumor and anti- phase and bind to purified tubulin prepatations. The lactone
viral activity and many ate cytotoxic (Ayres and Loike, ring of podophyllotoxin is involved in this binding (MacRae
1990; Davin et al., 1992; MacRae and Towers, 1984; Mac- and Towers, 1984).
Rae et al., 1989), although there ate no common structural A phenolic compound that resembles lignans in structure,
features to explain this activity. Podophyllotoxin (51) (Fig. platyphylloside (61), from Betula pendula, the Scandinavian
8.15), which has been isolated from a number of plant spe- birch, has been shown to account for 80% of the observed
cies, is a potent antineoplastic (i.e., tumor destructive) com- depression of in vitro organic matter digestibility in ruminant
pound. Although too toxic for practical value, synthetically test systems (Sunnerheim et al., 1988). This observation cor-
 Phenylpropanoids 121

relates with seasonal variation of phenolic compounds in the ture to compounds derived from acetate-malonate pathways
plant and with observed resistance to herbivores. Platyphyl- (e.g., 6-methylsalicylic acid) and to those derived from early
loside also occurs in B. platyphylla, the Japanese birch. intermediates in the shikimic acid pathway (often from shiki-
A number of other types of biological activity have been mic or quinic acid) (see Cbapters 6 and 7). Hydroxybenzoic
reviewed (MacRae and Towers, 1984). acids usually occur in plants as esters or as glucosides that
are widely distributed among higher plants. Salicin (63) was
the first glucoside to be found in nature; it is widely distrib-
1'IEOLiGNAi'lS uted in the Salicaceae, the willow family.
Related dimers in which the two C.-C, units are joined by Biosynthesis
dimerization of the aromatic rings instead of by the propa-
Hydroxybenzoic acids may arise by polyketide pathways
noid "tail" are called neolignans (Davin et aI., 1992; Got-
(primarily in bacteria and fungi), from shikimate and possi-
tlieb, 1978). Eusiderin (62) is an example of this type of
bly other intermediates of the sbikimate pathway, by l3-oxi-
compound (Fig. 8.17). Neolignans occur commonly in the
dation of cinnamic acid and related compounds, and by hy-
heartwood of species of the families Lauraceae, Magno-
droxylation of benzoic acid and other C6-C l compounds
liaceae, and Piperaceae. Neolignans also occur in the fruit
(Leon et aI., 1993; Yalpani et aI., 1993).
and seed of Ocotea veraguensis (Lauraceae). These fruits
In virus-inoculated tobacco, benzoic acid is converted by
are involved in a number of biological interactions (Dodson
the monooxygenase enzyme, benzoic acid 2-hydroylase, into
et aI., 1987).
salicylic acid. NADPH was required. Feeding of phenylala-
nine, cinnamic acid, or o-coumaric acid (all potential precur-
HYDROXYBENZOIC ACID CCs-C,) CO!IIPOUl'lDS sors) failed to induce the enzyme (Leon et aI., 1993).
L-Phenylalanine and cinnarnic acid serve as efficient pre-
A family of hydroxybenzoic acids (C.-C l ) and structurally cursors for salicin (63) (Fig. 8.18) (Pridliam and Saltmarsh
related compounds in plants is derived from cinnamic acid 1963; Zenk 1967). The initial steps of this process involve
and p-coumaric acid. These are sometimes similar in struc- l3-oxidation and removal of acetyl-CoA from the side chain

voJ:'
OCH,

OCH,
kobusin

: ,.
monoepoxylignanolide
0\

o
oE},
(I#HO
""'"
H·
OH~O
,H
CH,-O?,

CH,.O :::,...
OCH,
I 0
~

o asirinin (59) eusiderin (62)

OH

HO HO-Q--lYY'-O-OH

OCH, O-~.D.g1u<osyl

I
plalyphylloside (61)
OH
silycbristin
(a) a navono-lignan
Fig. 8.17. (continued)
122 Phenyipropanoids

(X1---+
o

~
C02-

~
CHO

I
~
H''''''
NH,+
- I""" OH_
# ~ OH

JOI --- ~ VI
phenylalanine (4) E-cinnamic acid (1) salicylaldehyde

HO~O~
OH

~
OH

HO HOHO 0
OH CHO OH CH,OH
H H

helicin salicin (63)

CHO

H"~ CH,OH
~~, OH

(b) (64) saligenin (83) vanillin (84)

Fig. 8.18. Proposed biosynthesis of salicin (modified from Zenk, 1967).

of the phenylpropanoid substrate. A related compound, 0- and Populus (Salicaceae) and in a number of other higher
hydroxybenzyl-[3-D-glucoside (64), was formed from sali- plants.
genin (83), but not salicin (63). It has been suggested that The methyl ester of salicylic acid is encountered com-
vanillin (84) (Fig. 8.18) results from the breakdown offerulic monly in essential oils and is especially abundant in the oil
acid in a similar manner (Gross, 1981; Haslam, 1974). This of wintergreen (Gaultheria procumbens, Ericaceae). Methyl
general process has been proposed by Zenk (1979) as a way salicylate occurs as a glycoside but is hydrolyzed upon injury
to explain the origin of many types of C 6 -C, compounds to the plant. This ester is widely used as a flavoring in can-
(Fig. 8.19). dies and as a perfumery ingredient. It is a major component
Ferulic, caffeic, and related acids, derived from cinnamic of liniments and is reputed to have antirheumatic properties.
and p-coumaric acid, give rise to a family of C6 -C, deriva- Anisaldehyde (66) (Fig. 8.19), a major component of the
tives. seed oil of anise (Pimpinella anisum, Apiaceae), also is used
widely as a flavoring.
Distribution of Hydroxybenzoic Acids and
Their Derivatives Biological Activity
Many hydroxybenzoic-acid-derived (C6 -C,) compounds C6 C,-Phenolics and their derivatives have a range of bio-
(acids, esters, aldehydes, alcohols, etc.) are found in the es- logical activities. One such compound, salicylic acid (65),
sential oils of plants. Several essential oils have commercial has extensive functions in plants (Leon et aI., 1993; Raskin,
importance, although many important components are read- 1992a, 1992b; Yalpani et aI., 1993). Others are involved in
ily synthesized. In many cases, the synthetic products have plant movements, play developmental roles, are participants
eclipsed the naturally occurring ones in importance. For ex- in plant-insect interactions and plant-plant interactions (al-
ample, vanilla extract contains vanillin (Fig. 8.18). As the lelopathy), or play defensive or pheromonal roles in insects.
user of synthetic vanilla will realize, however, synthetic sub- Salicylic acid has long been known to be an analgesic,
stitutes often lack minor components that make natural va- antipyretic, and anti-inflammatory compound. Willow bark
nilla preferable. has been used for thousands of years by peoples from various
In some fungi, salicylic acid (65) and related compounds, parts of the worlds as a pain reliever. Aspirin, the sodium
such as 6-methylsalicylic acid, are derived from acetate-ma- salt of acetylsalicylic acid, was introduced in 1898 by the
lonate pathways. In bacterial systems, similar compounds German chemical company Bayer as a medicinal. This com-
are derived from chorismic acid via isochorismic acid (see pound has become one of the world's leading drugs. Ameri-
Chapter 7). Salicylic acid often is found in species of Salix cans consume over 16,000 tons annually at a cost of $2
 Phenyipropanoids 123

billion (Raskin, 1992a, 1992b). In humans, aspirin blocks folding of leaves of plants such as Mimosa pudica (Faba-
the synthesis of prostaglandins involved in pain and bleeding ceae) (Schildknecht, 1983).
responses (see Chapter 2), as well as a variety of other ac- Acetosyringone (68) and a-hydroxyacetosyringone (69)
tions. Surprisingly, salicyclic acid has been found to be an activate the virulence (Vir) gene in Agrobacterium tumefa·
important endogenous substance in plants as well (Yalpani ciens. These molecules induce the entire vir regulon as well
et a!., 1993). Salicylic acid induces flowering in many plants. as the formation of T·DNA intermediate molecules (Fig.
In addition, this phenolic compound initiates a series of 8.20) (Stachel et a!., 1985). However, these compounds do
events in thermogenic plants, notably those of the family not appear to be common in plants. The lignin precursors,
Araceae, that lead to a rise in temperature of the reproductive sinapyl (43) (Fig. 8.1) and coniferyl alcohol (42) (Fig. 8.7)
parts as well as formation of amines involved in pollination have activity in the range of that of acetosyringone (Spencer
of the plants (Raskin, 1992a, 1992b). Salicylic acid also is and Towers, 1988).
involved in disease resistance of plants. This compound is Larvae of the butterfly Papilio glaucus glaucus feed
exported from the site of infection to uninfected tissues; the mainly on members of the Magnoliaceae, but rarely survive
highest concentrations are observed in and around hypersen· past the first instar when fed plant material of the genus
sitive lesions (Raskin, 1992a, 1992b). Salicylic acid also Populus (Salicaceae). In contrast, larvae of Papilio glaucus
has been found to occur in allelopathically active phenolic canadensis perform well on aspen and other Populus spe-
fractions and inhibits growth of surrounding vegetation. cies, but die when fed members of the Magnoliaceae (Lin·
Certain gallic acid glycosides such as K-PLMF 1 (67) droth et a!., 1988). The major compounds responsible for the
(turgorins) are involved in turgor movement in plants and lack of acceptability of Populus species to larvae of Papilio

o 0

~~~OH
~CO,.

H'''~H'+
V
() __
HO~ ...

j
~ p.coumaric acid (2)
phenylalanine (4)

~
(yCO,H

~OH -- V I ....::::-
COlH

HO~
(Y COH
1

salicylic acid (65) benzoic acid p-hydroxybenzoic acid (78)

o
Ho)(yCOlH HO~
I -- I ~ OH
HO""::::- HO""::::-

protocatechuic acid caffeic acid (12)

! o
CH3'O~
I "" OH
HO #
vanillic acid (77) ferulie acid (13)

Q CHO
CH"OXlCOlH

HO
I ""
#
OCH,

syringic acid (80)

--
CHjO~"'" CO,H
HO
I h
OCH,

sinapic acid (14)

anisaldehyde (66)

Fig. 8.19. Suggested route of biosynthesis for some C6 -C, compounds (modified from Haslam, 1974),
124 Phenylpropanoids

glaucus glaucus are four phenolic glycosides, salicin (63), Myrmica rubra show that salicylaldehyde is a more potent
salicortin (70), tremuloidin (71), and tremulacin (72) (Lin- deterrent that salicin (63) or its aglycone saligenin (83). The
droth et aI., 1987a). All four compounds occurred in P. trem- eggs of some of the beetles that feed on members of the
ulaides and P. grandidentata, but tremulacin was lacking Salicaceae contain salicin. Salicylaldehyde is detected in ne-
from P. deltaides (Lindroth et aI., 1987b). Special care must onate larvae, suggesting that salicin is quickly converted to
be used in studies of these compounds in Populus and Salix the aldehyde (Pasteels et aI., 1986; Rowell-Rabier and Pas-
tissues, however, because the amounts varied dramatically teels, 1986). This shift to members of the willow family
when leaves were freeze-dried or oven-dried. These changes appears to have occurred at least three times independently.
in composition were avoided only by working with fresh The species that shifted are able to exploit a food source,
plant tissue (Lindroth and Pajutee, 1987). yet are well protected. The larvae are provided with an addi-
Several simple phenols are found as defensive com- tional method of defense; the adults are able to sequester
pounds in insects (Fig. 8.21). sufficient salicin in the eggs to make them potentially lethal
Although many chrysomelid beetles synthesize and accu- to ants and to provide protection for the neonate larvae (Pas-
mulate iridoid monoterpenes (see Chapter 20), which ap- teels et aI., 1986, 1988, 1989; Rowell-Rabier and Pasteels,
pears to be a primitive defensive system within the family, 1986).
several species of the genera Chrysamela (subttibe The simple phenolic compounds 5-ethyl-2-methoxy-
Chrysomelina) and Phratora (subttibe Phyllodectina) have phenol, phenol (74), guaiacol, and veralrole (33) (Fig. 8.8)
shifted onto plants of the genus Salix (Pasteels et aI., 1988; have been proposed as sex pheromones for grasshoppers;
Rowell-Rabier and Pasteels, 1986). In this instance, the in- phenol (73), a-cresol, p-cresol (75), guaiacol, hydroquinone
sect larvae convert salicin (63) from the plant into salicylal- (76), catechol, p-benzoquinone, 4-methoxybenzaldehyde,
dehyde (73). Laboratory tests with a generalist predator ant 2,6,6-trimethylcyclohex-2-ene-1,4-dione, isophorone, and

acetosyringone (68) a;·hydroxyacetosyringone (69) K-PLMF 1 (67)

HO~O, -0
HO~O ~ j
HOH4o-o
~
OHH CH,OH O~o
salicin (63) salicortin (70) lJ

tremuloidin (71) tremulacin (72)

Fig. 8.20. Some biologically active C6~Cl compounds.

 Phenylpropanoids 125

¢ A A
OH

y OH
Y CO,H

p-cresol (75) hydroquinone (76) p-hydroxybenzoic acid (78)

ground-beetle water-beetle
Calasoma Dytiscus

6
OH
~

cx
CHO

#' OH

phenol (74) salicylaldehyde (73)

protocatechuic acid
methyl ester grass grub beetle water-boatman
water-beetle
Notonecta
Dytiscus

Fig. 8.21. Some simple phenols used as defensive agents by arthropods (modified from Harhorne, 1982).

2,5-dichlorophenol have been isolated from grasshopper de- teraceae), and Heterotheca subaxillaris (Asteraceae). This
fensive secretions (Whitman, 1990). The sex pheromone of compound had little effect on big blue stem (Andropogon
the soybean cyst nematode, Heterodera glycines, has been gerardii, Poaceae) (Fischer, et aI., 1988; Weidenhamer et
identified as vanillic acid (77) (Jaffe et aI., 1989). aI., 1988).
p-Hydroxybenzoic acid (78) (Fig. 8.20) has vitamin-like In the Mojave desert of California, the well-known plant
activity in some organisms. This growth factor requirement physiologist Fritz Went observed that the soil was usually
is associated with the formation of ubiquinone (coenzyme bare around the shrub EnceliaJarinosa (Asteraceae) (Went,
Q). The compound serves as an effective precursor for the 1970). He suggested that this situation was produced by the
benzoquinone ring of ubiquinone in mammals, bacteria, toxic effects of the root exudates. Exudates of the plant are
fungi, and plants (Haslam, 1974; also see preceding chapter). apparently not self-inhibitory, but they inhibit growth of
most annual plants. An active substance was later identified
as 3-acetyl-6-methoxybenzaldehyde (82) (Fig. 8.22). Al-
though primarily produced in the leaves, this toxin is re-
ALLELOPATHY leased when the leaves fall to the ground and decompose.
It is persistent and is not removed, except by very heavy
Compounds derived from shikimic acid pathways and espe- rains (Muller, 1953; Harborne, 1982).
cially phenolic substances have been encountered in a num- The Californian chaparral is a vegetational area of rela-
ber of studies of chemical interactions between plants. This tively low Mediterranean rainfall (i.e., wet cool winters and
"chemical warfare" is often called allelopathy. Although hot dry summers) along the coastal strip of southern Califor-
much controversy surrounds the importance of this phenom- nia. Many areas of chaparral are adjacent to areas of natural,
enon, allelopathy seems to be most important in seasonally uncultivated grassland. One of the most striking natural phe-
dry habitats. nomena of this shrubby grassland area is the zonation of
A number of phenylpropanoid-derived compounds are herbs around the thickets of shrubs which dominate this
capable of inhibition of seed germination and seedling flora. Two of the dominant shrubs in the chaparral are cham-
growth of other plants. Dihydrocinnamic acid (79), from ise, AdenostomaJasiculatum (Rosaceae) and Arctostaphylos
Ceratiola ericoides (Empetraceae), a common plant in the glandulosa (Ericacae). Both are widespread and reported to
Florida scrub, was shown to inhibit both germination and exert allelopathic effects on herbs. Adenostoma grows in
growth of Schizachyrium scoparium (little blue stem, pure stands on dry exposed slopes. Even though the soil is
Poaceae), Leptochloa dubia (Poaceae), Rudbeckia hirta (As- fully exposed to the sun and receives ample rain, no herbs
 cc"'
126 PhenyJpropanoids

o o

~OH ~OH
CO,H

b OH
l(AOH HO~
salicylic acid (6S) o-coumaric acid (81) p-coumaric acid (2)

o
CH'.O~OH
(yCO,H CH,·0 r(YCO,H

HO~
HO~ HO~
ferotic acid (13) p-hydroxybenzoic acid (78) vanillic acid (77)

))H'
YCHO
OCH,

dihydrocinnamic acid (79) syringic acid (SO) 3-acetyl-6-methoxybenza1dehyde

Enceliafarinosa (82)

Fig. 8.22. Phenylpropanoid-derived compounds with allelopathic activity.

grow in the vicinity of the lower slopes. Adjacent roadsides ybenzoic (78), and vanillic (77), syringic (80), o-coumarlc
produce a heavy crop of anouals. (81), cinnamic (1), salicylic (65), and ferulic acids (13) (Fig.
Experiments of McPherson and Muller (1969), McPher- 8.22). The effects of several of these compounds have been
son et al. (1971), and Muller and Chou (1972) have shown examined in other systems. Ferulic acid (13) is especially
that substances responsible for the observed allelopathic ef- toxic to several test species (Blum and Dalton, 1985a,b;
fects are water soluble and waterborne with both species. Blum et al., 1987; Toro et al., 1988).
One of the major climatic features of this vegetation is Periodic occurrence of fire removes the litter from the
coastal fog; even in the summer months and although rainfall ground. During the following rainy season, growth of herbs
is only moderate, there is a constant drip of moisture on to is again observed in the area around the shrubby plants. Most
the leaves of these shrubs and, hence, onto the surrounding of the shrubs survive the fires as crowns. Over a period of
soil. This dripping is sufficient to carry a regular supply of years, the shrubs again "poison" the ground around them
water-soluble inhibitors from leaves to the soil where they and the herbs gradually disappear.
remain in sufficient concentration to prevent growth or seed As is true for most allelopathy problems, one must con-
germination of any anoual species near these shrubs (Mc- sider many factors. Among these are nutrition, water, min-
Pherson and Muller, 1969). erals, soil oxygen, shading versus light, grazing effects, and
The leaves of both Adenostoma and Arctostaphylos spe- all of the above combined.
cies are relatively rich in water-soluble compounds and it is Several plants that normally grow in chaparral areas (Bro-
not surprising that the inhibitors turned out to be varying mus rigidus (introduced), Oenothera micrantha, Helian-
mixtures of phenols and phenolic derivatives (Fig. 8.22). A themum scoparium, Lotus scoparius, and Calindrinia cil-
similar, but not identical range of compounds, was isolated iata) were tested (in addition to tomato and other cultivated
by leaching the foliage and by extracting the surrounding plants). Seed germination was markedly decreased and plant
soil. Differences between leachate and soil probably occur growth strongly inhibited (McPherson and Muller, 1969).
because some leachate compounds are irreversibly bound to The allelopathic effects are cyclic and seem to involve
the soil or metabolized rapidly by microorganisms (Bar- the following series of events: Materials accumulate in the
borne, 1982; McPherson et aI., 1971; Muller and Chou, soil in the dry season. Rain carries the toxins into the soil
1972). All of the compounds involved were simple, wide- where they are absorbed into the top 3 cm of soil. The effec-
spread phenolics derived from shikimic acid. Among the tiveness of procedures for isolation of phenolic acids from
active compounds were hydroquinone (76) and p-hydrox- soil have been evaluated (Dalton et al., 1987). These biologi-
 Phenyipropanoids 127

cally active compounds inhibit germination of virtually all chemistry of Plants (P. K. Stumpf and E. E. Conn, eds.), Aca-
seeds, and when the seeds do germinate, they fail to develop. demic Press, New York, 1981.
Root growth is suppressed. Fire interrupts the cycle by re- CONN, E. E. (ed.), The Shikimic Acid Pathway, Recent Advances
moving the source of the phytotoxins and destroying some in Phytochemistry Vol. 20), Plenum Press, New York, 1986
of the accumulated compounds near the soil surface. The CUTLER, H. G., Herbicidal compounds from higher plants, in Phyto-
soil microorganisms decompose the compounds also, but chemical Resources for Medicine and Agriculture (H. G. Nigg
can work effectively only when the soil is wet (Harborne, and D. S. Seigler, eds.), 205-226, Plenum Press, New York,
1982; McPherson and Muller, 1969; McPherson et aI., 1992.
1971). DALTON, B. R., S. B. WEED, and U. BLUM, Plant phenolic acids in
soils: A comparison of extraction procedures, Soil Sci. Soc.
Am. J., 51,1515-1521 (1987).
DAVIN, L. B., N. G. LEWIS, and T. UMEZAWA, Phenylpropanoid
REFERENCES metabolism: Biosynthesis of monolignols, lignans and neolig-
nans, lignins, and suberins, in Phenolic Metabolism in Plants
AYRES, D. C. and J. D. LoIKB, Chemistry and Pharmacology of (H. A. Stafford and R. K. Ibrahim, eds.), Recent Advances in
Natural Products: Lignans-Chemical, Biological, and Clinical Phytochemistry Vol. 26, 325-375, Plenum Press, New York,
Properties, Cambridge University Press, Cambridge, 1990. 1992.
DEWICK, P. M., The biosynthesis of shikimate metabolites, Nat.
BERENBAUM, M. R., Brementown revisited: Interactions among al-
lelochemicals in plants in Chemically Mediated Interactions be- Prod. Rep., 1,451-469 (1984).
tween Plants and Other Organisms (G. A. Cooper-Driver, T. DoDSON, C. D., F. R. STERMITZ, O. CASTRO C., and D. H. JANZEN,
Swain and E. E. Conn, eds.), Recent Advances in Phytochemis- Neolignans from fruits of Ocotea veraguensis, Phytochemistry,
try Vol. 19) 139-169, Plenum, New York, 1985. 26, 2037-2040 (1987).
BERENBAUM, M. R. and 1. J. NEAL, Synergism between myristicin DOWNUM, K. R., Light-activated plant defence, New Phytol., 122,
and xanthotoxin, a naturally co-occurring plant toxicant, 1. 4011-420 (1992).
Chern. Bcol., 11, 1349-1358 (1985). Er.us, B. E., Natural products from plant tissue culture, Nat. Prod.
BERENBAUM, M. R. and J. J. NEAL, Interactions among alle10chemi- Rep., 5, 581-612 (1988).
cals and insect resistance in crop plants, in Allelochemicals: FISCHER, N. H., N. TANRlSEVER, and G. B. Wn.LlAMSON, Allelopa-
Role in Agriculture and Forestry (G. R. Waller, ed.), ACS Sym- thy in the Florida scrub community as a model for natural herbi-
posium Series 330, 417-430, American Chemical Society, cide actions, in Biologically Active Natural Products: Potential
Washington, DC, 1987. in Agriculture (H. G. Cutler, ed.), ACS Symposium Series 380,
233-249, American Chemical Society, Washington, DC, 1988.
BINNS, A. N., R. H. CHEN, H. N. WOOD, and D. G. LYNN, Cell
division promoting activity of naturally occurring dehydroconi- FISHBEIN, L., J. FAWKES, H. L. FALl< and S. THOMPSON, Thin-layer
feryl glucosides: Do cell wall components control cell division? chromatography of rat bile and urine following intravenous ad-
Proc. Natl. Acad. Sci., 84, 980-984 (1987). ministration of safrole, isosafrole, and dihydrosafrole, J. Chro-
matogr., 29, 267-273 (1967).
BLUM, U. and B. R. DALTON, Effects offerulic acid, an allelopathic
compound, on leaf expansion of cucumber seedlings grown in FRIEND, 1., Phenolic substances and plant disease, in Biochemistry
nutrient culture, 1. Chern. Bcol., 11, 279-301 (1985a). of Plant Phenolics (T. Swain, J. B. Harborne, and C. F. van
Sumere, eds.), Recent Advances in Phytochemistry Vol. 12),
BLUM, U., B. R. DALTON, and J. R. SHANN, Effects of various
557-588, Plenum Press, New York, 1979.
mixtures of ferulic acid and some of its microbial metabolic
GO'ITLlEB, O. R., Neolignans, Fortschr. Chern. Org. Naturst., 35,
products on cucumber leaf expansion and dry matter in nutrient
culture, J. Chern. Ecol., 11, 619-641 (1985b). 1-72 (1978).
GRAND, c., Ferulic acid 5-hydruxylase: A new cytochrome P450-
BLUM, U., S. B. WEED, and B. R. DALTON, Influence of various
dependent enzyme from higher plant microsomes involved in
soil factors on the effects of ferulic acid on leaf expansion of
lignin biosynthesis, Federation of European Biochemical Socie-
cucumber seedlings, Plant Soil, 98,111-130 (1987).
ties (FEBS), 169, 7-11 (1984).
BONNER, 1. and A. W. GALSTON, Toxic substances from the culture
GRlSEBACH, H., Lignins, in Secondary Plant Products (E. E. Conn,
media of guayule which may inhibit growth, Bot. Gaz., 106,
ed.), Vol. 7 of The Biochemistry of Plants (P. K. Stumpf and
185-198 (1944).
E. E. Conn, eds.), 457-478, Academic Press, New York, 1981.
BUTI, V. S. and C. J. LAME, Oxygenases and the metabolism of GROSS, G. G., Phenolic acids, in Secondary Plant Products (E. E.
plant products, in Secondary Plant Products (E. E. Conn, ed.), Conn, ed.), Vol. 7 of The Biochemistry of Plants (P. K. Stumpf
Vol. 7 of The Biochemistry of Plants (P. K. Stumpf and E. E. and E. E. Conn, eds.), 301-316, Academic Press, New York,
Conn, eds.), 627-665, Academic Press, New York, 1981.
1981.
CALIS, I., A. A. BASARAN, J. SARACOOLU, O. STICHER and P. RUBOI, GUERIN, P. M., E. STADLER, and H. R. BUSER, Identification of host
Phlinosides A, B, and C, three phenylpropanoid glycosides from plant attractants for the carrot fly, Psila rosae, J. Chern. Eco1.,
Phlomis linearis, Phytochemistry, 29, 1253-1257 (1990). 9,843-861 (1983).
CASTRO, 0., J. LoPEZ, A. VERGARA, and F. R. STERMITZ, Phenylpro- HANSON, K. R. and E. A. HAVlR, Phenylalanine ammonia lyase,
panoids in alkaloid-free species of Phoebe, J. Nat. Prod., 48, in Secondary Plant Products (E. E. Conn, ed.), Vol. 7 of The
640-641 (1985). Biochemistry of Plants (P. K. Stumpf and E. E. Conn, eds.),
CONN, E. E. (ed.), Secondary Plant Products, Vol. 7 of The Bio- 577-625, Academic Press, New York, 1981.
128 Phenylpropanoids

HARBORNE, J. B., Introduction to Ecological Biochemistry, Aca- LICHTENSTEIN, E. P. and J. E. CASIDA, Myristicin, an insecticide
demic Press. London. 1977; 2nd edition. 1982. and synergist occurring naturally in the edible parts of parsnips,
HASLAM. E.• The Shikimate Pathway. Wiley. New York. 1974. J. Agric. Food Chern.• 11. 410-415 (1963).
HAVIR. E. A. and K. R. HANSON. Phenylalanine ammonia lyase. LINDROTH. R. L. and M. S. PAJUTBE. Chemical analysis of phenolic
Evidence that the prosthetic group contains a dehydroolanyl glycosides: Art. facts. and artifacts. Oecologia. 74, 144-148
residue and mechanism of action, Arch. Biochem. Biophys., (1987).
141, 1-17 (1970). LINDROTII. R. L.• M. T. S. HsIA. and 1. M. SCRIBER. Characterization
HEllER, W. and T. KiiHNL, Elicitor induction of a microsomal 5- of phenolic glycosides from quaking aspen. Biochem. Syst.
0-(4-coumaroyl)shikimate 3'-hydroxylase in parsley cell sus- Bco!.. 15. 677-680 (1987a).
pension cultures. Arch. Biochem. Biophys .• 241. 453-450 LINDROTH. R. L.. M. T. S. HSIA. and 1. M. SCRIBER. Seasonal pat-
(1985). terns in the phytochemistry of three Populus species. Biochem.
JACKSON, D. E. and P. M. DEWICK, Cinnamic acid precursors of
Syst. Bcol.. 15, 681-686 (1987b).
podophyllotoxin in Podophyllum hexandrum. Phytochemistry. LINDROTH. R. L.. 1. M. SCRIBER. and M. T. S. HSIA. Chemical
23. 1029-1035 (1984). ecology of the tiger swallowtail: Mediation of host use by phe-
JAFFE. H.• R. N. HUEITEL. A. B. DEMILO. D. K. HAyES. and R. V. nolic g1ycosides. Bcology. 69. 814-822 (1988).
REaOIS, Isolation and identification of a compound from soy- LYNN. D. G.• R. H. CHEN. K. S. MANNING. and H. N. WOOD. The
bean cyst nematode, Heterodera glycines, with sex pheromone structural characterization of endogenous factors from Vinca
activity. J. Chern. Ecol..15. 2031-2043 (1989). rosea crown gall tumors that promote cell division of tobacco
JENSEN, R. A., Tyrosine and phenylalanine biosynthesis: Relation-
cells. Proc. Nail. Acad. Sci .• 84, 615-619 (1987).
ship between alternative pathways. regulation and subcellular MAcRAE. W. D. and G. H. N. TOWERS. Biological activities of
location. in The Shikimic Acid Pathway (E. E. Conn. ed.). Re- liguans. Phytochemistry. 23, 1207-1220 (1984).
cent Advances in Phytochemistry Vol. 20. 57-81. Plenum MAcRAE. W. D.• J. B. HUDSON. and G. H. N. TOWERS. The antiviral
Press. New York. 1986. action of liguans. Planta Medica. 55. 531-535 (1989).
JIMENEZ. C.• M. C. Vn.LAVERDE. R. RIOUERA. L. CASTEDO. and F. MANN. J .• Secondery Metabolism. Clarendon Press. Oxford. 1978;
R. STERMITZ. Three phenylpropanoid glycosides from Mussalia. 2nd edition. 1987.
Phytochemistry. 26. 1805-18\0 (1987). McPHERSON. 1. K. and C. H. MULLER. Allelopathic effects of Aden-
KAMIKADO. T .• C. CHANG. S. MURAKOSHI. A. SAKURAI. and S. ostoma fasciculatum, "chamise," in the California chaparral.
TAMURA, Isolation and structure elucidation of growth inhibitors Bcol. Monogr.• 39, 177-198 (1969).
on silkworm larvae from Magnolia kobus DC.• Agric. Bioi. McPHERSON. 1. K .• C. CHOU. and C. H. MUUJlR. Allelopathic con-
Chern .• 39. 833-836 (1975). stituents of the chaparral shrub Adenoslorna fasciculalum. Phy-
KINGHORN, A. D., Plants as sources of medicinally and phannaceut- tochemistry. 10. 2925-2933 (1971).
ically important compounds, in Phytochemical Resources for METCALF. R. L. and R. L. LAMPMAN. Estragole analogues as attrac-
Medicine and Agricultnre (H. N. Nigg and D. S. Seigler. eds.). IaDts for corn rootworms (Coleoptera: Chrysomelidae). J. Ecol.
Plenum Press. New York. 1992. Ent.• 82. 123-129 (1989).
KuCK. S. and K. liERRMANN. Glucosides and glucose ester of hy- METCALF. R. L.. W. C. MITcHELL. T. R. FuKUTO. and E. R. MET-
droxybenzoic acids in plants. Phytochemistry. 27. 2177-2180 CALF, Attraction of the oriental fruit fly, Dacus dorsalis, to
(1988). methyl eugenol and related olfactory stimulants. Proc. Natl.
KOHNL. T.. U. KOCH. W. Hm.ulR. and E. WELLMANN. Chlorogenic Acad. Sci .• 72, 2501-2505 (1975).
acid biosynthesis: Characterization of a light-induced microso- MffLGAARD, P. and H. RA VN, Evolutionary aspects of caffeoyl ester
mal 5-0-(coumaroY\)-D-quinateishikimate 3'-hydrolyase from distribution in dicotyledons. Phytochemistry. 27. 2411-2421
carrot (Daucus carota L.) cell suspension cultures, Arch. Bio- (1988).
chem. Biophys .• 258. 226-232 (1987). MULLER, C. H., The association of desert annuals with shrubs, Am.
LAMPMAN. R. L.. R. L. METCALF. and J. F. ANDERSEN. Semiochemi- J. Bot.• 40. 52-60 (1953).
cal attractants of Diabrotica undecimpunctata howardii Barber, MULLER. C. H. and C. CHou. Phytotoxins: An ecological phase of
southern com rootwonn, andDiabrotica virgifera virgifera Lec- phytochemistry. in Phytochemical Ecology (1. B. Harborne.
onte, the western com rootwonn (Coleoptera: Chrysomelidae), ed.). 201-216. Academic Press. London. 1972.
1. Chem. Bcol.. 13. 959-975 (1987). NEAL. J. 1.. Myristicin. saffrole. and fagarantide as phytosynergists
LAVIB. D .• E. C. LEVY. A. CoHEN. M. EBENARI. and Y. GUTIERMAN. of xanthotoxin. J. Chem. Bco!.. 15. 309-315 (1989).
New germination inhibitor from Aegilops ovata. Natnre. 249. PABST. A.• D. BARRON. J. AoDA. and P. SCHREIER. Phenylbutan-2-
388 (1974). one ~-D-g1ucosides from raspberry fruit. Phytochemistry. 29.
LEON. J .• N. YALPANI. I. RASKIN. and M. A. LAWTON. Induction of 3853-3858 (1990).
benzoic acid 2-hydroxylase in virus-inoculated tobacco, Plant PASTBELS. 1. M.• D. DALOZE. and M. ROWELL-RAHIER. Chemical
Physiol..l03. 323-328 (1993). defence in chrysomelid eggs and neonate larvae, Physiol. Ento-
LEVIN. D. A.. The chemical defenses of plants to pathogens and mol..n, 29-37 (1986).
herbivores. Annu. Rev. Bco!. Syst.. 7. 121-159 (1976). PASTEELS. J. M.• M. ROWELL-RAlilER. J. C. BRAEKMAN. D. DALOZE.
LEWIS, N. G. and E. YAMAMOTO, LIGNIN: Occurrence, biogenesis and S. DUFFEY, Evolution of exocrine chemical defenses in leaf-
and biodegradation. Annu. Rev. Plant Physiol. Plant Mo!. Bio!.. beelles (Coleoptera: Chrysomelidae). Experientia. 45. 295-300
41. 455-496 (1990). (1989).
 Phenylpropanoids 129

PASTBELS, J. M., M. ROWELL-RAHlER, and M. J. RAupp, Plant-de- TODD, G. W .. A. GETCHUM, and D. C. CRESS, Resistance in barley
rived defense in chrysomelid beetles. in Novel Aspects of In- to greenbug, Schizaphis graminum L. Toxicity of the phenolic
sect-Plant Interactions (P. Barbosa and D. Letourneau, eds.), and flavonoid compounds and related substances, Ann. Bota-
235-272, Wiley, New York, 1988. mo!' Soc. Am., 64, 718-722 (1971).
FELTER, A., Lignans: Some properties and syntheses, in The Shiki- TORO G. I. R., G. R. LEATHER, and F. A. EINHBLUG, Effects of
mic Acid Pathway, (E. E. Conn, ed.), Recent Advances in Phyto- three phenolic compounds on Lemna gibba G3, J. Chem. Ecol.,
chemistry Vol. 20, 201-241, Plenum Press, New York, 1986. 14. 845-853 (1988).
POULTON, J. E., Transmethylation and demethylation in the metabo- UMEZAWA, T., L. B. DAVIN, and N. G. LEWIS, Formation of the
lism of secondary plant products, in Secondary Plant Products tignan, (- )-secoisolariciresinol, by cell free extracts of For-
(E. E. Conn, ed.), Vo!' 7 of The Biochemistry of Plants (P. K. sythia intermedia, Bioehem. Biophys. Res. Commun., 171,
Stumpf and E. E. Conn, eds.), 667-723, Academic Press, New 1008-1014 (1990a).
York,1981. UMEZAWA, T., L. B. DAVIN, E. YAMAMOTO, D. G. I. KINGSTON, and
PruoHAM, J. B. and M. J. SALTMARSH, The biosynthesis of phenolic- N. G. LEWIS, Lignan biosynthesis in Forsythia species, Chern.
glycosides in plants, Biochem. J., 87, 218-224 (1963). Commun., 20, 1405-1408 (1990b).
RAsKIN, I., Salicylate, a new plant hormone, Plant Physio!., 99, UNITED STATES DEPARTMENT OF ENERGY, Lignin Research: Needs
799-803 (1992a). and Opportunities, U.S. Dept. Energy, Washington, DC. 1988.
RASKIN, I., Role of salicylic acid in plants, Annu. Rev. Plant Phys- WENT, F. W., Plants and the chemical environment, in Chemical
io!. Plant Mo!. Bio!., 43, 439-463 (1992). Ecology (E. Sondheimer andJ. B. Simeone, eds.), 71-82, Aca-
RAVN, H. and L. BRIMER, Structure and antibacterial activity of demic Press, New York, 1970.
plantamajoside, a caffeic acid sugar ester from Plantago major WBlDBNHAMER, J. D., G. B. WlLLlAMSON, N. H. FISCHER, and N.
subsp. major, Phytochemistry, 27, 3433-3437 (1988). TANRISEVER, Selective phytotoxic effects of hydrocinnamic
ROWELL-RAmER, M. and J. M. PASTEBLS, Economics of chemical acid, Abstracts, Phytochemical Society of North America, An-
defense in Chrysomelinae, J. Chern. Eco!., 12, 1189-1203 nual Meeting, 1988.
(1986).
WEINGES, K. and R. SPANIO, Lignans and cyclolignans. in Oxidative
SAXENA, B. P., O. KOUL, K. T1KKU, and C. K. ATAL, A new insect Coupling of Phenols (W. J. Taylor and A. R. Battersby, eds.),
chemosterilant isolated from Acorus calamus, Nature, 270, Marcel Dekker, Inc., New York, 1967.
512-513 (1977).
WENT, F. W.o Plants and the chemical environment, in Chemical
SCHll..OKNECHT, H., Turgorins, honnones of the endogeneous daily Ecology (E. Sondheimer and J. B. Simeone, eds.), 71-82, Aca-
rhythms of higher organized plants-detection, isolation, struc- demic Press, New York, 1970.
ture, synthesis and activity, Angew. Chern. Int. Ed., 22,
WHITING, D. A., Lignans and neolignans, Nat. Prod. Rep., 2,
695-710 (1983).
191-211 (1985).
SEIGLER, D. S., Role of lipids in plant resistance to insects, in Plant
Resistance to Insects (P. A. Hedin, ed.), ACS Symposium Series
WHITMAN, D. W.o Grasshopper chemical communication, in Biol-
ogy of Grasshoppers (R. F. Chapman and A. Joem, eds.),
208,303-327, American Chemical Society, Washington, DC,
357-391, Wiley, New York, 1990.
1983.
SHon, N. A., A. IWASA, T. TAKEMOTO, Y. ISHlO, and Y. OHlZUMl, Y ALPANl, N., J. LEON, M. A. LAWTON, and I. RASKIN, Pathway
Cardiotonic priociples of ginger (Zingiber officinale Roscoe), of salicylic acid biosynthesis in healthy and virus-inoculated
J. Pharm. Sci., 71, 1174-1175 (1982). tobacco, Plant Physio!., 103, 315-321 (1993).
SPENCER, P. A. and G. H. N. TOWERS, Specificity of signal com- YATES, S. G., Increased concentration of myristicin and 6-me-
pounds detected by Agrobacterium tumejaciens, Phytochemis- thoxymellein in carrot root upon irradiation with UV light, in
try, 27, 2781-2785 (1988). Allelochemicals: Role in Agriculture and Forestry (G. R. Wal-
ler, ed.), ACS Symposium Series 330, 295-299, American
STACHm., S. E., E. MEsSBNs, M. VAN MONTAGU, and P. ZAMBRYSKI,
Identification of the signal molecules produced by wounded Chemical Society, Washington, DC, 1987.
plant cells that activate T-DNA transfer in Agrobocterium tu- 7....ENK. M. H" Pathways of salicyl alcohol and salicin fonnation in
mefaciens, Nature, 318, 624-629 (1985). Salix purpurea L., Phytochemistry, 6, 245-252 (1967).
STADLER, E. and H. R. BUSER, Defense chemicals in leaf snrface ZENK. M. H.• Recent work on cinnamoyl-CoA derivatives, in Bio-
wax synergistically stimulate oviposition by a phytophagous chemistry of Plant Phenolics (T. Swain, J. B. Harbome and C.
insect, Experientia, 40, 1157-1159 (1984). F. van Sumere eds.), Recent Advances in Phytochemistry Vol.
SUNNERHBlM, K., R. T. PALO, O. THBANDER, and P. KNvTsSON, 12,139-176, Plenum Press, New York, 1979.
Chemical defense in birch. Platyphylloside: A phenol from Be- ZINSMEISTER, H. D. and R. MUES, Moose als Reservoir bemerkenw-
tula pendula inhibiting digestibility, J. Chern. Eco!., 14, erter sekundarer Inhaltsstoffe, GIT Fachzeitschr. Laboratorium,
549-560 (1988). 31,499-512 (1987).
9
Coumarins

Introduction Cownarin (I), fIrst isolated from Dipteryx (Coumarouna)

Biosynthesis odorata (tonka bean, Fabaceae) and a component of a num-
Biosynthesis of Cownarins ber of spices and herbs, is found in essential oils from many
Biosynthesis of Cownarins derived from p-coumaric plants. Cownarin from plant sources is still used to a certain
acid extent as a flavoring, but most coumarin comes from syn-
Biological Activity thetic sources. The use of syothetic coumarin as a flavoring
Furanocoumarins in food products is restricted in the United States.
Biosyothesis of Furanocoumarins Although free cownarins are known to occur in plants,
Systematic and Phylogenetic Importance of most simple cownarins occur as glucosides. The involve-
Furanocoumarins ment of such glucosides in the biosynthetic sequence is a
Biological Properties of Furanocownarins unique feature among higher plants. The ~-D-glucoside of
Effects of Furanocoumarins on Plants o-coumarinic acid (7) (Fig. 9.2) is found in plants such as
Furanocoumarins as Phytoalexins sweet clover (Melilotus alba) and lavender (Lavandula offi-
Furanocoumarins and Insects cinalis). Usually, free cownarin (1) is formed by hydrolysis
Toxicity of Furanocoumarins to Mammals of the glucoside and subsequent lactonization of the Z-o-
Furanocoumarins in Foods coumarinic acid (8). A similar situation probably is true for
Medicinal Uses of Furanocownarins umbelliferone and scopoletin (Haslam, 1974).
References Coumarins have been found in all plant parts; most have
been reported from seeds and roots. Concentrations as great
as 9% by weight have been observed. Generally, cownarins
Il'ITKODVCTlOI'l are located within specialized structures such as secretory
ducts called vittae (Berenbaum, 1991).
Simple cownarins (iactones of phenylpropanoic acids with
a 2H-l-benwpyran-2-one nucleus) are widespread in plants;
the four most widely distributed coumarins are coumarin BIOSYl'lTHESIS
(I), wnbelliferone (2), esculetin (aesculetin) (3), and scopo-
letin (4) (Fig. 9.1) (Brown, 1979). Coumarins have not been
Biosynthesis of Coumarin
reported from algae or mosses, although umbelliferone has
been reported from a liverwort (Berenbaum, 1991). Cou- L-Phenylalanine, a precursor for coumarin (1) in Melilo-
marin is both the name of a specific compound and the name tus alba, is first converted to E-cinnamic acid by the action
of a group of structurally related compounds. Variations of of phenylalanine ammonia lyase (PAL) and then to ortho-
these basic coumarin structures are encountered commonly coumaric acid (9) by the action of cinnamic acid o-hydroxyl-
(especially in the Apiaceae) and at least 300 simple coumar- ase (Fig. 9.2). Although this step of the pathway was initially
ins and a total of at least 1000 naturally occurring cownarins reported to occur in the chloroplast, at present none of the
are known (Berenbaum, 1991; Brown, 1986, personal com- steps of the reaction sequence appear to occur there. Coum-
munication). Many coumarins, such as xanthyletin (5) and aric acid (9) is converted into the corresponding ~-glucoside
seselin (6), are prenylated. (10) by UDP-glucose and an O-glucosyltransferase. This

130
 rrx
Coumarins 131

5 4

~ <?'
I
""", HO~ CH3-0~

~oAo HO 7"'"
8
0
2

0 Ho~oAo Ho~oAo
coumarin (1) umbelliferone (2) esculetin (3) scopoletin (4)

HO~"""
I gIUCOSYI-O~

HO""'OO
HO HO
~"A
0 0

our"
dicoumarol (13)
daphnetin esculin

OH
xanthyletin (5) warfarin (14) sesilin (6) columbianetin (28)

Fig. 9.1. Some important coumarin structures.

glucoside (10) undergoes a UV-light-mediated E-Z Biosynthesis of Coumarlns from P'

(trans-cis) rearrangement to produce the isomeric J3-gluco- Coumarlc Acid
side (7) that is accumulated in the plant. Examination of
Melilotus cells indicated that a mixture of the two isomers Although cinnamic acid does serve as a direct precursor
is present (Oba et aI., 1981). Under various conditions, espe- for coumarin (1) itself, most other coumarins are derived
cially injury, a co-occurring J3-glucosidase cleaves the gluco- tromp-coumaric acid and umbelliferone (2) (Fig. 9.3). Addi-
side (7) to yield glucose and coumarinic acid (8), which is tional hydroxyl functions usually are added after the cou-
spontaneously cyclized to coumarin (1). In this plant, the marin nucleus is constructed. There is some evidence for the
glucosides were shown to occur in the vacuoles. The 13- conversion of ferulic acid to scopoletin in tobacco, but this
glucosidase occurs in the sarne cells, but outside the vacuole, appears to be an exceptional case (Brown, 1986).
perhaps in the intercellular spaces or attached to the cell wall As in coumarin biosynthesis involving phenylalanine ox-
(Brown, 1986; Oba et aI., 1981; Poulton et aI., 1980). ygenation of p-coumaric acid in the ortho position is the key

o-coumaric acid (9)

O-glucosyl

o-coumaric acid
OH

--
poD-glucoside (10)

~C02H
~O.gIUCOSYI
o-coumarinic acid
- ~CO~
~OH
Z-o-coumarinic acid (8)
~
~oAo
coumarin (1)
P-D-glucoside (7)

Fig. 9.2. Biosynthesis of coumarin in plants (Luckner, 1972; used with pennission of the author).
132 Coumarins

reaction in this series. A number of feeding studies indicate actively inhibited 6-phosphogluconate dehydrogenase, an
that umbelliferone (2) is an efficient precursor for many cou- enzyme of the pentose phosphate pathway. Coumarin is
marins of this type. The 2' -hydroxylase responsible differs known to enhance IAA (Indole Acetic Acid) oxidase activity
from the one involved in the biosynthesis of coumarin itself in some plants and lAA synthetase in other systems. Effects
(see above) in its site specificity and the requirement for a on gibberellin and abscisic acid also have been observed
tetrahydropteridine cofactor (Berenbaum, 1991). Glycosyla- (Brown, 1981, 1986).
tion, followed by a light-mediated E-Z isomerization of the Most coumarins also have pronounced physiological ef-
side-chain double bond, occurs. Removal of glucose by a fects in mammals and insects. Coumarin (0.1 %), ferulic acid
i3-g1ucosidase leads to the acid (11) which immediately cy- (0.1 %), and p-coumaric acid (5%) were shown to be toxic to
clizes. Umbelliferone (2) then is formed by removal of the the larvae of the bruchid beetle, Callosobruchus maculatus
remaining sugar from umbelliferone glucoside (12). (Janzen et al., 1977).
In tobacco cell cultures, a UDP-glucose o-dihydroxycou- In fermenting hay, o-coumaric acid (8) is converted into
marin 7-0-glucosyltransferase mediates glucosylation of es- dicoumarol [3,3'-methylenebis(4-hydroxycoumarin)] (13), a
culetin (3) with strict positional specificity (Brown, 1986). compound with marked anticoagulant properties (Bellis et
Coumarins may be formed at various sites in plants. In a!., 1967; Davies and Ashton, 1964), that occasionally has
Melitotus alba, sweet clover, young actively growing leaves been responsible for the death of livestock (Sneader, 1985).
seem to be the principal site of synthesis (Brown, 1981). The synthesis of warfarin (14), a powerful rodenticide, was
based on these properties, and for a time, this compound
was effective in controlling populations of rats. Recently,
BIOLOOICAL ACTIVITY however, a strain of "super rats" has emerged which have
developed an enzyme system that allows them to cope with
A large number of experiments involving simple coumarins this warfarin (Marm, 1987; Sneader, 1985). Regular low
and growth of plants have been conducted (Brown, 1981). doses of warfarin can prevent strokes in humans (Singer,
Observed effects include growth inhibition or stimulation, 1990).
inhibition of pollen tube formation, modification of stomatal
action, rhizoid fonnation, inhibition of seed germination, and
petiole abscission. Scopcietin (4) (Fig. 9.1) is a potent germi- FURANOCOUMARll'lS
nation stimulant (active at concentrations of 2-20 ppm).
Growth of Chinese cabbage seedlings is strongly inhibited In addition to relatively simple coumarins, a second group
by both umbelliferone (2) and scopoletin (4) (Brown, 1986). of more complex coumarins is known in which the coumarin
Many effects on specific enzymes also have been observed structure is prenylated (i.e., a five-carbon unit derived from
(Brown, 1981). For instance, both scopoletin and esculetin mevalonic acid metabolism is attached). The five-carbon
have inhibitory effects in tobacco cultures. The glycosides mevalonate-derived unit of these compounds usually is re-

o ~O
~OR I "'" ~
0
~ __ 14 ~ "'"
-- ~
OR
OR
RO~ RO OR
glucosyl-- 0
#
O~glucosyl

-:OSYI-O cc:
I ""
# O-glucosyl
CO,R

--- glucosyl- 0 cc --
I ""
#
(11)
OR
CO,R

'"_~O~O - umbelliferone glycoside (12)

""~O
umbelliferone (2)

Fig. 9.3. Biosynthesis of simple coumarins derived from p-Coumaric acid in plants (Luckner, 1972; used with permission of the author),
 Coumarins 133

o ~
__
HO"'"
9'
I ""

0 0
-

umbelliferooe (2) demethylsuberosin (20)

HO~
0:0
9'

"'"
I
0
""
--
0
HO
~O
I 9' ""

(+)-(S)-marmesin (21)
--5'
ccexo
4'54

f 9'

•
I

psoraleo (17)
"'"

Fig. 9.4. Proposed biosynthetic pathway to linear furanocoumarins.

duced to two carbons which are part of a furan ring. At least Coumarin biosynthesis involves formation of PAL and 4-
200 structures of these furocoumarins or furanocoumarins coumarate CoA ligase which is light and elicitor inducible.
occur in plants (Ivie, 1987, Ivie et aI .. 1987). Two principal Cells of P. crispum respond by producing the linear furano-
types, linear and angular furanocoumarins, arise by prenyla- coumarins psoralen (17), bergapten (18), and xanthotoxin
tion at position C-6 or C-8, respectively, followed by cycliza- (19) (Fig. 9.6), as well as other furanocoumarins (Ellis,
tion and modification of the ring (Fig. 9.4). In contrast to 1988). The relative amounts offuranocoumarins in the mix-
simple coumarins, furanocoumarins are restricted in distri- ture was changed by use of different fungal elicitor sources.
bution. A substantial proportion of the furanocoumarins of Floss and Mothes (1964) established that two carbons of
many plants is found on the leaf surface (Zobel and Brown, the furan ring are derived from mevalonate. Experimentally,
1989). the overall incorporation of mevalonate is low, a phenome-
Most common furanocoumarins have characteristic reten- non observed in many other studies (see Chapter 19). In
tion properties on HPLC (High Performance Liquid Chro- Ruta graveolens (Rutaceae), the first step in linear furano-
matography) with both reversed-phase and conventional- coumarin formation is the prenylation of the C-6 position
phase columns (Spencer et aI., 1987). The assigument of of umbelliferone (2) to yield demethylsuberosin (20) (Fig.
ring-substitution patterns of plant-derived furanocoumarins 9.4). In this case, no intermediate that leads to the formation
can be made by negative-ion mass spectrometry (plattner of angular furanocoumarins is formed. In elicitor-treated
and Spencer, 1988). Ammi majus cell cultures two dimethylaIIyl diphosphate:
umbelliferone dimethylallyltransferase activities, one cata-
lyzing 6-prenylation and the other 7-0-prenylation were in-
BIOSl'l'lTHESlS OF FURANOCOUJllARll'lS duced (Hamerski et aI., 19900). Demethylsuberosin (20) was
rapidly metabolized. In a separate work, Brown and Steck
Cinnamic acid is a precursor of p-coumaric acid and of most (1973) demonstrated that (l-hydroxyisopropyldihydrofura-
furanocoumarins, just as it is for ordinary coumarins (Floss nocoumarins, such as ( + }-(S}-marmesin (21), are interme-
and Mothes, 1964). Umbelliferone (7-hydroxycoumarin) (2) diates in the biosynthesis of furanocoumarins. An as-yet un-
also is incorporated into most furanocoumarins. Coumarin identified "psoralen synthase," that appears to be a
(I) and scopoletin (6-methoxy-7-hydroxycoumarin) (4) are cytochrome P-450 monooxygenase, may be involved in the
much poorer precursors than umbelliferone for both angular formation of psoralen (17) from marmesin (21) (Berenbaum,
furanocoumarins [such as sphondin (IS) and pimpinellin 1991). Psoralen (17) then can be modified further to produce
(16) (Fig. 9.6)] and linear furanocoumarins, suggesting that a series of oxygenated furanocoumarins (Brown, 1986;
any additional oxygenation occurs after formation of the Grundon, 1983).
coumarin nucleus (Brown, 1986). In intact plants, this bio- In Ruta graveolens, two different methyltransferases, S-
synthetic system may represent a channeled process; incor- adenosylmethionine:xanthotoxol O-methyltransferase and
poration of probable intermediates is low. Suspension cul- S-adenosylmethionine:bergaptol O-methyltransferase, are
tures of PetroseUnum hortense (syn. P. hortense) contain involved in the formation of 8-hydroxybergapten (22) and 5-
only traces of furanocoumarins until challenged by prepara- hydroxyxanthoxin (23) from bergaptol (24) and xanthotoxol
tions of fungal elicitors. Regulation of coumarin biosyn- (25), respectively (Berenbaum, 1991). 5,8-Dihydroxypso-
thesis at the molecular level has been studied (Berenbaum, ralen (26) is a much more efficient precursor for the biosyn-
1991; Hahlbrock and Ragg, 1975; Hahlbrock et aI., 1981). thesis of isopimpinellin (27) than are the monomethylated
134 Coumarins

OCH,

~
\VoAo
OH
bergaptol (24)
S·bydroxybergapten (22)

i
$0-+--
OH OCH,

~
'o~OAo
~
\VoAo
OCH,

/~"'
psoralen (17)
isopimpinellin (27)

! t
OH

~
'o~oAo
~
\VoAo
OH OCH,

xanthotoxol (25) 5·bydroxyxanthotoxln (23)

Fig. 9.5. Proposed pathways from psoralen to isopimpineilin.

furanocoumarins bergapten (18) or xanthotoxin (19) (Fig. Although linear furanocoumarins occur in at least 19
9.5) (Brown, 1986; Dewick, 1984). plant families, they are common only in two, the Rutaceae
Angular furanocoumarins are derived by prenylation at and the Apiaceae (Berenbaum, 1991). Within the Apiaceae,
the 8·position of umbelliferone followed by cyclization via coumarins are only found in subfamily Apioideae (not in
similar intermediates, although experimental evidence is not the Hydrocotyloideae or Saniculoideae). Further, these com·
as good as for linear furanocoumarin biosynthesis. Columbi· pounds are not found in the closely related family Araliaceae
anetin (28) (derived by 8·prenylation) is involved in the bio· (Berenbaum, 1991). 1n contrast to linear furanocoumarins,
synthesis of angelicin and derived angular furanocoumarins angular furanocoumarins are restricted primarily to two, the
(Fig. 9.3). Apiaceae and Fabaceae. 1n the Apiaceae, angular furano·
The mRNAs induced by fungal elicitors and by light in coumarins are found in two advanced trihes, the Apieae and
parsley proved to be similar or identical, suggesting a •• com· the Peucedaneae. Although many plants synthesize linear
mon" defense response to different sources of plant stress furanocoumarins in the absence of angular furanocoumarins,
(Berenbaum, 1991). few, if any, produce angular furanocoumarins in the ahsence
of linear furanocoumarins suggesting that the biosynthesis
of angular furanocoumarins evolved more recently. Angular
SYSTEJllADC Al'ID PHYLOGEl'IEDC furanocoumarins appear to he less toxic to most organisms
IJI1POKTANCE OF FURAl'IOCOlJlllAKll'lS than linear furanocoumarins (Berenbaum and Feeny, 1981;
Ivie, 1987; Ivie et al., 1987).
Although simple coumarins are found in many plant fami·
lies, the distribution of furanocoumarins is restricted. Furan·
ocoumarins have been reported from the Apiaceae (Umbelli·
ferae), Asteraceae (Compositae), Moraceae (Brosimum, BIOLOGICAL PKOPEKnES
Dorstenia, Fatoua, and Ficus), Pittosporaceae, Rosaceae, OF FURAl'IOCOlJMARll'lS
Rutaceae, Solanaceae, and Tbymelaeaceae (Bilia et al.,
1992,1993; Murrayetal., 1982; Swain and Downum, 1990). Linear furanocoumarins are potent phototoxins capable of
Certain precursors to this group of compounds are found in killing or inhihiting the growth of viruses, bacteria, and fungi
the Cneoraceae (Murray, 1978). as well as affecting many higher organisms including both
 Coumarins 135

insects and nematodes. Photoactive furanocoumarins require formation of large amounts of umbelliferone in addition to
activation at UV-A wavelengths (320-400 nm). Under these isopimpinellin (27), (S)-marmesin (21), (R)-ammirin, as
conditions, linear furanocoumarins can form cross-linkages well as other furanocoumarins (Harnerski et a!., 1990b).
between the two strands of the DNA double helix by joining Xanthotoxin (19) has been identified as a phytoalexin in
pyrimidine bases, resulting in a partial loss of template activ- parsnip. The ED,o for xanthotoxin against Ceratoeystis fim-
ity for RNA synthesis as well as inhibition of DNA replica- briatum is 21.6 flog/ml (KuG, 1992).
tion (Berenbaum, 1991; Downum, 1992; Downum and However, strains of the fungus Gibberefla pulicaris (Fu-
Nemec, 1987). sarium sambucinum) isolated from both furanocoumarins
and non-furanocoumarin-containing plants were tolerant to
Effects of Furanocoumarins on Plants and able to metabolize 16 furanocoumarins and furanocoum-
arin precursors (Desjardins et a!., 1989).
As is true for simple coumarins, many furanocoumarins
affect various growth and developmental processes in plants Furanocoumarins and Insects
and specific enzymes (see above) (Brown, 1981). For exam-
ple, a number of furanocoumarins such as bergapten (18) and Insect herbivory is an important selective force in the
sphondin (15), vaginidiol, isopimpinellin (27), pimpinellin ecology and evolution of plants. The linear furanocoumarin,
(17), and isobergapten (Fig. 9.6) are known to either stimu- xanthotoxin (19), found in many plants of the Apiaceae and
late or inhibit growth of Chinese cabbage seedlings (Brown, Rutaceae is toxic to the larvae of Spodoptera eridania, a
1986). Psoralen (17) is a powerful germination inhibitor and generalist insect herbivore. This toxicity was shown to be
can inhibit growth of plants (Downum, 1992). There is some increased in the presence of UV light.
evidence that coumarins interact with other growth regula- Photons can alter the electronic configuration and form
tors. a highly reactive excited state that can interact directly with
biomolecules such as DNA, proteins, or membrane lipids
with concomitant toxic effects (Berenbaum, 1987). The lar-
Furanocoumarins as Phytoalexins
vae of many of the herbivores of apiaceous plants live in
Cell suspension cultures of Ammi majus propagated con- rolled leaves or in webbed inflorescences, possess physical
tinuously in the dark produced only the coumarin umbelli- resistance in the form of body pigments that shield the insect
ferone (2). The addition of fungal cell wall fractions induced from damaging wavelengths of light, or quench excited

CH30~
p- I "'"
o ~ 0 0

angelicin (30) spbondin (15) imperatorin (29)

bergapten (18)
~
\yoA
OCH3
<We OCH 3
xanthotoxin (19)
khellin (33) (a 'Y-pyrone)

00,0 Yo
{)cr 0

osthol (31)

Fig. 9.6. Some important furanocoumarins.

136 Coumarins

states (biochemical resistance) by enzymatic degradation of (Fig. 9.6), an angular furanocoumarin found in only a few
phototoxic molecules (Berenbaum, 1978, 1987). relatively advanced tribes of the Apiaceae, reduces both
Xanthotoxin (19), imperatorin (29), and bergapten (18) growth rate and fecundity in this insect. This finding sug-
(linear furanocoumarins) occur in all above ground parts gested that, in this instance, plants that produce angular fu-
of Pastinaca sativa (Apiaceae), whereas angelicin (30) and ranocoumarins may be favored over those that make only
sphondin (15) (angular furanocoumarins) occur only in um- linear furanocoumarins (Berenbaum, 1983).
bels of some individuals (Fig. 9.6). The total furanocoumarin Midgut and body tissues of the black swallowtail (Papilio
content is highest in reproductive parts, particularly buds polyxenes) possess high enzymatic activity that catalyzes
and seeds (Berenbaum, 1981a). Even in the seeds, furano- the detoxification of linear furanocoumarins, explaining the
coumarins are not distributed in a homogeneous manner. tolerance of this species for these compounds. The detoxifi-
The vittae or oil glands, which contain these compounds, on cation of angular furanocoumarins occurs at a slower rate
the commissural side of the mericarp contain over 90% of (Ivie et aI., 1987). Other Papilio species, for example, P.
the furanocoumarin content of the seed (Berenbaum and brevicauda, feed exclusively on plants of the genera Her-
Zanger!, 1986). Generalist insects tend to feed on plants or acleum, Angelica, Ligusticum, and Petroselinum. The first
plant parts low in furanocoumarin content, whereas parsnip three contain angular furanocoumarins (Berenbaum and
specialists, notably Depressaria pastinacella (Lepidoptera: Feeny, 1981). As few insects can utilize plants with angular
Oecophoridae), feed exclusively on umbels (Berenbaum, furanocoumarins, plants that possess pathways that permit
1981a). angular attachment of the furan ring may have been favored
Individual plants of wild parsnip (Pastinaca sativa) vary in some instances. However, Papilio species that can feed
in both content and composition of furanocoumarins. Fur- on linear furanocoumarins may have become even more spe-
ther, much of this variation is genetically based. The effects cialized herbivores by adapting to feeding on plants with
of light and nutrients on furanocoumarin production have angular furanocoumarins.
been evaluated (Zanger! and Berenbaum, 1987). Wild par- There was no effect on larval growth or survivorship
snip also varies in resistance to damage by Depressaria when angelicin (30) was included in the diet of parsnip web
pastinacella which feeds on flowers and developing seeds. worms (Depressaria pastinacella) (Nitao, 1989).
Approximately 75% of the variation in resistance was attrib- Furanocoumarins such as isopimpinellin (27), bergapten
utable to four furanocoumarin characteristics. Resistance is (18), and kokusagin have antifeedant activity against
correlated positively to the proportion of bergapten (18) and Spodoptera litura. A number of similar compounds from
amount of sphondin (15) in seeds and correlated negatively umbelliferous plants have been demonstrated to be active
with the amount of bergapten and proportion of sphondin antifeedants against Spodoptera litura, Blattela germanica,
in leaves. In greenhouse studies, however, several of the and Stylopyga rhombifolia (Munakata, 1977; Yajima and
resistance factors were negatively correlated with potential Munakata, 1979).
seed production, and ostensibly, highly resistant plants, in A mixture of volatiles from carrot (Daucus carota, Apia-
the absence of herbivory, would be at a competitive disad- ceae) is responsible for oviposition by females of the carrot
vantage in the field (Berenbaum et aI., 1986). fly (Psila rosae). Among these are the phenylpropanoids
When parsnip webworm larvae were fed radioactively E-methyleugenol, E-asarone, a prenylated coumarin, osthol
labeled xanthotoxin (19), approximately 95% was metabo- (31), bergapten (18), xanthotoxin (19), and an acetylenic
lized, indicating that metabolic detoxification is important compound, falcarindiol (Harhorne, 1986). No single compo-
in tolerance of the insect to furanocoumarins. Excretion of nent of the mixture is responsible for the behavior of the
the remaining xanthotoxin and metabolites occurred both in fly.
the frass and through the silk glands. The silk glands con-
tained half as much tritium as the rest of the body (Nitao, Toxicity of Furanocoumarins to Mammals
1989, 1990). Furanocoumarins are rapidly catabolized in mammals. In
Species of Apiaceae that possess furanocoumarins typi- rodents, xanthotoxin (19) is metabolized by O-demethyl-
cally are those of open habitats (swamps or thickets) or dis- ation, aryl hydroxylation at position 5, oxidation of the 5,8-
turbed areas such as roadsides or old fields. This is in keeping dihydroquinone to the quinone, hydrolysis of the pyrone
with a role of light in the effects of these plants on herbivores ring, oxidative opening of the furan ring, and sulfate and
(Berenbaum, 1981 b). glucuronide conjugation (lvie, 1987; Ivie et aI., 1987).
Butterflies of the genus Papilio (Lepidoptera) often are Plants that contain furanocoumarins have been implicated
associated with members of the Apiaceae. Xanthotoxin (19), in several problems of livestock poisoning. For example, in
a linear furanocoumarin, occurs in many plants of the Apia- the western United States, Cymopterus watsoni (Apiaceae)
ceae. This compound is not appreciably toxic to the larvae has been involved in the poisoning of sheep (Ivie, 1978).
of Papilio polyxenes which normally feed on umbelliferous Starved individuals of the rock hyrax, Procavia capensis
plants. Most of these insects feed only on plants that have syriaca, which is notable for its ability to consume toxic
linear furanocoumarins and, in general, do not consume plants, were killed when they consumed shoots of Pitur-
plants that make angular furanocoumarins. Angelicin (30) anthos triadiatus (Apiaceae), which contain furanocoumar-
 Coumarins 137

ins. This animal nonnally eats the young shoots of the plant, REFERENCES
which have much lower concentrations of these compounds
(Harborne, 1986). The toxicity offuranocoumarins to mam- ANON., Vegetable health risk?, Chern. Br., 203 (1988).
mals has been reviewed (Ivie, 1987; Ivie et aI., 1987; Seigler, BEIER, R. C. and H. N. NIGG, Natural toxicants in foods, in Phyto-
1983). chemical Resources for Medicine and Agriculture (H. N. Nigg
and D. S. Seigler, eds.), 247-367, Plenum Press New York,
Furanocoumarins in Foods 1992.
BELLIS, D. M., M. S. SPRING, and J. R. STOKER, The biosynthesis
Parsnips and perhaps celery and parsley contain apprecia- of dicoumarol, Biochem. J., 103, 202-206 (1967).
ble levels of the linear furanocoumarins psoralen (17), xan- BERENBAUM, M. R.. Toxicity of a furanocoumarin to armywonns:
thotoxin (19), and bergapten (18). Consumption of 100 g of A case of biosynthetic escape from insect herbivores, Science,
parsnip root would expose an individual to 4-5 mg of these 201,532-534 (1978).
compounds, which are known to be phytotoxic, mutagenic, BERBNBAUM, M. R.o Patterns of furanocoumarin production and
and photocarcinogenic. Furanocoumarins are not destroyed insect herbivory in a population of wild parsnip (Pastinaca sat-
by nonnal cooking (Ivie, 1981). iva L.), Oecologia, 49, 236-244 (1981a).
Contact with plants containing linear furanocoumarins by BERBNBAUM. M. R., Patterns of furanocoumarin distribution and
humans and subsequent exposure of the area to sunlight re- insect herbivory in the Umbelliferae: Plant chemistry and com-
sults in painful, vesicated areas on the skin. This problem munity structure, Ecology, 62, 1254-1266 (198Jb).
has been observed in field workers and others who handle BERENBAUM, M. R., Coumarins and caterpillars: A case for coevo-
celery (Anon., 1988; Beier and Nigg, 1992). A similar prob- lution, Evolution, 37, 163-179 (1983).
lem has been observed in bartenders and others who handle BERENBAUM, M. R., Charge of the light brigade: Phototoxicity as
limes and later are exposed to sunlight. The coumarin con- a defense against insects, in Light-Activated Pesticides (J. R
tent of lime oils is about 7%. Figs (Ficus carica, Morace3e) Heitz and K. R. Downum, eds.), ACS Symposium Series 339,
also are known to cause photodennatitis; fig leaves contain 206-216, American Chemical Society, Washington, DC, 1987.
large quantities of psoralen (17) (1.65 mg/g fresh weight) BERENBAUM, M. R., Coumarins, in Herbivores: Their Interactions
and bergapten (18) (Beier and Nigg, 1992). with Secondary Plant Metabolites, Vol. 1 (G. A. Rosenthal and
The production of phytoalexins in celery plants infected M. R. Berenbaum, eds.), 221-249, Academic Press, San Diego,
CA,1991.
with the fungus Sclerotinia sclerotiorum caused workers to
develop severe initation. These plants contain xanthotoxin BERENBAUM, M. R. and P. FEENY, Toxicity of angular furanocoum-
(19) and 4,5' ,8-trimethylpsoralene. Uninfected plants of ce- arins to swallowtail butterflies: Escalation in a coevolutionary
anns race, Science, 212, 927-929 (1981).
lery do not contain these coumarins in significant quantities,
although there is evidence that grocery workers have experi- BERENBAUM, M. R. and A. R. ZANGERL, Variation in seed furano-
coumarin content within the wild parsnip (Pastinaca sativa),
enced phytophotodennatitis after handling apparently
Phytochemistry, 25, 659-661 (1986).
healthy celery plants. Some of these lines, which contain
BERENBAUM, M. R, A. R. ZANoERL, and J. K. NITAo, Constraints on
levels of furanocoumarins 10-15 times higher than more
chemical coevolution: Wild parsnips and the parsnip webwonn,
usual cu1tivars of celery, had been selected for disease and Evolution, 40, 1215-1228 (1986).
insect resistance (Beier and Nigg, 1992).
Bll.IA, A. R., G. FLAMINI, L. PISTELLI, and I. MoRELI.I, New constit-
uents from Pyracantha coccinea leaves, J. Nat. Prod., 56,
Medicinal Uses of Furanocoumarins 1741-1747 (1992); errata, 56, 984 (1993).
The ability to bind to DNA has been used to provide BROWN, S. A., Biochemistry of the coumarins, in The Biochemistry
of Plant Phenolics (T. Swain, J. B. Harbome, and C. F. van
an effective treatment for the stubborn and disfiguring skin
Sumere, eds.), Recent Advances in Phytochemistry Vol. 12
disease, psoriasis, a disorder characterized by the prolifera-
249-286, Plenum Press New York, 1979.
tion of epidennal cells. Oral doses of xanthotoxin (19) fol-
BROWN, S. A., Coumarins, in Secondary Plant Products (E. E.
lowed by controlled exposure to ultraviolet irradiation have
Conn, ed.), Vol. 7 ofTbeBiochemistry of Plants (P. K. Stumpf
shown remarkable success in controlling this problem. An- and E. E. Conn, eds.), 269-300, Academic Press, New York,
gular furanocoumarins do not bind to DNA in the same man- 1981.
ner and have little effect (Beier and Nigg, 1992; Brown, BROWN, S. A., Biochemistry of plant coumarins, in The Shikimic
1986; Ivie, 1987; Ivie et al., 1987). Acid Pathway (E. E. Conn, ed.), Recent Advances in Phyto-
Visnadine (32), an angular pyranocoumarin, has pro- chemistry Vol. 20, 287-316, Plenum Press, New York, 1986.
nounced vasodilatory effects (Berenbaum, 1991). BROWN, S. A. and W. STECK, 7-Demethylsuberosin and osthenotas
Khellin (33), an antispasmodic drug that comprises ap- intennediates in furanocoumarin biosynthesis, Phytochemistry,
proximately 1% of the dry weight of plants of Ammi visnaga 12, 1315-1324 (1973)
(Apiaceae), has a pyrone structure reminiscent of that of DAVIES, E. G. and W. M. ASHTON, Coumarin and related com-
many linear furanocoumarins (Fig. 9.6). This compound acts pounds of Anthoxanthum puelii and Meli/otus alba and dicoum-
directly on smooth muscle, is a bronchial dilator, and is used arol fonnation in spoiled sweet vernal and sweet clover hay, J.
in the treatment of asthma (Sneader, 1985). Sci. Food Agric., 15, 733-738 (1964).
138 Coumarins

DESJARDINS, A. E., G. F. SPENCER, and R. D. PLA'ITNER, Tolerance compounds to the seed-eating larvae of the bruchid beetle Callo-
and metabolism of furanocoumarins by the phytopathogenic sobruchus maculatus, Phytochemistry, 16, 223-227 (1977).
fungus Gibberella pulcharis (Fusarium sambucinum), Phyto- Kuc, J., Antifungal compounds from plants, in Phytochemical Re-
chemistry, 28, 2963-2969 (1989). sources for Medicine and Agriculture (H. N. Nigg and D. S.
DEWICK, P. M., The biosynthesis of shikimate metabolites, Nat. Seigler, eds.), 159-184, Plenum Press, New York, 1992.
Prod. Rep., I, 451-469 (1984). LUCKNER, M., Secondary Metabolism in Plants and Animals, Aca-
DOWNUM, K. R., Light-activated plant defence, New Phytol., 122, demic Press, New York, 1972.
401-420 (1992). MANN, J., Secondary Metabolism, Clarendon Press, Oxford, 1978;
DOWNUM, K. R. and S. NEMEC, Light-activated antimicrobial chem- 2nd edition, 1987.
icals from plants: Their potential role in resistance to disease- MUNAKATA, K., Insect antifeedants of Spodoptera litura in plants,
causing organisms, in Light-Activated Pesticides (J. R. Heitz in Host Plant Resistance to Insects (P. A. Hedin, ed.), ACS
and K. R. Downum, eds.), ACS Symposium Series 339, Symposium Series 62, 185-196, American Chemical Society,
281-294, American Chemical Society, Washington, DC, 1987. Washington, DC, 1977.
ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod. MURRAY, R. D. H., Naturally occurring plant coumarins, Fort.
Rep., 5, 581-612 (1988). Chern. Org. Naturst, 35, 199-429 (1978).
FLOSS, H. G. and U. MOTHES, Zur Biosynthese von Furanocumari- MURRAY, R. D. H., J. MENDEZ, and S. A. BROWN, The Natural
nen in Pimpinella magna, Z. Naturforsch. B.: Anorg. Chern., Coumarins: Occurrence. Chemistry, and Biochemistry, Wiley,
Org. Chern. Biochem. Biophys. Bioi., 19, 770-771 (1964). New York, 1982.
GRUNDON, M. F., Aspects of the biosynthesis of coumarins and NITAo, J. K., Enzymatic adaptation in a specialist herbivore for
quinoline alkaloids in the Rutaceae, in Chemistry and Chemical feeding on furanocoumarin-containing plants, Ecology, 70,
Taxonomy of the Rutales (p. G. Waterman and M. F. Gruodon, 629-635 (1989).
eds.), Anoual Proceedings of the Phytochemistry Society, Eu- NITAO, 1. K., Metabolism and excretion of the furanocoumarin
rope, No. 22, 9-30, Academic Press, London, 1983. xanthotoxin by parsnip webwonn, Depressaria pastinacella, J.
HAHLBROCK, K. and H. RAGG, Light-induced changes of enzyme Chern. Ecol., 16,417-428 (1990).
activities in parsley cell suspension cultures, Arch. Biochem. OBA, K" E. E. CONN, H. CANUT, and A. M. BOUDEr, Subcellular
Biophys., 166, 41-46 (1975). localization of 2-(j3-o-glucosyloxy)-cinnamic acids and their re-
HAHLBROCK, K., C. LAMB, C. PuRWlN, J. EBEL, E. FAUTZ, and E. lated j3-glucosidase in the leaves of Melilotus alba Desv., Plant
SCHAFER. Rapid response of suspension-cultured parsley cells Physiol.,68, 1359-1363 (1981).
to the elicitor from Phytophthora megasperma var. sojae, Plant PLA'ITNER, R. D. and G. F. SPENCER, Assigument of ring substitu-
Physiol., 78, 768-773 (1981). tion pattern for substituted furanocoumarins by negative ion
HAMERSKI, D., D. SCHMI1T, and U. MATERN, Induction of two pre- mass spectrometry, Org. Mass Spect., 23, 624-626 (1988).
nyltransferases for the accumulation of coumarin phytoalexins POULTON, J. E., D. E. McREE, and E. E. CONN, Intracellular locali-
in elicitor-treated Ammi majus cell suspension cultures, Phyto- zation of two enzymes involved in coumarin biosynthesis in
chemistry,29, 1131-1135 (1990a). Melilotus alba, Plant Physiol., 65, 171-175 (1980).
HAMERSKI, D, R. C. BBIER, R. E. KNEUSEL, U. MATERN, and K. SEIGLER, D. S., Role of lipids in plant resistance to insects, in Plant
HIMMELSPACH, Accumulation of coumarins in elicitor-treated Resistance to Insects (P. A. Hedin, ed.), ACS Symposium Series
cell suspension cultures of Ammi majus, Phytochemistry, 29, 208,303-327, American Chemical Society, Washington, DC,
1137-1142 (1990b). 1983.
HARBORNE,1. B.. Recent advances in chemical ecology, Nat. Prod. SINGER, D. E., The effects of low-dose warfarin on the risk of stroke
Rep., 4, 323-344 (1986). in patients with nonrheumatic atrial fibrillation, New Engl. J.
HASLAM, E., The Shikimate Pathway, Wiley, New York, 1974. Med., 323, 1505-1511 (1990).
IVIE, G. W.. Toxicological significance of plant furanocoumarins, SNEADER, W., Drag Discovery: The Evolution of Modem Medi-
in Effects of Poisonous Plants on Livestock (R. F. Keeler. K. cines, John Wiley & Sons, Ltd., Chichester, 1985.
R. van Kampen, and L. F. James eds.), 475-485, Academic SPBNCER, G. F., L. W. TJARKS, andR. G. POWELL, Analysis of linear
Press, New York, 1978. and angular furanocoumarins by dual-column high-performance
IVIE, G. W., Natural toxicants in human foods: Psoralens in raw liquid chromatography, J. Agric. Food Chern., 35, 803-805
and cooked parsnip root, Science, 213, 909-910 (1981). (1987).
IVIE, G. W., Biological actions and metabolic transfonnations of SWAIN, L. A. and K. R. DOWNUM, Light-activated toxins of the
furanocoumarins, in Light-Activated Pesticides (J. R. Heitz and Moraceae, Biochem. Syst. Ecol" 18, 153-156 (1990).
K. R. Downum, eds.), ACS Symposium Series 339, 217-230, YADMA, T. and K. MUNAKATA, Phloroglucinol-type furocoumarins,
American Chemical Society, Washington, DC, 1987. a group of potent naturally occurring insect antifeedants. Agric.
IVIE, G. W., D. L. BULL, R. C. BEIER, and N. W. PRYOR, Metabolic BioI. Chern., 43, 1701-1706 (1979).
detoxification of linear and angular furanocoumarins by cater- ZANoERL, A. R. and M. R. BBRENBAUM, Furanocoumarins in wild
pillars of the black swallowtail butterfly, in Allelochemicals: parsnip: Effects of photosynthetically active radiation, ultravi-
Role in Agriculture and Forestry (G. R. Waller, ed.), ACS Sym- olet light, and nutrients, Ecology, 63, 516-520 (1987).
posium Series 330, 455-462, American Chemical Society, ZoBEL, A. M. and S. A. BROWN, Histological localization of furano-
Washington, DC, 1987. coumarins in Ruta graveolens shoots, Can. J. Bot., 67, 915-921
JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary (1989).
10
2-Pyrones, Stilbenes,
Dihydrophenanthrenes, and Xanthones

Introduction Similar compounds are found in kawa root, kavakava,

2-Pyrones and Styrylpyrones kawakawa or yangona (Piper methysticum, Piperaceae),
Biosynthesis which is used to prepare a beverage widely consumed in the
Systematic usefulness Pacific islands. The beverage is an integral part of the reli-
Biological activity gious and social life and has tranquilizing and sleep-inducing
Stilbenes properties (Lebot, 1991; Lewin, 1931; Schery, 1972).
Biosynthesis The stem bark of Goniothalmus giganteus (Annonaceae)
Biological activity contains the related compounds, goniothalenone (altholac-
Dihydrophenanthrenes and Phenanthrenes tone) (1) and goniothalamin (2), in addition to a series of
Biosynthesis acetogenins (Fig. 10.1) (see Polyketides) (McLaughlin,
Biological activity 1991).
Benzophenones
Xanthones Biosynthesis
Distribution of Xanthones The mechanism for addition of two carbon units is like
Biological Activity that of polyketide synthesis (Birch and Donovan, 1953). The
References starter unit is first converted to the CoA ester and C 2 units
added from malonyl-CoA (Fig. 10.2). The chain then cy-
elizes in several ways which may be determined by the num-
Il'ITKODUCTION ber of units added, the enzyme systems present, and other
factors. Thus, one portion of the molecule is derived from
The enzymatic mechanisms for lengthening of the side chain shikimic acid and the other from acetate. In some cases, it
of cinnamic acids by one or more C 2 units followed by cyeli- is not possible to tell if a C6 -C3 precursor is extended by
zation are widespread in higher plants. Although by far the two acetate units or if a C 6 -C, precursor is extended by
largest group of compounds formed by this route is the flavo- three units (Fig. 10.2). For example, paracotoin (3) [found
noids, several other important structural entities are known in paracoto bark from Aniba pseudocoto (Lauraceae)] can
to arise in this manner. Among these are the 2-pyrones, stil- be derived by either pathway (Fig. 10.3) (Manitto, 1981).
benes, dihydrophenanthrenes, and xanthones. After the chain is extended to six carbon atoms, cyclization
occurs to produce a 6-membered lactone ring.

2·P¥KONES AND STYKYLPYKONES Systematic Usefulness

The trunk woods of many species of Aniba (Lauraceae)
The bark of the plants Nectandra coto (syn. Aniba coto), contain phenylpropanoid compounds that serve as precur-
coto, and Ocotea pseudocoto (Aniba pseudocoto), paracoto, sors to neolignans (see Chapters 8 and II); other Aniba spe-
(as well as other species of the Lauraceae) have long been cies contain pyrones and benzophenones, whereas yet others
known to contain biologically active secondary compounds possess benzylisoquinoline alkaloids and benzyl benzoates.
(Geissman and Crout, 1969). The bark ofthese plants is used It is possible to divide Aniba species into those that contain
medicinally in South America. pyrones and those that contain neolignans (Gottlieb, 1980).

139
140 2-Pyrones, Stilbenes, Dihydrophenanthrenes, and Xant/tones

kawain (5) yangonin (4) methysticln (6)

anlbine 4-methoxyparacotoin paraeotoin (3) pbenylcoumalin

o OH o OCH3
o OH

~ ~ ~
VHO~OH VHO~OCH3 VHO~OCH3
cotoln bydrocotoln metbylbydrocotoin

~:::,..
I H '.:'
• '<::::

H i 0 0
OH H

goniotbalenone (1) gonlothalamln (2)

Fig. 10.1. 2-Pyrones and styrylpyrones.

Biological Activity sides are known from higher plants and more than 80 from
liverworts (Gorham, 1980, 1989). They are especially impor-
Both the beverage, kava or yangona, and the dried root
tant in the heartwood of trees of the genera Pinus (Pinaeeae),
of Piper methysticum are used as sources of a serles of py- Eucalyptus (Myrtaceae), and Madura (Moraceae) (Hart,
rones with biological activity. Kavalactones in this plant are
1981; Weiss and Edwards, 1980). Stilbenes are common
diuretic, soporific, anticonvulsant, spasmolytic, local anes- by-products of paper manufacture (Kindl, 1985). Although
thetic, and antimycotic (Lebot, 1991). The dried root con- stilbene aglycones are common in heartwood, living tissue
tains about 5-6% resin from which six related pyrones [yan- often contains small amounts of stilbene glycosides (Hart,
gonin (4), desmethoxyyangonin, kawain (5), dihydrokawain, 1981). In several woody plants, stilbenes are accompanied
methysticin (6), and dibydromethylsticin] have been iso- by dihydrostilbenes or bibenzyl compounds, dibydrophe-
lated. All are more or less potent, centrally acting skeletal nanthrenes, and phenanthrenes (Kindl, 1985).
muscle relaxants that also possess antipyretic and local anes- The para-hydroxylated compound, resveratrol (7), is the
thetic properties (Tyler et al., 1981). most widespread stilbene in nature. This compound occurs
in Picea, Pinus, the Fabaceae, Myrtaeeae, and the Vitaeeae
among others (Kindl, 1985). Pinosylvin (8) and its deriva-
STlLBEl'IES tives are mostly restricted to the genus Pinus. The distribu-
tion of stilbenes in plants has been reviewed (Gorham, 1980,
Stilbenes are a relatively small, but widely distributed, group 1989) (Fig. 10.4).
of plant secondary metabolites found mostly as heartwood Oxidative dimers of stilbenes called goetins occur in both
constituents in a heterogenous assembly of plant species. of the unusual gymnosperms Gnetum and Welwitschia (Fig.
Over 200 naturally occurring stilbenes and stilbene glyco- 10.5) (Gottlieb and Kubitzki, 1988).
 2-Pyrones, Slilbenes, Dihydrophenanlhrenes, and Xanthones 141

hOH
co~
° r- malonyl-eoA

°y0
},.. CoASH + CO, 0H

~OH ° ,,-'
"-
COW' . --+
OH

° V° malonyl~CoA
dihydropyrone

J
} . CoASH + CO,
OH

CoA
, ,...1

° ° ° styrylpyrone
V-maI0nYloCOA ~OH
I:
~
~COASH+CO' r OH HO r ".

CoAS
, "- 1 a1_dO_I_co_n_de_n_
...~tion ~ I

OH

° ° ° ° ~condensation OH

HO r

OH °
chalcone
Fig. 10.2. Biosynthesis of flavonoids and stilbenes from acetate and cinnamate (modified from Grisebach, 1985; used with pennission of the copyright
owner, Academic Press, Orlando, Fl.,).

OCH3

• .
--
CH3CO,H
C.-C,
°
CH3CO,H

kawain(5)

.
( :oD
~o

--
CH3CO,H
I 0

I ~ • ° °
° b
paracotoin (3) paraeotoln (3)

Fig. 10.3. Labeling of kawain and paracotoin by radioactively labeled precursors (modified from Manitto, 1981).
142 2-pyrones. Sti/benes. Dihydrophenanthrenes. and Xanthones

ooyO HO

resveratrol (7) piceatannol (11)

pinosylvin (8)

OH
OH

HO
HO
HO

HO

oxyresveratrol (10) 4 -prenyloxyresveratrol

broussonin A (14) broussonin B (15)

Fig. 10.4. Some important stilbenes and related compounds.

Lunularic acid (9) is found in most liverworts, and a series malonyl-CoA (Kind!, 1985). Mter three units of acetate (via
of structurally related derivatives are found in mosses and malonyl-CoA) are added, cyclization occurs via an aldol con-
liverworts (Fig. 10.6) (Zinsmeister and Mues, 1987). Earlier densation. Stilbenes differ from the better known and more
reports of lunularic acid in algae need to be confirmed. common flavonoids in that the latter suffer a Claisen-type
Stilbenes and related compounds can be separated with condensation (Fig. 10.2). In genera!, stilbenes have hydroxyl-
a number of techniques such as thin-layer (TLC), gas liquid ation at positions 3 and 5. The resveratrol-forming enzyme
(GLC), and high-performance liquid (HPLC) chromatogra- is a dimer with a molecular weight of 90,000. The levels of
phy (Gorham, 1989). The detection and identification of stilbene synthase in grape leaves is increased 100-fold by
these compounds can best be accomplished by nuclear mag- treatment with fungal or artificial elicitors (Kindl, 1985). 1-
netic resonance (NMR) spectroscopy and mass spectrometry 14C-Acetate and 14C-p-coumaric acid were incorporated into
(Gorham, 1980; 1989). oxyresveratrol (10) during feeding experiments in Morus
alba (Moraceae). Labeling studies with phenylalanine and
Biosynthesis acetate indicate that both are incorporated into pinosylvin (8)
Stilbenes are derived from a pathway similar to that of fla- in Pinus resinosa and P. sylvestris and into resveratrol (7) in
vonoids (Birch and Donovan, 1953). These phenolic com- Eucalyptus sideroxylon (Myrtaceae) (Kind!, 1985; Weiss and
pounds are of mixed biosynthetic derivation with one ring Edwards, 1980). In studies of oxyresveratrul (10) biosyn-
from polyketide metabolism and the other from shikimic acid. thesis, 1- l4C-acetate was incorporated into carbons 3 and 5
Three units of malonate are added to a shikimate-derived and !3_14C-p-coumaric acid into carbon 8. Piceatannol (11) in
starter unit, probably cinnamic or p-coumaric acid and then Laburnum anagyroides (Fabaceae) was labeled in a similar
cyclized via an aldol condensation (Gorham, 1980). Instead manner (Fig. 10.8). In contrast to chalcone synthesis, the hy-
of undergoing a Claisen-type condensation (as in the case of droxylation pattern of the B ring generally is determined at
flavonoids), that portion of the molecule undergoes an aldol- the phenylpropanoid level in stilbene biosynthesis (Stafford,
type condensation (Fig. 10.2). Considerable homology of an- 1991). Formation of compounds such as pinosylvin (8) pre-
giosperm chalcone synthases with stilbene synthases has been sumably involves decarboxylation of the aldol-derived por-
reported (Stafford, 1991). This condensation is catalyzed by tion of an intermediate (Fig. 10.7).
the enzyme stilbene synthase which has a high Km value for Compounds found in the genus Hydrangea (Hydran-
 2-Pynme.f, Srilhelles, Dihydrol'heJulIllhref1es, and XOllrJwlles 143

OH OH

HO

HO

Gnetum HO

OH

OH

Welwitschio

HO

HO

Fig. 10.5. Gnetins from Gnetum and Welwitschia.

geaceae) (e.g., 12) resemble probable intermediates in the nularic acid (9) during extraction (Dewick, 1984; Ellis,
biosynthesis of stilbenes (Fig. 10.8) (Gorham, 1980). 1988) (Fig. 10.6).
Prelunularic acid (13) (Fig. 10.6) is the major compound
accumulated by cell cultures of Marchantia polymorpha, Biological Activity
Jungermannia subulata, Lophocolea heterophylla, and Ca- Many of the woods from which stilbenoids have been
lypogeia tosana. This compound is converted rapidly to lu- isolated are highly resistant to decay (Gorham, 1989). These

o OH

,rl-n~~ ~"" """ OH

HO~
o h CO,H
HO

prelnnularic acid (13) lunularic acid (9) (+)·(S)-abscisic acid (19)

Fig. 10.6. Lunularic acid, prelunularic acid and abscisic acid.

144 2~Pyrones, Stilbenes, Dihydrophenanthrenes, and Xanthones

o
HO,C
o

--
o

d
SCoA

+ 3 malonyl-CoA - o

OH

OH

pinosylvin (S)

Fig. 10.7. Biosynthesis of pinosylvin (Haslam, 1974).

~OH
E-cinnamic acid
o

o
- "Or HO pinosylvin (8)

HO~
~OH
-- ""~""
1 OH
HO resveratrol (7)

~
OCH3

HO,:? ~
I ,:::; OH

HO
CO,H
hydrangeic acid (12)
~HO
I .haponligenin

HO~OH
Y HO
-- piceatannol (11)
resveratrol (7)

Fig. 10.S. Biogenetic origin of selected stilbenes (modified from Kindl, 1985; used with pennission of the copyright owner, Academic Press, Orlando.
FL).
 2-Pymlle.I". Sli/helle.I·, Dihydroph('/1(1l1fltrenes. and Xanthones 145

compounds cause numerous difficulties in paper manufac- viniferin (17) (Fig. 10.10), are formed following infection
ture from a number of trees. Pinosylvin (8) is a phytoalexin by Botrytis cinerea in grapes (Vitis l'inijera). These phyto-
in young pine seedlings formed only after the plants have alexins are highly antifungal (Gorham, 1980).
suffered microbial infection. but it is a constitutive resistance A similar stilbene (18), that inhibits wood decay fungi,
factor in older trees. Z- and E-Resveratrol (7) are phytoalex- has been isolated from Diphysa robinioides (Fabaceae),
ins in the peanut, Arachis hypogaea (Fabaceae) (Kuc, 1992); Schotia brachypetala (Fabaceae), Vouacapoua species (Fa-
several C-alkylated isoprene derivatives of resveratrol also baceae), and Maclura pomijera (Moraceae) (Castro el aI.,
are found in legumes such as the peanut (Kindl, 1985). 1986).
Synthesis of a number of antifungal stilbenoids can be The stilbene piceatannol (11) (Fig. 10.4) is produced
induced by elicitation with fungal preparations or other fac- when slalk tissues of sugar cane, Saccharum officinarum, are
tors such as UV light. A family ofphytoalexins from mulber- inoculated with the red rot fungus, Colletotrichum[alcatum
ries (Moms spp., Moraceae) possess stilbene structures (Fig. (Brinker and Seigler, 1991). Piceatannol has antitumor activ-
10.9) (Coxon, 1982). The moracins were isolated from ity, is fungicidal and algicidal, and inhibits plant growth and
shoots of Moms alba infected with Fusarium solani f. sp. photosynthesis. Piceatannol proved to be the most active
mori and were not present in detectable quantities in unin- compound from Scirpus maritimus against P-388 lympho-
fected tissue. Two additional compounds, oxyresveratrol cytic murine leukemia, and brine shrimp (Artemia salina),
(10) and 4'-prenyloxyresveratrol (Fig. 10.4), were isolated fall army worm (Spodoptera [rugiperda), and duckweed
from fungus-infected xylem extracts of mUlberry. Although (Lemna minor) (Powell et aI., 1987). Piceatannol (11) may
oxyresveratrol (10) (Fig. 10.4) occurs in heartwood of mul- act as a specific inhibitor of tyrosine kinase in developing
berry, this compound is formed in the sapwood as a phyto- leukemic cells (McLaughlin, 1991). In these and other assays
alexin. Two similar compounds, broussonin A and B (14, for biological activity, piceatannol has greater activity than
15), are found in the shoots of paper mulberry (Broussonetia most other stilbenes.
papyrijera, Moraceae) infected wilh the same fungus The stilbenoid compound lunularic acid (9) occurs in all
(Coxon, 1982; Kuc, 1992). liverworts examined thus far and appears to fulfill the same
Oligomers of resveratrol, such as a-viniferin (16) and €- growth-regulating function in these plants that abscisic acid

OCH, HoWy
I I
CH, 0~7'I9' I
0
I
OH
CH,O '" 0 P

""- I
OH
HO

'" OCH,
HO HO
moracinA moracinB moracinC

HO CH'O~ I I
CHO'"
, 0
P
I OH
""-
HO
moracinD moracin E moracin F

OCH,

OH
OH

HO
HO
moracin G moracin H

Fig. 10.9. Phytoalexins from mulberries.

146 2-Pyrones, Srilbenes, Dihydrophenanthrenes, and Xanthones

OH

OH

~
YY
OHHO CH iHO
"'CH "" OH

HO~ (18)
OH

HO

OH
OH OH

a·vinlferin (16) E·vlnlferin (17)

Fig. 10.10. Ct- Viniferin and e-viniferin.

(19) (Fig. 10.6) does in higher plants (Gorham, 1980; Man- DIHYDKOPHEl'IANTIIRENES Al'ID
dava, 1979; Zinsmeister and Mues, 1987) (see Chapter 23). PHEl'IANTIIRENES
Lunularic acid controls drought resistance and dormancy in
Lunularia. However,lunularic acid had no significant effects A number of dihydrophenanthrenes and phenanthrenes pos-
on transpiration in the higher plants Lycopersicon, Phaseo- sess antifungal activity and are important as phytoalexins.
Ius, or Apium or on the stomatal aperture in epidermal strips Although many of these and related compounds are derived
of Commelina (Gorham, 1980). However, at 10-30 ppm, from pathways discussed in this chapter, others that coinci-
lunularic acid does inhibit the auxin-induced elongation of dentally possess phenanthrene stroctures are derived from
Avena coleoptile segments. isoflavonoid precursors and are discussed under that topic.

o o
~OH ~OH
Y - y---..-~
rnro®
E-cinnamic acid OH OH m·coumaric acid (22)

oo-QO~0 OHHO (23) OCH,HO OCH,HO

~
rn,o-Qb-oo
hircinol (21) loroglossol (24)

~OCH'

o---Q-OH
OCH,
OCH,
orebinol (20) aucuparin (32)

Fig. 10.11. Proposed biosynthetic origin of orchinol and hircinol (Dewick, 1984; modified and used with pennission of the copyright owner, the Royal
Society of Chemistry, Cambridge).
 2-Pyrones, Stilbenes, Dihydrophenanthrenes. and Xanthones 147

batatasin I (24) batatasi. IV (25) batalasi. m (26)

Fig. 10.12. Batatasi. I. IV. and ill.

9,IO-Dihydrophenanthrenes are characteristic of the Or- poration of m-coumaric acid (22) proceeds by a pathway
chidaceae, Dioscoreaceae, and Combretaceae. Phenan- similar to that of stilbene formation (Fig. 10.11) (Dewick,
threnes and phenanthrene derivatives also have been found 1984). Wounding of the rhizomes of the orchid Epipactis
in liverworts (Gorham, 1989). palustris caused induction of a bibenzyl synthase which con-
verts m-hydroxyphenylpropionyl-CoA and malonyl-CoA
Biosyntbesls into 3,5,3'-trihydroxybibenzyl (23) (Gehlert and Kindl,
1991). Production of this bibenzyl compound parallels the
Biosynthesis of dihydrophenanthrenes is similar to that formation of a dihydrophenanthrene. When the bibenzyl
of stilbenes but appears to involve dihydrocinuamic acid and compound was fed to tuber slices of Orchis militaris, orchi-
the enzyme bibenzyl synthase, whereas the biosynthesis of nol (20) was produced (Brooks and Watson, 1985).
phenanthrenes iuvolves the corresponding unsaturated acids
(Gorham, 1989). Biological Activity
In a series of feeding experiments, it was found that phe-
nylalanine, cinuamic acid, meta-coumaric acid, and dihydro- Dormancy in yam bulbs (Dioscorea hatatas, Dioscorea-
m-coumaric acid were good precursors for orchinol (20) and ceae) is induced by three growth inhibitors, batatasins I (24),
hircinol (21) in Orchis militaris, but 0- and p-coumaric acids IV (25), and III (26). Batatasin I is 6-hydroxy-2,4,7-trimeth-
were not. The production of both meta-substituted com- oxyphenanthrene and batatasin III is 3,3'-dihydroxy-5-me-
pounds in this orchid was confirmed (Dewick, 1984). Incor- thoxybibenzyl (Mandava, 1979) (Fig. 10.12).

OH OH
OH
CH,'O CH,'O
CH,O

CH,.O CH,-O

CH,'O CH,-O
OH OH OH
(27) (28) (29)

OH OH
HO

CH,-O CH,-O

OH OCH,

(30) (31)

Fig. 10.13_ Dihydrophenanthrenes and phenanthrenes from Oncidium cebolleta.

 ,:7
148 2-Pyrones, Sti/benes, Dihydrophenanthrenes, and Xanthones

HO~
~
CO,H

3malonyl-CoA o--.n ~ ~
'~O~__
0

OH
OH tco,:7
HO"'"
OH

1
0

OH"'"
1-
OH

HO OH 1,3,S,6-tetrahydroxyxanthone (34)

OH
mangostin (35)
mangiferin (37)
H
HO o

OH

jacarubin (36) OH

Fig. 10.14. Proposed biosynthetic paths to some naturally occurring benzophenones and xanthones (Modified from Manitto, 1981).

The compounds orcinol (20) and hircinol (21) are phyto- 8ENZOPIfflNONES
alexins in Orchis militaris and in other species of Orchis
(Orchidaceae) when infected with Rhizoctonia repens Benzophenones are derived from several pathways. In higher
(Brooks and Watson, 1985; Coxon, 1982; Dewick, 1984; plants, most benzophenones arise by cyclization of a polyke-
Kuc, 1992). These compounds are active as phytoalexins tide chain of three malonates added to a hydroxybenzoic acid
against a range of organisms at 10-4 M (Gorham, 1989). precursor (ManJttu, 1981). Cyclization occurs via a Claisen
Hircinol has also been isolated from tubers of yam (Diosc- condensation mechanJsm (Fig. 10.14). These compounds
orea rotundata, Dioscoreaceae) where it is a preformed anti- serve as precursors for xanthones in many plants. In some
fungal compound. instances [e.g., in the heartwood of Symphonia globulifera
One species of the Orchidaceae, Oncidium ceboletta, is (Clusiaceae)], the benzophenone and the corresponding xan-
used by the Tarahumara Indians of Mexico as a replacement thone co-occur [in this case maclurin (33) and 1,3,5,6-tetra-
for peyote, a hallucinogenic species of cactus. The whole hydroxyxanthone (34)] (Weiss and Edwards, 1980).
green leaf is crushed in water and the mixture consumed.
This plant contains 2,7-dihydroxy-3.4,6-trimethoxyphe-
nanthrene (27), 2,7-dihydroxy-3.4,6-trirnethoxy-9,lO-dihy-
drophenanthrene (28), 2,3-dihydroxy-4,7 ,8-trimethoxyphe- XANTHONES
nanthrene (31), 2,7-dihydroxy-3,4-dimethoxyphenanthrene
(29), and 2,7-dihydroxy-4,8-dimethoxyphenanthrene (30) About 150 xanthones have been discovered. Many are po-
(Fig. 10.13) (Stermitz et al., 1983). lyketide derived, although others are formed from combined
Bibenzyls also have antifungal activity. Aucuparin (32) shikimic acid pathways combined with acetate-malonate
(Fig. 10.12) was induced in tissues of loquat (Eriobotrya units. Three units of malonate react with a hydroxybenzoic
japonica, Rosaceae) when challenged with the fungus Col- acid (Co-C 1 ). Benzophenones may be converted by oxida-
letotrichum lindemuthianum (Brooks and Watson, 1985). tive ring closure into xanthones (Fig. 10.14) (Weiss and Ed-
 2-Pyrones. Stilbenes, Dihydrophenanthrenes, and Xanthones 149

wards, 1980). Alternatively, two units of malonate can react OEHLERT, R. and H. KINDL, Induced formation of dihydrophe-
with a C6 -C3 precursor to yield other xanthones. Cinnamic nanthrenes and bibenzyI synthase upon destruction of orchid
acid, benzoic acid, m-hydroxybenzoic acid, malonic acid, mycorrhiza, Phytochemistry, 30. 457-460 (1991).
and an intermediate benzophenone all proved to be precur- GElSSMAN, T. A. and D. H. O. CROUT, Organic Chemistry of Sec-
sors for mangostin (35) (Hostettmann and Hostettmann, ondary Plant Metabolism. Freeman Cooper, San Francisco,
1989). This route seems most likely for compounds such as 1969.
jacarubin (36) and mangiferin (37) (Hostettmann and Hostet- GORHAM, J., The stilbenoids, in Progress in Phytochemistry, Vol.
tmann, 1989; Mannito, 1981). Direct coupling of the oxy- 6 (L. Reinhold, J. B. Harborne, and T. Swain, ed.), 203-252,
Pergamon, Oxford, 1980.
gens seems more likely for xanthones of the lichexanthone
type produced by microorganisms (Mannito, 1981). GORHAM, J., Stilbenes and phenanthrenes, in Plant Phenolics I (J.
Maclurin (33) and two related cydization products that B. Harbone, eds.), Vol. 1 of Modem Methods of Plant Biochem-
istry (p. M. Dey and J. B. Harborne, eds.), 159-196, Academic
are found in the heartwood of Symphonia globulifera (Clusi-
Press, London, 1989.
aceae) possibly arise by ring closure of a benzophenone pre-
cursor (Weiss and Edwards, 1980). GOTILma, O. R., Towards a scientific status for micromolecular
systematics, Acta Amazonica, 10. 845-862 (1980).

Distribution of Xanthones OOTfLlliB, O. R. and K. KUBITZ!(], A chemical link between Gnetum

and Welwitschia. Naturwissenschaften, 75. 575-577 (1988).
Xanthone aglycones are found in eight families: Clusia- GRISEBACH, H., Biosynthesis of flavonoids, in Biosynthesis and
ceae, Fabaceae, Gentianaceae, Loganiaceae, Lythraceae, Biodegradation of Wood Components (T. Higuchi, ed.),
Moraceae, Polygalaceae, and Rhamnaceae. Aglycones and 291-324, Academic Press, Orlando, FL, 1985.
their corresponding O-glycosides are found only in the Gen- HART, J. H., Role of phytostilbenes in decay and disease resistance,
tianaceae and Polygalaceae (Hostettmann and Hostettmann, Annu. Rev. Phytopathol., 19, 437-458 (1981).
1989). However, C-glycosides, such as mangiferin (37), are HASLAM, E., The Shikimate Pathway, Wiley, New York, 1974.
widespread among plants. Among the glycosidic sugars, HOSTETTMANN, K. and M. HOSTE"ITMANN, Xanthones, in Plant Phe-
only l3-n-glucose is found (Vermes and Wagner, 1986). nolics (J. B. Harborne, eds.), Vol. 1 of Modem Methods of Plant
Biochemistry (P. M. Dey and J. B. Harborne, eds.), 493-508,
Biological Activity Academic Press, London, 1989.
Several xanthones that possess antidepressant activity in- KINoL, H., Biosynthesis of stilbenes, in Biosynthesis and Biodegra-
hibit monoamine oxidases. These compounds have in vitro dation of Wood Compunents (T. Higuchi, ed.), 349-377, Aca-
demic Press, Orlando, FL, 1985.
cytotoxicity and in vivo antitomor activity (Hostettmann and
Hostettmann, 1989). The xanthone psorospermin (38) from Kuo:, J., Antifungal compounds from plants, in Phytochemical Re-
sources for Medicine and Agriculture (H. N. Nigg and D.
Psorospermumfebrifugum (Clusiaceae) exhibited antileuke-
Seigler, eds.). 159-184, Plenum, New York, 1992.
mic activity in several test systems. Other xanthones have
tuberculostatic effects (Vermes and Wagner, 1986). LEBOT, V., Kava (Piper methysticum forst. f.): The Polynesian dis-
persal of an Oceanian plant, in Islands, Plants, and Polynesians
(P. A. Cox andS. A. Banack, eds.), 169-201, DioscoridesPress.
Portland, OR, 1991.
REFERENCES
LEWIN, 1.., Phantastica, Dutton, New York, 1931.

BIRCH, A. J., and F. W. DONOVAN, I. Some possible routes to deriva- MANnAVA, N. B., Natural products in plant growth regulation, in
tives of orcinol and phloroglucinol, Aust. J. Chem. 6 360-368 Plant Growth Substances (N. B. Mandava, ed.), ACS Sympo-
(1953). sium Series 111, 135-213, American Chemical Society, Wash-
ington, DC, 1979.
BRINKER, A. M. and D. S. SEIGLER, Isolation and identification of
piceatannol as a phytoalexin from sugarcane, Phytochemistry, MANrrro, P .• Biosynthesis of Natural Products, Ellis Horwood,
30.3229-3232 (1991). Chichester, 1981.
BROOKS, C. J. W. and D. G. WATSON. Phytoalexins, Nat. Prod. McLAUGIILlN, J. L., Crown gaU tumours on potato discs and brine
Rep., 2. 427-459 (1985). shtimp lethality: Two simple bioassays for higher plant screen-
ing and fractionation, in Assays for Bioactivity (K. Hostett-
CASTRO, 0., J. LoPEZ, A. VERGARA, F. R. STERMITZ, and D. R.
mann, ed.), Vol. 6 of Modem Methods in Plant Biochemistry
GARDNER, Isoflavans and a stilbene from wood of the decay-
(P. M. Dey and J. B. Harborne, eds.), 1-32, Academic Press,
resistant tropical tree Diphysa robinoides, J. Nat. Prod.• 49.
London, 1991.
680-683 (1986).
COXON, D. T., Phytoalexins from other families, in Phytoalexins POWELL, R. G., R. BAJAJ, and J. L. McLAUGHLIN, Bioactive stil-
(J. A. Bailey and J. W. Mansfield, eds.), 106-132. Wiley, New benes of Scirpus maritimus, J. Nat. Prod., 50.293-296 (1987).
York,1982. SCHERY, R. W., Plants for Mao, 2nd ed., Prentice-Hall, Englewood
DBWICK, P. M.. The biosynthesis of shikimate metabolites, Nat. Cliffs, NJ, 1972.
Prod. Rep., 1. 451-469 (1984). STAFFORD, H. A., Flavonoid evolution: An enzymic approach, Plant
ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod. Physiol., 96, 680-685 (1991).
Rep., 5. 581-612 (1988). STIlRMITZ, F. R., T. R. SUESS, C. K. SCHAUER, O. P. ANDERSON,
ISO 2-Pyrones, Sti/benes. Dihydrophenanthrenes, and Xanthones

and R. A. BYE, JR., New and old phenanthrene derivatives from Gabor, and F. KaJlay, eds.), 63-73, Akademiai Kiada, Buda-
Oncidium cebolleta. a peyote-replacement plant, J. Nat. Prod., pest, 1986.
46,417-423 (1983). WEISS, U. and J. M. EowARDS, The Biosynthesis of Aromatic Com-
TYLER, V. E., L. R. BRADY, and J. E. ROBBERS, Phannacognosy, pounds, Wiley, New York, 1980.
8th ed., Lea and Febiger, Philadelphia, 1981. ZINSMEISTER, H. D. and R. MUES, Moose als Reservoir bemerken-
VERMES, B. and H. WAGNER, Natural xanthones and their glyco- swerter sekundiirer Inhaltsstoffe, GIT Fachzeitschr. Laborato-
sides, in Flavonoids and Bioflavonoids, 1985 (L. Farkas, M. rium, 31, 499-512 (1987).
11
Flavonoids

Introduction Isoflavans
Biosynthesis of Flavonoids Pterocarpans
Initial Steps of Flavonoid Biosynthesis Phytoalexins
Chalcones as Intennediates in Flavonoid Biosynthesis Rotenoids
Flavanones Neoflavonoids
Flavones Isolation, Purification, and Characterization of Flavonoids
Dihydroflavonols References
Flavonols
Substitution Patterns of Flavonoids
Flavone and Flavonol Aglycones Il'ITRODVCTIOI'l
Glycosylation and Methylation of Flavones and
Flavonols At least 4000 naturally occurring flavonoids have been de-
Biosynthesis of C-Glycosyl Flavonoids scribed; many are common in higher plants (Harborne,
Biosynthesis of Anthocyanins 1991). Most of the flavonoid compounds reported in the
Systematic Studies with Flavone and Flavonol Glycosides literature are glycosides of a relatively small number of fla-
Biological Activity of Flavone and Flavonol Aglycones vonoid aglycones which often are accumulated in the vacu-
and Glycosides oles of plant cells. These compounds frequently serve as
Biological Activity of Aglycones pigments in plants and are involved in many biological inter-
Biological Activity of Glycosides actions (Harborne, 1991). Flavonoids are based on a CIS
Anthocyanins skeleton and include a chroman ring bearing an aromatic
Factors Responsible for Color ring in position 2,3, or 4 (Hahlbrock, 1981).
Biological Activity of Anthocyanins Of the several hundred flavonoid aglycones (approxi-
Commercial Importance of Anthocyanins mately 200 flavones and 300 flavonols) that have been iso-
C-Glycosylflavones lated from plants (Wollenweber, 1982; Wollenweber and
Biflavonoids Dietz, 1981; Wollenweber and Jay, 1988), only 8 are distrib-
Chalcones uted widely (Figs. 11.1 and 11.2). One or some combination
Aurones of these eight flavonoid aglycones may be expected to occur
Dihydrochalcones in the hydrolyzed extract of most higher plants. All eight
Flavanones aglycones have the same basic hydroxylation pattern in the
The Role of Flavonoids in Pollination and Propagule A ring and differ mainly in the oxidation level of the central
Dispersal Mechanisms pyran ring and in the number of hydroxyl groups in the B
Isoflavones ring. Although all eight are known to occur free in plants,
Biosynthesis these compounds most commonly are found as a part of
Distribution water-soluble glycosides (Harborne, 1991).
Biological Properties Flavonoids may be divided conveniently into 14 classes
Derivatives of Isoflavones according to the oxidation level of the central ring (ring C)
Coumestans (Fig. 11.2). The most common of these are anthocyanins

151
152 Flal'onoids

(such as 1-3), flavones (such as 4 and S), and flavonols luteolin (4), are common; these compounds are widespread,
(such as 6-8), Some flavonoids are strongly pigmented; oth- especially in herbaceous plants, and occur both as C-glyco-
ers are colorless (Harborne, 1991), sides and O-glycosides. Flavonoids that differ in oxygena-
Three of these common flavonoid compounds, pelargoni- tion pattern from these eight compounds have limited distri-
din (4), cyanidin (3), and delphinidin (2), are anthocyanidins. butions and are uncommon. Other representative
These colored pigments are found widely in flowers· and (compounds) are illustrated in Fig, 11.3.
fruits and frequently are involved in the attraction of pollina- Methylated flavonoids corresponding to the eight com-
tors and in fruit and seed dispersal, but they also occur in mon structures and compounds with unusual hydroxylation
vegetative portions of plants. patterns are known; most of these also are restricted in distri-
The flavonols, kaempferol (6), quercetin (7), and myri- bution.
cetin (8), have increasing B-ring hydroxylation. These com- The presence of methylation, alkylation, or ester groups
pounds occur as copigments in flowers and fruits, but also makes flavonoids more lipid-like in nature (Fig. 11.4). in
they are found commonly in leaves, stems, and roots. in a many cases, aglycones and glycosides are additionally sub-
sample of 1000 plant species, over 60% had 1 or more of stituted and occur as esters (acetate, malonate, butyrate, cin-
these compounds as glycosides, Myricetin tends to be found namate, p-coumarate, femlate, etc.) or ethers. in contrast to
in association with tannins in woody plants. water-soluble glycosides that are usually accumulated in the
Flavones lack the 3-hydroxy group of flavonols and an- vacuole, flavonoid esters and ethers usually are associated
thocyanins. Although only two flavones, apigenin (S) and with waxes or found in the cytoplasm.

OH
OH OH

HO
"
OH
0 OH 0
OH OH
OH OH

HO HO
OH

OH 0 OH 0

OH OH

HO HO

OH OH
apigenin (5) pelargonidin (1) OH
OH
OH

HO

OH
OH

delphinidin (2) cyanidin (3)

Fig. 11.1. The eight most common flavonoid nuclei.

 Flavonoids 153

o CO,H HO CO,H HO 0
/O~~ OH _ _ h ~ ~ OH_ _ ~0'lco.~
~ ~ ~OH
o I "'" HO OH ~OH - OH
• H " 1
HO
O--=--O-OH
HO
HO"I

/OHO
~ ~
HO"I°'"
=--.
OHO

«pt
a stilbene / Ja chalcone (9) a flavanone (10)

H0l"'ir0~
~ VOH
HO ~ HO::;;." 0
"1 OH
~
OH 0 ,.. 1 1

an auraDe (11) OH OH 0
an isoflavone (46) a flavone (5)

HO_ ~~OH--O/H
TYV. HO
+ HO
OH

OH 0
a dihydrocbalcone (12)
a rotenoid
/ OH 0

OH a dihydronavonol (13)
HO 0 HO
i OH
OH
OH HO
a navan-3-ol (46)
a pterocarpan (95)
OH
OH OH 0
HO
a flavonol (6)

OH
an anthocyanidin (1) a navan-3,4-diol (33)

Fig. 11.2. Major classes of flavonoids (modified from Weiss and Edwards, 1980; used with pennission of the copyright owner, Copyright© 1980, John
Wiley & Sons, Inc., New York).

Flavonoids vary widely in their biological properties; it is required for the synthesis of flavonoids in this system. An
is among the colorless flavonoids that most compounds with irradiated culture of parsley petiole tissue produces a high
significant physiological activity have been reported. concentration of both types of flavonoids and high levels of
enzyme activity. The intermediate compounds and enzymes
involved in many of the biosynthetic steps have been charac-
BIOSYNfHESIS OF FLAVOI'lOmS terized (Fig. 11.5). Use of flower cultivars with defined ge-
notypes has proven valuable for correlations of genes with
Many aspects of the biosynthesis of flavonoids have been particular enzymes, as well as for elucidation of some steps
elucidated, but major progress in sorting out certain reaction of the biosynthetic pathway (Grisebach, 1985).
steps and the enzymes involved in flavonoid biosynthesis The enzymes of flavonoid biosynthesis are compartment-
was not possible before tissue and cell culture of suitable alized by membranes that surround the organelles and those
plants were developed (Hutchinson, 1986). The use of flow- which separate cytoplasmic regions into different microcom-
ers as sources of enzymes has provided the means for pro- partments. In addition, the association of biosynthetic en-
ducing 14C-labeled substrates (Heller and Forkmann, 1988). zymes as loose aggregates, as well as interaction between
Cell cultures of Petroselinum hortense (parsley, Apiaceae) constituent enzymes, permit direct transfer of substrates
contain both flavone and flavonol glycosides; most of the from one enzyme to another and channeling of intermediates
approximately 13 enzymes that catalyze the formation of and final products to the different sites of accumulation
flavonoid glycosides have been isolated and studied. Light (Hrazdina, 1992; Ibrahim, 1992). Flavonoid derivatives that
 w
154 Flavonoids

OCH,

o "" 1
: 1 1 CH,'O HO

OH 0
OH 0 OH 0
5~hydroxyt1avone digicitrin isorhamnetin
OH OH OR
OH
OR
OHd '
HO HO V 0 "" 1

R-O "" 1 1
OH 0 OH 0
OH 0
gossypetin quercetagetin, R=H chrysoerioJ, R=CH;Jt R'=H (38)

#
patuletin, R=CH3 diosmetin, R=H, R'=CH 3

OH OH

HO V

"" 1
0

1
:1 OH OH

OH
OH 0

#
luteolinldin
methoxyluteolin

OH
OH
OH OH

CH,-O HO r 0 : 1

""I
OH 0 OH 0

rhamnetin (51) eriodietyol (17)

Fig. 11.3. Other representative flavones and flavonols.

OH

OH

OH 0

sideroxylin mulberrin
Fig. 11.4. Alkylated !lavonoids.
 Flavonoids ISS

3 malonyl CoA

4-coumaroyl CoA
-+
,
HO
OH
HO~ ycfJ""I
oH",,1
0H

OH 0

/ a chalcone (9) ;/ a navanone (10)

OH

W
+~
HO~~-!!HO
OH OH
HO 0 "" 1
1 - ~ 1 1
""
OH 0
""
OH 0 ~ OH 0

an aurone (13) a dihydroflavonol (13)

'- o~ a flavone (S)
HO ~ OH OR

OH
an isoflavone (46)
a flavonol (6) a catechin (20)
or flavon-3-ol
OR OR

Hoo:rP~-!!
HO

I -
:::,...
OH
o
an anthocyanidin (3) sulfuretin (73)

a chalcone synthase b chalcone isomerase c flavanone 3-hydroxylase

Fig. 11.5. Proposed biosynthesis of major groups of flavonoids and major enzymes (modified from Grisebach, 1985; used with pennission of the
copyright owner, Academic Press, Orlando, FL).

are hydrophilic usually accumulate in the vacuole, whereas The enzymatic conversion of cinnamic acid to 4-hydrox-
lipophilic compounds are found in epidermal glandular cells ycinnamic acid (p-coumaric acid) in peas is associated with
or on plant surfaces, or are exuded from plant roots (Ibrahim, a microsomal fraction. This enzyme is a mixed-function oxi-
1992). Many of the enzymes of flavonoid biosynthesis are dase that requires molecular oxygen, NADPH, and mercap-
found in the cytoplasm, but are membrane associated. A toethanol for activity. p-Coumaric acid has a strong regula-
model system has been proposed (Stafford, 1990). tory effect on the enzyme at 3 X 10-7 to 10- 6 M
concentration (Hahlbrock, 1981).
Initial Steps of Flavonoid Biosynthesis The next major enzyme in the series, p-coumarate : CoA
ligase, was first isolated from illuminated cell suspension
Most flavonoids are derived from phenylalanine via cin- cultures of PetroseUnum crispum (syn. P. hortense) and
namic and p-coumaric acids by the addition of three malo- shown to be specifically related to flavonoid and stilbene
nate units and subsequent cycJization (Hahlbrock and biosynthesis. The enzyme requires p-coumaric acid,
Grisebach, 1975) (see Chapters 7 and 8). Phenylalanine am- CoASH, ATP, and Mg2+ for activity (Hahlbrock, 1981;
monia lyase (PAL), which catalyzes the first step of the Hutchinson, 1986). Three malonyl-CoA units are added and
biosynthetic process, is found in parenchyma cells of many cyclized via a elaisen condensation (see Chapter 5) to pro-
plants. PAL activity in vivo may be regulated by end-product duce a chalcone intermediate (9). Condensation of the same
inhibition by flavonoids. intermediates via an aldol condensation yields pyrones and
156 FJavonoids

OH HO

~ ~ -- ~ -
3 malonyl eoA
A" ~
NH3+

CO,- CO,-
CO,-
0 SCoA
phenylalanine E-cinnamate 4-coumarate 4·coumaroyl·CoA

OH

HO

OH o OH o

naringenin chalcone (9) naringenin (10)

Fig. 11.6, Steps common to the biosynthesis of all flavonoids (Hahlbrock, 1981; modified and used with pennission of the copyright owner, Academic
Press, Orlando, Fl.).

stilbenes [see Fig. 10.2 (Chapter 10)] which co-occur with to form the A ring may be similar to that of the J3-ketoacyl-
flavonoids in some plants. acyl carrier protein offatty acid synthases, that latter enzyme
may be the ancestral type (Heller and Forkmann 1988,
CbaIcones as Intermediates In Flavonoid Hutchinson, 1986; Stafford, 1991).
Biosyntbesls Mutants that lack chalcone isomerase, the next enzyme
in the sequence, accumulate chalcones, not flavanones, and
As described above, the fIrst enzyme general to all flavo- establish that the initial product of the reaction is a chalcone
noid biosynthesis, chalcone synthase (naringenin-chalcone (Grisebach, 1985). Recessive genotypes of certain flowers
synthase), catalyzes the cyclization of a precursor formed that lack chalcone synthase accumulate cinnamic acid gluco-
from p-coumaryl-CoA and three units of malonyl-CoA sides (Grisebach, 1985).
(Fig. 11.6) (Dewick, 1989; Gerats and Martin, 1992; Heller
Chalcones (e.g., 9) are key intermediates in the formation
and Forkmann, 1988). The enzyme, usually found in plant
of several major groups of flavonoids (see Fig. 11.2). In
epidermal cells, has a molecular weight of about 42,000,
some plants, they are converted into aurones (such as 11).
requires no cofactors, and has been isolated from several
In other instances, chalcones undergo reduction of the exocy-
plant cell cultures such as French bean (Phaseolus vulgaris),
clic double bond to produce dihydrochalcones (such as 12).
parsley (Petroselinum crispum), and the flowers of the car-
Chalcones and flavanones (e.g., 10) exist in eqUilibrium in
nation (Dianthus caryophyllus, Caryophyllaceae) (Hutchin-
son, 1986). p-Coumaric acid and malonyl-CoA are the pre- in vitro systems. However, as only flavanones with a (2S)-
ferred precursors. Malonyl-ACP will not serve. configuration are known to occur naturally, this interconver-
Chalcone synthase from parsley culture cells has a molec- sion appears to be enzymatically controlled and does not
ular weight of about 77,000 and is composed of two identical involve racemization. Desaturation of flavanones can yield
subunits of about 42,000 MW. The cDNA complementary flavones (such as 4 and 5), whereas introduction of oxygen
to the mRNA of chalcone synthase from irradiated parsley at the 3-position gives dihydroflavonols (flavanonols) (such
cell cultures has been cloned in Escherichia coli (Grisebach, as 13) which, in tum, are the precursors of flavonols (e.g.,
1985). 6-8), anthocyanins (e.g., 1-3), and condensed tannin precur-
Considerable homology of chalcone synthases with stil- sors (Ebel and Hahlbrock, 1982; Hahlbrock, 1981; Haslam
bene synthases has been found (Melchior and Kind!, 1990). 1974) (Figs. 11.7 and 11.8).
Because the mechanism of the stepwise addition of C-2 units In plants that have both 2',4'-dihydroxychalcones and
 Flavonoids 157

~OH

COASy I c:r OH

O~/O­
~C ~
cl
E-CH,COSCoA
"'-'"'=~' " ~~,yYI~ +E+CoASH
o 0
co,
Fig. 11.7. Proposed mechanism for fonnation of chalcone (modified from Grisebach, 1985; used with pennission of the copyright owner, Academic
Press, Orlando, FL).

OH

HO OH HOn;x: O1 OH
OH ------" I OH
~ ""-
both isolable, stable 0
o
butein butin

OH

HO

---------
o
2',4',6' ,4-tetrahydroxychalcone (9) naringenin (10)
(nnstable) (stabilized by hydrogen bonding)
OH
OH

HO
HO
4'
'>

isoliquiritigenin (15) liquiritigenin (14)

o a 5·deoxyflavonoid

HO
~
=--
g>Ki"
~ 1 O""""--...'·'----" •••
H,q

'H" ~
(-H2S)-7,4'-dihydroxyflavanone

OH

Fig. 11.8. Chalcone-isomerase-controlled isomerization (modified from Geissman and Crout, 1969).
158 Flavonoids

2',4',6'-trihydroxychalcones, the chalcone isomerase is ac- liquiritigenin (14) has been isolated from seedlings of Amer-
tive with both precursors (Heller and Forionann, 1988). pha fruticosa (Fabaceae) (Dewick, 1989).

Flavanones Flavones
Flavanones, by virtue of their role as biosynthetic inter- The approximately 650 known flavones arise from flava-
mediates, occur in most plants, but they are also accumulated nones by oxidative processes (Harbome, 1991). Most of
widely (Grayer, 1989). Most of the approximately 320 these are glycosides of 200 flavone aglycones. Both mo-
known flavanones possess the ( - )-(2S)-configuration. Fla- nooxygenase and dioxygenase types of flavone synthases
vanones are common in the Asteraceae and Fabaceae, and have been found (Stafford, 1991). With a soluble enzyme
the genus Citrus of the Rutaceae, but they have been reported preparation of irradiated cell suspension cultures of parsley,
in at least 60 other families (Bohm, 1988). However, com- conversion of naringenin (10) or eriodictyol (17) (Fig. 11.3)
pounds of this type are overlooked frequently in surveys of to the corresponding dihydroflavonol, flavonol, and flavone
flavonoid composition of plants, and representatives of this was observed; all these reactions require 2-oxoglutarate,
group of compounds may occur more widely in nature. Fe2+, and ascorbate as cofactors (flavone synthase I)
As described above, chalcones and flavanones are easily (Grisebach, 1985; Heller and Forkmann, 1988). An enzyme
interconverted under mild conditions and the original isom- from parsley that catalyzes the oxidation of naringenin (10)
erization in plants may have been nonenzymatic (Stafford, to the corresponding flavone, apigenin (5), in the presence
1991). However, because most hydrolyases utilize the (2S)- of Fe2+ and a nonproteinaceous cofactor has been demon-
isomer, enzymatic control would be more efficient. An en- strated. In parsley, 2-oxoglutarate (a-ketoglutarate), Fe2+,
zyme capable of catalyzing this interconversion, chalcone and ascorbate are required as cofactors for conversion to
isomerase, has now been demonstrated to be present in a both flavones and flavonols (Britsch et al., 1981).
number of plants. The enzyme has no cofactor requirements. However, the flavanone naringenin (10) [but not dihydro-
A proton is introduced specifically into the axial position at kaempferol (13)1 is the substrate for flavone formation in
C-3. The effect of illumination suggests that the enzyme snapdragons, Antirrhinum majus (Scrophulariaceae) (fla-
is related to and probably essential for the biosynthesis of vone synthase m (Fig. 11.10). In this plant, flavones arise
flavonoids. from dehydrogenation of flavanones and not from dehydra-
The chalcone isomerase from parsley is specific for tion of dihydroflavonols (Britsch et al., 1981). A similar
4,2',4',6'-tetrahydroxychalcone substrates (such as 9). ~uly enzyme system converts dihydroflavonols to flavonols
5,7-dihydroxyflavonoids (such as 14) are found in the plant. (Britsch et al., 1981). In other work, the enzyme responsible
Chalcone isomerase activity is present in anthocyanin or fla- for oxidation offlavanones to flavones in snapdragon (Antir-
vonol-containing genotypes of the china aster, Callistephus rhinum majus) was isolated from a microsomal fraction and
chinensis (Asteraceae), but completely absent from a yellow shown to require NADPH and molecular oxygen (Britsch et
line that accumulates the chalcone glucoside isosalipurpuro- al., 1981; Dewick, 1989; Forkmann and Stotz, 1981). The
side (16) (Fig. 11.9) (Grisebach, 1985). An enzyme capable system appears to be a cytochrome P-450-dependent mo-
of converting the deoxychalcone isoliquiritigenin (15) into nooxygenase. This system also is known from Glycine max

ROW: OR
p 1 1
""-
OR °
,
.
isoliquiritigenin (15) liquiritigenin (14)

ROy:;:o0-gIUOOSYIOR ,. ,I "'"
P "'" .' PI"
""- 1 ::::,.. 1 "'" S' ""-

OR ° "
°
chalcone
isosBlipurpuroside (16)
marein 2, 131,4' ,4·pentahydroxy·4'·glucoside
coreopsin 2,'3 ,4',3,4·pentahydroxy-4'·glucoside
1

Fig. 11.9. Representative chalcones. flavanones, and deoxychalcones.

 FIQ)'onoids 159

HO~OWOH HO

W--
H 0 y r : P ' :OH
~, ,
""-
OH 0 OH 0 OH 0
(2S)-naringenin (10) 2-hydroxynarlngenin apigenin (51)

~
OH "'" OH
HO &

OH
OH 0
OH 0

dihydrokaempferol (13) kaempferol (6)

Fig. 11.10. Biosynthesis of apigenin and kaempferol (Heller et at., 1985a; modified and used with pemlission of the copyright owner, Springer-Verlag,
Heidelberg).

(Fabaceae), Verbena hybrida (Verbenaceae), and Taraxa- cyanins, and catechins (such as 20) (flavan-3-0Is; see Chap-
cum officinale (Asteraceae) (Dewick, 1989). Chalcones do ter 12) has been demonstrated by tracer studies. To date, the
not appear to be substrates (Ebel and Hahlbrock, 1982; HahI- origin of the oxygen of dihydroflavonols in position 3 is not
brock, 1981). clear, although an enzyme !hal introduces oxygen into that
position in naringenin has been isolated. The same protein
Dihydroftavonols extract described above for the synthesis of flavones also
has the ability to hydroxylate naringenin (10), yielding dihy-
Certain dihydroflavonols are intennediates in the biosyn- drokaempferol (13), and the ability to oxidize dihydrokaem-
thesis of flavonols, but approximately 110 other dihydrofla- pferol to kaempferol (6). Dihydrokaempferol serves as an
vonols and dihydroflavonol glycosides are found in a variety efficient precursor of cyanidin (an anthocyanidin) and quer-
offamilies, especially the Asteraceae and Fabaceae (Grayer, cetin (7) (a flavonol) in buckwheat (Fagopyrum esculentum,
1989). Polygonaceae) (Kerscher and Franz, 1988).
Dihydrokaempferol (13) appears to arise by direct hy-
droxylation ofnaringenin (10) at the 3-position, catalyzed by Substitution Patterns of F1avonoids
a dioxygenase, flavanone 3-hydroxylase (Grisebach, 1985;
Heller and Forkmann, 1988; Stafford, 1991). The enzyme The substitution pattern of flavonoids is detennined by
requires 2-oxoglutarate, Fe2+, and ascorbate as cofactors. several factors. Hydroxylation at the 4'-position of the B
(2S)-Naringenin [but not the (2R)-enantiomer] is a substrate ring is derived from the p-coumaric acid precursor involved
for the enzyme. The product fonned has been identified as in the fonnation of most flavonoids. Compounds that lack
(2R, 3R)-dihydroquercetin. Although the intennediacy of a this substitution are comparatively tare. Additional hydroxyl
chalcone 2,3-epoxide (19) for the synthesis of dihydroflavo- groups appear to be added after the flavonoid is fonned (i.e.,
nols such as taxifolin (18) has been proposed, this synthesis at least at the chalcone stage) (Stafford, 1991). Although
may occur by hydroxylated intennediates (Fig. 11.11). 4-coumaryl-CoA is the most efficient precursor, chalcone
synthase from some plants can also use caffeoyl-CoA and
feruloyl-CoA as substrates; in which case, the correspond-
F1avonols
ing substituted chalcones are formed. Hydroxylation of the
Approximately 1030 flavonols are known (Harborne, 3'- and 5'-positions is the most common type ofB-ring oxy-
1991). Most of these are glycosides derived from approxi- genation. A microsomal flavonoid 3'-hydroxylase from par-
mately 300 flavonoid aglycones. Flavonol biosynthesis sley cell cultures has been demonstrated. This enzyme will
probably occurs via a 2-hydroxy intennediate with subse- convert naringenin (10) to eriodictyol (17) and will accept
quent dehydration, in a manner similar to that proposed for dihydrokaempferol (13), kaempferol (6), and apigenin (5)
fonnation of flavones. Flavonol fonnation with extracts from as substrates. The enzyme appears to be a cytochrome P-450-
flowers of Matthiola and Petunia requires a soluble 2-oxo- dependent monooxygenase (Grisebach, 1985). ER-Iocalized
glutarate-dependent dioxygenase (Heller and Forkmann, cytochrome P-450 monooxygenases, which probably arose
1988). from cinnamic hydrolyase of the phenylpropanoid pathway,
The conversion of dihydroflavonols into flavonols, antho- are responsible for 3' and 5' hydroxylation (Stafford, 1991).
160 Flavonoids

R
OR

RO
R'

OR 0
R=OH, R'=H; eriodictyol (17)
R = OH, R' =H; dihydroquercetin (18)
R =R' =OH; dihydromyricetin (29)
R
OR

~
OR
RO

",,' ,
R0 7 0 '""" R'

OR 0
R=OH, luteolin (4) R=OH, R'=H; quercetin (7)

Fig. 11.11. Proposed biosynthetic schemes leading to taxifolin (Britsch et aI., 1981; modified and used with pennission of the copyright owner, Vedag
der Zeitschrift fUr Naturforschung, Tiibingen).

The actual pattern of oxidation observed may depend on Glycosylation and Methylation of Flavones
the nature of the enzyme complex involved in a particular and Flavonols
situation. The eventual substitution of the A ring is deter-
mined by the substitution pattern at the chalcone stage. 5- A large number of flavones and flavonols occur naturally
Deoxyflavonoids are derived from chalcones without a hy- as glycosides. Although there are about 475 flavone and
droxyl group in the corresponding position (the numbering flavonol aglycones, 720 flavone and flavonol O-glycosides
system of chalcones is different from that of cyclized flavo- and 214 C-glycosides (and their O-glycosides) (see below)
noids) formed by the action of soluble NADPH-dependent have been reported (Markham, 1989). A number of highly
reductases (Fig. 11.8) (Stafford, 1991). specific flavone and flavonol glucosyltransferases are pres-
An enzyme from the flowers of Sinningia cardinalis ent in parsley cell cultures as well as a number of other plant
(Reichsteinia cardinalis, Gesneriaceae) has hydrolase activ- sources (Ebel and Hahlbrock, 1982; Grisebach, 1985; Hosel,
ity associated with microsomal fractions and requires 1981). In Pisum sativum, three distinct glucosyltransferases
NADPH as an essential cofactor (hydrolyase activity II). catalyze the stepwise glucosylation of kaempferol (6) and
This enzyme converts naringenin (10) and apigenin (5) to quercetin (7) to produce a series of triglucosides all of which
eriodictyol (17) and luteolin (4), respectively (Dewick, have a 3-0-glucoside substitution.
1989). The flavone synthase activity of this enzyme was Enzymes that catalyze methylation of specific hydroxyl
abolished completely by treattnent with the cytochrome P- groups in flavonols have been isolated (Grisebach, 1985;
450 inhibitor ancymidol, but the flavonoid 3'-monooxygen- Heller and Forkmann, 1988). A quercetin 3-0-methyltrans-
ase activity was not altered. ferase has been isolated from several plants (Poulton, 1981),
Other enzymes that introduce oxygen into the 3'- and 5'- and in one plant, at least three distinct flavonol O-methyl-
positions have been isolated (Dewick, 1984, 1989; transferases were found. These enzymes catalyzed the step-
Grisebach, 1985). wise methylation of quercetin to mono-, di-, and tri-O-
methyl derivatives (Ebel and Hahlbrock, 1982).
Flavone and Flavonol Aglycones Glycosides of flavonoid sulfates occur in a number of
Although the majority of flavones and flavonols in plants plants (Fig. 11.12) (Harhorne, 1991). More than 100 flavone
occur as glycosides or other "bound" forms, small amounts and flavonol sulfates from more than 250 species and 30
of aglycones frequently are present and occasionally repre- families have been characterized; compounds of this type
sent a sizable proportion of the total flavonoid compounds may be recognized by examination of their UV spectra be-
present in or on the plant. Flavonoid aglycones frequently fore and after addition of HeI and aryl-sulfatase (Barron and
are methylated or esterified and the mixture is lipophilic. In Ibrahim, 1988; Harhorne, 1991; Varin, 1992).
many plants, these compounds are the products of epidermal A number of flavonoids possess a malonyl unit linked to
glandular trichomes (Rodriguez et aI., 1984; Wollenweber the sugar moieties. Others with malic, oxalic and succinic
and Dietz, 1981). Some highly methylated flavonoids are acid have been reported (Harborne, 1991).
toxic to mammals and other animals. The techniques for isolation and study of flavone and
 Flavonoids 161

OSO,- OR
OR OCR,

-O,S-O "" CR,O

OR
OR 0 OR 0

luteolin 7,3 1·disulfate tamarixetin 3·sulfate

Chap. 1l,Fig. II

Fig. 11.12. Sulfates of flavonoids.

OR

RO

OR 0
2-hydroxynaringenin (23)
C-glucosyltransferase

RO OR

RO
II OR 0

OR

""
§

OR 0
RO
RO
! ""
OR

OR o
vitexin (21)
Wessely-Moser isovitexin (24)
0
1
O-glucosylaHon

isosmerization
C·glycosylflavone 7·0-g1ucosides

Fig. 11.13. Proposed biosynthesis of flavone C-glycosides (modified from Dewick, 1989 and Chopin and Boulliant, 1975; llsed with permission of the
copyright owners, the Royal Society of Chemistry, London and Academic Press, New York).
162 Flavonoids

flavonol glycosides have been reviewed (Harborne, 1973; sylation occurs after conversion to flavones (Kerscher and
Mabry et aI., 1970; Markham, 1982, 1989) (also see below). Franz, 1988).

Biosynthesis of OGlycosyJ F1avonoids Biosynthesis of Anthocyanins

Biosynthetic studies in Spirodela polyrhiza demonstrated The biosynthesis of anthocyanidins has been elucidated
that apigenin (5) and luteolin (4) served as efficient precur- more recently than most other flavonoids. These positively
sors of the corresponding O-glycosylated compounds but charged compounds are derived from flavan-3,4-diols via
were not incorporated into the corresponding C-glycosides dihydroflavonols. The hydroxyl group of the 3-position of
(Wallace and Grisebach, 1973; Wallace et aI., 1969). In con- anthocyanidins is introduced by flavanone 3-hydroxylase
trast, naringenin (10) was incorporated in a parallel manner which converts flavanones to dihydroflavonols (see above).
into both 7-0-glycosides and into vitexin (21) and orientin (2S)-F1avanone 3-hydroxylase from flowers of Petunia hy-
(22) (Figs. 11.13 and 11.14) (Wallace and Grisebach, 1973). brida catalyzes the conversion of (2S)-naringenin (10) [but
2-Hydroxyflavanones are intennediates in the biosynthesis not (2R)-naringeninJ to (2R,3R)-dihydrokaempferol (13).
(Kerscher and Franz, 1988). A C-glucosyltransferase prepa- The hydrolase is a soluble enzyme that requires oxygen, 2-
ration from buckwheat, F agopyrum esculentum (Poly- oxoglutarate, Fez+, and ascorbate as cofactors (Britsch and
gonaceae), catalyzes the transfer of glucose from UDP-glu- Grisebach, 1986). (2S)-Eriodictyol (17) is converted to
cose or ADP-glucose to 2-hydroxynaringenin (23) (2R,3R)-dihydroquercetin.
producing a mixture of vitexin (21) and isovitexin (24) and Subsequent reduction of dihydroflavonols, catalyzed by
to 2,5,7-trihydroxyflavanone (25) giving 8- (27) and/or 6- dihydroflavonol 4-reductase, leads to flavan-3,4-diols (Fig.
C-glucosylchrysin and (26) (Figure 11-14). Thus, C-glyco- 11.15) (leucoanthocyanidins), which are intennediates in an-

S·C·glucosylchrysin (27) 6-C·gJucosylchrysin (26)

OD
OR
OD

OD 0

orientin (22) isoorientin

DO
OD
DO

rhamnosyl-4-ketofucosyl
OD 0
OD 0

2,5,7-trihydroxyflavanone (25) maysin (71)

Fig. 11.14. Some important C-glycosylflavones.

 Flavonoids 163

OH
OH
o 0 HO

~'"
1~ OH
_~'"
'" OH _ _

«pI:
HO ~

OH
0H
OH ~OH
HO '"
",I
0
_
HOWO
1
~R HO
R

'" OH
OHO OHO
eriodletyol (17)
R=H,(18)
(+)·2,3·/ran,·dihydroquercetin R=H
R=OH
OH
OH
OH

HO HO R

OH 0 OH
R =H, (+)-catechin R =H, a tlavan-3,4-dlol (33)
2,3-cts-dihydroquerc:etin R =OH, (+)-gallocatechln leucocyanidin
R=OH~leUCOd~:hlnldin
OH
~ '" OH
HO 0+ I",
HO
'" ~

"" OH OH
I
h OH OH an anthocyanidin (2)
O~OH
OH OH
procyanidins
cyanldin·3·Q·g1ucoside (34)

Fig. 11.1S. Biosynthetic pathway for the origin of anthocyanins.

thocyanin biosynthesis with flower extracts from Matthiola the flowers of Sinningia (syn. Reichsteinia) cardinalis (Ges-
incana (Heller and Forkmann, 1988; Heller et al., neriaceae) catalyze an NADPH-dependent reaction of (2S)-
1985a,1985b; Porter, 1988; Stich and Forkmann, 1988) (see naringenin to apiforol (leucoapigeninidin) (31) and (2S)-eri-
also Chapter 12). The patterns of anthocyanidins in flowers odictyol (17) to luteoforol (leucoluteolinidin) (32) (Stich and
of Petunia hybrida (Solanaceae) are attributable to the sub· Forkmann, 1988) (Fig. 11.16). All the steps leading to 3,4-
strate specificity of dihydroflavonol 4·reductases. The en- diols are shared with condensed tannin biosynthesis (Staf-
zyme preparation catalyzes NADPH-dependent reduction of ford, 1989). A series of enzymes involved in the reduction
dihydroflavonols to 2,3-trans-3,4-cis-flavandiols (leucoan- of dihydroflavonols to flavan-3,4-diols (such as 33) have
thocyanidins). In particular, dihydroquercetin (18) is con- been reviewed in regard to proanthocyanidin biosynthesis
verted into leucocyanidin (28) and dihydromyricetin (29) (Stafford, 1989) (see Chapter 12).
into leucodelphinidin (30). Dihydrokaempferol (13) is con- Dihydroflavonols or their glycosides that accumulated in
verted into pelargonidin (1), but Petunia does not contain white flowered mutants were supplied to acceptor mutants
anthocyanins of that series. At least one of the steps is light that were blocked in the synthesis of dihydroflavonols (Stich
controlled (Ebel and Hahlbrock, 1982; Hrazdina. 1982). Re- and Forkmann, 1988). This led to anthocyanin synthesis in
duction of the flavanones naringenin (10) and eriodictyol the acceptor mutant. For example, white-flowered mutants
(17) leads to the biosynthesis of 3·deoxyanthocyanins (Stich of Petunia hybrida accumulate dihydroquercetin 7-0-gluco-
and Forkmann, 1988). Soluble enzyme preparations from side, dihydroquercetin 4'-O-glucoside, dihydroquercetin.
164 Flal'onoids

OR OR
OR OR

RO RO
OR

# #
OR
OR OR OR OR

I:
leucodelphinidin (30)

I:
lencocyanidin (28)
OR OR

RO "'" 0 RO 0
~ 1 OR
::::,..1
::::,..
OR OR OR OR

apiforol (31) luleoforol (32)

Fig. 11.16. Selected leucoanthocyanidins.

and dihydrokaempferol 7-0-glucoside. When dihydroquer- of flavonoid glycosides. Flavonoid chemistry was especially
cetin (17) was supplied to these plants, cyanidin 3-glucoside useful for analysis of the many hybrid swarms encountered,
(34) was formed as the major pigment (Grisebach, 1985). some of which involved as many as four different species.
In contrast to flavones and flavonols, anthocyanidins are In general, it was possible to recognize hybrid plants because
rarely, if every, found free in nature (Heller and Forkmann, they had simply additive patterns (i.e., the hybrids often had
1988). 3-0-G1ycosylation is a likely prerequisite for antho- a combination of all the compounds found in the putative
cyanin accumulation because of the instability of the flavyl- parental types). In some hybrids, however, a few compounds
ium cation. UDP-Glucose flavonoid 3-0-glucosyltransfer- failed to appear and, in others, new hybrid compounds were
ase is involved in formation of anthocyanidin glycosides. observed.
In hybrids of Baptisia leucophaea and B. spherocarpa, the
appearance of a hybrid compound could be explained when
SYSTEl'IAllC S11JDms WITH FLAVOrm the chemical nature of the compound was ascertained. Com-
AM> FLAVONOL GLYCOSIDES bination of the enzymes that produced a 3-0-rutinoside (rutin)
(35) in the former and the 7 -O-glucoside (36) in the latter pro-
No other group of compounds has been as useful as flavone duced a 7-0-glucoside-3-0-rutinoside (37) in the hybrids
and flavonol glycosides for the study of problems in plant (Fig. 11.17) (Alston and Turner, 1963; Alston et al., 1965).
systematics (Harborne, 1975; Harborne and Mabry, 1982; In a survey of the flavonoids of the genus, some com-
Harhorne and Turner, 1984). Although many flavonoids are pounds proved to be common to all species, some were found
widely distributed, characteristic patterns of flavonoids are only in one species, some only in hybrids, and some ap-
found in many plant groups which permit recognition of peared sporadically (Alston et aI., 1962; Alston and Turner,
particular taxa. 1963; Alston et aI., 1965; Cranmer and Mabry, 1966; Har-
Study of flavonoids has proven useful for examination borne, 1975; Markham and Mabry, 1968).
of many problems at the species and genus level for several In other instances, flavones and flavonols with particular
reasons. Most plants contain several major compounds (usu- patterns of oxidation are of sufficiently restricted distribution
ally glycosides) that tend to differ in the various taxa (forms, among plant groups to be useful for systematic purposes.
subspecies, varieties, or species) which are to be examined. For example, in addition to the usual flavones [apigenin (5),
Analysis of these mixtures is reasonably straightforward; luteolin (4), and chryseriol (38) (Fig. 11.3), usually as the 7-
techniques for identification of the compounds have been O-glucuronidesl, plants of the Lamiaceae, Scrophulariaceae,
reviewed (Mabry et aI., 1970; Markham, 1982). The flavo- and related families frequently contain 8-hydroxyflavones,
noid compounds present in hybrids often represent an addi- 6-hydroxyflavones, or 6-methoxyflavones in glycosidic
tive combination of those of the parental types. combination. 8-Hydroxyflavones based on apigenin and lu-
The systematics of the genus Baptisia (Fabaceae) has teolin are restricted to the Lamiaceae (Tomas-Barbenin et
been studied extensively, and the genus is well known with al., 1988).
respect to the morphology, genetics, and phytochemistry. The distribution of flavones and flavonols and their gly-
Almost all species of Baptisia have distinctive complements cosides has been reviewed (Harborne and Williams, 1982).
 Flavonoids 165

OH
OH

HO

OH
OH 0 OH

~o
rutin (35)
Baptisia leucophaea

O-rutinosyl
OH OH 0

OH
hybrid compound
glucosyl- 0 quercetin-7-0-glucoside-3-0-rutinoside (37)

OH 0

quercetin 7-D-glucoside (36)

Baptisio spherocarpa

Fig. 11.17. The origin of a hybrid compound in Haptisia hybrids.

For example, the validity of Dahlgren's proposed Liliiflorae In the United States, humans consume about 1 g offlavo-
was appraised by an examination of flavonoids (Williams noids daily; in some cultures, this may be as high as 2-3
et aI., 1988). The flavonols quercetin (7) and kaempferol (6) glday (Beier and Nigg, 1992). About 40% comes from
occurred in about half the plants tested; the flavones apigenin cocoa, cola, coffee, beer, and wine. Fruit juices and tea also
(5) and luteolin (4) were found less frequently. The limited are major contributors. The digestive systems of animals are
diversity of flavonoid patterns argues for a more conserva- capable of metabolizing many flavonoids. p-Hydroxyben-
tive treatment at the family level (Williams et al., 1988). zoic acid, cinnamic acid, phenylacetic acid, and phenylpropi-
Other applications of flavonoid data to the understanding onic acid are observed as breakdown products of rutin (35)
of plant phylogeny include bryophytes, ferns and fern allies and apigenin (5) (Beier and Nigg, 1992). However, certain
(Markham, 1988), gymnosperms (Niemann, 1988), monoco- other flavonoids, such as epicatechin, are excreted un-
tyledons (Williams and Harborne, 1988) and dicotyledons changed (Fig. 11.17).
(Giannasi, 1988). Of 70 flavonoids tested, 33 were positive in the Ames
assay. Other flavonoids have been associated with carcino-
genic activity. However, flavonoids also have been shown
BIOLOGICAL ACTIVITY OF FLAVONE AI'ID to have antimutagenic effects (Beier and Nigg, 1992).
FLAVONOL AOLYCONES AI'ID OLYCOSIDES
Biological Activity of Aglycones
Because flavonoids are found in virtually all plants, some
animals, and a few fungi, these compounds are found in Most flavone and flavonol aglycones occur on the surface
many different types of biological interactions (Beier and of plants or associated with secretory structures. The isola-
Nigg, 1992; Cody et aI., 1986, 1988; Harborne, 1991). Fur- tion and characterization of flavone and flavonol aglycones
ther, plant flavonoid content may be influenced by light, have been reviewed (Wollenweber and Jay, 1988).
water, temperature, mineral nutrition, sugars, mechanical Flavonoids probably play major roles as internal physio-
damage, pathogens, plant growth regulators, and other fac- logical regulators and chemical messengers in plants. This
tors. Flavonoids may serve as antioxidants, enzyme inhibi- may have been their major role in the early evolution of land
tors, pigments for light absorbance, and visual attractants plants (Stafford, 1991).
for pollination,light screens, promotors of inhibitors of plant At physiological concentrations (10- 5 M), flavonoids
growth, plant growth regulators, chemical signals in root may either stimulate or inhibit IAA oxidase (an enzyme
nodulation gene induction, phytoalexins, and other functions which regulates the amount of the plant-growth-regulating
(Beier and Nigg, 1992; McClure, 1975). hormone indole acetic acid (IAA or auxin) activity in peas
166 Flavonoids

in in vitro experiments. Stenlid (1970) showed that, in gen- compounds acted primarily as electron transport inhibitors.
eral, 4'-hydroxy compounds stimulate and 3', 4'-dihydroxy Inhibition of substrate oxidation appears to result from alter-
compounds inhibit IAA oxidase activity. In general, cin- ations and perturbations induced in the inner membrane, as
namic acids and their derivatives were ineffective. evidenced by interference with carrier-mediated transport
The process of auxin transport can be inhibited by a group processes (Moreland and Novitzky, 1987). Thirteen flavones
of synthetic compounds which bind to a specific cell mem- (of 21 tested) are selective inhibitors of the external NADH
brane receptor. Several flavonoids, including quercetin (7), dehydrogenase of the inner membrane of the mitochondria
apigenin (5), and kaempferol (6), can specifically compete of potato tubers (Ravanel et al., 1990).
with one of the synthetic compounds, naphthylphthalamic Quercetin (7), morin (39), myricetin (8), and fisetin (40)
acid, at micromolar levels for binding to the receptor and, (Fig. 11.18) were active inhibitors of induced lipid peroxida-
thereby, perturb auxin transport in a variety of plant tissues tion in the presence of ferrous ion. Isovitexin (24) is an
(Jacobs and Rubery, 1988). Although quercetin (7) was effective antioxidant-being more effective than BHA (bu-
highly active, a common quercetin glycoside, rutin (35) tylated hydroxyanisole) and a-tocopherol. Epicatechin 3-0-
proved inactive. gallate (41) is a potent oxygen free-radical scavenger (Beier
Certain flavonoids can interrupt the electron transport and Nigg, 1992).
chain of respiration and photosynthesis (Arntzen et al., Nitrogen fixation by legumes is an essential component
1974). In chloroplast thylakoids, the primary effect was on of many natural and agricultural ecosystems. In this process,
the ATP-generating pathway. The flavonoids tested did not atmospheric nitrogen is converted into ammonia, a fonn usa-
appear to be decouplers; in mung bean mitochondria, the ble by plants and other organisms. Species of the bacterium

OCH, OCH,
OCH,
CH,'O
CH,'O

CH,-O
OH
OCH, 0 OH 0
tangeretin (48) ermanin (49)
(5,6,7,8,4'-pentamethoxyflavone)
(7,4'-di·O-methylkaempferol)
OH OH
OH
HO
glucosyl-O

OH 0
4,4'-dihydroxy-2'-methoxychalcone (45) luteoHn 7-0-glucoside (42)

HO~O~OH HO~O~OH
Uy' o
~-
o
7,4'.dihydroxyflavanone (43) 7,4'-dihydroxyflavone (44)
OH
OH OH
HO
HO

OH
OH 0
o
(al isoetin (47) fisetin (40)

Fig. 11.18 (a & b). Selected flavonoids with biological activity.

 Flavonoids 167

OH
OH
HO HO

isoquercitrin (51) morin (39)

«p
OH OH
OH
rhamnosyl- 0:;.-' 0 I /.: oHxylosyl- 0 I'"
1 1 '"
CH3 0 :::-.... OH
OH 0 OH 0
6-methoxyluteoUn '·rhamnoside (57) catechin 7-xyloside (58)

W ,V-
OH
HO '"/- HO,.::0'.J O

OH
OH 0

o«Pl:
OH 0
dihydrokaempferol (13R = H pinocembrin (5)
taxifolin (17) = OH OH

rhamnoglucosyI- OCH,
""I
hesperidin (61 ~

o 0 OH 0

HO
)t.,J
:o~
HO~
_ _ o«P
o=O,
I I
I
OH
OH

Ollfl "-
(b) luteotin '·O-(6"-malonyl)glucoside (62) OB 0

Fig. 11.18 (continued)

Rhizobium are responsible for the nodulation of many spe- 3-0-galactoside, does not induce nod genes, but both luteolin
cies; several stages are involved in the nodulation process. (4) and quercetin (7) increase growth of Rhizobium meliloti
Host-symbiont signaling occurs at an early stage in nodule (Hartwig and Phillips, 1991; Hartwig et aI., 1990a, 1990b).
development Expression of nodulation (nod) genes is essen- Seed exudates are less active than root exudates. Alfalfa
tial for nodulation, and this gene is induced by host plant- roots exude three major nod gene inducers; 4',7-dihydrox-
derived flavonoids (Harbome, 1988a, 1988b; Phillips, 1992; yflavanone (43), 4',7-dihydroxyflavone (44), and 4,4'-dihy-
Towers et aI., 1989). After this induction, rhizobial products droxy-2'-methoxychaIcone (45) (Maxwell et aI., 1989). Re-
induce root hair curling, cortical cell divisions, and produc- lease of these compounds is associated with concurrent
tion of a receptor-like protein (Hartwig et aI., 1990a, 1990b). biosynthesis (Hartwig et aI., 1990a, 1990b; Maxwell and
The symbiotic interaction of Rhizobium meliloti and al- Phillips, 1990). The flavonoid active in Trifolium repens,
falfa results in the formation of nitrogen-fixing nodules. white clover, is 7,4'-dihydroxyflavone (44) (Towers et aI.,
Seed coats of alfalfa are the major source of the two major 1989). Both daidzein (7,4'-dihydroxyisoflavone) and gen-
inducer molecules, chryseriol (38) and luteolin (4), for no- istein (46) (Fig. 11.26) from soybeans induce nod genes in
dABC gene expression in Rhizobium meliloti during nodule Bradyrhizobium japonicum. Certain other plant flavonoids
formation (Hartwig et aI., 1990b; Phillips, 1992). Luteolin and phenols inhibit nod germination (Towers et aI., 1989).
7-0-glucoside (42), released from seed exudates, is con- Flavonoid aglycones frequently serve as pigments in
verted to luteolin (4), apparently by enzymes from either flowers. For example, isoetin (2'-hydroxyisoetin) (47) is a
the host plant or the bacterium (Hartwig and Phillips, 1991; yellow pigment in Heywoodiella oligocephala (Asteraceae)
Peters et aI., 1986). The major flavonoid released, quercetin (Harbome, 1991).
168 Flavonoids

Flavonoid aglycones (especially highly methylated ones) guminosarum bv. phaseolii, are 3-0-glycosides of 10 antho-
often are found as resinous exudates on plants (Harbome, cyanidins and flavonols; most important among these were
1991; Seigler, 1983). These compounds have higher insect delphinidin, petunidin, and malvidin. In these cases, the cor-
and mammalian toxicity than the corresponding nonmethyl- responding aglycones were less active (Hungria et aI.,
ated derivatives. Tangeretin (5,6,7,8,4'-pentamethoxyfla- 1991a). Three other flavonoids, eriodictyol (17), naringenin
vone) (48), which occurs in tangerine peel, is cytotoxic to (10), and a 7-0-glycoside of genistein (46) were the most
humans and causes neonatal death in rats (Harborne, 1991). active compounds from root exudates of the same plant
The lipophilic material found on the surface of Larrea (Hungria et al., 1991b).
species (Zygophyllaceae) is composed of several methylated The major flavonoid, quercetin 3-0-galactoside from al-
flavonoid aglycones and lignans such as nordihydroguaiare- falfa seeds, promotes spore germination of two fungi respon-
tic acid (see Chapter 8). This resinous material was shown sible for the formation of vesicular-arbuscular mycorrhizal
to act as an antiherbivore substance and appeared to reduce (VAM) interactions, G/omus etunicatum and Glomus ma-
digestibility of the plant for several herbivores (Seigler, crocarpum, in vitro. Quercetin (7) produced the maximum
1983). effects in spore germination, hyphal elongation, and hyphal
Ermanin (49), 7,4'-di-O-methylkaempferol, from the res- branching in Glomus etunicatum at 1-2.5 tJM concentrations
inous material on the surface of leaves of Passiflorafoetida, (Tsai and Phillips, 1991).
is deterrent to feeding by the larvae of Dione juno. This Most flavone and flavonol glycosides are not particularly
insect eats many other Passiflora species (Echeverri et aI., toxic to man or other higher animals (McClure, 1975). One
1991). compound of this type, rutin (35), was once considered to
Pinocembrin (50) has been implicated as an attractant to be a vitamin, but is generally not at present (Harborne, 1991).
a bark beetles feeding on fruit trees (Harborne, 1991). In Compounds of this general structure may be required in the
contrast, the flavonoid aglycone, morin (39) [as well as iso- human diet, but it is difficult to imagine anyone suffering
quercitrin (51) and several essential oils1, is involved in feed- from such a deficiency, as flavonoids are found in most of
ing of silkworm larvae (Bombyx mori) on mulberry plants the plant materials that we eat.
(Morus nigra) (Harborne, 1991). Certain flavonoids have antiulcer properties in humans.
These compounds raise prostaglandin levels in gastric mu-
Biological Activity of Glycosides cosa by stimulating cyclooxygenase or by inbibiting 15-hy-
At least 330 flavone and 650 flavonol glycosides are droxyprostaglandin dehydrogenases (Lewis, 1992). Hypo-
known (Harborne and Williams, 1988). Among the sugars laetin 8-0-glucoside (57), from Sideritis mugronensis
involved in flavonoid glycosides are o-apiose, L-arabinose, (Lamiaceae), stimulates prostaglandin synthetase. The most
L-rhamnose, D-xylose, D-allose, D-galactose, D-glucose, 0- widely used antiulcer flavonoid is solon, 2-carbomethoxy-
galacturonic acid, and o-glucuronic acid. Glycosides may 4,4-bis(3-methyl-2-butenyloxy)chalcone. This compound is
include a variety of disaccharides and trisaccharides. Both based on the structure of sophoradin (53) from a Chinese
the sugars and the aglycones may be acylated, or modified traditional drug gnangdougen, from Sophora subprostrata
with other substituents. Techniques for the isolation and (Fabaceae). Solon increases prostaglandin levels by inhibit-
characterization of flavonoid glycosides have been reviewed ing 15-hydroxyprostaglandin dehydrogenase (Lewis, 1992).
(Harborne and Williams, 1988). Other flavonoids such as catechin (20) and naringenin (10)
Flavonoids (both glycosides and aglycones) have been appear to inhibit histidine decarboxylase and limit tlie forma-
suggested to serve as sunscreens which absorb harmful fre- tion of the gastric acid stimulator histamine (Lewis, 1992).
quencies of UV light (UV-C less than 280 nm, UV-B A series of flavonoids from the milk thistle, Si/ybum mar-
280-320 nm, UV-A 320-400) and prevent destruction of ianum (Asteraceae) have antihepatotoxic activity. The indi-
important compounds in plants as well as permit entry of vidually active compounds are silybin (54) (the major com-
useful wavelengths (Caldwell et al., 1983; McClure, 1975; pound), silycbristin (55), and silydianin (56), collectively
Stafford, 1991). Indeed, the epidermal layer where many called silymarin (Fig. 11.19). Silymarin is used in Germany
flavonoids are accumulated absorbs more than 90% of the for treatment of human liver disorders (Vogel, 1976, 1982).
UV-B radiation athninistered (Robberecht and Caldwell, The elimination half-life of silybin is about 2 h (Beier and
1978). Most flavone and flavonol aglycones and glycosides Nigg, 1992).
absorb in the range 330-350 nm, the same area of the spec- Many flavonoids appear to be toxic or repellent to insects
trum where NAD and NADP absorb (McClure, 1975). Fla- (Hedin and Waage, 1986). Flavone and flavonol glycosides
vonoids also have a strong absorbance in the area 250-280 may serve as both allomones and kairomones (Fig. 11.18).
nm, where proteins and nucleic acids absorb. Probably the A flavone glycoside, 7-a-L-rhamnosyl-6-methoxyluteolin
most sensitive part of the photosynthetic machinery to be (57), from alligator weed (A/ternanthera philexeroides, Am-
protected is photosystem II (PSII) (Stafford, 1991). aranthaceae) serves as a feeding stimulant for Agasic/es bee-
The signals from a number of plants involved in nodula- tles. Kaempferol 3-0-xylosylgalactoside stimulates feeding
tion by Rhizobium bacteria are now known (see above). in the monophagous flea beetle, Phyllotreta armoraciae, on
Those of Phaseo/us vulgaris, black bean, for Rhizobium /e- horseradish, Armoracia rusticana (Brassicaceae) (Nielsen et
 Flavolloids 169

HO

silychristin (55)

HO

HO OCH,
HO OH
HO HO
OH
OH
HO

OH 0
.-7
sophoradin (53)
hypolae~in 8-glucoside (52)

Fig. 11.19. SHybin, silychristin, silydianin, and other medicinally useful flavonoids.

aI., 1979). A catechin xyloside (58) is responsible for feeding properties of flavonoids. These include alkaloids, glucosino-
of the smaller European elm bark beetle, Scolytus multistria- lates, cucorbitacins, and cardenolides (Harbome, 1988b).
tus, on elm (Hedin and Waage, 1986). Silkworms prefer Foor flavonoids [vicenin-2, hesperidin (60), narirutin, and
food plants with rutin (35), but this compound is repellent rutin (35)] from Citrus unshiu are oviposition stimulants for
to tobacco budworm (Heliothis virescens) and to the boll- Papilio xuthus (Nishida et aI., 1987; Ohsugi et aI., 1985).
worm (H. zea). When fed at low concentrations, quercetin However, these compounds are only weakly active alone or
3-rharnnoside, quercetin 3-glucoside, and rutin (35) repelled in mixtures. When mixed with two bases, adenosine and 5-
feeding by the tobacco budworm, the bollworm, and Ptecti- hydroxy-N-methyltryptarnine, which are inactive by them-
nophora gossypiella; however, at concentrations of 0.2% or selves, 100% of the activity of the plant was observed (Har-
more, these common flavonoids killed the larvae (Harbome, borne, 1988b).
1991). Many other common flavonoid glycosides are toxic The flavonoid glycosides hesperidin (60) and naringin
to these insects at the 0.2% level in artificial diets (Harborne, (61) (Fig. 11.25) from Citrus natsudaidai stimulated ovipo-
1988c, 199 I). Toxicity seems to require vicinal hydroxyl sition by Papilio protenor (Honda, 1986). Again, the com-
groups [i.e., ortho-dihydroxybenzene groups (catechol)], pounds were inactive alone, but were active when combined
and this is a necessary, but not sufficient, condition for bio- with a more polar extract of the plant.
logical activity. Quercetin (7) is toxic to Spodoptera eri- Oviposition by females of the black swallowtail butterfly,
dania; rharnnetin (59) (Fig. 11.3) is not (Elliger et aI., 1980; Papilio polyxenes, was stimulated by tarsal coutact with
Lindroth and Peterson, 1988). ethanolic extracts of carrot foliage, Daucus carota. Two of
Many other groups of compounds reinforce the repellent the stimulants were identified as E-chlorogenic acid and lute-
170 Flavonoids

olin 7-0-(6"-O-malonyl)-j3-o-glucopyranoside (62) (Feeny, Cyanidin (3) (Fig. 11.1) is especially widespread and is
1991; Feeny et al., 1988). Although each compound was responsible for the purple coloration of most vegetative parts
inactive alone, in admixture they accounted for 70% of the of plants; this anthocyanidin is also common in fruits and
ovipositional response of the original extract. flowers. The purple pigments of red cabbage have been
Several flavonol glycosides accumulate in the wings of shown, by chromatography on polyvinylpolypyrrolidone
certain butterflies. Differences in the compounds seques- (PVPP), to be a complex mixture of eight acylated cyanidin
tered and those of the host plants indicate that the butterflies glycosides that correspond in properties to the previously
metabolize the compounds before sequestration (Harbome, reported "rubrobrassicins." One of the major compounds
1989, 1991; Wilson, 1987). is cyanidin-3-(6"-O-p-coumaruylsophoroside)-5-glucoside
Many flavonoids (especially 3-methoxyflavones) are (63) (Fig. 11.21) (Hrazdina et al., 1977). White cabbage is
known to serve as antiviral (Selway, 1986; Vanden Berghe et known to have a block in the pathway to these pigments.
ai., 1985; Vlietinck et al., 1986), anti-inflammatory (Vogel, Increased accumnlation of anthocyanins is observed in
1976, 1982), and antitumor compounds (Cassady et al., many plants in response to stress. This includes fall color-
1990; Mabry and Ulubelen, 1980). ation of leaves, response to nltraviolet radiation, attack by
pathogens. deficiency of some minerals, mechanical dam-
age, water stress, and many other factors (Hrazdina, 1982).
ANTHOCYANINS
Factors Responsible for Color
Anthocyanins, glycosides of anthocyanidins, such as (1-3),
are primarily responsible for the red, blue, and violet colors At least seven different factors seem to be important in
of flowers and fruits. More than 260 naturally occurring flower pigmentation: pH, the nature of the aglycone, the
anthocyanins have been described (Harborne, 1991); the extent of glycosylation, the concentration of the anthocya-
techniques for isolation, purification, and characterization of nin, complexation with metals, the presence of pectins and
these compounds have been reviewed (Harborne and Grayer, sugars, and complex anthocyanins which are linked through
1988; Strack and Wray, 1989). their 4-positions with tannins and other phenols (Brouillard,
Anthocyanins and anthocyanidins are widely distributed 1988; Harborne and Grayer, 1988; Harborne et ai., 1975).
among gymnosperms and both monocotyledonous and most Anthocyanins occur as salts within the cell. Extraction
dicotyledonous angiosperms (except in many members of under very mild conditions yields "genuine" anthocyanins
the order Caryophyllales or Centrospermae). Six relatively that have been studied electrophoretically. NMR spectros-
common aglycones (pelargonidin, cyanidin, delphinidin, copy and mass spectrometry (especially fast atom bombard-
peonidin, petunidin, and malvidin) are found in most cases; ment or FAB) have been extremely useful for the determina-
almost all others are rare. Only about 18 naturally occurring tion of structures of relatively intact anthocyanins
anthocyanin aglycones are known (Fig. 11.20) (Harborne (Brouillard, 1988).
and Grayer. 1988; Hrazdina, 1982; Timberlake and Bridle, Both metallic and nonmetallic pigments have been iso-
1975). Most of the numerous derivatives are variants pro- lated. At low pH values, anthocyanins usually are red,
duced by glycosylation, acylation, and methylation. whereas at high pH values, they usually shift to blue colors.

OCH, OR OCH,
OH OH

HO
I'"
HO ~
OCH, OCH,

OH OH OCH,
peonidin petunidin R = H
malvidin R = OCH3 capenslnidin
R
R
OH

CH,O

OH
OH OH

hirsutidin pelargonidin (I) R=R =H apigenlnidin R = H

cyanidin (2) R= OH, R = H luteolinidin R =OH
= =
delphinidin (3) R R OH
Fig. 11.20. Some representative antbocyanidin chromopbores.
 Flavonoids 171

cyanidin-3-poocoumaroylsophoroside-S-glucoside (63)

Fig. 11.21. Cyanidin-3-(6"-O-p-coumaroylsophoroside)-5-g1ucoside. the major pigment of red cabbage.

The pH of cell sap of most flowers is such that many antho- In the superorder Caryophyllidae (or Centrospermae),
cyanin-containing flowers should be colorless, yet the pres- only the families Caryophyllaceae and Molluginaceae pos-
ence in nature of colorless anthocyanins is rare. Anthocy- sess anthocyanins. In the other families of the superorder,
anidins undergo a series of complex color changes in water anthocyanins appear to have been replaced by alkaloidal be-
at varying pH (Brouillard, 1988). The flavylium cation of talain pigments. These compounds have nearly identical
natural anthocyanins behaves as a weak diacid, whereas a spectral properties to those of anthocyanins and also serve
neutral quinonoid base is at the same time a weak acid and in pollination and fruit and seed dispersal. This phenomenon
a weak base. The pH of crude extracts of flowers varies from will be discussed more fully under betalain pigments (see
2.8 to 6.2. Chapter 37).
Anthocyanin and anthocyanidin anhydro bases are known
to be stabilized by formation of highly colored complexes Commercial Importance of Antbocyanins
with metals, such as aluminum, iron, or magnesium, which
can be susceptible to pH changes within narrow limits. A number of anthocyanins are used as food additives
Copigmentation plays an important role in pigmentation (Harborne and Grayer, 1988; Strack and Wray, 1989; Tim-
by anthocyanidins (Brouillard, 1988). Most anthocyanins in- berlake and Henry, 1988). Anthocyanins also are important
volve copigmentation when present in suitable concentra- in that they control the marketability and taste properties of
tions. Copigmentation may involve flavones or other pheno- many commercial products. The sale price of many fruits
lic compounds such as chlorogenic acid. The blue pigment and vegetables is dependent on full color development.
of Hydrangea macrophyl/a flowers is comprised of del- Some major commercial sources of these natural pigments
phinidin 3-glucoside, aluminum, and 3-caffeoylquinic acid. are grapes (Vilis vinifera), roselle (Hibiscus sabdariffa),
In another complex case that has been studied, the blue pig- blueberries (Vaccinium spp.), and red cabbage (Timberlake
ment of Comme/ina communis involves a dimagnesium and Henry, 1988).
complex in which six anthocyanin and six flavone molecules Anthocyanins are especially complex in members of the
are linked either covalently or by hydrogen-bonding (Har- Vitaceae, the grape family. In this family, five or six agly-
borne and Grayer, 1988). cones are known and most occur as monoglycosides and
diglycosides. The sugars of anthocyanins often have acyl
groups attached. No wild or cultivated grape has less than
Biological Activity of Antbocyanlns
5 and most have at least 10 anthocyanins. These compounds
Anthocyanins typically absorb at 520-560 urn, a region are important, as they are modified in the aging process of
of the spectrum to which mammalian eyes are most sensitive. wine and are partly responsible for the color and the taste
These compounds are important pigments in flowers and properties of wines. In some instances, anthocyanins may
fruits and play major roles in pollination mechanisms and impart bitterness to the wines (Pierpoint, 1986).
fruit dispersal (Harborne, 1991). Chlorophyll, carotenoids, Tea leaves (Camellia or Thea sinensis, Theaceae) contain
and phytochrome have minimal absorption in this region. cyanidin 3-glucoside (34) and other anthocyanins. Phenolic
Cyanidin 3-0-glucoside (34) (Fig. 11.15) is an important materials make up as much as 30% of the dry weight of
factor of resistance in cotton leaves to feeding of tobacco fresh tea leaves (Pierpoint, 1986; Sanderson, 1972). The five
budworm (Hedin and Waage, 1986). major catechins in "green tea" are (+ )-catechin (64) (Fig.
172 Flal'onoids

11.15}, (- }-gallocatechin (68), (+ }-epicatechin (66), (+)- OGLYCOSYLfLAVONES

epigallocatechin (65), and (- }-epigallocatechin (67) (see
Chapter 12). Kaempferol and quercetin glycosides are also Glycosides of flavones in which a linkage through a car-
present. To make black tea, the leaves are fermented and bon-carbon bond involving the anomeric carbon of the sugar
concurrent oxidation of the flavonoid compounds occurs. and position 6, 8, or both, of a flavone are found in many
These phenolic compounds are substrates for oxidation and groups of plants (Chopin et aI., 1982). Approximately 310
condensation into products desirable for tea quality. The an- of these compounds are known (Harborne, 1991). C-Glyco-
thocyanidins cyanidin (3), delphinidin (2), and tricelinidin sides involving glucose, galactose, xylose, arabinose, and
(69) are found in black teas (Fig. 11.22). Two of the major rhamnose have been isolated (Chopin and Dellamonica,
phenolic fractions after fermentation are theaflavins (such as 1988). C-Glycosy1flavones often possess O-glycosides in
70) and thearubigins (such as 71). These polymeric fractions addition to the carbon-carbon-linked sugar moiety. Methods
confer on brewed tea much of its color, palate, and some for the isolation and characterization of C-glycosylflavones
of its odor (Pierpoint, 1986; Sanderson, 1972). Theaflavins have been reviewed (Chopin and Dellarnonica, 1988).
(such as 70) are dimeric compounds containing a 7-memb- C-Glycosyl flavones have been reported from green
ered tropolone ling (Fig. 11.22). The thearubigins (71) are algae, mosses, ferns, and both monocotyledonous and dicot-
polymeric materials which are responsible for much of the yledonous angiosperms (Chopin and Dellarnonica, 1988;
color of black tea (Sanderson, 1972). Markham, 1988); records from algae are questionable

OH OH OH
OH OH
HO
OH

OH OH OH (-j-gallocalechin (68) OH ,~,

OH
OK " (65)'
(+)~epigallocatechin

OH
HO
HO
OH

OH
OH
(+)-eplcatechin (66) (-)-epigallocalechin (67)
OH
OH
OH
HO

HOWO~....""'f ~ OH
I - OH
"" OR OH R=galloyl
OH H a structural component of a

"'
thearubigin (71)
a theaflavin (70) OH
R may be H or galloyl
OH ~OH
HO
yyv,(,~ 0
OH
HO

OH - - ~0-v°
IOH
OH

OH 6' OH
(69) epigallocatechin gallate OH

Fig. 11.22. Phenolic materials in tea.

 Flavonoids 173

(Markham, 1988). This type of flavonoid is most commonly CHALCOl'IES

found in vegetative parts of plants.
Under acidic conditions, interconversion of 6- and 8-sub- Although chalcones, such as (9) (Fig. 11.2), are intennedi-
stituted 5-hydroxy-6-C-glycosylflavones occurs. Attempts ates in the fonnation of all flavonoids, they usually are not
to hydrolyze mixtures of C- and O-glycosides in the labora- accumulated. Most are conspicuously colored yellow com-
tory often result in isomerization of C-glycosylflavones and pounds and, although found in many plant organs, chalcones
mixtures of both 6- and 8-substituted 5-hydroxy-6-C-glyco- are especially common in flowers. These phenolic com-
sylflavones (Fig. 11.14) (Markham, 1989). pounds are encountered frequently in certain ferns, and in
C-Glycosylflavones have been isolated as probing stimu- plants of at least 37 families, including the Acanthaceae,
lants of planthoppers feeding on rice plants, and vicenin-2 Asteraceae, Fabaceae, Gesneriaceae, Liliaceae, Oxalida-
ceae, and Scrophulariaceae. Approximately 20 aglycone
is an oviposition stimulant of Papilio xuthus (Chopin and
structures with varied patterns of hydroxylation and, in some
Dellamonica, 1988). Vitexin deters feeding of Schizaphis
cases, methylation and prenylation, are known (Bohm, 1975,
graminum and Myzus persicae on wheat (Chopin and Della-
1982, 1988). Although many chalcones occur as glycosides,
monica, 1988). Maysin (71) (Fig. 11.14), a C-glycoside from
the majority are found as free aglycones. Chalcones are iso-
com, Zea mays, is toxic to the com earworrn (Waiss et 31.,
merized to flavanones in plants by the enzyme chalcone iso-
1979).
merase, but are readily isomerized in vitro in the presence
of acid (see the earlier discussion of flavonoid biosynthesis).
When the chalcone is 2',6'-dihydroxylated (the numbering
system of chalcones is distinct from that of other flavonoids
BIFLAVOI'lOIDS
that have a C-ring system), the isomeric flavanone bears a
5-hydroxy group. The stabilizing influence of 4-carbonyl-5-
The majority of naturally occurring biflavonoids are flavone hydroxy hydrogen-bonding causes the chalcone-flavanone
and flavanone dimers with a simple 5,7-4'- or, less com- equilibrium to lie largely on the side of the flavanone. Thus,
monly, a 5,7,3',4'-oxygenation pattem. The monomers may chalcones that occur in nature usually possess 2',4'-hydroxyl
be of the same or of different structural types (Geiger and groups or 2'-hydroxy-6'-glycosyloxy groups (Fig. 11.9).
Quinn, 1988; Williams and Harbome, 1989a). The petals of flowers of Oenothera hookeri ssp. venusta
The fonnation of biflavonoids may be explained in tenns contain only carotenoids in the distal ends, but the basal
of oxidative coupling of two chalcone units and subsequent portions contain the chalcone isosalipurposide (16) (Dement
modification of the central C3 units (Fig. 11.23) (Geiger and and Raven, 1974).
Quinn, 1975).
Biflavonoids are characteristic of gymnospenns (includ-
AUROl'lES
ing several cycads and Ginkgo bi/oba), the Psilotales (Psi/o-
tum), and Selaginallales (Selaginella), but have been identi- Aurones, (such as sulfuretin, 73) represent a small group
fied from six moss species (Williams and Harborne, 1989a), of conspicuously colored pigments that are often, but not
including as species of Dicranum and two ferns, Osmunda exclusively, found in flowers. Many of these compounds are
japonica and Cyathea spinulosa (Geiger and Quinn, 1988; golden-yellow in color. Aurones also are encountered in
Williams and Harborne, 1989a) and several flowering plants. bark, wood, or leaves of certain plants. These compounds
Biflavonoids are not found in the Pinaceae or the Gnetales. are isomeric with flavones (Fig. 11.5). They possess Z-ster-
The distribution of biflavonoids is limited and disjunct in eochemistry at the double bond.
angiospenns, these compounds being found in 15 families, Aurones are found in at least 10 plant families but are
including the Anacardiaceae, Caprifoliaceae, Casuarinaceae most common in those plants with yellow flowers; they are
(Casuarina), Euphorbiaceae, Clusiaceae (Guttiferae or Hyp- especially frequent in members of the Asteraceae, Gesneria-
ericaceae), Ochnaceae, and Rharnnaceae (Geiger and Quinn, ceae, and Scrophulariaceae (see tables in Bohm, 1982,
1982). 1888). Most, but not all, aurones occur as glycosides in the
Biflavonoids accumulate in large amounts in a number plant.
of plants. Their most important functions appear to be as Chalcones and aurones collectively are sometimes called
fungitoxins and insect feeding deterrents. For example, anthochlors (Harbone, 1991). Techniques for the isolation
amentoflavone (72) 4'"-mono and 7",4'"-dimethyl ethers have and characterization of chalcones and aurones have been
been shown to be effective against insects browsing on reviewed (Bohm, 1988, 1989).
leaves of Decussocarpus gracilior (Podocarpaceae) and
amentoflavone has been shown to inhibit the growth of sev- DIHYDKOCHALCOl'IES
eral fungi (Fig. 11.24) (Williams and Harborne, 1989a).
Techniques for isolation and analysis of biflavonoids The approximately 75 known dihydrochalcones are accumu-
have been reviewed (Geiger and Quinn, 1988; Williams and lated in at least 20 families, but are especially common in
Harborne, 1989a). the Asteraceae and Fabaceae (Bohm, 1988). Compounds of
174 Flal'onoids

HO~OH
1 1
'<::: OH -O~H OH
~ '<:::
0yY0H NOH
'" "',&

HO 0
- ",I '" 1,& _
HO 0 ~
HO 0 b
naringenin chalcone (9)

(a)

&omc+d :qfX0H
HOrOH r OH 10H
'(YVD
I
,.. (1.0 0no H
HO '? I
0 ~
0 7
-----+- ochnaflavone
group biOavones

~'RW",H'"
000 1 1 _ ~ I 0 00

W
'" '" OH 0 ~ 1 '"
0 0 0 , &

I:
o OH

fromc+b HO 9" 0

HO~H
l O~-WO
r~I r 0 OH ",OHHO 0 ",I
OHOhlnokillavone
~ ~ ~ I::... 1 h r 1 - groupbiftavonea
OHO::'" ~ ",OH
OHO OHO

fromd+b
HO

(b)

Fig. 11.23 (8 & b). Proposed biosynthesis of biflavonoids.

this type have been reported from a fungus (Phallus impu- Phloridzin (74) was first found in 1835. This dihydrochal-
dicus), a liverwort (Radula variabilis), several ferns (Pityro- cone occurs in the bark of apple trees, apparently confers
gramma, Notholaena, and Adiantum), a conifer (Podocar- disease resistance on the plants, and has been involved in
pus nubigena), and from 17 angiosperm families (Grayer, some a1lelopathic problems. Phloridzin is a feeding stimu-
1989). lant to some insects (Stadler, 1986). When ingested by hu-
Dihydrochalcones [e.g., (12) (Fig. 11.2)] are derived from mans, phloridzin impairs reabsorption of glucose by the kid-
chalcones by reduction of the a,l3 double bond. As in the neys and produces a diabetes-like condition (Le., excess
case of chalcones, the A ring is derived from acetate and sugar in the blood and urine) (McClure, 1975).
has a phloroglucinol oxidation pattern. Similarly, the B ring Although most phytoalexins in the Fabaceae are of isofla-
is derived from a phenylpropanoid precursor. These com- vonoid origin, a-hydroxydihydrochalcone (75), odoratol
pounds are comparatively rare; they occur as both aglycones (76), and methylodoratol (77) are formed de novo as stress
and as glycosides. Dihydrochalcones are colorless and are compounds in the pods and cotyledons of Lathyrus odoratus
not easily detected (Fig. 11.25). (Fuchs et aI., 1984).
 Flavonoids 175

The dihydrochalcone ceratiolin (78), from the foliage of F1avanones are accumulated sporadically but they are
Ceratiola ericoides (Empetraceae), has little phytotoxic ac- most commonly encountered in the Fabaceae and Rosaceae
tivity, but decomposes slowly to yield hydrocinnamic acid (see table in Bohm, 1982).
and other products which are inhibitory to seed germination In the genus Citrus (Rutaceae), a balance between bitter-
and radicle growth of test organisms (Tanrisever et al., ness and palatability is important. Naringin (61) (Fig. 11.25),
1987). one-fifth as bitter as quinine, produces an intensely bitter
taste in some grapefruits (Citrus paradisii). Naringenin 7-
rutinoside (79), an isomer of naringin, possesses a I - 6
FLAVANONES linkage of the sugar to the aglycone instead of a I - 2
linkage. Curiously, naringenin 7-rutinoside (79) has no taste.
Conversion of naringin to the corresponding dihydrochal-
F1avanones are widely distributed (at least 60 families) but
cone (80) yields a compound 500 times sweeter than sucrose
they are accumulated in few plants (Bohm, 1982, 1988).
These flavonoids have a center of asymmetry at C-2 (see (Fig. 11.25) (Harborne, 1988c). The dihydroflavonol, dihy-
earlier discussion under flavone and flavonol biosynthesis). droquercetin 3-acetate (81), from the young shoots ofTessa-
The absolute configuration of a number of these compounds ria dodoneifolia (Asteraceae) is 80 times sweeter than su-
has been established. Dihydroflavonols, or 3-hydroxyflava- crose, whereas (+ )-dihydroquercetin (18) was devoid of
nones, that have two asymmetric carbons, C-2 and C-3, also sweetoess (Nanayakkara et al., 1988).
are intermediates in the synthesis of several major types of Many flavanones and dihydroflavonols have fungistatic
flavonoids such as flavonols and anthocyanins (see above). or fungitoxic properties. Naringenin has been established as
Although the majority of dihydroflavonols possess (2R,3R)- a growth inhibitor in dormant peach flowers. Eriodictyol
stereochemistry, compounds with (2S,3S)-stereochemistry (17), dihydroquercetin (18), and dihydroquercetin 3-0-
are known. rhamnoside inhibited growth in Helicoverpa zea larvae when

5'

KO OK

~x;x:P"
amentotlavone (72) OK

KO#' :1' ~I ,I
~ I I
hinokiftavone
OK 0

OR

glnkgetin R = K OK 0
scladopitysin R = CK,

FIg. 11.14. Some representative biflavonoids.

 W WI:
176 Ffal'onoids

OH OH

rhamnosyl-O-g1ucosyl-O 9" ° I: rhamnosyl-O-glucosyl-O 9" °

.,1 ....2 I .,1-+6 1
"'-
"'-
OH ° OH °

y:;P i fI:
oaringin (61)

I: 0H
naringenin 7-rutinoside (79)

OCH3
rhamnosyl-O-glucosyl-O 9" OH
OH
.,1~ 1
rhamnosyl-O-glucosyl-O
9" OH
"'- .,1 __2 "'- 1
OH ° OH °
naringin dibydrocbalcone (SO) neobesperidin dibydrocbalcone

OH OH
OH
HO
HO

OH

W
(a) eriodictyol (17) dihydrochalcone (U) pbloridzin (74)

OH

H0w»oH
9"1 I"" HO
OH
HO
OH
° 1 ""
.&
OH
HO

"'- .& 1
.&
HO ° OH °
a-bydroxydibydrochalcone (75) ceratiolin (78) dibydroquercetin 3-acetate (81)

odoratol (76) metbylodoratol (77)

(b)

Fig. 11.25 (a & b). Some bioactive dibydrocbalcones and flavanones (modified in part from Harbome, 1982).

added to artificial diets, but naringeniu (10), naringin (61), (with male and female flowers on separate plants) and self-
hesperetiu, and neohesperidiu did not (Elliger et aI., 1980). iucompatible plants. When iusects, bats, or birds visit flow-
ers to feed or collect pollen or nectar, they usually pollinate
the plant and both participants benefit from the iuteraction.
TIlE KOLE OF FLAVONOmS IN POLLINATION Compounds that mediate iuteractions of this type are called
AND PKOPAGULE DISPERSAL JllECHANlSJIIS synomones. These compounds may provide scent and odor
or color, or they may contribute to the nutritional value of
Although many plants are pollinated by wiud or mechanical the pollen and/or nectar (Harbome, 1988c). Animals other
agents, the majority of angiosperms require outside agents than pollinators may visit flowers as well. Many come to
for pollination. This is especially true of dioecious plants "steal" nectar and pollen. Ants are notorious nectar thieves
 Flavonoids 177

that are small enough to enter and leave flowers without often are produced by the presence of pure peonidin. Copig-
touchiog the reproductive organs, but they may inadvertently mentation with flavones and complexation with metals cou-
be pollioators, pled with mytiad other effects alters the color of anthocyanin
Animals live in a world of chemical communication and pigments in flowers (Harborne, 1988c).
receive both olfactory and visual cues that iodicate appropri- The color of many yellow flowers is due to the presence
ate host plants. When the animal has located the flower of carotenoids, although chalcones and aurones are responsi-
against the generally green background, it may be attracted ble for the color in others, especially those of the family
to the nectar by nectar guides on the petals. These are derived Asteraceae. Orange flowers are produced by carotenoids
from differential distribution of pigments withio the flower alone, or by pelargouidins and aurones (Harborne, (1988c).
tissue. The biochemistry of plant pollination mechanisms Flowers that appear white or cream colored to the human
has not been studied extensively. eye often contain pigments that bees and other pollinators
In general, pollinators visit one or a limited number of can see but humans cannot. Many of these pigments are
species at one time. Such fidelity is guaranteed by the plant's flavonols and flavones (both aglycones and glycosides)
morphology, odor, and color. Olfactory cues will be dis- (Harborne, 1988c).
cussed later under terpenes (Chapter 19). Many plants are Pelargonidio (1), which occurs often io orange-red flow-
pollinated only by one vector and are characteristically ers, is common among trupical plants, but rarely found in
known as "bee flowers" or butterfly flowers or hummiog- temperate ones. Many of the plants with pelargonidin are
bird flowers. This mutual coadaptation has many benefits hummingbird pollinated. In the West Indies, where there
for both the plant and the animal. are many hummiogbirds, approximately 17% of the flowers
In general, plants pollioated by bats are white to drab have pelargonidio, whereas io Australia, where humming-
(pale purple to green), but bats and certain other pollioators birds are lackiog, only 2% of the plants have this pigment
are not especially sensitive to color. Flowers attractive to (Harborne, 1988c). Other types of bird pollinators are pres-
bees usually are yellow or blue, but some are white. Al- ent io both Africa and Australia, however.
though bees apparently cannot see red, some bee-pollioated Delphinidio (2) and its derivatives are found frequently
flowers are red. Bees and some other iosects can see a portion io bee-pollioated plants and are typical of families such as
of the ultraviolet spectrum not seen by humans. Beetle-polli- the Boraginaceae, Hydrophyllaceae, Polemoniaceae, and
nated flowers are usually cream to greenish; many beetles Scrophulariaceae. Sometimes the color of flowers of a spe-
appear to be color blind. In general, bird-pollioated flowers cies apparently varies in response to the type of pollioator
are scarlet or have bicolors, such as red and yellow. Butter- present. As an example, one species of Penstemon, a blue,
flies tend to prefer vivid colors, iocludiog reds and purples, carpenter bee-pollinated species, hybridizes naturally with
whereas nocturnal moths usually pollinate white or pale piok a red, hummiogbird-pollioated species to produce a purple-
flowers. Flowers pollinated by flies are generally brown, flowered hybrid. Although normally this hybrid would fall
purple, or green, some with checkered patterns. Similar color outside the limits that the pollioators of the parental species
preferences are observed for wasps (Harborne, 1988c). The would pollinate, this purple flowered hybrid is, fortuitously,
color preferences of many other pollinators have not been pollioated by a wasp (Harborne, 1988c).
studied. Honey or nectar guides are part of the pigmentation of
Flower color is largely due to the presence of pigments many flowers, but these features are particnlarly common
present io cbromoplasts or cell vacuoles of floral tissues. The in bee flowers.
colors produced by refraction of light are not as important io Many floral features are not visible to the human eye, but
plants as they are io animals. The most important group of can only be seen in ultraviolet light. These markiogs often
compounds in floral pigmentation is the flavonoids which are produced by local concentration of flavonoid pigments
provide cyanic colors (orange and red to blue) as well as io the petals. In Rudbeckia hirta, the petals are almost uni-
yellow and white. Carotenoids principally provide yellows, formly yellow io daylight and appear yellow to humans io
along with a few oranges and reds. Other less important normal daylight, but, under ultraviolet light, have dark areas
floral and fruit pigments are anthraquinones, betalains, and (ultraviolet nectar guides) at the base. Bees apparently see
chlorophylls. the base of the petals as bright yellow and other parts as
Anthocyanios are especially important in the production purple (Thompson et aI., 1972). Many variations on this
of cyanic colors. Three compounds, cyanidin (3), pelargoni- theme occur io nature. However, bees (and a number of other
din (1), and delphioidio (2), usually occur siogly, or io com- insects) have trichromatic vision and perceive floral patterns
bination, to provide the range of colors from pink, orange, in a more complex way than is visualized (by humans) by
scarlet, and red to violet (mauve), and purple to blue. Factors photography under ultraviolet light. The problems involved
which modify the color of anthocyanios were discussed pre- io visualization of flower colors by honeybees have been
viously. Often a flavone andlor flavonol which forms a weak reviewed and a photographic visualization for humans devel-
complex with the anthocyanidin and shifts the color pro- oped (McCrea and Levy, 1983).
duced is involved. The color of delphioidin is shifted from In the family Polemoniaceae, anthocyanin chemistry cor-
mauve to pure shades of blue by this mechanism. Pink colors relates more strongly with pollination ecology than with tax-
178 Flavonoids

onomic affinities of the species examined (Harbome and netin (82) (Fig. 11.26) (Barz and Welle, 1992). The distribu-
Smith, 1978). tion of radioisotope was demonstrated by degradation. These
experiments show that a phenyl migration takes place in the
course of the biosynthesis, and experiments with chalcone
ISOFLAVOrms glycosides indicate that the aryl rearrangement occurs after
the formation of the C6 -C3-C6 intermediate. An enzyme,
isoflavone synthase, that requires NADPH and molecular
Isoflavones differ structorally from flavones in that the B
oxygen, has been isolated from microsomal preparations of
ring is attached to the 3-position; the spectral properties of
cell suspension cultores of soybean after these cells were
the compounds differ markedly from those of regular fla-
challenged with an elicitor of phytoalexins from Phytophth-
vones as the natore of the conjugated system is altered. Most
of the 234 isoflavone aglycones (out of 630 known isoflavo-
ora megasperma f. sp. glycinea. A microsomal preparation
from elicitor-challenged soybean cell suspension cultores or
noids) are found in the Fabaceae (Dewick, 1988; Williams
soybean seedlings catalyzes an NADPH and dioxygen-de-
and Harbome, 1989b). Many isoflavonoids have marked
physiological effects in biological systems and are often in- pendent rearrangement of (2S)-naringenin (10) to genistein
volved in interactions with other organisms. Isoflavones (5,7,4'-trihydroxyisoflavone) (46) (Barz and Welle, 1992).
commonly occur in the plants as glycosides and many have (2S)-Liquiritigenin (14) was converted to daidzein (83). La-
prenyl groups attached to the phenolic rings. A large number beling stodies confirm that the rearrangement is an intramo-
of other isoflavonoids are derived secondarily from isofla- lecular process (Dewick, 1985; Grisebach, 1985; Heller and
vones. These derivatives typically occur as aglycones and Forkmann, 1988). Rearrangement to the isoflavone seems
only rarely as glycosides (about 48 glycosides have been to take place with the flavanone and not with the chalcone
reported). Isoflavonoids are of considerable interest because (Grisebach, 1985). Two enzymatic steps seem to be involved
of their physiological properties (Williams and Harbome, in the transformation of naringeoin to genistein. The first
I 989b). step comprises oxidation and rearrangement of naringenin
(10) to 2-hydroxy-2,3-dihydrogenistein (84) (Barz and
Welle, 1992). This step requires NADPH, 0,., and a mem-
Biosynthesis
brane-bound cytochrome P-450 monooxygenase. The stable
L-Phenylalanine variously labeled with 1"(; at C-2 and C- 2-hydroxyisoflavanone intermediate is then converted by a
3 and in the carboxyl group is incorporated into formono- soluble dehydratase to genistein (46) (Heller and Forkmann,

80

08

oaring.nln (10) R = 08 (84) genistein (46) R = 08

liquiriligenin (14) R = 8 daidzein (83) R =8

80%",,1°1
80 <?

OC83
° ~ 1
08
formononetin (82) R = H 6,7,4'.trihydroxyisonavone (85)
blocbanin A (112) R =08

08

luteone (86) wighteon. (87)

Fig. 11.26. Proposed biosynthesis of isoflavones (modified from Kochs and Grisebach, 1986) and some representative isoflavones.
 F/lII'olioids 179

1988). A radical mechanism seems to be involved (Barz and The isoflavones luteone (86) and wighteone (87) occur
Welle, 1992). on the leaf surfaces of Lupinus species and appear to be
Many isoflavone derivatives are formed from precursors preformed antifungal defenses; both compounds are fungi-
lacking the 5-hydroxyl group. Studies with "c labeling con- toxic to He/minthosporium cO/'bonum (Harbome, 1986,
firm that loss of the hydroxyl group usually occurs before 1991).
cyclization of the A ring (i.e., at the chalcone stage) (Dewick,
1984). The enzyme chalcone reductase coacts with chalcone Derivatives of lsonavones
synthase and NADPH as a cofactor in the formation of
Several other important groups of secondary metabolites
4,2',4'-trihydroxychalcone (Barz and Welle, 1992).
are derived from isoflavones (Fig. 11.27) and are discussed
An isoflavone O-methyltransferase from elicitor-treated
below. Many of these compounds have pronounced biologi-
cell suspension cultures of Medicago sativa exhibited activ-
cal properties.
ity with 6,7,4'-trihydroxyisoflavone (85), as well as a num-
ber of other isoflavones, isoflavans, and pterocarpans (Ed-
wards and Dixon, 1991). Both a 4'-O-methyltransferase and
COUMESTA1'IS
a 7-0-methyltransferase occur (Barz and Welle, 1992).

Distribution Coumestans, such as (88), which possess a coumarin struc-

ture, are derivatives of isoflavones (Fig. 11.28); most of the
In contrast to other groups of flavonoids, isoflavones 31 known compounds of this structural type are restricted to
mostly are restricted to one group of plants, the subfamily the Fabaceae (Dewick, 1982, 1988; Williams and Harborne,
Papilionoideae of the Fabaceae (Leguminosae), although 1989b). Study of the biosynthesis of coumestans has been
they occur occasionally in the subfamilies Caesalpinioideae accomplished because elicitors stimulate synthesis of these
and Mimoisoidae of the Fabaceae and in the families Ama- compounds in plant cell cultures (Grisebach, 1985). The only
ranthaceae, Asteraceae, Brassicaceae, Chenopodiaceae, Iri- known coumestan glycoside occurs in EcUpta alba (As-
daceae, Menispermaceae, Moraceae, Myristicaceae, Podo- teraceae).
carpaceae, Rosaceae, Scrophulariaceae, Stemonaceae, and Coumestrol (89) from alfalfa, Medicago sativa, and clo-
Zingiberaceae (Dewick, 1982, 1988; Williams and Har- ver, Trifolium repens, is a more active estrogenic compound
borne, 1989b; Wong, 1975). Isoflavonoids also have been than simple isoflavones. Although it is 30 times more active
isolated from Juniperus (Cupressaceae) and Podocarpus than geuistein (43) or formononetin (82), coumestrol occurs
(Podocarpaceae) and the moss, Bryum capi/lare (Dewick, at lower concentrations and has less total effect (McClure,
1988). 1975; Wong, 1975).
Isoflavonoids do not have the same UV spectral patterns
as other flavonoids and often are not observed in routine
studies of plant flavonoids. It is quite possible that isoflavo- ISOFLAV A1'IS
noids will be encountered in additional families. The meth-
ods for isolation and purification of isoflavonoids have been Isoflavans, such as (90), represent the most reduced struc-
reviewed (Dewick, 1988; Williams and Harborne, 1989b). tures of isoflavonoids; about 51 structures have been re-
The best methods for characterization of these compounds ported (Dewick, 1988; Williams and Harborne, 1989b). The
are NMR and MS (mass spectrometry). animal flavonoid metabolite, equol (91), is a representative
of this class of compounds. Equol is formed in animals from
Biological Properties the metabolism of compounds such as formononetin (82).
Most isoflavones are weak estrogens (McClure, 1975; Other isoflavans are known to occur in plants; for example,
Wong, 1975). The presence of these compounds in forage ( - )-5'-methoxysativan is found in alfalfa foliage (Fig.
legumes, such as subterranean clover (Trifolium subter- 11.29) (Dewick, 1982; Miller et al., 1989).
raneum) and red clover (Trifolium pratense), has been rec- Most phytoalexins from the family Fabaceae are either
ognized as the cause of infertility problems in animals graz- isoflavans or pterocarpans (Barz and Welle, 1992; Dewick,
ing on these plants (Williams and Harborne, 1989b). In years 1982, 1988; Kuc, 1992). More than 400 species of legumes
of good rainfall, the plants are relatively low in isoflavones, bave been found to have the ability to produce phytoalexins
but in years of drought, the concentration is proportionately (Williams and Harborne, 1989b) (see below). (- )-Vestitol
higher. The population size of certain birds (quail) that feed (92) and sativan (94) (Fig. 11.35), isoflavans from Lotus
in pastures rich in these legume seeds may be affected by species, are phytoalexins. (- )-(3R)-Vestito1 from the resis-
the presence of isoflavones. In dry seasons, the birds lay tant pasture legume, Lotus pedunculatus, is a feeding deter-
fewer eggs. rent to larvae of Costelytra zealandica (Coleoptera: Scara-
Two isoprenylated derivatives of ( - )-maackiain from an baeidae), a serious agricultural pest in New Zealand.
unidentified South American plant had high antidote activity Pastures containing perennial rye grass (Lolium perenne)
toward venom of the snake Bothrops atrox (Nakagawa et and as little as 20% Lotus peduncularus were relatively resis-
aI., 1982; Williams and Harborne, 1989b). tant to attack by this insect (Dewick, 1982).
 oc:o
180 Flavonoids

OH

o
1

chalcones
O!o
'"
IOH

o
'"
1 ""
0=60
~1 0

o I'"
h
- ",I
o
HO
I'"
h

0+ / a-methyldeoxybenzoim pterocarpam '-....... 2'-hydroxyisoflavans

0(1 ~ _ ex:lt ~_
~- -~Q -LJ)
* "'-.
h

%0 0100
0
isoflavones isoflavanones dehYdroPterocarpan~ 6a-hydroxypterocarpans

o
CJ(j(~
-- K U ~ Jr
",I
O
HO""
1
1 '"
-+ ",I
oH O
I'
"
h
",I

0
""
1 '"
h
2-hydroxyisoflavones 2'-hydroxyisoflavones 2'-hydroxyOavanones coumestans

Jf ~ ~ o
o 1 OCH'_~IO
0
""/-"..--.,, r '" r
"I 0
I "I

--
HO

3-aryl-4-hydroxycoumarins coumaranochromones
2 1-methoxyisoflavones
0
rotenoids
0
+

dehydrorotenoids 12a-hydroxyrotenoids

Fig. 11.27. Major groups of compounds derived from isoflavone precursors (modified from Wong, 1975;

HO~"

-<
1 0 1 _ _H o1U
0 1f u "
OH '" 1 '"
HO 0 "IOH °HO ""OH

daidzein (83) J~ (103)

o
isoliquiritigenin (15) HO%O
:-...1 HO~O
----...~I

o "" I 0 '"
1 ""
OH HO OH

coumestan (8S) 10

CH,O

wedelolactone trifolio. medicagol

Fig. 11.28. Probable biosynthetic pathways to coumestrol and some representative coumestans (modified from Haslam. 1974).
 Flavonoids 181

HO
HO~O HO 0

~ I
""0 aVl~
H

~oH~~D
""

OH OH
OCH,

(3S)-isoflavan (90) equol (91) lonchocarpin

HO~IO ~
HOSO
I
CH,-O

;7 I ~ ;7 OCH,

HO ~ OCH, '>- I HO
CH,-O OCH,
HO OH

vestitol (92) (-)-S'-methoxysativan (93) licoricidin

Fig. 11.29. Selected isoflavans.

PTEROCARPANS co-occurring 3'-hydroxylases) are NADPH-dependent mi-

crosomal monooxygenases (Barz and Welle, 1992; Stafford,
The second largest group of isoflavonoids is the pterocarpans 1991). A flexible isoflavanone intermediate, rather than an
[e.g., (95)]. Approximately 139 compounds have been re- isoflavone, may be required for formation of a fused furan
ported (Williams and Harborne, 1989b). The numbering sys- ring (Stafford, 1991). In eicer arietinum, the (6aR; llaR)
tem of plerocarpans is different from that of isoflavones (Fig. pterocarpans ( - )-medicarpin (96) and ( - )-maackiain (97)
11.30). (Fig. 11.31) arise from formononetin (82) by 2'- oxidation
As is true for coumestans and other related compounds, (or 5f -oxidation in some isomers) followed by conversion
elicitation often stimulates the production of pterocarpans to the corresponding isoflavanones vestitone (98) and 'i'-
in plant cell cultures and facilitates biosynthetic studies baptigenin (99) (Barz and Welle, 1992; Dewick, 1989). The
(Grisebach, 1985). enzymes that catalyze these reductions require NADPH and
Oxygenation at the 2'-position of isoflavones is requisite are specific for 2' -hydroxyisoflavones. The entire enzyme
for formation of pterocarpans. 2'-Hydroxylases (as well as system is specific for 4'-O-methylisoflavones, such as (82)

o H

o
)
o
H-(6aR,llaR)'pterocarpan (95) preferred configuration (94) (-)·pisatin (101)
CH,-O o
o
CH3-0~1
0 CH,-O p

~.o ~

I~
o ..0
OCH, OCH,

6a-hydroxyphaseollin a dimethoxy- 6a,lla-dehydropterocarpan variabilin

Fig. 11.30. Some common pterocarpans.

182 Flavonoids

HO
NADPH
Hr OI I%
r 0, '" r OH

HO 0 !
NADPHIO ".. I o "..1

HO%
0, OCH, OCH,
caJycosin
formononetin (82)

,:>' I 1

'" °
OCH, 'P
° :
-baptigenin (99) ! 1
°>
O HO%IOI
HO% H I
'" '" °
to 1° : >
7
o ".. I I
NADPH HO 0
.estitone (98) OCH, (U3)

HO 0
H°)X(01
'"
°H °
/
°HO ~ 7 1
0
>
OCH, HO
(-)-medicarpin (96)

°;;
(-)-maackiain (97)

Fig. 11.31. Biosynthesis of (- )-maackiain (Dewick, 1989; modified and used with pennission of the copyright owner, the Royal Society of Chemistry,
Cambridge).

(Barz and Welle, 1992). In both instances, an NADPH: this hydroxyl present (kievitone, 106) leads to six acetate
isoflavone oxidoreductase reduces a 2'- or 5'-hydroxyisofla- 13C couplings (Fig. 11.32) (Dewick, 1984).
vone to the corresponding isoflavanone. This reaction is fol- Seedlings and pods of Glycine max, the soy bean, synthe-
lowed by the fonoation of pterocarpans catalyzed by ptero- size three major phytoalexins, glyceollin I, II, and III
carpan synthase. The pathways in soybean, pea (pisatin), and (107-109). These compounds have been shown to share a
alfalfa (medicarpin) all reflect a high degree of homology common pathway with phaseollin as far as the pterocarpan
(Barz and Welle, 1992). inteonediate (105). Glycinol (110) was incorporated into all
Pterocarpans [e.g., (95)] occur primarily in legumes, but three glyceollins. Prenylation occurs late in the scheme (Fig.
are widely distributed in that family (Barz and Welle, 1992; 11.33) (Dewick, 1984). Most of these enzymes are mem-
Dewick, 1982, 1988). These compounds generally occur in brane-bound cytochrome P-450 monooxygenases. A prenyl-
the free state, although some glucosides are known (Fig. transferase enzyme has been isolated and studied (Welle and
11.30). Grisebach, 1991).
The biosyntheses of phaseollin (100) and (- )-pisatin A similar series of feeding experiments with Pisum sari-
(101) have been examined and pathways proposed (Figs. vum, pea, indicated that isoliquiritigenin (15), isoflavones
11.32 and 11.33) (Dewick, 1984). In the biosynthesis of (83, 99, 111-113), and (-)- and (+ )-maackiain (97, 114)
phaseollin, the chalcone, isoliquiritigenin (15), daidzein (but not methyl ethers of 83, 97, or 114) were incorporated.
(83), and (103), and the pterocarpans (105) and phaseollini- ( + )-6a-Hydroxymaackiain (115) also proved to be an effec-
din (104) were all incorporated, suggesting that these com- tive precursor (Fig. 11.33) (Dewick, 1984).
pounds may represent a logical sequence leading to phaseol- An additional enzyme, 6a-hydroxylase, is responsible for
lin (100) (Dewick, 1984). Furthenoore, use of 13C-Iabeled the foonation of many pterocarpan derivatives (Barz and
acetate in cell cultures of Glycyrrhiza echinara yielded only Welle, 1992; Heller and Forkmann, 1988). 6a-Hydroxypter-
three pairs of acetate couplings in the product phaseollin. ocarpans [such as (- )-pisatin (101) and 6a,lla-dehydropt-
This indicates that loss of the 5-hydroxyl group occurs early erocarpan derivatives are well-known antifungal compounds
in the biosynthesis. Biosynthesis of similar compounds with (see the section on phytoalexins, below).
 FllII'CJl/oitis 183

(- )-Medicarpin (96) and methoxymedicarpin, from al- of fonnation of the phytoalexin. The defense mechanism is
falfa roots, Medicago .\'CI/il'll, inhibit seed gennination (at confined to the tissue colonized by the fungus and its imme-
10- 1 and 10-" M, respectively) and growth of alfalfa seed- diate neighborhood. The resistant state is not inherited; it is
lings (Cutler, 1992; Miller et aI., 1988). Medicarpin occurs developed after the fungus has attempted infection (Brooks
in alfalfa roots as the 3-0-glucoside, medicocarpin. and Watson, 1985; Harborne, 1988c). The first phytoalexin
to be chemically characterized was ( - )-pisatin (101), a pter-
ocarpan, from Pisum sativum. This compound was induced
PHYTOALEXlI'IS by the presence of conidia of the brown rot fungus, Monilinia
fructicola. Plants of Phaseolus vulgaris also produce a ptero-
Millier and Borger (1941) defined phytoalexins as com- carpan phytoalexin, phaseollin (100).
pounds that inhibited the growth of fungi, but emphasized Most of the phytoalexins studied to date are from the
that these bioactive compounds were not produced or acti- Fabaceae and the Solanaceae (Kuc, 1992), a fact that proba-
vated until the host came into contact with the parasite. This bly reflects the economic importance of these families but
defensive reaction occurs only in living cells. The inhibitory may also reflect the stability and ease of isolation and charac-
compound(s) is a discrete chemical substance, a product of terization of the phytoalexins. At least 200 phytoalexins are
the host cell. Most phytoalexins inhibit growth of fungi at known (Brooks and Watson, 1985). Although it is widely
10- 3 _10- 5 M (Kuc, 1992). The phytoalexin is nonspecific accepted that phytoalexins playa major role in disease resis-
in its action toward fungi; however, fungal species may be tance, their role in disease resistance is less well understood
differentially sensitive to it. The basic response in both resis- than their production and metabolism (Dixon et aI., 1983;
tant and susceptible cells is the same, the basis of differentia- Kuc, 1992). Interestingly, in some susceptible reactions be-
tion between resistant and susceptible hosts being the rate tween fungi and plants, phytoalexins accumulate to higher

H0'f"l(0H("yOHHO~O HO~O
~",II
0: I----. 0: I
",11
o --+ ----+
OH HO OH
isollquiritigenin (15) daidzein (83)
(103)

H0 0 Cfu
'O
"I H _ _HO
,....
~O " I OH OH

HO
HO

! OH
glycinol (110)

OH
HO

HO

OH
glyeeollin m (109) glyeeollin I (107)

Fig. 11.32. Proposed biosynthesis of phaseollin and glyceollins I-m (Dewick. 1984; modified and used with pennission of the copyright owner, the
Royal Society of Chemistty, Cambridge),
184 Flavonoids

HO~ HO
: 1 1
"
°HO" 1
........
OCH, OCH,

H0'f"lr0H NoH HO%j HO (~_):6aR~IaR)_medicarpin (96)

~ "I I" "I 1 OH
o ---+ 0 ~I ----.
OCH,
0:'-' OCH
isoUquiritigenin (15) rormononelin (83) (Ill) ,

HO~O.
HO
'" 1
°1 trans HO
reduction
°
Ufu' H
~I I ~ ~ ~ ~ 0
° ° °
:;..0"

",-baptlgenin (99)
"I>
"o/..
/red:~On?
HO
(113)
"I>
° *
HO
"I>
°
H0I""'Ir 0 , HO HO
~H
" °OO~O
~O>- °J °J
(+)-(6a5,llaS)-maackiain (114)
HO
/ HO

°)
°
CH,O CH,O

°) °>
(+)-(6aR,l1aR)-pisatin (102) ° (-)-pisatin (101) °
Fig. 11.33. Proposed biosynthetic pathways to pisatio (Dewick. 1984; modified and used with pennission of the copyright owner, the Royal Society of
Chemistry. Cambridge).

concentrations than in resistant interactions. This effect Several phytoalexins are known to inhibit the growth of in-
probably is due to the timing of production of the fungitoxic sect larvae and nematodes (Dewick, 1982).
compounds (Kuc, 1992). The largest group of phytoalexins studied to date are pter-
The ability of cell cultures from many different species ocarpans and related isoflavones and isoflavans from the
of plants to produce flavonoid phytoalexins upon induction Fabaceae (Fig. 11.34) (Friend, 1979; Harborne, 1982,
with elicitors has been of major importance in elucidation 1988c). For example, the leaves of Anthy/lis vulneraria and
of the mechanisms and control of biosynthesis (Ellis, 1988). five Tetragonolobus species inoculated with Helminthospor-
The amounts of mRNA for chalcone synthase and PAL in ium carbonum produce 7,4'-dihydroxy-2'-methoxyisoflavan
[isovestitol (116)] and 7,2',4'-trihydroxyisoflavan [demeth-
Phaseolus vulgaris suspension culture cells increase within
ylvestitol (117)]. Those of Lotus corniculatus produce de-
2 h of treatment of the cells with an elicitor made from
methylvestitol (117), vestitol (92) and sativan (94). Lotus
Colletotrichum lindemuthianum cell walls. The amount of
uliginosus produces demethylvestitol (117) and vestitol (92).
translatable mRNA for these enzymes reaches a maximum The leaves of the European licorice (Glycyrrhiza glabra)
4 h after elicitor treatment. In the case of chalcone synthase produce 7,2'-dihydroxy-3,4'-dimethoxyisoflavan. Members
mRNA, this is probably due to in vitro transcription. In elici- of the genus Trigonella produce a variety of isoflavans or
tor-treated cells, 50 proteins increased and 10 decreased pterocarpan phytoalexins. Seedlings of soybean (Glycine
when compared to untreated cells (Kuhn, 1987). max) produce the pterocarpan phytoalexins, glyceollins I-III
In some cases, conditions other than fungal attack can (107-109) (Barz and Welle, 1992; Friend, 1979). Normal
induce the formation of phytoalexins. Bacterial, viral, insect, tissues accumulate mostly daidzein (83) and genistein (46).
or nematode attack, stress, and so forth may also do so. Chickpea plants and cell cultures synthesize medicarpin and
 Flavonoids 185

HO

HO CH,-O

isoflavone OCH, o>

Cajanus
(Lupinus, preinfectional) pisatin (101) pbaseollin (100)

HO

-OCH,
HO~IO HO~IO
"'"
o
I~
h
-"'"
OCH, HO
I ~
h
OCH,

isoflavanone pterocarpan isoflavan

Cajanus Baptisia, Cicer, Melilotus, Loteae:
Pisum, many genera Anthyllis, Lotus, Tetragonolobus

Both types found in: Medicago, Trifolium, Trigonella

Fig. 11.34. Phytoalexin phyletic sequence (modified from Harhorne, 1988c);

maael"aln (97) medicarpin (96) isomucronuIatol

OH
glyceollin n (108) glyceoltin m (109)
HO

HO
o
)
o

kievitone hydrate (106) 6a-hydroxymedicarpin R=H (118) 6a-hydroxymaackiain R=H (115)

6a,7-dihydroxymedlcarpin R=OH (119) 6a,1-dihydroxymaackiain (120)

CH,O HO

HO'(}(°l,;

~ CH,O
r~J~) OH
(-)-pisatin (101)
lsoveslitol (116)

HO~"
I ° H O a 01 ~ HO~"
I 0

HO"
~ I OH
~ :00
HO OCH, CH,O
:1 OCH,

demetbylveslitol (117) vestitol (92) sativan (94)

Fig. 11.35. Representative isoflavone-derived phytoalexins from the Fabaceae.
186 Flavonoids

maackiain in response to the fungus Ascochyta rabiei. Nor- hydroxylated derivatives 6a-hydroxymedicarpin (118), 6a,
mal tissues primarily accumulate formononetin (82) and bio- 7-dihydroxymedicarpin (119), 6a-hydroxymaackiain (115),
chanin A (112) (Fig. 11.35) in the vacuoles, mostly as the and 6a,7-dihydroxymaackiain (120) when challenged with
7-0-glucosides-6"-malonate derivatives (Barz and Welle, Colletotrichum coffeanum, a nonpathogen (Friend, 1979).
1992). In Cicer cell suspension cultures, labeled formono- Degradation of biochanin A (112) from chickpea with Fu-
netin was shown to be quantitatively converted to pterocar- sarium javanicum involves formation of dihydrobiochanin
pans upon elicitation and PAL inhibition, strongly suggest- A, 8-hydroxydihydrobiochanin A, and a dioxygenase cleav-
ing that these isoflavones serve as precursors to phytoalexins age to a diketodihydropyran (Barz and Welle, 1992).
and that de novo flavonoid biosynthesis is not required (Barz The microbial transformations of isoflavonoids have been
and Welle, 1992). reviewed (Dewick, 1988).
Resistance to fungal atrack has been rationalized by the
rapid accumulation of phytoalexins. Susceptibility has been
explained by the ability of virulent pathogens to detoxify
these compounds. These detoxification reactions frequently KOTEI'IOIDS
produce a more hydrophilic product. The susceptibility of
French bean hypocotyls to Fusarium solani f. sp. phasiolli About 60 isoflavone derivatives that incorporate a prenyl
has been partly explained by the metabolism of the phyto- group into the structure are insecticidal and piscicidal (De-
alexin kievitone (106) by the fungus. Although the phyto- wick, 1988; Harborne, 1991). Plants that contain rotenone
alexin initially accumulates, it is rapidly converted to (121) (Derris and Lonchocarpus spp.) have long been used
phaseollin (100), 2'-methoxyphaseollin-flavan, and phaseol- as natural insecticides and piscicides. These compounds are
linisoflavan. All of these compounds accumulate when the best known from the fabaceous genera Amorpha, Derris,
hypocotyl is completely infected. l-Hydroxyphaseollone, Lonchocarpus, Milletia, Mundulea, and Tephrosia (Dewick,
the detoxification product of phaseollin (100), also was pres- 1988; Harborne, 1991).
ent (Friend, 1979). Rotenoids are derived from isoflavones and occur in the
Both pathogenic and nonpathogenic fungi may detoxify same plants that have isoflavones (Fig. 11.36) (Dewick,
phytoalexins either in vivo or in vitro (Barz and Welle, 1992; 1982). The cis-B/C ring coupling of rotenone was estab-
Friend, 1979; Ku6, 1992). Red clover (Trifolium pratense) lished by NMR spectroscopy (Carlson et al., 1973). Prenyla-
produces primarily, medicarpin (96) and maackiain (97) tion of the isoflavone nucleus is known to occur late in the
[formononetin (82) is an intermediate] when inoculated with biosynthesis of rotenoids. Rotenoids are isoflavanones that
Helminthosporium carbonum, but only the nonfungitoxic have been modified with an "extra" carbon atom. The extra

OH HO

:
~ H 0-"-'
HO 7'

:,... -- • SAM

. .
H,N CO,H
OCH,
o OCH3 •
OCH,
H
HO

.',. .
OCH3
rotenone (121)
OCH,
•

tephrosin (122)

Fig. 11.36. Proposed biosynthesis of rotenone (modified from Crombie. 1984; the Royal Society of Chemistry. Cambridge).
 Ffal'onoids 187

carbon has been shown to come from S-adenosyl methio-

nine.
Rotenoids are effective inhibitors of oxidative phosphory-

~
lation in animal mitochondria; this effect is responsible for
the insecticidal properties of this group of compounds (Wil-
liams and Harborne, 1989b). This effect may occur at con-
R
centrations as low as 24 nmol/g of mitochondrial tissue. The HO OH
lethal dose of rotenone in the silkworm is 0.003 mg/g body I HO~OH ~
weight (Harborne, 1991). This series of compounds also in-
hibits other enzymes (McClure, 1975).
HO
~
The rotenoid tephrasin (121) from Tephrosia elata roots
deters insect feeding activity (Williams and Harborne,
1989b). Although generally considered to have low mamma-
lian toxicity, rotenone-containing roots have been used as a
CH'O~O
'" I
source of arrow poisons in Sumatra (Harborne, 1991). o '" H

Methods for the isolation and purification of rotenoids "'I

have been reviewed (Williams and Harborne, 1989b). Most '"
(R)-4-methoxydalbergione
rotenoids can be characterized by means of their NMR spec-
tra (Carlson et aI., 1973).
CH'O~"'I
0", 0
HO

rmoFLAVOl'lOIDS "'I
"
dalbergin
Neoflavonoids resemble flavonoids in their overall structure R=HorOCH3
and properties and arise by processes similar to those leading

Vt
to isoflavones (Fig. 11.37). These compounds are found in • H CH30 o
'COzH
the Clusiaceae (Hypericaceae), Fabaceae, Passifloraceae,
1 --+
Polygonaceae, and Rubiaceae; several structural types and '" NH, I
subtypes are known (Donnelly, 1975; Donnelly and Sheri-
dan, 1988; Gottlieb et al., 1970). The similarity of oxygena-
tion pattern in the dalbergiones, dalbergiquinols, and the 4-
caliophyllolide (123)
arylcoumarins suggests that they are related. There are many
difficulties in the biosynthetic stndy of heartwood constitn- Fig. 11.37. Biosynthesis of neoflavonoids (Gottlieb et aI., 1970;
ents and the biosynthetic pathway for this group of com- modified and used with permission of the copyright owner, Academia
Brasileira de Ciencias, Rio de Janeiro).
pounds is not entirely clear.
The notable resistance to decay of rosewoods (usually
from the genera Dalbergia and Machaerium) is due to the al., 1970; Markham, 1976, 1982; 1989; Markham et aI.,
presence of several types of phenolic compounds; the neofla- 1978, 1982; Ternai and Markham, 1976). Many flavonoids
vonoids are major among these (Donnelly, 1982; Donnelly can be separated by HPLC (Banwart et aI., 1985; Porter et
and Cannon, 1986). al., 1985, 1986).
In the compound calophyllolide (123) from Calophyllum
inophyllum (Clusiaceae or Hypericaceae), phenylalanine
was incorporated into the 2,3,4-position and the aromatic
REFERENCES
ring attached at the C-4 position. Acetate served as a precur-
sor for the aromatic ring with the phloroglucinol substitntion.
ALsTON, R. E. and B. L. 'fuRNER, Natural hybridization among four
Compounds from the Fabaceae generally have a 6,7-oxygen-
species of Baptisia (Leguminosae), Am. J. Bot., 50, 159-173
ated pattern rather than that of calophyllolide. The possibility
(1963).
of different pathways cannot be excluded.
ALsTON, R. E., B. L. 'fuRNER, R. N. LESTER, and D. HORNE, Cbro-
matographic validation of two morphologically similar hybrids
of different origins, Science, 137. 1048-1050 (1962).
ISOLATIOl'L PURlFICATIOl'l Al'ID ALsTON, R. E., H. ROSLER, K. NAIFEH. and T. J. MABRY, Hybrid
CHARACTERIZATIOl'l OF FLAVOl'lOIDS compounds in natural interspecific hybrids, Proe. NatI. Acad.
Sci.. 54. 1458-1465 (1965).
Techniques for the isolation, purification, and characteriza- ARNTZEN. C. J., S. V. FALKENTHAL. and S. BOBICK, Inhibition of
tion of flavonoids have been reviewed (Hostettmann and photophosphorylation by kaempferol, Plant Physio!. 53.
Hostettmann, 1982; Mabry and Markham, 1975; Mabry et 304-306 (1974).
188 Flavonoids

BANWART, W. L., P. M. PORTER, T. C. GRANATO, and J. J. HASSETT, CODY, V., E. MIDDLETON, JR., and J. B. HARBORNE, (eds.), Plant
HPLC separation and wavelength area ratios of more than 50 Flavonoids and Medicine: Biochemical, Phannacologicai, and
phenolic acids and flavonoids, 1. Chern. Beo!., 11, 383-395 Structure-Activity Relationships, Liss, New York, 1986.
(1985). CODY, V., E. MIDDLETON, JR., J. B. HARBoRNE, and A BEREI'Z
BARRON, D. and R. K. iBRAHIM, Hydrochloric acid and aryl-sul- (eds.), Plant Flavonoids and Medicine II: Biochemical, Cellular,
fatase as reagents for UV-spectral detection of 3- and 4'-sul- and Medicinal Properties, Liss, New York, 1988.
phated flavonoids, Phytochemistry, 27, 2335-2338 (1988). CRANMER, M. F. and T. J. MABRY, The lupine alkaloids of the
BARZ, W. and R. WELLE, Biosynthesis and metabolism of isofla- genus Baptisia, Phytochemistry, 5, 1133-1138 (1966).
vones and pterocarpan phytoalexins in chickpea, soybean, and CROMBIE, L., Rotenoids and their biosynthesis, Nat. Prod. Rep., 1,
phytopathogenic fungi, in Phenolic Metabolism in Plants (H. 3-19 (1984).
A Stafford and R. K.lbrahim, eds.), Recent Advances in Phyto- CUTLER, H. G., Herbicidal compounds from higher plants, in Phyto-
chemistry Vol. 26, 139-164, Plenum Press, New York, 1992. chemical Resources for Medicine and Agriculture (H. N. Nigg
BEIER, R. C. and H. N. NIGG, Natural toxicants in foods, in Phyto- and D. Seigler, eds.), 205-226, Plenum Press, New York, 1992.
chemical Resources for Medicine and Agriculture (H. N. Nigg DEMENT, W. A and P. H. RAVEN, Pigments responsible for ultravi-
and D. Seigler, eds.), 247-367, Plenum Press, New York, 1992. olet patterns in flowers of Oenothera (Onagraceae), Nature
BOHM. B. A .• FIavanones and dihydroflavonols. in The Flavonoids (London), 252, 705-706 (1974).
(J. B. Harborne, T. J. Mabry and H. Mabry,.eds.), 560-631, DEWICK, P. M., Isoflavonoids, in The Flavonoids: Advances in
Academic Press, New York, 1975. Research (J. B. Harborne and T. J. Mabry, eds.), 535-640,
BOHM, B. A., The minor flavonoids, in The Flavonoids: Advances Chapman & Hall, London, 1982.
in Research (J. B. Harborne and T. J. Mabry, eds.), 313-416, DEWICK, P. M., The biosynthesis of shikimate metabolites, Nat.
Chapman & Hall, London, 1982. Prod. Rep., 1, 451-469 (1984).
BOHM, B. A., The minor flavonoids, in The Flavonoids: Advances DEWICK, P. M., The biosynthesis of shikimate metabolites, Nat.
in Research since 1980 (J. B. Harborne, ed.), 329-388, Chap- Prod. Rep., 2, 495-511 (1985).
man & Hall, London, 1988. DEWICK, P. M., Isoflavonoids, in The Flavonoids: Advances in
BOHM, B. A, Chalcones and aurones, in Plant Phenolics (J. B. Research since 1980 (J. B. Harborne, ed.), 125-209, Chap-
Harborne, ed.), Vol. 1 of Methods in Plant Biochemistry (P. man & Hall, London, 1988.
M. Dey and 1. B. Harborne, eds.), 237-282, Academic Press, DEWICK, P. M., The biosynthesis of shikimate metabolites, Nat.
London, 1989. Prod. Rep., 6, 263-290 (1989).
BRITSCH, L. and H. GRISEBACH, Purification and characterization DIXON, R. A, P. M. DEy, and C. J. LAMB, Phytoalexins: Enzymol-
of (2S)-flavanone 3-hydrolase from Petunia hybrida, Eur. J. ogy and molecular biology, Adv. Euzymo!., 55,1-136 (1983).
Biochem., 156, 569-577 (1986).
DoNNELLY, D. M. X., Neoflavonoids, in The Flavonoids (J. B.
BRITSCH, L., W. HELLER, and H. GRISEBACH, Conversion of flava- Harborne, T. J. Mabry and H. Mabry, eds.), 801-865, Academic
none to flavone, dihydroflavonol and flavonol with an enzyme Press, New York, 1975.
system from cell cultures of parsley, Z. Naturforsch., 36c,
DONNELLY, D. M. X., Metabolites of Dalbergia: New structural
742-750 (1981).
variants of the neoflavonoids, in Proc. InternatI. Bioflavonoid
BROOKS, c. J. W. and D. G. WATSON, Phytoalexins, Nat. Prod. Symposium, 1981 (L. Farkas, M. Gabor, F. Kallay, and H. Wag-
Rep., 2, 427-459 (1985). ner, eds.), 263-278, Elsevier, Amsterdam, 1982.
BROUILLARD, R, Flavonoids and flower color, in The Flavonoids: DONNELLY, D. M. X. and P. G. M. CANNON, The synthesis and
Advances in Research since 1980 (J. B. Harborne, ed.), biological activity of the latinones, in Flavonoids and Bioflavo-
525-538, Chapman & Hall, London, 1988. noids, 1985 (L. Farkas, M. Gabor, and F. Kallay, eds.), 1-13,
CALDWELL, M. M., R. ROBBERECHT, and S. D. FLINT, Internal filters: Akademiai Kiad6, Budapest, 1986.
prospects for UV-acclimitation in higher plants, Physio!. Plant., DoNNELLY, D. M. X. and M. H. SHERIDAN, NEOFLAVONOIDS, in
58, 445-450 (1983). The Flavonoids: Advances in Research since 1980 (J. B. Har-
CARLSON, D. G., D. WElSLEDER, and W. H. TALLENT, NMR investi- borne, ed.), 211-232, Chapman & Hall, London, 1988.
gations of rotenoids, Tetrahedron, 29, 2731-2741 (1973). EBEL, J. and K. liAm.BROCK, Biosynthesis, in The Flavonoids: Ad-
CASSADY, J. M., W. M. BAIRD, and C. CHANG, Natural products vances in Research (J. B. Harborne and T. J. Mabry, eds.),
as a source of potential cancer chemotherapeutic and chemo- 641-679, Chapman & Hall, London, 1982.
preventative agents, 1. Nat. Prod., 53, 23-41 (1990). EcHEVERRl, F., G. CARDONA, F. ToRRES, C. I'ELABZ, W. QUINONBS,
CHOPIN, J. and M. L. BoulLLANT, C-Glycosylflavonoids, in The and E. RENwRlA, Ennanin: An insect deterrent flavonoid from
Flavonoids (J. B. Harborne, T. J. Mabry, and H. Mabry, eds.), Passiflorafoetida, Phytochemistry, 30, 153-155 (1991).
632-691, Academic Press, New York, 1975. EDWARDs, R. and R. A DIXON, Isofl.vone O-methyltransferase
CHOPIN, J. and G. DELLAMONICA, C-Glycosylflavonoids, in The activities in elicitor-treated cell suspension cultures of Med-
Flavonoids: Advances in Research since 1980 (J. B. Harborne, icago sativa, Phytochemistry, 30, 2597-2606 (1991).
ed.), 63-97, Chapman & Hall, London, 1988. ELUoER, C. A, B. C. CHAN, and A. C. W AISS, JR., Flavonoids
CHOPIN, J., M. L. BoulLLANT, and E. BESSON, C-Glycosylflavo- as larval growth inhibitors, Naturwissenschaften, 67, 358-360
noids, in The Flavonoids: Advances in Research (J. B. Harborne (1980).
and T. J. Mabry, eds.), 449-503, Chapman & Hall, London, ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod.
1982. Rep., 5, 581-612 (1988).
 Flal'onoids 189

FEENY, P., Chemical constraints on the evolution of swallowtail The Flavonoids (J. B. Harborne, T. J. Mabry, and H. Mabry,
butterflies, in Plant-Animal Interactions. Evolutionary Ecology eds.), 1056-1095, Academic Press, New York, 1975.
in Tropical and Temperate Perspectives (P. W. Price, T. M. HARBORNE, J. B., Recent advances in chemical ecology. Nat. Prod.
Lewinsohn, W. W. Benson, and G. W. Fernandes, eds.), Rep., 4, 323-344 (1986).
315-340, Wiley, New York, 1991.
HARBORNB, J. B. (ed.), The Flavonoids: Advances in Research since
FEENY, P., K. SACHDBV, L. ROSENBERRY, and M. CARTER, Luteolin 1980, Chapman & Hall, London, 1988a.
7-0-(6"-O-malonyl)-~-D-glucoside and trans-chlorogenic acid:
IiARBORNB,1. B., Flavonoids in the environment: Structure-activ-
Oviposition stimulants for the black swallowtail butterfly, Phy-
ity relationships. in Plant Flavonoids and Medicine II: Biochem-
tochemistry, 27, 3439-3448 (1988).
ical, Cellular, aod Medicinal Properties (V. Cody, E. Middleton,
FORKMANN, G. and G. ST01Z, Genetic control of flavanone 3-hy- Jr., J. B. Harborne, and A. Beretz, eds.), 17-27, Liss, New York,
droxylase activity and flavonoid 3'-hydroxylase activity in An- 1988b.
tirrhinum majus (snapdragon), Z. Naturforsch., 36c, 411-416
(1981). HARBORNB, J. B., Ecological Biochemistry, 2nd ed., Academic
Press, New York, 1982; 3rd edition, 1988c.
FRIEND, J., Phenolic substances and plant disease, in Biochemistry
of Plant Phenolics (T. Swain, J. B. Harborne, and C. F. van HARBORNB. J. B., Recent advances in chemical ecology. Nat. Prod.
Sumere, eds.), Recent Advances in Phytochemistry Vol. 12, Rep., 7, 85-109 (1989).
557-588, Plenum Press, New York, 1979. HARBORNE,1. B.. Flavonoid pigments. in Herbivores: Their Interac-
FUCHS, A., F. W. DE VRIES, C. A LANDHEER, and VAN VELDHUIZEN, tions with Secondary Plant Metabolites, Vol. 1 (G. A Rosenthal
Odoratol and methylodoratol, two ex-hydroxydihydrochalcone and M. R. Berenbaum, eds.), 389-429, Academic Press, San
stress metabolites from Lathyrus odoratus, Phytochemistry, 23, Diego, CA, 1991.
2199-2201 (1984). HARBORNB, J. B. and R. J. GRAYER, The anthocyanins, in The Flavo-
GEIGER, H. and C. QUINN, BlFLAVONOIDS, in The Flavonoids (J. B. noids: Advances in Research since 1980 (J. B. Harborne, ed.),
Harborne, T. J. Mabry, and H. Mabry, eds.), 692-742, Aca- 1-20, Chapman & Hall, London, 1988.
demic Press, New York, 1975. HARBORNB, J. B. and T. J. MABRY (eds.), The Flavonoids: Advances
GEIGER, H. and C. QUINN, Biflavonoids, in The Flavonoids: Ad- in Research, Chapman & Hall, London, 1982.
vances in Research (J. B. Harborne and T. J. Mabry, eds.), HARBORNB, J. B. and D. M. SMITH, Correlations between anthocya-
505-534, Chapman & Hall, London, 1982. nin chemistry and pollination ecology in the Polemoniaceae,
GEIGER, H. and C. QUINN, Biflavonoids, in The Flavonoids: Ad- Biochem. Syst. Ecol., 6, 127-130 (1978).
vances in Research since 1980 (J. B. Harborne, ed.), 99-124, HARBORNE, J. B. and B. L. TuRNER, Plant Chemosystematics, Aca-
Chapman & Hall, London, 1988. demic Press, London, 1984.
GBRATS, A. G. M. and C. MARTIN, Flavonoid synthesis in Petunia HARBORNB, J. B. and C. A. WILLIAMS, Flavone and flavonol glyco-
hybrida; genetics and molecular biology of flower color, in sides, in The Flavonoids: Advances in Research O. B. Harborne
Phenolic Metabolism in Plants (H. A Stafford and R K. and T. J. Mabry, eds.), 261-311, Chapman & Hall, London,
Jbrahim, eds.), Recent Advances in Phytochemistry Vol. 26,
1982.
165-199, Plenum Press, New York, 1992.
HARBORNE, J. B. and C. A. Wn..LIAMS, Flavone and flavonol glyco-
GIANNASI, D. E., Flavonoids and evolution in the dicotyledons.
sides, in The Flavonoids: Advances in Research since 1980 (J.
in The Flavonoids: Advances in Research since 1980 (J. B.
B. Harbome, ed.), 303-328, Chapman & Hall, London, 1988.
Harborne, ed.), 479-504, Chapman & Hall, London, 1988.
HARBORNB, J. B., T. J. MABRY, and H. MABRY (eds), The Flavo-
GOTI'LIBB, O. R, S. MAGESWARAN, W. D. OLLIS, R. J. ROBERTS,
noids, Academic Press, New York, 1975.
and I. O. SUTHERLAND, Recent developments in neoflavonoid
chemistry, An. Acad. Brasil. Cienc., 42 (Suppl.), 417-423 HARTWIG, U. A and D. A PmLLlPS, Release and modification of
(1970). nod-gene-inducing flavonoids from alfalfa seeds, Plant Physiol.,
95, 804-807 (1991).
GRAYER, R J., Flavanoids, in Plant Phenolics (J. B. Harborne, ed.),
Vol. 1 of Methods in Plant Biochemistry (p. M. Dey and J. B. HARTWIG, U. A., C. A. MAxwBu., C. M. JOSEPH, and D. A PHlLUPS,
Harborne, eds.), 283-323, Academic Press, London, 1989. Effects of alfalfa nod gene-inducing flavonoids on nodABC
GRISEBACH, H., Biosynthesis of flavonoids, in Biosynthesis and transcription in Rhizobium meliloti strains containing different
Biodegradation of Wood Components (T. Higuchi, ed.), nodD genes, J. Bacteriol., 172, 2793-2773 (1990a).
291-324, Academic Press, Orlando, Fl., 1985. HARTWIG, U. A, C. A MAxwELL, C. M. JOSEPH, and D. A PHlLuPS,
HAHLBROCK, K., FLAVONOlOS, in Secondary Plant Products (E. E. Chryseriol and 1uteolin released from alfalfa seeds induce nod
Conn, ed.), Vol. 7 of The Biochemistry of Plants (p. K. Stumpf genes in Rhizobium meliloli, Plant Physiol., 92, 116-122
and E. E. Conn, eds.), 425-456, Academic Press, New York, (1990b).
1981. HASLAM, E., The Shikimate Pathway, John Wiley & Sons, Ltd"
HAHLBROCK, R. and H. GRISEBACH. Biosynthesis of flavonoids. in Chichester, 1974.
The Flavonoids (J. B. Harborne, T. J. Mabry, and H. Mabry, HEDIN, P. A and S. K. WAAGE, Roles of flavonoids in plant resis-
eds.), 866-915, Academic Press, New York, 1975. tance to insects, in Flavonoids and Medicine: Biochemical,
HARBORNB, J. B., Flavonoids, in Phytochemistry, Vol. 2 (L. P. Pharmacological, and Structure-Activity Relationships (V.
Miller, ed.), 344-380, Van Nostrand-Reinhold, New York, Cody, E. Middleton, Jr., and J. B. Harborne, eds.), 87-100,
1973. Liss, New York, 1986.
HARBORNB, J. B., The biochemical systematics of flavonoids. in HELLER, W. and G. FORKMANN. Biosynthesis, in The Flavonoids:
190 ,..ftll'l!lltlid\

Advances in Rese.m:h siIH..'e 19X() (J. B. Hurborne. ed. ).99-124. LEWIS. D. A.. Antiulcer drugs from plants. Chern. Br.. 141-144
Chapm.m & Hall. London. Il)XX. (1991).
HELLER. W.. L. BRITS{,II. G. FORK\It\:./S. and H. GRISEBACH. Leu- LINDROTH. R. and S. S. PETERSON. Effects of plant phenols on
cmmthocyanidins as intermediates in anthocyanidin biosyn- perfonnance of southern annyworm larvae. Oecologia. 75.
thesis in flowers of Matthio/a illcalltJ. Planta. 163. 191-196 185-189 (1988).
(19850). MABRY, T. J. and K. R. MARKHAM, Mass spectrometry of flavo-
HELLER. W.. G. FORKMANN. L. BRITSCH. and H. GRISEBACH, Enzy- noids, in The Flavonoids (J. B. Harbome, T. J. Mabry. and H.
matic reduction of ( + }-dihydroflavonols to flavan-3,4-ds-diols Mabry, eds.), 78-126, Academic Press, New York, 1975.
with flower extracts from Matthio/a incana and its role in antho- MABRY, T. J. and A UWBELEN, Chemistry and utilization of phen-
cyanin biosynthesis. Planta. 165. 284-287 (l985b). ylpropanoids including flavonoids. coumarins and lignans, J.
HONDA. K.. Flavanone glycosides as oviposition stimulants in a Agric. Food Chern., 28, 188-196 (1980).
papilionid butterfly Papilio prolenor. J. Chem. &01.. 12, MABRY, T. 1., K. R. MARKHAM, and M. B. THOMAS, The Systematic
1999-2010 (1986). Identification of Flavonoid., Springer-Verlag, Berlin, 1970.
HasEL. W.o Glycosylation and glycosidases, in Secondary Plant MARKHAM, K. R, 13C_NMR offlavonoids II. Flavonoids other than
Products (E. E. Conn. ed.). Vol. 7 of The Biochemistry of Plants flavone and flavonol aglycones, Tetrahedron, 32. 2607-2612
(P. K. Stumpf and E. E. Conn. eds.). 725-753. Academic Press. (1976).
New York. 1981.
MARKHAM, K. R, Techniques of Flavonoid Identification, Aca-
HOSTETIMANN. K. and M. HOSTE'ITMANN, Isolation techniques for
demic Press, London, 1982.
flavonoids, in The Flavonoids: Advances in Research (J. B.
Harbome and T. J. Mabry. eds.). 1-18, Chapman & Hall, Lon- MARKHAM, K. R.. Distribution of flavonoids in the lower plants
don, 1982. and its evolutionary significance, in The Flavonoids, Advances
in Research since 19800. B. Harborne, ed.), 427-468, Chap-
HRAZDINA, G., Anthocyanins, in The Flavonoids: Advances in Re-
man & Hall, London, 1988.
search (J. B. Harbnme and T. J. Mabry, eds.), 135-188, Chap-
man & Hall, London, 1982. MARKHAM, K. R., Flavones, f1avonols, and their g1ycosides, in Plant
Phenolics (J. B. Harbome, ed.), Vol. I of Methods in Plant
HRAZDINA, G., Compartmentation in aromatic metabolism, in Phe-
Biochemistry (p. M. Dey and J. B. Harborne, eds.), 197-235,
nolic Metabolism in Plants (H. A. Stafford and R. K. Ibrahim,
Academic Press, London, 1989.
eds.), Recent Advances in Phytochemistry Vol. 26, 1-23,
Plenum Press, New York, 1992. MARKHAM, K. R. and T. J. MABRY, The identification of twenty-
three 5-deoxy- and ten 5-hydroxy-f1avonoids from Baptisia (e-
HRAZDINA, G., H.IREDALE, and L. R. MATTICK, Anthocyanin com-
contei (Leguminosae), Phytochemistry, 7, 791-801 (1968).
position of Brassica oleracea cv. Red Danish, Phytochemistry,
16.297-299 (1977). MARKHAM, K. R., B. 'fBRNAt, R. STANLEY, H. GEIGER, and T. J.
HUNGRIA, M., c. M. JOSEPH, and D. A PHILLIPS, Anthocyanidins MABRY, Carbon-13 NMR studies of f1avonoids Ill. Naturally
and flavonols, major nod-gene inducers from seeds of a black- occurring flavonoid glycosides and their acylated derivatives,
seeded common bean (Phaseolus vulgaris 1..), Plant Physiol., Tetrahedron, 34, 1389-1397 (1978).
97,751-758 (199Ia). MARKHAM, K. R., V. M. CHARI, and T. J. MABRY, Carbon-13 NMR
HUNGRIA, M., C. M. JOSEPH, and D. A. PHILLIPS, Rhizobium nod- spectroscopy of flavonoids. in The Flavonoids: Advances in
gene inducers exuded naturally from roots of common bean Research 0. B. Harbome and T. J. Mabry, eds.), 119-134,
(Phaseolus vulgaris 1..), Plant Physiol., 97, 759-764 (199Ib). Chapman & Hall, London, 1982.
HUTCHINSON, C. R., Biological method. for studying the biosyn- MAxwELL, C. A. and D. A i'HILuPs, Concurrent synthesis and
thesis of natural products, Nat. Pmd. Rep., 4, 133-152 (1986). release of nod-gene-inducing flavonoids from alfalfa roots,
Plant Physiol., 93, 1552-1558 (1990).
IBRAHIM, R. K.• Immunolocalization ,of flavonoid conjugates and
their enzymes in Phenolic Metabolism in Plants (H. A. Stafford MAXWELL, C. A, U. AlIARTWIG, C. M. JOSEPH, and D. A. PHILLIPS,
and R. K. Ibrahim, eds.), Recent Advances in Phytochemistry A chalcone and two related f1avonoids released from alfalfa
Vol. 26, 25-61, Plenum Press, New York, 1992. roots induce nod genes of Rhizobium meliloti, Plant Physiol.,
JACOBS, M. and P. H. RUBERY, Naturally occurring auxin transport 91.842-847 (1989).
regulators, Science, 241, 346-349 (1988). MCCLURE, J. W., Physiology and functions of f1avonoids, in The
KERSCHER, F. and G. FRANZ. Isolation and some properties of an Flavonoids (J. B. Harbome, T. J. Mabry, and H. Mabry, eds.),
UDP-g1ucose: 2-Hydroxyflavanone-6(or 8)-C-g1ucosyltransfer- 970-1055, Academic Press, New York, 1975.
ase from Fagopyrum esculemum M. cotyledons, J. Plant Phys- McCREA, K. D. and M. LEVY, Photographic visualization of floral
iol.,132. 1I0-U5 (1988). colors as perceived by honeybee pollinators, Am. J. Bot., 70,
KoeHs, G. and H. GRISEBACH, Enzymic synthesis of isoflavones, 369-375 (1983).
Eur. J. Biochem., 155. 311-318 (1986). MELcmoR, F. and H. KINnL, Grapevine stilbene synthase eDNA
Kuc, J., Antifungal compounds from plants, in Phytochemical Re- only slightly differing from chalcone synthase eDNA is ex-
sources for Medicine and Agriculture (H. N. Nigg and D. pressed in Escherichia coli into a catalytically active enzyme,
Seigler, eds.), 159-184, Plenum Press, New York, 1992. FEBS Lett., 268, \7-20 (1990).
KUHN, D. N., Plant responses to stresses at the molecular level, in MaLER, R. W., R. KLEIMAN, R. G. POWELL, and A R. PUTNAM,
Plant-Microbe Interactions, Molecular and Genetic Perspec- Germination and growth inhibitors of alfalfa, J. Nat. Pmd., 51.
tives, Vol. 2 (T. Kosuge and E. W. Nester, eds.), 414-440, 328-330 (1988).
Macmillan, New York, 1987. MILLER, R. W., R. G. POWELL, and A R. PuTNAM, (- )-5'-Me-
 Flavonoids 191

thoxysativan, a new isoflavan from alfalfa, 1. Nat. Prod., 52, RAVANEL, P., S. CREUZET, and M. TJsSUT, Inhibitory effect of hy-
634-636 (1989). droxyflavones on the exogenous NADH dehydrogenase of plant
MORELAND, D. E. and W. P. NovfIZKY, Effects of phenolic acids, mitochondrial inner membranes, Phytochemistry, 29, 441-445
coumarins, and flavonoids on isolated chloroplasts and mito- (1990).
chondria, in Allelochemicals: Role in Agriculture and Forestry ROBBERECIIT, R. and M. M. CALDWELL, Leaf epidermal transmit-
(G. R. Waller, ed.), ACS Symposium Series 330, 247-261, tance of ultraviolet radiation and its implications for plant sensi-
American Chemical Society, Washington, DC, 1987. tivity to ultraviolet-radiation induced injury, Oecologia, 32,
Mih.LER, K. O. and H. BORGER, Experimentelle Untersuchuogen 277-287 (1978).
fiber Phytophthora-Resistenz der Kartoffel. Zug1eich ein Beitrag RODRiGUEZ, E., P. L. IlEALEy, and L MEHTA (eds.) Biology and
zum Problem der "erwobener Resistenz" im Pflanzenreich, Chemistry of Plant Trichomes, Plenum Press, New York, 1984.
Arb. BioI. Anst. (Reichsamst.), Beriin, 23, 189-231 (1941). SANDERSON, G. W., The chemistry of tea and tea manufacturing,
NAKAGAWA, M., K. NAKANISHI, L. L. DARKO, and J. A. VICK, in Structural and Functional Aspects of Phytochemistry (V. C.
Structures of cabenigrins A-I and A-II, potent anti-snake ven- Runeckles and T. C. Tso, eds.), Recent Advances in Phytochem-
oms, Tetrahedron Lett., 23, 3855-3858 (1982). istry Vol. 5, 247-316, Academic Press, New York, 1972.
NANAYAKKARA, N. P. D., R. A HUSSAlN, J. M. PEzzUTO, D. D. SEIGLER, D. S., Role of lipids in plant resistance to insects, in Plant
SOEJARTO, and A D. KINGHORN, An intensely sweet dihydrofla- Resistance to Insects (P. A Hedin, ed.), ACS Symposium Series
vonol derivative based on a natural product lead compound, J. 208, 303-327, Arnerican Chemical Society, Washington, DC,
Med. Chern., 31, 1250-1253 (1988). 1983.
NIELSEN, J. K., L. M. LARSEN, and H. SORENSEN, Host plant selec- SELWAY, J. W. T., Antiviral activity of flavones and flavans, in
tion of the horseradish flea beetle Phyllotreta armoraciae (Col- Plant Flavonoids in Biology and Medicine: Biochemical, Phar-
eoptera: ChIysomelidae). Identification of two flavonol glyco- macological, and Structure-Activity Relationships (V. Cody,
sides stimulating feeding in combination with glucosinolates, E. Middleton, Jr., and J. B. Harborne, eds.), 521-536, Liss,
Ent. Exp. Appl., 26, 40-48 (1979). New York, 1986.
NmMANN, G. 1., Distribution and evolution of the flavonoids in STADLER, E., Oviposition and feeding stimuli in leaf surface waxes,
gymnosperms, in The Flavonoids: Advances in Research since in Insects and the Plant Surface (B. Juniper and R. Southwood,
1980 (J. B. Harborne, ed.), 469-478, Chapman & Hall, London, eds.), 105-121, Arnold, London, 1986.
1988.
STAFFORD, H. A., The enzymology of proanthocyanidin biosyn-
NISHIDA, R., T. OHSUGI, S. KOBUBO, and H. FUKAMI, Oviposition thesis, in Chemistry and Significance of Condensed Tannins
stimulants of a citrus-feeding swallowtail butterlly, Papilio xu- (R. W. Hemingway and J. J. Karchesy, eds.), 47-70, Plenum
thus, Experientia, 43, 342-344 (1987). Press, New York, 1989.
OHSUGI, T., R. NISHIDA, and H. FuKAMI, Oviposition stimulant of STAFFORD, H. A, Flavonoid Metabolism, CRC Press, Boca Raton,
Papilio xunthus, a citrus feeding swallowtail butterfly, Agric. FL,1990.
Bioi. Chern., 49, 1897-1900 (1985).
STAFFORD, H. A., Flavonoid evolution: An enzymic approach, Plant
PETERS, N. K., J. W. FROST, and S. R. LoNG, A plant flavone, Physiol., 96, 680-685 (1991).
luteolin, induces expression of Rhizobium meliloti nodulation
STENLID, G., Flavonoids as inhibitors of the formation of adenosine
genes, Science, 233, 977-980 (1986).
triphosphate in plant mitochondria, Phytochemistry, 9,
PHILLIPS, D. A., Flavonoids: Plant signals to soil microbes, in Phe- 2251-2256 (1970).
nolic Metabolism in Plants (H. A Stafford and R. K. Ibrahim,
STICH, K. and G. FORKMANN, Biosynthesis of3-deoxyanthocyanins
eds.), Recent Advances in Phytochemistry 26, 201-231,
with flower extracts from Sinningia cardinalis, Phytochemistry,
Plenum Press, New York, 1992.
27,785-789 (1988).
PIERPOINT, W. S., Flavonoids in the human diet, in Flavonoids and
STRACK, D. and V. WRAY, Anthocyanins, in Plant Phenolics (J. B.
Medicine: Biochemical, Phannacological. and Structure-Activ-
Harborne, ed.), Vol. 1 of Methods in Plant Biochemistry (P.
ity Relationships (V. Cody, E. Middleton, Jr., and J. B. Har-
M. Dey and J. B. Harborne, eds.), 325-356, Academic Press,
borne, eds.), 125-140, Liss, New York, 1986.
London, 1989.
PORlER, L. J., Flavans and proanthocyanidins, in The Flavonoids:
Advances in Research sinoe 1980 (J. B. Harborne, ed.), 21-62, TANRISEVER, N., F. R. FRONCZEK, N. H. FISCHER, and G. B. WIL-
Chapman & Hall, London, 1988. LIAMSON, Ceratiolin and other flavonoids from Ceratiola eri-
coides, Phytochemistry, 26,175-179 (1987).
PORTER, P. M., W. L. BANWART, and J. J. HASSETT, HPLC isolation
and GC-MS identification of genistein, daidzein, and coumes- TERNAI, B. and K. R. MARKHAM, I3C-NMR Studies of Flavo-
trol from unhydrolyzed soybean root extracts, Env. Exp. Bot., noids-1. Flavones and Flavonols, Tetrahedron, 32, 565-569
25, 229-232 (1985). (1976).
PORTER, P. M., W. L. BANWART, and J. J. HASSETT, Phenolic acids THOMPSON, W. R., J. MElNwALD, D. ANESHANSLEY, and T. EISNER,
and flavonoids in soybean root and leaf extracts, Env. Exp. Bot., Flavonols: Pigments responsible for ultraviolet absorption in
26,65-73 (1986). nectar guide of flower, Science, 177, 528-530 (1972).
POULTON, J. E., Transmethylation and demethylation reactions in TIMBERLAKE, C. F. andP. BRIDLE, Anthocyanins, in The Flavonoids
the metabolism of secondary plant products, in Secondary Plant (1. B. Harborne, T. J. Mabry, and H. Mabry, eds.), 214-266,
Products 7 (E. E. Conn, ed.), Vol. 7 of The Biochemistry of Academic Press, New York, 1975.
Plants (P. K. Stumpf and E. E. Conn, eds.), 668-725, Academic TIMBERLAKE, C. F. and B. S. HENRY, Anthocyanins as natural food
Press, New York, 1981. colorants, in Plant Flavonoids and Medicine II: Biochemical,
192 Flavonoids

Cellular, and Medicinal Properties (V. Cody, E. Middleton, Jr., antibiotic activity toward the com earwonn, J. Econ. Entomo!.,
J. B. Harborne, and A. Beretz, eds.), 107-121, Liss, New York, 72, 256-258 (1979).
1988. WALLACE, J. W. and H. GRISEBACH, The in vivo incorporation of
TOMAS-BARBERAN, F. A., R. J. GRAYBR-BARKMEUER, M. I. GIL, a flavanone into C-glycosylflavones, Biochim. Biophys. Acta,
and J. B. HARBORNE, Distribution of 6-hydroxy-, 6-methoxy- 304, 837-841 (1973).
and 8-hydroxyflavone glycosides in the Labiatae, the Scrophu- WAllACE,J. W., T.J. MABRY,andR.E.ALsToN,Onthebiogenesis
lariaceae, and related families, Phytochemistry, 27, 2631-2645 of flavone O-glycosides and C-glycosides in the Lemnaceae,
(1988). Phytochemistry, 8, 93-99 (1969).
TOWERS, G. H. N., P. A. SPENCER, and E. RODRiGUEZ, Recent topics WEISS, U. and J. M. EDWARDS, The Biosynthesis of Aromatic Com-
in phytochemical ecology research, in Phytochemical Ecology: pounds, Wiley-lDterscience, New York, 1980.
Allelochemicals, Mycotoxins, and Insect Pheromones and Allo- WELLE, R. and H. BRISBEACH, Properties and solubilization of the
mones (C. H. Chou and G. R. Waller, eds.), Academia Sinica prenyltransferase of isoflavonoid phytoalexin biosynthesis in
Monograph Series, 8 1-17, Institute of Botany, Taipei, 1989. soybean, Phytochemistry, 30, 479-484 (1991).
TSAl, S. M. and D. A. PHILLIPS, Flavonoids released naturally from WnLIAMS, C. A. and J. B. lIARBORNE, Distribution and evolution
alfalfa promote development of symbiotic Glomus spores in of flavonoids in the monocotyledons, in The Flavonoids: Ad-
vitro, App. Env. Microbiol., 57, 1485-1488 (1991). vances in Research since 1980 (J. B. Harborne, ed.), 505-524,
V ANDBN BERGHE, D. A., A. J. VLIBTNlNCK, and L. V AN HOOF, Chapman & Hall, London, 1988.
Present status and prospects of plant products as antiviral agents, WILLIAMS, C. A. and J. B. HARBORNE, Biflavonoids, in Plant Pheno-
in Advances in Medicinal Plant Research (A. J. Vlietinck and lics (J. B. Harborne, ed.), Vol. I of Methods in Plailt Biochemis-
R. A. Dommisse, eds.), 47-99, Wissenschaftliche Verlagsges. try (P. M. Dey and J. B. Harborne, eds.), 357-388, Academic
mbH, Stuttgart, 1985. Press, London, 1989a.
VARIN, L., Flavonoid sulfation: Phytochemistry, enzymology, and WlLLlAMS, C. A. and J. B. HARBORNE, Isoflavonoids, in Plant Phe-
molecular biology, in Phenolic Metabolism in Plants (H. A. nolics (J. B. Harborne, ed.), Vol. 1 of Methods in Plant Bio-
Stafford and R. K. Ibra1tim, eds.), Recent Advances in Phyto- chemistry (P. M. Dey andJ. B. Harborne, eds.), 421-450, Aca-
chemistry Vol. 26, 233-254, Plenum Press, New York, 1992. demic Press, London, 1989b.
VLffiTINCK, A. J., D. A. VANDEN BBRGHE, L. M. V AN HOOF, R. WlLLlAMS, C. A., J. B. HARBORNE, and B. MATHEW, A chemical
VRUSEN, and A. BOBYE, Antiviral activity of 3-methoxyfla- appraisal via leaf flavonoids of Dahlgren's Lilliflorae, Phyto-
vones, in Plant Flavonoids in Biology and Medicine: Biochemi- chemistry, 27, 2609-2629 (1988).
cal, Pharmacological, and Structure-Activity Relationships (V. WasON, A.• Flavonoid pigments in chalkhill blue (Lysandra cori-
Cody, E. Middleton, Jr., and J. B. Harborne, eds.), 537-540, don Poda) and other Iycaenid butterflies, J. Chem. Ecol., 13,
Liss, New York, 1986. 473-493 (1987).
VOGEL, G., Natural substances with effects on the liver, in New WOLLENWEBER, E., Flavones and flavonols, in The Flavonoids:
Natural Products and Plant Drugs with Pharmacological or Advances in Research 0. B. Harborne and T. J. Mabry, eds.},
Therapeutic Activity (H. Wagner and P. Wolff, eds.), 249-265, 189-259, Chapman & Hall, London, 1982.
Springer-Verlag, New York, 1976. WOLLENWEBER, E. and V. H. DIElZ. Occurrence and distribution
VOGEL, G., A peculiarity among the flavonoids-Silymarin, a com- of free flavonoid aglycones in plants, Phytochemistry, 20,
pound active on the liver, in Proc. International Bioflavonoid 869-932 (1981).
Symposium, Munich, F. R. G., 1981 (L. Farkas, M. Gabor, F. WOllENWEBER, E. and M. JAY, Flavones and flavonols, in The
K3llay, and H. Wagner, eds.), 461-474, Elsevier, Amsterdam, Flavonoids: Advances in Research since 1980 (1. B. Harborne,
1982. ed.), 233-302, Chapman & Hall, London, 1988.
WAISS, A. C., JR., B. G. CHAN, C. A. ELUGER, B. R. WISEMAN, WONG, E.. Isoflavonoids, in The Flavonoids (1. B. Harborne, T. J.
W. W. McMtuAN, N. W. WIDSlROM, M. S. ZUBER, and A. J. Mabry, and H. Mabry, eds.), 743-800, Academic Press, New
KEAsTER. Maysin. a flavone glycoside from com stalks with York,1975.
12
Tannins

Introduction (1), chlorogenic acid (2), and ( + )-catechin (3), for example,
Hydrolyzable tannins have been referred to in the literatore as tannins, but they
Biosynthesis do not possess the ability to precipitate proteins (nonetheless,
Distribution of Hydrolyzable Tannins these compounds may bind or interact with proteins). The
Proanthocyanidins or Condensed Tannins C.-C 1 tribydroxy compound, gallic acid (1) appears to be
Flavan-3,4-diols the biological equivalent of the C.-C3 compounds, cinnamic
Flavan-3-ols acid, p-coumaric acid, caffeic acid, and so forth. (Bate-
Procyanidin Dimers Smith, 1962) (see Chapter 8); many types of gallate esters
Condensed Units Derived from Proanthocyadin Units are found in natore. Among these are compounds such as
Biological Activity of Tannins the turgorins from Acacia !<aroo (4-6) (Fig. 12.1), that are
Binding of Tannins to Protein involved in leaf movement in plants (Haslam and Lilley,
Tannins in the Diets of Animals 1986).
The Cost of Tannin Defenses and Resource Allocation Tannins usually are located in the vacuoles of plant cells
Other Proposed Functions of Tannins and, in many cases, make up a sizable proportion of the dry
Medicinal Properties of Tannins weight of plants. Tannins often are concentrated in epider-
Correlation of Tannin Consumption and Throat Cancer mal tissues and in the bark of woody plants, but these pheno-
Tannins and the Production of Leather lic compounds may be found in leaves, roots, stems, fruits,
Other Types of Tannins bark, wood, or other plant parts. Tannin content may vary
Analysis of Tannins widely during the growing season in a particular plant part.
References There are two major structural types of tannins-conde-
nsed tannins and hydrolyzable tannins. Condensed tannins,
or proanthocyanidins, are compounds that should produce
INTRODUCTION anthocyanidins on hydrolysis (although, in practice, many
recently isolated compounds of this group are known not to
Plant polyphenols, which have the ability to precipitate pro- yield anthocyanidins) (Hemingway, 1989). These tannins
tein, collectively, are called tannins. These compounds have are derived from flavonoid precursors. Hydrolyzable tannins
been used for millenia to convert raw animal hides into (or gallotannins and ellagitannins) are derived from the shik-
leather. In this process, tannin molecules cross-link the pro- imic acid pathway intennediate, dehydroshikimic acid (7)
tein and make it more resistant to bacterial and fungal attack. (Fig. 12.3). Ellagitannins appear to be much more common
Molecules in the molecular weight range of 500-2000 that gallotannins. Furthennore, ellagitannins generally differ
(3000) are most effective, but the ability to bind effectively from other types of tannins in that individual compounds
varies with different tannin structures. Today, however, can be isolated as a comparatively stable entity that can be
many substances considered to be tannins by virtue of their handled as a single compound of defined structure. Gallotan-
structure and biosynthetic origin have limited, if any, ability nins usually occur as complex mixtores (Okuda et ai.,
to make leather (Hagennan and Butler, 1981). 1989b).
Although vegetable tannins are invariably polyphenolic Evolutionarily, condensed tannins preceded hydrolyzable
substances, not all phenolic materials are tannins. Gallic acid tannins; these flavonoid-derived phenolics are found in

193
194 Tannin.f

H o y q0: 8
'. ">...
HO
~ ~I
OH

H o y : ) : 0 ">...
J8(HOOH
~
I
HO

iIY
CO,H

I "I" OHHOAf
.">... .'OH ">... OH OH
OH OH

(-)-epicatechin (30) (-)-epigallocatechin (34) gallic acid (1)

a tlavan-3-o1
HO
HO

0
CO,H
OH

HO
OH
HO , OH
•
OH
OH OH
(+ )-catechin (3) (+ )-gallocatechin (33) shikimic acid (12)

~ o~OH
OSO'H OSO,H

H~

(S)r
HSO~O O~
HOHO 0 OH
OH 7 OH
H I H
7
I
HO '" CO,H HO '" CO,H
(4) HO OH
71

o
'"
~
CO'H 0 OH 1

HO HO 0 ~
OH 7 I OH

HHO '" CO,H

HO,e. ~OH
~rr chlorogenicacid (2)
(a) (6) OH OH

Fig. 12.1 (a & b). Nontannin phenolic compounds and {3-pentagaIloylglucose.

ferns, gymnospenns, and angiospennous plants, whereas hy- HYDKOLYZABLBT~

drolyzable tannins are found only in a number of relatively
advanced dicotyledonous plant families. Hydrolyzable tan- Complex esters of gallic acid with a carbohydrate (usually
nins are abundant in the leaves, fruits, pods, and galls (some- glucose), called gallotannins, are widely distributed among
times bark and wood) of a number of dicotyledonous fami- dicotyledonous plants. The most common gallotannin is 13-
lies (Lewis and Yamamoto, 1989). Plant extracts may pentagalloylglucose (8). Additional galloyl units can be
contain both hydrolyzable and condensed tannins. For exam- added via depside (ester) linkages, usually at the C-3 position
ple, in some Acacia species, leaves contain primarily hydro- of gallic acid. Esters involving a dimer of gallic acid, hexa-
lyzable tannins, whereas bark contains mostly condensed hydroxydiphenic acid (9) [nonnally isolated after hydrolysis
tannins. Hydrolyzable tannins also predominate in the imma- as the dilactone, ellagic acid (10)] are called ellagitannins
ture fruits of some of the same species (Readel et aI., unpub- (Fig. 12.2). Hexahydroxydiphenic acid occurs in two opti-
lished data). cally active fonns in tannins. Ellagitannins (such as II, cori-
Treatment of hydrolyzable tannins with acid or alkali and, lagin) often co-occur with gallotannins, but are especially
in some cases, hydrolytic enzymes splits them into sugars important in oak, eucalyptus, chestnut, some Caesalpinia
and phenolic carboxylic acids. Condensed tannins do not species, and Terminalia chebula. Collectively, the two
break down readily in this manner nor do they contain sugar groups are called hydrolyzable tannins. As the name implies,
moieties as a part of their structure (Haslam, 1979). How- these compounds may be degraded into simpler fragments
ever, gallic acid units are sometimes associated with con- under hydrolytic conditions. Hydrolyzable tannins usually
densed tannins (Lewis and Yamamoto, 1989). are classified by the phenolic acid(s) liberated on hydrolysis.
 Tannin I 195

'r1
OH

OH
r OH
OH

quinic acid (lJ) scyllo-quercitol (14) (+)-proto-quercito) (15)

"O~
::'0 o~o 0>-QoH
ro.

°
~° ° ~
10H
fi

Hoi[
° ° ° OH

O~O"
HO OH VOH
HO

rrOH
j3-pentagalloylglucose (8)

Hoyyoi'·'·O'V
yY° OH

naringenin (31) liquiritigenin (32)

(b)

Fig. 12.1. (continued)

Tannins involving (- )-shikimic acid (12), quinic acid the corresponding oxygenated acids, Conn and Swain (1961)
(13), scyllo-quercitol (14), and proto-quercitol (15) (Fig. showed that 14C-phenylalanine was incorporated into both
12.1) cores are known. Most of the unusual types that have quercetin and myricetin, but not into gallic acid in Geranium
been studied are from the genus Quercus (oaks) and other pyrenaicum. However, 14C-glucose significantly labeled
members of the family Fagaceae (Porter, 1989). gallic acid. Subsequent studies demonstrated that [14C]shiki-
Hydrolyzable tannins involving depsides are known from mic acid was a more efficient precursor of gallic acid than
the Aceraceae, Anacardiaceae, Combretaceae, Ericaceae, [14C]phenylalanine in Rhus typhina and in Camellia sinensis
Geraniaceae, Hamamelidaceae and Paeoniaceae (Porter, (Dewick and Haslam, 1969; Saijo, 1983). Further, incorpora-
1989). The major sources of commercial ellagitannins are tion of the label did not appear to be specific in feeding
myrabolans, divi-divi, algabobilla, valonea, and the bark of experiments with phenylalanine. Additional supporting evi-
oak and Spanish chestnut (Haslam, 1981). Unfortunately, few dence for C6C I origin of gallic acid has been obtained by
structures of hydrolyzable tannins other than those from a studies with specific inhibitors for the respective pathways.
small number of economically important plants have been The addition of L-2-aminooxy-3-phenylpropionic acid
studied. (AOPP, inhibits PAL activity) to cell suspensions of Quer-
cus rabur (Fagaceae) had no effect, whereas N-(phosphono-
Biosynthesis methyl)glycine (glyphosate), which inhibits enzymes rela-
tively late in the skimimic acid pathway, resulted in
Although gallic acid has been proposed to arise by 13- accumulation of additional gallic acid (Lewis and Yama-
oxidation of C6-C3 compounds such as p-coumaric acid or moto, 1989). Thus, a body of current evidence seems to
196 Tannins

OH

HOllCO,H ,<:::
HO

HO 0 ./ OH
/''-1
1
HO OHHO OH

__"';, ~ .o~oo·'" OH
bexabydroxydipbenic acid (9)

:v~OOOO HO
OH

OH
o
OH OH
corilagin (11) (myrobalans, divi-divi)
OH
an eIlagitannin

Fig. 12.2. Coriiagin, an ellagitannin, gallic acid, hexahydroxydiphenic acid and eUagic acid.

favor gallic acid being formed directly from shikimate (Fig. two galloyl units in gallotannins of Liquidambar formosana
12.3). However, in other plants (e.g., Lithospermum), proba- seem to be replaced by a hexahydroxydiphenyl group later
bly also depending on the stage of development, these com- in the growing season (Okuda et al., 1989b). Liquidambin
pounds may be derived via pheny1alanine-cinnamate (17) (Fig. 12.5), a compound with an aldehydic function, is
(Gross, 1992). possibly an intermediate in the synthesis of C-glucosidic
Enzyme preparations :rom Quercus robur leaves indicate ellagitannins (Okuda et al., 1989b).
that the fIrst product is J3-0-g1ucogallin (16). Gallic acid is Other tannins of this type (Fig. 12.6) contain modifIed
activated via the UDP-glucose not the CoA derivative (Gross, acids such as chebulagic acid (18) and peduncu1agin (19), and
1992; Lewis and Yamamoto, 1989). J3-0-G1ucogallin serves the brevilagins I (20) and 2 (21) which are normally consid-
as both donor and acceptor in the sequential biosynthetic steps ered ellagitannins (Haslam, 1979). When the galloyl groups
(Fig. 12.4). An enzyme responsible for the formation of 1,6- are in the diaxial position, radical coupling can occur to pro-
digalloylglucose also was isolated from this source (Gross, duce compounds such as chebulinic acid (22) (Fig. 12.7).
1992), an enzyme for the formation of 1,2,6-trigalloylglucose Some plants produce simple esters of gallic acid and glu-
from Rhus typhina, and another for the formation of 1,2,3,6- cose (Fig. 12.8). Others make J3-penta-O-gal10yl-o-glucose
tetragal10ylglucose, followed by another enzyme that forms (8), a "biosynthetic watershed" from which three broad and
1,2,3,4,6-pentagalloylglucose (Gross, 1992). distinctive pathways diverge. One pathway (a) leads to the
The hexahydroxydiphenyl group of ellagitannins appears gallotannins, by the addition of further gallic acid molecules
to arise by coupling of galloyl units in gallotannins. Indeed, linked as meta-depsides (complex esters with gallic acid)

HOllCO'H
HO
1'<:::
0

OH OH

n
dehydroshikimic acid (7) gallic acid (1)

Fig. 12.3. Biogenesis of gallic acid.

 Tannins 197

OH OG

~
) P'Q.gluC'ogallin~) p·O-gluC'ogallin
HO IIHO II
OG OG
HO OH HO OH
H H
p-O-glurogallin (16)
OG

~
P-O-glucogallin
HO •
OG
GO OG
H

o
G~OG
0
H,'<::::
Oif
OG G=galloyl
GO OG HO ~
H
OH
p-penta-O-galloyl-D-g1ucose (8)

Fig. 12.4. Biosynthesis of j3-pentagalIoylglucose (Gross, 1989; modified and used with permission of the copyright owner, the American Chemical
Society, Copyright 1989).

(Fig. 12.8). Two further pathways (b) and (c) diverge to geniin, telemagrandin I (23), casuarictin (24), pedunculagin
ellagitannins by oxidative coupling of appropriately dis- (19), and potentillin (25), arise in this manner.
posed galloyl groups (Fig. 12.8). Enzyme extracts of Rhus Io a relatively small group of plants, more complex oxida-
typhina leaves catalyze the galloylation of 1,2,3,4,6-penta- tive coupling occurs via pathway (c) to produce distinctive
galloylglucose, yielding a mixture of numerous hexa-, metabolites such as those found in some Geranium, Euphor-
hepta-, octa-, and nonagalloylglucoses (Gross, 1992). bia, andAcer species (Fig. 12.10) (Haslam and Lilley, 1986).
~-GlucogalIin is the principal source of galloyl units. Geraniin (26), widespread in plants, is one of these (Porter,
Compounds belonging to the fIrst of these ellagitannin- 1989).
forming groups (pathway b) involve relatively simple link-
age of the galloyl groups attached to a sugar nucleus (Fig.
12.9). Hydrolyzable tannins, such as telemagrandin II, eu- Distribution of Hydrolyzable Tannins
Hydrolyzable tannins occur in a series of relatively ad-
HO vanced families mostly in the Rosales and Myrta1es (Bate-
Smith, 1984; Dahlgren et aI., 1981; Haddock et aI., 1982a,

o oJ
1982b, 1982c). Among these families are the Aceraceae, Ana-
cardiaceae, Betulaceae, Casuarinaceae, Cercidiphyllaceae,
Cistaceae, Clusiaceae, Combretaceae, Coriariaceae, Coma-
ceae, Cunoniaceae, Dilleniaceae, Dipterocarpaceae, Drosera-
o 0 0/ ~-OH

O~CHO
ceae, Ebenaceae, Ericaceae, Euphorbiaceae, Fabaceae (Cae-
HO salpiniodeae), Fagaceae, Fouquieraceae, Geraniaceae,
OH
Guoneraceae, Haloragaceae, Hamamelidaceae, Juglanda-
o 0
ceae, Lecithydaceae, Loranthaceae, Lythraceae, Melastoma-
taceae, Myricaceae, Myrtaceae, Nympheaceae, Nyssaceae,
Onagraceae, Plumbaginaceae, Polygonaceae, Rbizophora-
ceae, Rosaceae, Santaiaceae, Simaroubaceae, and Theaceae
OH (Bate-Smith, 1984). The distribution of complex ellagitan-
HO
OH nins also is restricted (Haslam, 1986). Wolter-Filho et aI.
(1989) have used the presence of ellagitannins in Rhabdoden-
dron (Rbabdodendraceae) species to argue for placement of
liquidambin (17)
this enigmatic family in the Rosiflorae or Myrtiflorae and not
FIg. 12.5_ Liquidambin. in the Caryophyllales or Rutales, as others have argued.
198 Tannins

OH

HO~O ~ I peduncuJagin (19)

HO
HO
OO~
0
I~
~ 0
00 0 00
HO 0

HO OH
OH OH

HO~"H""~"
HO ~ I O~O
o
0J-QoH
Ii OH
o 2 0 ~

HO){O "~k""OH'
HOVo - orO: OH
HO ~ /,
"H
OH
tannic acid, Chinese gallotannin 0 ~ /;
OH 0
n =0,1, 2; C-I galloyl group may be absent OU
(a)

0
0
0
0

0 0

OH OH

OH
OH OH

(b) brevilagin I (20) brevilagin 2 (21)

Fig. 12.6 (a-c). Some representative hydrolyzable tannins.

PKOAl'ITHOCYAl'IIDIl'IS OK COl'lDEl'ISED between flavonoid units, hydroxyation patterns, stereochem-

TAl'Il'IIl'IS istry of carbons 2, 3, and 4 of the pyran ring, and the presence
of additional substituents (such as gallic acid) (Lewis and
Proanthocyanidins are widely distributed, but are particu- Yamamoto, 1989). More than 100 proanthocyanidins with
larly common in conifers. The structures are based on oligo- subtle, but well-defined, structural variations have been iso-
meric flavonoid precursors and vary in the type of linkages lated and characterized. Most interflavonoid linkages occur
 Tannins 199

HO

o
~
H

o '"
\ h OH
OH

o
cbebulagic acid (18)

Ho,e

o
(e)

Fig. 12.6. (continued)

RO

OR RO
N~
R.O~O
RO~
" , - : : ; - H ROR
ORYOR H
o - o 0 0

OR OR
(3·pentagalloylglucose (8)

HO OH
RO

N~ terchebin

o 0 0 OH
HO
eo,H

HO
OH OH
chebulinic acid (22) OH
(9) ~
OH
eO,H HO

eO,H

HO HO~ OH

OH OH
HOY ellagic acid (10)
chebulic acid
OH
R =galloyl

Fig.!2.7. Biogenesis of chebul"lnlC .

aCId (modified f rom Haslam, 1974).
200 Tannins

at C-4, C-6, and C-8, but those at C-2 and C-7 also are other cyclic systems. The mopanols (such as 28) and pelto-
known (Lewis and Yamamoto, 1989). The flavonoid skeleta gynols (such as 29) are representatives of this sort of modifi-
can be divided into two broad categories, those containing cation (Fig. 12.11).
either phloroglucinol or resorcinol (5-deoxy) substitution The reduction of 3-hydroxyflavones (dihydroflavanols)
patterns. to 3,4-diols (Ieucoanthocyanidins) was first demonstrated in
Colorless compounds that yield anthocyanidins upon cell-free extracts of Pseudotsuga menziesii and Ginkgo bi-
heating with acid are called proanthocyanidins. Compounds loba, by the reduction of ( + )-dihydroquercetin with dihy-
that yield cyanidin are called procyanidins and so forth. droflavonol 4-reductase in the presence of NADPH to a 2,3-
Monomeric proanthocyanidins are often referred to as leu- trans-3,4-cis-diol which differs from the product of nonen-
coanthocyaninidins, but the term leucoanthocyaninidin zymatic reduction, 2,3-trans-3,4-trans-diol (2R,3S,4R)-
sometimes has been used ambiguously in the literature. Most (+ )-3,4,5,7,3',4'-hexahydroxyflavan (Fig. 11.15 of Chapter
important in this group of compounds are flavan-3,4-diols 11). This result has now been confirmed in several other
and flavan-4-0Is. plant systems (Heller and Forkmann, 1988; Stafford, 1989).
Condensed tannins arise from monomeric flavan-3-01 A similar dihydroquercetin reductase activity has been
(catechins) and flavan-3,4-diol units. shown to be a key step in the biosynthesis of anthocyanidins
(Stafford, 1989). The enzyme from Dahlia has a molecular
F1avan·3.4·diols weight of about 41,000. However, there is evidence that the
The presence of monomeric proanthocyanidins or flavan- specificity of 3-hydroxyflavanone reductases of anthocy-
3,4-diols in plants has long been known. In hot acid, almost anidin biosynthesis are distinct from those of flavan-3,4-diol
all flavan-3,4-diols are converted to the corresponding an- biosynthesis (Stafford, 1989) (see Chapter 11).
thocyanidin. The first structure determined for a member of The conversion of (+ )-dihydromyricetin to the corre-
this class of compounds was that of melacacidin (27) from sponding 3,4-cis-isomer by an NADPH-dependent reductase
Acacia melanoxylon (Fig. 12.11). Flavan-3,4-diols do not has been demonstrated in cell cultures of Ginkgo bi/oba that
possess an affinity for collagen substrates and, hence, lack accumulate large quantities of prodelphinidins (Stafford,
tanning properties. Some flavan-3,4-diols are modified to 1989).

OH

gallotannin. O~:: OH

HO~ ~~O, 0 &OH

~ o.,.O.O~O~OH
additiOnOf
HO
;~~O_ 0 zo H 0 &OH
HO

gallic acid HO:\ I. ..... 0 ~OH

as m-depsldes HO OH "I ~O
./ HO 0 -
OR /_, HO \ , OH
.... <
R·O '\~~ ' \
~O
OR
(a) OH HO

R~YH (b) coupling of 4 and 6

olig~:'~l~~on Jr
and/or 2 and 3 gaIloyl groups
OH

H'
ellagitannins OH
oxidative coupling of pedunculagin (19)
vicinal galloyl ester o
OH
groups

OR OR
coupling of 1 and 6,
p-penta-O-galioyl-D-g1ucose (8) 3 and 6 and/or 2 and 4
o ~
HO~=
galIoyl groups
gslloyl geraniin (26)
H O Y R,
OH
OH HO
OH
Fig. 12.8. The overall patterns of metabolism of j)-pentagalloyl-n-glucose in plants.
 Tannins 201

G =galloyl H O i f I ~
HO b
OH OH
OH

G~O,
-G~
OG
HO
H

!
p-penta-O-gaIloyl-D-glucose (8)
G -G = hexabydroxydipbenoyl-
coupling of 4 und 6 galloyl group•

(Y'O~'
'0'
0
GO OG
H
OG
-- ..t'0~' '0 •
GO
0
OG
H
OH

telemagrandin H, eugeniin telemagrandin I (23)

~ coupling of 2 and 3 gaUoyl groups

G 6 G.... 6

G'O~
...... 0
4 1 --+- G,o~
0 4 2

o
\
3
__
G
-0 H
OG
0, __ G-O
G
3

H
OH

pedunculagln (19)
casuarictin (24)

'"
G •
I
G':O~lO
0, __ G-O 3 H

G OG
potentillin (25)

Fig. 12.9. Ellagitannins fonned by simple linkage of galloyl groups (modified from Gupta et aI .• 1982; modified and used with pennission of the
copyright owner, the Royal Society of Chemistry, Cambridge).

F1avan-3-ols pounds give similar color reactions to Ihose produced by

flavan-3,4-diols and, occasionally, have been calledleucoan-
Flavan-3-0Is, often known as calechins (Fig. 11.16 of thocyanins.
Chapter 11), are widely distributed. The most widely distrib- Flavan-3-0Is are derived from flavanones, via 3-hydroxy-
uled members of this class of compounds are catechin [( + )- flavanones (or dihydroflavonols). Naringenin (31) gives rise
(2R,3S)-calechinj (2,3-trans) (3) and epicatechin [( -)- to 5,7-dihydroxyproanthocyanidins, whereas Jiquiritigenin
(2R,3R)-epicalechinj (2,3-cis) (30) (Fig. 12.1) which usually (32) (Fig. 12.11) appears to be Ihe precursor for 5-deoxy-
occur as free aglycones and nol as glycosides. The known proanthocyanidins, both via the intermediacy of flavan-3-
(2R,3S)-proanthocyanidins are labulated by Porter (1988). ols and the same basic pathway as in Fig. 11.16 (Heller
The enantiomeric compounds are ent-calechin [( - )- and Forkmann, 1988; Lewis and Yamamolo, 1989; Stafford,
(2S,3R)-calechinj (2,3-trans) and ent-epicalechin [( +)- 1989).
(2S,3S)-epicalechinj (2,3-cis). Some 2S-flavan-3-0Is are The conversion of flavan-3,4-diols 10 flavan-3-0Is and to
found free and as gallale eslers in plants (Slafford, 1989). their oligomeric proanthocyanidin forms is the beginning of
Tissues that contain flavan-3-0Is oflen contain procyanidin the palhway unique 10 the biosynthesis of these secondary
dimers and trimers (Haslam, 1975). Some flavan-3-01 com- products (Stafford, 1989). Two basic types of hydroxylases
202 Tannins

::x;T'
o

R=
OH
R_R=
HO~QO
t~ 0
H:R-o l o
HO OH OH

HO~O
-
'" OH
I""
"1
O"'OHOR OR
R-R = HO ~ I - OH davidiin
aU' Davidia involucrata
f
'd
~0 g :.:;:cose 1 to 6 gaUoyl coupling

R
\
+
OR 0
"""""'- <OR --,0 /(c)
jI HO
~O, (a) RO\~OR
~R~OR - RO ORH
R H
Rhus, Acer, Pe/argonium speci.. (b) p-penta-O-galloyl-D-glucose (8)

+(b)
4 to 6 gaUoyl COUPling"/
/0--....J'r
' 0
.~~~OR /~o-l
o 0 6~0'1/
OH 0 0 0 OR' R
H090H 0 0 OH
1 '"
~ 0
'" 1 OH" 1
:-... -HO':-'" OH
O~OH'
I ~ vescalagin
o OIi:OH HO OH""
o
~ OH 0 0

Z
• OH
.. 0 OH 4to6and2to3 ~O O ...... R
OR _ _ it gaUoyl coupling 0 1
rugosin D (51) sanguin H·6 0 'R HO HO OH
Rosa sp., FiUpendula ulnwria Rubus, Geum and OR _ _ ~ OH OH
Poumtilla species Quercus species

Fig. 12.10. More complex ellagitannins (Haslam and Lilley, 1986; modified and used with pennission of the copyright owner, Akademiai IGado,
Budapest).

are involved in the synthesis of 2,3-trans-flavan-3-ols_ One The NADPH-dependent reduction of the 2,3-trans-3,4-cis-
is a microsomal cytochrome P-450 monooxygenase requir- diol with a dihydroxy B ring to the flavan-3-01, ( + )-catechin
ing oxygen and NADPH for 3' and 5' hydroxylations of the (3), was first shown in soluble extracts of Douglas fir cell
B ring_ The soluble 3-hydroxylase from Petunia has been cultures. The comparable reduction to (+ )-gallocatechin
purified and has a molecular weight of 74,000_ In addition, (33) from the corresponding 3,4-diol produced by dihydro-
a "soluble" cytosolic dioxygenase that requires Oz, 2-oxog- myricetin reductase activity was demonstrated with a soluble
lutarate, Fez+, and ascorbate is involved in stereospecific enzyme requiring NADPH in extracts of Ginkgo hi/aha
insertion of oxygen at C-3 to produce the common 2,3-trans- (Stafford, 1989).
isomer (2R,3R) of 3-hydroxyflavanones (Stafford, 1989). The pathways to 2,3-cis-flavan-3-0Is, such as (- )-epica-

HO
OH
~ OR

HO
~I OH yOH
HO I
~
OH
H0'O(X0
I OH

OH ~ , 0
! OH
melacaddin (27) OH
• peltogynol (29)
a mopanol (28)

Fig. 12.ll. Melacacidin, a flavan~3.4-diol. and related compounds.

 Ttl/millS 203

techin (30) (Fig. 11.1) and epigallocatechin (34) are not well C-3 and the flavan substituent at C-4 in the "upper" flavan
known (Stafford, 19X9). unit (Hillis, 1985 J.
A number of anomalous features in NMR spectra are
I'rocyanidin Dimers observed because rotation around the interflavan bond is
hindered. in most cases, one preferred conformation proba-
In addition to monomeric flavan-3-0Is and flavan-3,4- bly exists, but the energy barriers are not sufficient to permit
diols, many plants contain mixtures of dimeric procyanidins isolation of the different conformational forms of proantho-
such as (35-38) (Fig. 12.12). Procyanidins are common in cyanidins.
fruit, fruit pods, seed, and shells, either as their peracetates incorporation of labeled phenylalanine and cinnamic acid
or as the free phenolic forms. into dimeric proanthocyanidins occurs asymmetrically, with
The structures of procyanidins 35-38 were assigned on differences in the amount of precursors incorporated into the
a basis of lH-NMR spectra of the decaacetates. The absolute two portions of the molecule (Stafford, 1989). Further, the
stereochemistry has been determined as (4S) for compounds pool of free flavan-3-01 monomers including ( + )-catechin
(38) and (36) and (4R) for compounds (37) and (35). Mix- (3) and ( - )-epicatechin (30) contained much higher levels
tures of these particular dimers have not been found, but of radioactive label than the comparable initiating (or termi-
other dimers and trimers have been detected with them nating) unit. Tritium label at the C-2 position is retained,
(Hillis, 1985; Porter, 1988). 13C_NMR spectra are especially whereas that at the C-3 position is lost in the dimers (Staf-
useful for analyses of these compounds because a pro- ford, 1989).
nounced 'Y effect ( - 4.5 ppm) is observed and the chemical The proton H. at C-2 and C-2' is retained in both flavan
shift of C-2 is dependent on the orientation of the substituent units of the dimers. Proton He from the cinnamic acid precur-
at C-4 of the flavan system. All major natural procyanidin sor is lost. Differential labeling is observed in the "upper"
dimers possess a trans-orientation of the hydroxyl group at and "lower" part of the dimers. When cinnamic acid labeled

OR

BO BO

OR

OR OR

procyanidin B~2 (35) enl·epicatec:hin·(4~ ~8)·ent·catechin (36)

apple, hawthorn, cocoa bean, raspberry, blackberry
Cotoneaster, quince, cherry,
horse chestnut

BO

OR

OR
procyanidin D·3 (38)
epjcatechjn·(4~ ..... 8)·catechjn (B.l) (37) willow and poplar catkins,
grape, sorghum, cranberry, strawberry, rose hips, hops
pine cones

Fig. 12.12. Common dimeric proantbocyanidins (modified from Haslam, 1981; used with pennission of the copyright owner, Academic Press, New
York).
204 Tannins

R R

HO HO

OH 0
3-andS-

!
OH,3-0H
"upper" extension unit
NADPH R
OH HO

HO quinone methide
R
or carbocation intermediates
OH
NADPH
n=Ot08
or more
3,4-cis·diol OH
reductases
OH R
"lower" initiating unit
HO
R HO

oligomers
OH
navan-3-ols
Fig. 12.13. Proposed biosynthesis of procyanidin oligomers.

in the benzyl carbon is fed to plants, and the catechin and includes stereochemical and NMR analysis of the products,
procyanidin dimers produced are isolated, the label is found most of which also occur in nature. It is evident that in
in C-2 of the catechin and in the corresponding positions of vivo condensations follow the path of least resistance, being
both portions of the dimeric compounds (Fig. 12.13) (Has- regulated by the stability of the carbocation arising from the
lam, 1981; Porter, 1988). It is known that the oligomeric flavan-3,4-diol, by steric factors contributed by both elec-
forms of proanthocyanidins are synthesized by the sequential trophile and nUcleophile, 2,3-trans or 2,3-cis-stereochemis-
addition of an intermediate derived from the flavan-3,4-diols try of the electrophile, and the relative concentrations of the
to a preexisting chain or to a flavan-3-01 that initiates (or reactants (Roux and Ferreira, 1982).
terminates) a chain, rather than being self-condensation Treatment of procyanidin dimers with acid gives the ap-
products of the diols. Although the condensation step can propriate catechin unit corresponding to the lower half of
be done nonenzymatically in so-called biomimetic synthe- the molecule and a carbocation that normally collapses to
ses, the presence of enzymes is probable. A model for en- cyanidin (an anthocyanidin) under the reaction conditions.
zymic control of synthesis of these compounds has been
proposed (Stafford, 1989). Condel1!iled Units Derived from
If the supply of NADPH is rate limiting, intermediate Proanthocyanldin Units
C-4 carbocations with either (2S)- or (ZR)-stereochemistry
could react with either ( + )-catechin (3) or ( - )-epicatechin Most of the tannins important in the leather industry today
(30) (Fig. 12.1) to produce dimers or oligomers of procyanid- are condensed tannins. Tannins from quebracho (Schinopsis
ins. The nucleophilic character at C-6 or C-8 can be used lorentzii, S. balansae), gambier (Uncaria gambier), cutch
with either cation. The reactions would be regulated by the (Acacia catechu), mangrove (Rhizophora mangle), myrtan
stability of the cations arising from flavan-3,4-diols. Further- (Eucalyptus spp.), and wattle (Acacia meamsii) are all of
more, substitution occurs more readily at C-8 than at C-6. the condensed tannin type. Tannins from quebracho primar-
Substitution at nucleophilic centers is highly sensitive to ste- ily are (2S)-profisetinidins (39) (Fig. 12.14), whereas those
ric factors and reactions occur preferenially at the least hind- of wattle are (ZR)-profisetinidins (Hemingway, 1989). Of
ered sites (Hillis, 1985). these, the tannins of wattle have been the most studied. Most
A number of aspects of the biomimetic synthesis of di- of the proanthocyanidins isolated from plants have ZR abso-
mers and trimers based on flavan-3,4-diols and flavan-3-0Is lute-stereochemistry (Hemingway, 1989). Major exceptions
have been reviewed (Roux and Ferreira, 1982). This analysis are the proanthocyanidins of monocotyledonous plants, and
 Tannins 205

~OH

Q ".",'V
HO
HO
OH

OH
HO
y¥01-""""'"I
~
(foH
HO OH

o
, OH
HO'
Y¥l 0 1"".,,""'-' ' '
v
I
V00H ~OH HO
a proguibourtinidin (40)

H°Yy°1"",J,J
~ OH YOH
OH
a profisetinidin (39)

(XI oH
HO

'(XXI

"Ow""
HO 0 "..-,,''''-.
"'-. OH
OH

OH OH

a prorobinetinidin (41)

(a) a promelacacidin (42)

HO
.)-y0H

HO
"""V
OH

o
HO""'"

~OH
o
a procyanidin (43)

a prodelphinidin (44)

(b) OH
Fig. 12.14 (a & b). Selected proanthocyan'd'
1 InS.
206 Tannins

those of Uncaria, Polygonum, Rhapheolepis, Rhus, and que- f"'y0H

bracho (Schinopsis). 5-Deoxyproanthocyanidins mostly are OH o
based on the proguibourtinidin (40), profisetinidin (39), pro- ~
HO "'~OH
robinetinidin (41), and promelacacidin (42) skeleta and have H°Yy°'(~OH OH
2,3-trans-stereochemistry in the chain extender units. Most ~OH OH
OH
plants, other than those of the Fabaceae and Anacardiaceae,
contain 2,3-cis-(2R,3R)-procyanidins (Hemingway, 1989). mollisacacidin (45)

The vast majority of tannins in the procyanidin (43) and

~
HO ....·
prodelphinidin (44) classes are of mixed stereochemistry
with 2,3-cis- to 2,3-trans-ratios between 90: 10 and 50: SO.
Almost all natural and synthetic 2,3-cis-proanthocyanidins
HO~Oy.OOH
OH
A
VOH

have 3,4-trans-stereochemistry. In procyanidins and prodel- ~OH HO

O
phinidins, C-4 to C-8 and C-4 to C-6 linkages (in a ratio of both 4R and 4S
OH
about 3 : 1) are the most common. All known polymers of this "I
type are of the "linear" type. Linkages involving oxygen are HOYjfOi·""-'::-'" OH

N
o:;
rare. The terminal units of most natural proanthocyanidins ~OH ~OH
contain flavan-3-01s (Hemingway, 1989). H°Ylr°'l. . "~OH OH
In almost all tissues that contain proanthocyanidin di-
VV'OH "I
mers, oligomers and higher polymers also exist. Polymers OH
up to about 7000 MW usually are soluble, whereas higher- HO ,
o
(+)-catechin (3)
molecular-weight compounds will not dissolve. Polymers
with molecular weights as high as 20,000 are thought to
f)
VOH
occur. It has been suggested that procyanidins are covalently OH
bound to a saccharide matrix within plant cells (Haslam and
Fig. 12.15. Biogenesis of wattle condensed tannins (modified from
Lilley, 1985). Polymeric forms predominate over the soluble Haslam. 1981; used with pennission of the copyright owner, Academic
forms by 10: 1 or 20: 1 (Haslam and Lilley, 1985). Press. Orlando, FL).
Many structural types occur; these have been reviewed.
Most are based on hydroxylation variations in the starter
and extender units. In addition, there are variations in 0- properties. Condensed proanthocyanidins, however, do pos-
methylation, C - and O-glycosylation, and O-galloylation sess these properties. Dimers, trimers, tetramers, and other
(Hemingway, 1989). oligomers are capable of precipitating proteins. When tan-
Elaboration of oligomeric forms of procyanidins by the nins are consumed by man (and presumably other organ-
addition of flavan-3-ol units based on (- )-epicatechin (30) isms), they often have astringent properties. This property
or ( + )-catechin (3) leads to formation of two different heli- is caused by the cross-linkage of proteins and glycoproteins
cal structures. Those based on ( - )-epicatechin (procyanid- and a corresponding loss of lubrication in the mouth and
ins 35 and 37) produce a left-handed helix, whereas those throat. Maximum tanning and astringency occur with con-
based on (+ )-catechin (procyanidin 38) form right-handed densed proanthocyanidins with a molecular weight range of
helices. 500-2000 (3000). Proanthocyanidins are common in fruits,
Other proanthocyanidins involve flavans as one of the fruit pods, seeds, and shells, either as peracetates or in free
condensing units (Porter, 1988). phenolic forms. The loss of astringency that occurs during
Roux and his collaborators have isolated a group of profi- the ripening of fruit may be a reflection of the increased
setinidins from the wood of Acacia mearnsii. These com- degree of polymerization and, hence, molecular size of the
pounds were accompanied by their apparent precursor 2,3- associated tannins (Hagerman and Butler, 1989).
trans-3,4-( + )-mollisacacidin (45) (fisetinidol-4u-ol) (a fla- Although tannins are extracted in many types of solvents,
van-3,4-diol). Furthermore, these workers were able to con- methanol-water and acetone-water mixtures are often em-
dense ( + )-mollisacacidin (45) and ( + )-catechin (3) to give ployed (Hagerman, 1988). Hot water is usually used for com-
good yields of procyanidins that are naturally occurring in mercial extraction. For many laboratory procedures, 70%
black wattle or "mimosa extract" from Acacia mearnsii. acetone-water is an excellent solvent, although condensed
These observations support the view that flavan-3,4-diols tannins are less stable in this solvent than in aqueous metha-
(via their derived 4-carbocations) and flavan-3-01s condense nol (Cork and Krockenberger, 1991; Hagerman and Butler,
to form proanthocyanidins in the wood and bark of many 1991). Procedures for extraction of tannins from fresh and
trees (Fig. 12.15) (Haslam, 1982). preserved leaves have been reviewed recently (Cork and
Proanthocyanidins are those compounds that produce an- Krockenberger, 1991; Hagerman, 1988). Ellagitannins are
thocyanidins on hydrolysis. As previously observed, mono- less soluble than other types and are hard to extract from
meric proanthocyanidins (leucoanthocyanidins) do not pos- tissues (Hagerman and Butler, 1991). Condensed tannins are
sess an affinity for collagen substrates and lack tanning sensitive to both temperature and light during extraction pro-
 Tannins 207

cedures. The plant tissue should be preextracted with a lipo- bility, and ease of oxidation of the tannin. Generally, proteins
philic solvent to remove lipids and chlorophyll (Porter, are precipitated most readily by proanthocyanidins at pH
1989). The yield of tannins can be increased 30-75% by the values near the isoelectric point of the protein. Proline-rich
addition of antioxidants such as ascorbic acid or sodium proteins bave a very high affirtity for tannins. Tightly coiled
metabisulfite to 50% aqueous methanol solvents (peng and globular proteins have lower affinities than confonnationally
Jay-Allemand, 1991). Differences in tannin assays can be loose proteins (Hagennan, 1989; Hagennan and Butler,
due to differences in tannin extractibility as well as to dif- 1981; Mole and Watennan, 1987c). Less attention has been
fences in the amount of taunin present (Hagennan and But- paid to protein precipitation by hydrolyzable tannins, but
ler, 1991). binding to protein is, in part, a function of molecular size.
In the galloyl D-glucose series, association with protein is
enhanced by the addition of each galloyl group and reaches
BIOLOmCAL ACTIVITV OF TANNII'IS a maximum in the flexible disk-like structure of (3-penta-O-
galloyl-D-glucose (7) (Haslam and Lilley, 1985; Spencer et
In tenns of function, condensed taunins apparently act al., 1988). Confonnational mobility and flexibility is as sig-
mainly against the action of microbes, whereas hydrolyzable nificant as molecular size. When vicinal galloyl groups are
tannins defend primarily against chewing phytophagous in- constrained by the fonnation of intramolecular biphenyl
sects or animals (Lewis and Yamamoto, 1989). Because tan- linkages and the generation of hexahydroxydiphenoyl
nins bind salivary proteins, most tannins are astringent to groups, the loss of confonnational freedom is reflected in a
mammals. Many insects, as well as other animals, avoid reduced capacity to bind to protein (Haslam and Lilley,
plants rich in tannins. On the other hand, tannins may also 1985; Spencer et al., 1988). The effects of water of solvation
serve as phagostimulants and feeding cues (Schultz, 1989). are also important (Haslam et al., 1989).
Tannins playa central role in the fonnation of soil humus
in the development of soil profiles by producing complexes Tannins in the Diets of Animals
with proteins and carbohydrates that are resistant to micro-
In instances when tannins are consumed in the diet, it
bial decay (Ya et aI., 1989).
has been postulated that reduction in fituess occurs because
Although many of the properties of taunins are linked to
taunins bind to proteins and make the prorein less available
their ability to bind proteins, this role has been over empha-
to the animal. Plant proteins are the ultimate source of almost
sized (Hagennan and Butler, 1991), and individual tannin
all dietary amino acids for herbivores, carnivores, and detri-
molecules may exhibit more specific biological activity
tus-feeding animals; any factors that reduce the levels of
(Zucker, 1983). Many taunins are toxic to animals (Hag-
these nutritive compounds would be predicted to have dra-
ennan and Butler, 1991). This activity may be due to several
matic effects on the whole ecosystem. In excess, taunins
effects such as binding to biologically active molecules (pro-
should reduce the effectiveness of the digestive process in
teins, carbohydrates, and nucleic acids) or complexation of
the animals's gut by inhibiting digestive enzymes. These
nutrients or cofactors required for growth of the animal (Has-
factors have lead some to suggest that tannins may serve as
lam, 1989; Haslam et al., 1989; Okuda et aI., 1989b; Spencer
all-purpose, dose-dependent, digestibility-reducing defen-
et al., 1988; Zucker, 1983). Only recently, have scientists
sive substances. In practice, none of these has been conclu-
begun to view the general properties of the collective taunin
sively demonstrated (Mole and Watennan, 1987a).
mixtures independently of the biological activity of specific
When tannins (either hydrolyzable or condensed types)
tannin molecules produced by a plant (Okuda et al., 1989b).
are added to artificial diets, the fitness and growth rate of
animals (especially mammals) eating these diets is usually
Binding of Tannins to Protein
reduced (Mole and Watennan, 1987a; Schultz, 1989). The
Tannins (as discussed above) are a heterogeneous group percentage of tannins in plant materials often is in the range
of compounds that vary widely in physical and chemical 5-10%, but can be much greater. The rejection limit of most
properties. Many enzymes, such as (3-glucosidases, are deac- vertebrates is about 0.05% for hydrolyzable tannins. Many
tivated by the addition of tannins, but others are unaffected. animals refuse to eat diets containing sizable quantities of
Depending on a number of factors, these tannin-protein tannins.
complexes may precipitate or remain in solution. Both solu- Tannins from deciduous browse stems for mule deer
ble and insoluble complexes are stabilized by reversible, (Odocoileus hemionus), white-tailed deer (0. virginianus),
noncovalent bonds between protein and taunin. Although moose (Alces alees), carboulreindeer (Rangifer tarandus),
few studies have been catried out with single, purified tan- and elk (Cervus elephus) did not lower protein availability,
nins and single, purified proteins, it is clear that the ability but taunins from flowers, forbs, and tree and shrub leaves
of tannins to precipitate proteins varies widely (Blytt et aI., did. Tannins must be taken into account in understanding
1988; Hagennan, 1989, Harbome, 1989; Mole and Water- the defensive strategies of flowers and leaves (Robins et al.,
man, 1987c). The ability of a particular tannin to complex 1987). Furthennore, lesions are found in the gut of animals
proteins depends on the molecular size and shape, relative that nonnally do not consume taunins in quantity, whereas
number and orientation of hydroxyl groups, structure, solu- they are absent from animals that do. For example, Papilio
208 Tannins

polyxenes (which feeds on herbs of the family Apiaceae) despite the fact that there is sufficient tannin in leaves of
develops lesions and Papilio glaucus (which eats tanninifer- some oak species to fully precipitate all RuBPC present
ous trees of several families) does not develop lesions when (Martin and Martin, 1982, 1983). Further, the rate of hydrol-
tannins are included in artificial diets fed to the animals ysis of ribulose-1,5-bisphosphate carboxylase (RuBPC) in
(Steinly and Berenbaum, 1985). Usually, hydrolyzable tan- enzymatically active gut fluid of Manduca sexta larvae is
nins are responsible for gut lesions in insects (Schultz, 1989). very rapid and is unchanged by addition of tannic acid (Mar-
In contrast, many coadapted animals have the ability to tin et al., 1987).
eat plants containing large amounts of tannins. Some of these Purified samples of tannins from bitterbrush (Purshia tri-
animals even require tannins in the diet (Bernays and Wood- dentata) and blackbrush (Coleogyne ramosissima) (both
head, 1982; Harborne, 1986; Schultz, 1989). Tannic acid, Rosaceae) differ in the acceptability to snowshoe hares
gallic acid, and certain 3,4'-dihydroxyphenols were shown (Lepus americanus). The two series of tannins differ in their
to increase the growth rate of Anacridium melanorhodon overall stereochemistry at C-3 and C-4. Because the tannins
(Acrididae) when added to a diet deficient in them. These were highly purified, these differences in stereochemistry of
phenols appear to replace phenylalanine in the production the condensed tannins themselves must be important in these
of cuticular tanning agents (Bernays et al., 1983). Other ani- preferences (Clausen et al., 1990; Zucker, 1983).
mals, especially mammals, produce saliva with large num- Geraniin (26) (Fig. 12.10), a hydrolyzable tannin from
bers of proline residues which bind to tannins and presum- the leaves of Geranium species, releases ellagic acid on hy-
ably prevent them from binding other proteinaceous drolysis. Ellagic acid (10) is an antinutritional factor because
materials (Austin et ai., 1989; Hagerman and Butler, 1989; of its ability to bind essential metals (Harborne, 1989).
1991; Mehansho et al., 1983). The affmity and specificity Tannins also have been reported to bind to carbohydrates,
of binding is associated with the presence of oligosaccharide and, in contrast to binding with proteins, this binding is rela-
moieties in these proteins. The sugar portion (up to 40%) also tively pH independent. Molecular size, flexibility, and water
increases the solubility of the resulting tannin-carbohydrate solubility are important factors. In instances when the poly-
complex (Harborne, 1989). saccharide can sequester the hydrophilic aryl residues of the
Although there is an increase in excreted nitrogen when polyphenol (such as in holes in a crystal lattice or in hydro-
tannins are added to the diet of animals, this is probably phobic cavities), complexation is substantially enhanced
associated with a reduction in the utilization of digested and (Bilgener, 1988; Hagerman, 1989; Haslam and Lilley, 1985;
absorbed nutrients, rather than availability of protein (Hag- Spencer et ai., 1988; Ya et al., 1989). This binding can be
erman and Butler, 1991). Further, some components of con- strong; for example, the hydrolyzable tannins rugosin D (51)
densed tannin appear to be absorbed and produce systemic (Fig. 12.10) and sanguin H-6 bind tightly to Sephadex G-
effects (Hagerman and Butler, 1989). In any case, binding 25 and G-50 [a n-(l,6)-linked polymeric glucan (90-95%)]
of many digestive enzymes, such as trypsin, is lower than and do not desorb from those materials (Haslam et al., 1989;
that for many other proteins. Also, formation of tannin-pro- Spencer et al., 1988; Ya et al., 1989). The nature of binding
tein complexes often does prevent utilization of the protein of tannins to plant carbohydrates has yet to be ascertained.
in animal digestive systems. With concentrations of tannins
and proteins and pH conditions that might be expected to The Cost of Tannin Defenses and Resource
occur in insect herbivores and approaching those of the small Allocation
intestine of mammals, soluble tannin-protein products are Calculations by Lewis and Yamamoto (1989) indicate
formed (Hagerman and Robbins, 1987; Mole and Waterman, that the cost of production of tannins, lignins, and lipids are
1985, 1987a). Additionally, some enzymes are still active, among the highest for any plant-produced compounds. The
even though they are complexed with tannins (Mole and actual cost of lipids is less than calculated because these
Waterman, 1987a). The addition of the bile constituents cho- energy-reserve compounds are recycled within the piant, but
lic acid and glycocholate dissolves many tannin- protein tannins and lignins represent a uuidirectional flow of energy.
complexes at pH 6.2. The addition of cholic acid substan- The cost of condensed tannins is almost double that of hydro-
tially diminishes the inhibitory effect of tannins on the tryptic Iyzable tannins, if one assumes no reutilization of the latter
proteolysis of several proteins (Mole and Waterman, 1985), by the plant (Lewis and Yamamoto, 1989). The cost for
1987a). Although the digestive enzymes, alkaline phospha- proanthocyanidins and lignin is similar to that of protein.
tase and 5'-nucleotide phosphodiesterase, are strongly inhib- Synthesis of proanthocyanidins and lignin require major
ited by condensed tannins from sorghum seeds and quebra- commitments to the synthesis of phenylalanine, the second
cho, this inhibition was reversed by the addition of most expensive amino acid (after tryptophan). Further, be-
detergents, soluble polyvinylpyrrolidone, or by phophatidyl- cause large quantities of proanthocyanidins and lignin are
choline (a membrane component) (Blytt et al., 1988). deposited in plant tissues, the products of the phenylala-
Tannic acid and quebracho precipitate many times their uine-cinnamic acid pathway serve as a major sink for assim-
weight of ribulose-1 ,5-bisphosphate carboxylase (RuBPC), ilated carbon. Because of the importance of these processes,
the most abundant protein in most plant leaf tissues. At most vascular plants have high phenylalanine requirements.
mildly alkaline pH, RuBPC is not precipitated by tannins, One can argue that a ml\ior function of activation of the
 Tannins 209

phenylalanine-cinnamic acid pathway is to provide nitrogen densed tannins were found to be more common in nutrient-
for other metabolic processes (Lewis and Yamamoto, 1989). poor forests in Africa (McKey et aI., 1978). In general, these
When plants are grown under limited nitrogen supply, they theories predict that carbon-based defenses will be most im-
often respond by increased synthesis of phenolic com- portant in habitats in which nitrogen is limiting in accord
pounds, many of which are based on phenylalanine precur- with the conclusions of Lewis and Yamamoto (1989), based
sors. In this way (via dearnination of phenylalanine), optimal on plant biochemistry (see above and Chapter I).
use of plant nitrogen is made (Lewis and Yamamoto, 1989). Additionally, the type of defense used is determined by
Several theories relating the cost of plant defensive com- the phylogeny of parricular plants. Because the vegetation
pounds, availability of nutrients, and resource allocation of any system is determined by its past history and a number
have been proposed. A correlation of the type of defensive of phytogeographic effects, it seems unlikely that any gen-
compounds present and "apparency" or "nonapparency" eral theory will suffice for all instances.
to plant herbivores was noted by both Feeny (1976) and
Rhoades and Cates (1976). Feeny considered two fundamen- Other Proposed Functions of Tannins
tally different types of chemical defenses in plants. He con-
sidered tannins, which represented the major chemical de- Although many of the properties of tannins are linked to
fense of mature oak leaves, present in large amounts (up to their ability to bind proteins, individual tannins also may
exhihit biological activity. For example, tannic acid is a hep-
5% dry weight or more), to belong to a "quantitative" type
atotoxin (Zucker, 1983). Tannins may inhibit pathogens by
of defense and suggested that the potential for counteradap-
complexing with extracellular enzymes that the pathogens
tation by insects is limited (Feeny, 1976). He considered
produce or by interfering with metabolism of the pathogen
low-molecular-weight compounds, such as glucosinolates,
itself.
to act as "qualitative" barriers to herbivory and to be sus-
Tannins often possess molluscicidal activity against the
ceptible to rapid detoxication by coadapted insects. Further,
schistosomiasis-transmitting snail, Biomphalaria glabrata,
he considered plants that produce the latter type to be ephem-
.an intermediate host of Schistosoma mansoni. The active
eral, characteristic of early stages of community succession
molluscicidal and piscicidal compounds from Mammea sia-
and likely to be hard to find by their adapted enemies, relying
mensis (Clusiaceae), Polygonum stagninum, and Diospyros
primarily on escape in time and space. Oaks, in contrast, are
diepenhorstii are all proanthocyanidin polymers (Baiza et
climax forest dominants and are "bound to be found" by
al., 1989).
their enemies. In such plants, allocation of energy and nu-
trients required for quantitative defense would be likely to
bring increases in fitness. At almost the same time, Rhoades Medicinal Properties of Tannins
and Cates (1976) proposed that tissue availability and pre- Tannins are components of many traditional herbal medi-
dictability were major determinants of the type of defensive cines. Among these are tree paeony root bark (Paeonia lac-
compound used by the plant. They suggested that more toxic tiflora), bearberry (Arctostaphylos uva-ursi), agrimony
substances were associated with ephemeral tissues and di- (Agrimonia spp.), geranii herba (Geranium maculatum),
gestibility reducing compounds were characteristic of pre- meadowsweet (Filipendula ulmaria), and raspberry (Rubus
dictable tissues. However, in terms of cost to the plant, most idaeus) (Haslam et al., 1989; Okudaet aI., 1989b). The outer
compounds are actively turned over. This process requires portion of the root of the tree paeony (Paeonia lactiflora)
energy. Condensed tannins tum over very slowly, if at all, has been used to treat disorders of the bloodstream for at
and the relative metabolic cost for making them may com- least 2000 years. Although many medicinal uses are based
pare favorably to that of synthesizing compounds involved on the astringent properties of the plants, others appear to
in active turnover (Swain, 1979). Further, many predictable involve specific effects of tannin molecules (Haslam et al.,
plants contain qualitative or toxin-type defenses and a large 1989).
number contain tannins in addition to other defenses, such Many tannins are cytotoxic to cell cultures. Tannins may
as alkaloids. exhibit antibiotic activity by complexation of extracellular
After determining that herbivory and defense of tropical enzymes that the pathogens produce or by interference with
plants in Panama were not correlated with apparency, Coley the metabolism of the pathogen itself.
(1987) observed that slow-growing plants invest more heav- Several ellagitannins inhibit lipid peroxidation in rat liver
ily in defense because herbivory of a given amount would mitochondria and microsomes. Inhibition of 5-lipoxygenase
represent a proportionately greater loss than for rapidly in arachidonate metabolism in rats also was observed with
growing plants. In general, growth rates of plants have comparatively low levels of the ellagitannins geraniin (26)
evolved in response to resource availability of the habitat. (Fig. 12.10) and corilagin (11) (Fig. 12.2). Geraniin and tan-
The limiting resource may be light, water, nitrogen, or a nic acid strongly inhibit autoxidation of ascorbic acid. The
number of other factors. Defenses may be physical (such as reduction of several metallic ions (Cu2+, Fe3+, and Cr6+)
toughness ofleaves) or chemical. Several other workers have occurs in the presence of tannins at room temperature
found greater commitment to defensive substances in re- (Okuda et al., 1989b).
source-limited habitats (Coley, 1987). For example, con- Many tannins have weak antiviral and antitomor activity.
210 Tannins

However, a number of oligomeric hydrolyzable tannins ex- TAI'Ii'IIl'IS AND THE PRODUCTION OF LEATHER
hibit marked antitumor and anti-HIV activity. The dimeric
ellagitannins coriariin A (48), rugosin E (46), and oenothein Tannins have been used for millenia to convert raw animal
B (47) show the greatest antitumor activity (Fig. 12.16). hides into leather (Schmidt and Mayer, 1956). In this pro-
Orally administered oenothein B also has activity against cess, tannin molecules cross-link the protein and make it
certain solid tumors, such as the Ehrlich ascites tumor. These more resistant to bacterial and fungal attack. Molecules in
antitumor activities are thought to be due to enhancement the molecular weight range 500-2000 (3000) are most effec-
of the immune response of the host animals (Okuda et aI., tive, but the ability to bind effectively varies with different
1989b). tannin molecules. Many naturally occutring galloyl esters
The same dimeric ellagitannins inhibited replication of and gallotannins do not have tanning ability.
human immunodeficiency viruses (HIV). Oenothein B (47) Binding to protein is, in part, a function of molecular size.
inhibited vitus growth at concentrations of I and 10 fLg/ml. In the galloyl n-glucose series, the association with protein
The inhibitory effect may be due to the blocking of adsorp- is enhanced by the addition of each galloyl group and reaches
tion of mv on the cells and also inhibition of reverse tran- a maximum in the flexible disk-like structure of j3-penta-O-
scriptase activity (Okuda et aI., 1989b). A variety of other galloyl-n-glucose (8) (Haslam and Lilley, 1985). Effective
gallotannins and eliagitannins had similar activity. Gallotan- hydrolyzable tannins for tanning often have a chain of dep-
nins with a quinic acid core were especially active (Nonaka sidicaliy linked galloyl units in addition to simple galloyl
et aI., 1990). Geraniin (26, from Spondias mombin leaves, ester groups. Conformational mobility and flexibility is as
has pronounced antiviral activity (50 fLg/ml) against Cox- significant as molecular size. In the galloyl n-glucose series,
sackie and Herpes simplex viruses (Corthout et aI., 1991). where vicinal galloyl groups are constrained by the forma-
Several eliagitannins, for example, geraniin (26), inhibit tion of intramolecular biphenyl linkages and the generation
of hexahydroxydiphenoyl groups, the loss of conformational
mutagenicity of carcinogens. Ellagic acid (10), derived from
freedom is reflected in a reduced capacity to bind to protein
ellagitannins in plants, also has anticarcinogenic activity
(Haslam and Lilley, 1985).
(Okuda et aI., 1989b).

Correlation of Tannin Consumption and OTHER TYPES OF TAI'Ii'IIl'IS

Throat Cancer
In addition to these well-kuown types of tannins, other mate-
A correlation between consumption of beverages contain-
rials are kuown to precipitate proteins and have tanning ac-
ing large amounts of tannins and increased incidence of tivity. Among these are some lignans, such as nordihydrogn-
esophageal cancer has been noted (Morton, 1972, 1978, aiaretic acid (NDGA) from Larrea divaricata (Rhoades and
1989). This has been noted in particular with the consump- Cates, 1976) (see Chapter 8), and phlorotannins from brown
tion of hot tea (Japan, China, Soviet Asia, etc.), kaffir beer algae (Glombitza et aI., 1973, 1977; Koch and Gregson,
(made from Sorghum spp.) in southern Africa, some folk 1984). The latter group is found in brown algae of the family
medicines (such as Krameria spp. in Curacao), dry red wines Cystoseiraceae and consists of polymeric phloroglucinol de-
(western Europe and the United States), cider (the Normandy rivatives.
peninsula of France-where cider is made from apples too
astringent to eat), users of betel in southeastern Asia, and
khat (Catha edulis, Celastraceae) in Djibouti. As mentioned Al'IALYSIS OF TAI'Ii'IIl'IS
previously (see Chapter 11), approximately 30% of the dry
matter of tea leaves (Thea sinensis) is phenolic in nature. Tannins can be assayed by either determination of specific
The leaves are rich in flavan-3-0Is, principally as their 3-0- functional groups associated with particular structures or
gallate esters, but the level of procyanidin is minimal (Has- protein-binding assays (Hagerman and Butler, 1978, 1989).
lam and Lilley, 1985; Porter, 1989; Sanderson, 1972). Thear- Numerous types of assays have been carried out, often de-
ubigins are primarily prodelphinidins and their gallate esters pending on the goals and need for the information. The hide-
(Hemingway, 1989). Theaflavins in black tea result from powder method is still the standard of the tannin industry
oxidative coupling of thearubigins (see Chapter II). Interest- (Okuda et aI., 1989a). Many workers have utilized oxidative
ingly, in England, no elevated incidence is noted. Milk is analyses for phenols, either as a sole measure of tannins, or
usually added to tea and presumably precipitates the tannins before and after precipitation of tannins with protein. The
present. Many wines contain tannins, which are responsible most commonly used Folin-Denis assay is subject to inter-
for the astringent taste especially of red wines (see Chapter ference from phenolic groups present in protein and ascorbic
11). Wines without enough tannins seem flat and insipid, acid, if present. Hagerman and Butler (1989) recommend
but those with too much have a harsh and rough taste (Has- the Prussian Blue assay for this purpose, as it is less sensitive
lam, 1981). Similar considerations apply to cider and several to interferences, although some other limitations have been
other fruit-derived beverages. noted (Mole and Waterman, 1987b). Hagerman and Butler
 Tannins 211

(a)

OH~.o0H0 "
~
HO OH

,
H j : ( 1 00 0 0

00
0"'_' :~o ~-o"~o
"".
~O~ \ '~" o~ "'"~O"
HO 0 0 ,\
OH

0?:t"
ret}-(.
OH

~
HO

I fi OH "- I OH 0 HO 0'
HO OH HO OH

(b)
coriariin A (48) ble tannins.
Fig. 12.16 (a & b). Some biologically active hydrolyza
212 Tannins

suggest that the acid-butanol (or proanthocyanidin) assay anthocyanidin structures, in Chemistry and Significance of Tan-
is relatively specific for proanthocyanidins and is quite sensi- nins (R. W. Hemingway and J. J. Karchesy, eds.), 175-195
tive. Another method, the vanillin assay is useful for the Plenum Press, New York, 1989.
determination of flavanols (Hagerman and Butler, 1991; BATE-SMITH, E. C., The phenolic constituents of plants and their
Mole and Waterman, 1987b; Porter, 1989). Although often taxonomic significance. I. Dicotyledons. Linn. Soc. London
used, the iodate method for gallotannins is subject to many Bot. J., 58, 95-173 (1962).
interferences, and Hagerman and Butler suggest use of a BATE-SMITH, E. c., Age and distribution of galloyl esters, iridoids,
newer assay with rhodauine (Inoue and Hagerman, 1988). and certain other repellents in plants, Phytochemistry, 23,
A general, rapid, and highly reproducible method for deter- 945-950 (1984).
mination of tannins involving an agarose-gel-containing pro- BERNAYs, E. and S. WOODHEAD, Plant phenols utilized as nutrients
tein (called the radial diffusion method) is useful for the by a phytophagous insect, Science, 216, 201-202 (1982).
examination oflarge numbers of samples (Hagerman, 1987). BERNAYS, E., D. J. CHAMBERLAIN, and S. WOODHEAD, Phenols as
Difficulties and flaws in the analyses of tannins have been nutrients for a phytophagous insect Anacridium melanorhodon,
J. Insect Physiol., 29, 535-539 (1983).
reviewed (Schultz, 1989).
The enzyme tannase, produced by Aspergillus niger, spe- Bn.oENER, M., Chemical components of howler monkey (Alouatta
palliata) food choice and kinetics of tannin binding with natural
cifically cleaves the galloyllinkage to yield gallic acid and
polymers, Ph.D. Dissertation, Boston University, 1988.
a polyol, but it does not cleave hexahydroxydiphenyl-con-
taining esters. This reaction can be used to estimate the num- BLYIT, H. J., T. K. GUSCAR, and L. G. BUTLER, Antinutritional
effects and ecological significance of dietary condensed tannins
ber of galloyl units per polyol. The same enzyme cleaves
may not be due to binding and inhibiting digestive enzymes, J.
condensed tannins to yield flavonoids (porter, 1989). Chem. Ecol., 14, 1455-1465 (1988).
Purification of tannins is laborious but can be accom-
CLAUSEN, T. P., F. D. PROVENZA, E. A. BURRITT, P. B. REICHARDT,
plished by a variety of techniques (Hagerman and Butler, and J. P. BRYANT, Ecological implications of condensed tannin
1991). structure: A case study, J. Chem. Ecol., 16, 2381-2392 (1990).
Characterization of hydrolyzable tanuins can be accom- COLEY, P. D., Interspecific variation in plant anti-herbivore proper-
plished by hydrolysis and identification of the fragments ties: The role of habitat quality and rate of disturbance, New
by standard techuiques such as cochromatography on paper, Phytol., 106,(Suppl.), 251-263 (1987).
TLC or HPLC and NMR, UV, and MS (Karchesy et al., CONN, E. E. and T. SWAIN, Biosynthesis of gallic acid in higher
1989; Okuda et al., 1989a; Porter, 1989). Condensed tannins plants, Chem. Ind., 592-593 (1961).
can be cleaved in mild acidic solution to form a flavan-3-01 CORK, S. J. and A. K. KROCKENBERGER, Methods and pitfalls of
and a quinone methide. The quinone methide can be captured extracting condensed tannins and other phenolics from plants:
with a suitable nucleophiJe (e.g., -SCH2Ph) to form 4-thio- msights from investigations on Eucalyptus leaves, I. Chern. Ed.,
benzyl derivatives and permit determination of the constitu- 17, 123-134 (1991).
ent flavonoid units (Porter, 1989). The 4-thiobenzyl deriva- CORTI<OUT, J., L. A. PIETERS, M. CLAEYS, D. A. VANDEN BEROHE,
tives may be reduced with Raney-nickel to yield flavan-3- and A. J. VLlETNICK, Antiviral ellagitannins from Spondias
ols (Porter, 1989). mombin, Phytochemistry, 3D, 1129-1130 (1991).
The characterization of condensed tannins has been DAHLGREN, R. M. T., S. R. JENSEN, and B. J. NIELSEN, A revised
greatly aided by structural and conformational analysis by classification of the angiosperms with comments on correlation
NMR spectroscopy (Ferreira and Brandt, 1989). Both 'H_ between chemical and other characters, in Phytochemistry and
and 13C_NMR spectroscopy has been used to assign atoms Angiosperm Phylogeny (D. A. Young and D. S. Seigler, eds.),
in these complex structures. Fast atom bombardment (FAB) 149-204, Prseger, New York, 1981.
mass-spectral analysis permits observation of the parent ion DEWICK, P. M. and E. HAsLAM, Phenol biosynthesis in higher
as well as fragments of these compounds (Barofsky, 1989; plants. Gallic acid, Biochem. J., 113, 537-542 (1969).
Haslam and Lilley, 1986; Okuda et al., 1989a; Porter, 1989). FEENY, P. P., Plant apparency and chemical defence, in Biochemi-
cal Interaction between Plants and Insects (J. W. Wallace and
R. L. Mansell, eds.), Recent Advances in Phytochemistry Vol
REFERENCES 10, 1-40, Plenum Press, New York, 1976.
FBRRERIA, D. and E. V. BRANDT, New NMR experiments applicable
AUSTIN, P. J., L. A. SUCHAR, C. T. ROBBINS, and A. E. HAGERMAN, to structure and conformation analysis, in Chemistry and Signif-
Tannin-binding proteins in saliva of deer and their absence in icance of Tannins (R. W. Hemingway andJ. J. Karchesy, eds.),
saliva of sheep and cattie, J. Chem. Ecol., 15, 1335-1347 153-173, Plenum Press, New York, 1989.
(1989). GLOMBITZA, K. W., H. U. ROSENER, H. Vn.TER, and W. RAuwALD.
BALZA, F., Z. ABRAMOWSKI. G. H. N. TOWERS, and P. WIRIYACHI- Phloroglucin aus Braunalgen, Planta Medica, 24, 301-302
TRA, Identification of proanthocyanidin polymers as the piscici- (1973).
dal constituents of Mammea siamensis, Polygonum stagninum, GLOMBITZA, K. W., E. SATILER, F. W. WEHRLI, and G. EcKHARDT,
and Diospyros diepenhorstii, Phytochemistry, 28, 1827-1830 Polyhydroxyphenylether aus der Phaeophycee Ha/idrys sili·
(1989). quosa, Tetrahedron, 33, 1239-1244 (1977).
BAROFSKY, D. F., FAB-MS applications in the elucidation of pro- GROSS, G. G., Enzymology of gallotannin biosynthesis, in Plant
 Tannins 213

Cen Polymers: Biogenesis and Biodegradstion (N. G. Lewis (T. Swain, 1. B. Harborne, and C. F. van Sumere, eds.), Recent
and M. G. Paice, eds.), 108-121, American Chemical Society, Advances in Phytochemistry Vo!. 12,475-523, Plenum Press,
Washington, DC, 1989. New York, 1979.
GROSS, G. G., Enzymatic synthesis of gallotannins and related com- HASLAM, E., Vegetable tannins, in Secondary Plant Products (E.
pounds, in Phenolic Metabolism in Plants (H. A. Stafford and E. Conn, ed.), Vol. 7 of The Biochemistry of Plants (P. K.
R. K. Ibrabim, eds.), 297-324, Plenum Press, New York, 1992. Stumpf and E. E. Cona, eds.), 527-556, Academic Press, New
GUPTA, R. K., S. M. K. AL-SHAFI, K. LAYDEN, and E. HASLAM, York,1981.
The metabolism of gallic acid and hexabydroxydiphenic acid HASLAM, E., Proanthocyanidins, in The FIavonoids: Advances in
in plants. Part 2. Esters of (S)-hexabydroxydephenic acid with Research (J. B. Harborne and T. J. Mabry, eds.), 417-447,
D-glucopyranose(4C,), 1. Chem. Soc., Perkin Trans. I, Chapman & Hall, London, 1982.
2525-2534 (1982). HASLAM, E., Hydroxybenzoic acids and the enigma of gallic acid,
HADDOCK, E. A., R. K. GUPTA, S. M. K. AL-SHAFI, and E. HASLAM, in The Shikimic Acid Pathway (E. E. Cona, ed.), Recent Ad-
The metabolism of gallic acid and hexabydroxydiphenic acid vances in Phytochemistry Va!. 20, 163-200, Plenum Press,
in plants. Part I. Introduction. Naturally occurring galloyl esters, New York, 1986.
J. Chem. Soc. Perkin Trans. I, 2515-2524 (1982a). HASLAM, E., Plant Polyphenols, Cambridge University Press, Cam-
HADDOCK, E. A., R. K. GUPTA, and E. HASLAM, The metabolism bridge, 1989.
of gallic acid and hexabydroxydiphenic acid in plants. Part 3. HASLAM, E. and T. H. Ln.LEY, New polyphenols for old tannins,
Esters of (R)-and (S)-hexabydroxydiphenic acid and dehydro- in The Biochemistry of Plant Phenolics (C. F. van Sumere and
hexabydroxydiphenic acid with D-glucopyranose, J. Chern. Soc. P. J. Lea, eds.), 237-256, Clarendon Press, Oxford, 1985.
Perkin Trans. I, 2535-2545 (1982b). HASLAM, E. and T. H. LILLEY, Polyphenol complexation, in F1avo-
HADDOCK, E. A., R. K. GUPTA, S. M. K. AL-SHAFI, K. LAYDEN, noids and Bioflavonoids, 1985 (L. Farkas, M. Gabor, and F.
E. HASLAM, and D. MAGNOLATO, The metabolism of gallic acid Klillay, eds.), 113-138, Akademiai Kiade, Budapest, 1986.
and hexabydroxydiphenic acid in plants. Part 4. Biogenetic and HASLAM, E., T. H. LILLEY, Y. CAl, R. MARTIN, and D. MAGNOLATO,
molecular taxonomic considerations, Phytochemistry, 21, Traditional herbal medicines-The role of polyphenols, Planta
1049-1062 (1982c). Medica, 55, 1-8 (1989).
HAGERMAN, A. E., Radial diffusion method for determining tannin
HELLER, W. and G. FORKMANN, Biosynthesis, flavans and proantho-
in plant extracts, 1. Chern. Eco!., 13, 437-449 (1987).
cyanidins, in The Flavonoids: Advances in Research since 1980
HAGERMAN, A. E., Extraction of tannin from fresh and preserved (J. B. Harborne, ed.), 399-425, Chapman & Hall, London,
leaves, J. Chem. Bco!., 14,453-461 (1988). 1988.
HAGERMAN, A. E., Chemistry of tannin-protein complexation, in HEMINOWAY, R. W.o Structural variations in proanthocyanidins and
Chemistry and Significance of Tannins (R. W. Hemingway and their derivatives, in Chemistry and Significance of Tannins (R.
J. J. Karchesy, eds.), 323-333, Plenum Press, New York, 1989. W. Hemingway and J. J. Karchesy, eds.), 83-107, Plenum
HAGERMAN, A. E. and L. G. BUTLER, Protein precipitation method Press, New York, 1989.
for the quantitative determination of tannins, J. Agric. Food HlLLIS, W. E., Biosyothesis oftannins, in Biosynthesis and Biodeg-
Chern., 26, 809-812 (1978). radation of Wood Components (T. Hignchi, ed.), 325-347, Aca-
HAGERMAN, A. E. and L. G. BUTLER, The specificity of proanthocy- demic Press, Orlando, FL, 1985.
anidin-protein interactions, J. Bioi. Chem., 256, 4494-4497
INOUE, K. H. and A. E. HAGERMAN, Detennination of gallotannin
(1981).
with rhodanine, Anal. Biochem., 169, 363-369 (1988).
HAGERMAN, A. E. and L. G. BUTLER, Choosing appropriate methods
KARCHESY, J. J., Y. BAE, L. CHAi.KER-SCO'IT, R. F. HELM, and L.
and standards for assaying tannin, J. Chem. Eco!., 15,
Y. Foo, Chromatography of proanthocyanidins, in Chemistry
1795-1810 (1989).
and Significance of Tannins (R. W. Hemingway and J. J. Karch-
HAGERMAN, A. E. and L. G. BUTLER, Tannins and Iignins, in Herbo- esy, eds.), 139-151. Plenum' Press, New York, 1989.
vores: Their Interactions with Secondary Plant Metabolites. KOCH, M. and R. P. GREaSON, Brominated phlorethols and nonha-
Vol. I (G. A. Rosenthal and M. R. Berenbaum, eds.), 355-388,
logenated phlorotanoins from the brown alga Cystophora cong-
Academic Press, San Diego, CA, 1991.
esta, Phytochemistry, 23, 2633-2637 (1984).
HAGERMAN, A. E. and C. T. ROBBINS, Implications of soluble tan-
LEWlS, N. G. and E. YAMAMOTO, Tannins-Their place in plant
nin-protein complexes for tannin analysis and plant defense
metabolism, in Chemistry and Significance of Condensed Tan-
mechanisms, J. Chem. Bco!., 13, 1243-1259 (1987).
nins (R. W. Hemingway and J. J. Karchesy, eds.), 23-46,
HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod. Plenum Press, New York, 1989.
Rep., 4, 323-344 (1986).
MARTIN, J. S. and M. M. MARTIN, Tannin assays in ecological
lIARaORNE, J. B., Recent advances in chemical ecology, Nat. Prod. studies: Lack of correlation between phenolics, proanthocyanid-
Rep., 7, 85-109 (1989). ins. and protein precipitating constituents in mature foliage of
HASLAM, E., The Shikimate Pathway, John Wiley and Sons, New six oak species, Oecologia (Berlin), 54, 205-211 (1982).
York,1974. MARTIN, J. S. and M. M. MARTIN, Tannin assays in ecological
HASLAM, E., Natural proanthocyanidins, in The Flavonoids (1. B. studies. Precipitation of ribulose-I ,5-biphosphate carboxylase!
Harborne, T. J. Mabry, and H. Mabry eds.), 505-559, Academic oxygenase by tannic acid, quebracho. and oak foliage extracts,
Press, New York, 1975. J. Chem. Eco!., 9, 285-294 (1983).
HASLAM, E., Vegetable tannins, in Biochemistry of Plant Phenolics MARTIN,1. S., M. M. MARTIN, and E. A. BERNAYS, Failure of tannic
214 Tannins

acid to inhibit digestion or reduce digestibility of plant protein PORTER, L. J., TANNINS, in Plant Phenolics (J. B. Harbome, ed.),
in gut fluids of insect herbivores, J, Chern. Ecol., 13, 605-621 Vol. I of Methods in Plant Biochemistry (P. M. Dey and J. B.
(1987). Harbome, eds.), 389-419, Academic Press, New York, 1989.
McKEY, D. G., P. G. WATERMAN, C. N. MEr, J. S. GARTLAN, and RHOADES, D. F. and R. G. CATES, Toward a general theory of plant
T. T. STRUSAKER, Phenolic content of vegetation in two African antiherbivore chemistry, in Biochemical Interaction between
rain-forests: Ecological implications, Science, 202, 61-64 Plants and Insects (J. W. Wallace and R. L. Mansell, eds.),
(1978). Recent Advances in Phytochemistry, Vol 10), 168-213, Plenum
MEHANSHO, H., A HAGERMAN, S. CulMENrs, L. BUTI.ER, J. Press, New York, 1976.
RooLER, and D. M. CARLSON, Modulation of proline-rich pro- ROBBINS, C. T., T. A. HANLEy, A E. HAGERMAN, O. H1EL:FORD,
tein biosynthesis in rat parotid glands by sorghums with high D. L. BAKER, C. C. SCHWARTZ, and W. W. MAUTZ, Role of
tannin levels, Proc. Natl. Acad. Sci. USA, 80, 3948-3952 tannins in defending plants against ruminants: Reduction in pro-
(1983). tein availability, Ecology, 68, 98-107 (1987).
MOLE, S. and P. G. WATERMAN, Stimulatory effects of tannins Roux, D. G. and D. i'ERREIRA, "Angular" and "linear" condensed
and cholic acid on tryptic hydrolysis of proteins: Ecological tannins. Chemical parameters which direct the course of biomi-
implications, J. Chern. Ecol., 11, 1323-1332 (1985). metic condensations, in Flavonoids and Bioflavonoids, 1981 (L.
Farkas, M. Gabor, F. Kallay, and H. Wagner, eds.), 81-103,
MOLE, S. and P. G. WATERMAN, Tannins as antifeedants to mamma- Elsevier, Amsterdam, 1982.
lian herbivores-Still an open question? in Allelochemicals:
SADO, R.o Pathway of gallic acid biosynthesis and its esterification
Role in Agriculture and Forestry (G. R. WalJer, ed.), ACS Sym-
with catechins in young tea shoots, Agric. BioI. Chern., 47,
posium Series 330, 572-587, American Chemical Society,
455-460 (1983).
Washington, DC, 1987a.
SANDERSON, G. W., The chemistry of tea and tea manufacturing,
MOLE, S. and P. G. WATERMAN, A critical analysis of techniques (V. C. Runeckles and T. C. Tso, eds.), Recent Advances in
for measuring tannins in ecological studies. I. Techniques for Phytochemistry, Vol. 5 247-316, Academic Press, New York,
chemically derming tannins, Oecologia, 72, 137-147 (l987b). 1972.
MOLE, S. and P. G. WATERMAN, A critical analysis of techniques SCHMIDT, O. T. and W. MAYER, Natiirliche Gerbstoffe, Angew.
for measuring tannins in ecological studies. II. Techniques for Chern., 68, 103-105 (1956).
biochemically derming tannins, Oecologia, 72, 148-156 SCHULTZ, J. C., Tannin-insect interactions, in Chemistry and Sig-
(1987c). nificance of Tannins (R. W. Hemingway and J. J. Karchesy,
MORTON, J. F., Further associations of plant tannins and human eds.), 417-433, Plenum Press, New York, 1989.
cancer, J. Crude Drug Res., 12, 1829-1841 (1972). SPENCER, C. M., Y. CAr, R. MARTIN, S. H. GAFFNEY, P. N.
MORTON, J. F., Economic botany in epidemiology, Econ. Bot., 32, GoULDING, D. MAGNOLATO, T. H. LnLEY, and E. HAsLAM, Pol-
111-116 (1978). yphenol complexation-Some thoughts and observations, Phy-
MORTON, J. F., Tannin as a carcinogen in bush-tea, tea, mare, and tochemistry, 27, 2397-2409 (1988).
khat, in Chemistry and Significance of Tannins (R. W. Heming- STAFFORD, H. A, The enzymology of proanthocyanidio biosyn-
way and J. J. Karchesy, eds.), 403-416, Plenum Press, New thesis, in Chemistry and Significance of Condensed Tannins
York,1989. (R. W. Hemingway and J. J. Karchesy, eds.), 47-70, Plenum
Press, New York, 1989.
NONAKA, G., 1. NISHIOKA, M. NISHIZAWA, T. YAMAGISHI, Y. KASHI-
WADA,G. E. DUTSCHMAN, A J. BODNER, R. E. Kn.KUSKlE, Y. STBlNLY, B. A and M. BERENEAUM, Histopathological effects of
CHENG, and K. LEE, Anti-AIDS agents, 2. Inhibitory effects of tannins on the midgut epithelium of Papilio polyxenes and Papi-
tannins on mv reverse transcriptase and ffiV replication in H9 lio glaucus, Entomol. Exp. Appl., 39, 3-9 (1985).
lymphocyte cells, J. Nat. Prod., 53, 587-595 (1990). SWAIN, T., Phenolics in the environment, in The Biochemistry of
Plant Phenolics (T. Swain, J. B. Harbome, and C. F. van Sumere,
OKUDA, T., T. YosHIDA,andT.HATANo,Newmethodsofanalyzing
eds.), Recent Advances in Phytochemistry, Vol. 12,617-640,
tannins, J. Nat. Prod., 52, 1-31 (1989a).
Plenum Press, New York, 1979.
OKUDA, T., T. YOSHIDA, and T. HATANO, Ellagitannins as active WOLTER-FILHO, W., A I. DA ROCHA, M. YOSHIDA, and O. R. GOT-
constituents of medicinal plants, Planta Medica, 55, 117-122 1LIEB, Chemosystematics of Rhabdodendron. Phytochemistry,
(1989b). 28, 2355-2357 (1989).
PENO, S. and C. JAy-ALLEMAND, Use of antioxidants in extraction YA, C., S. H. GAFFNEY, T. H. LILLEY, and E. HASLAM, Carbohy-
of tannins from walnut plants, J. Chern. Ecol., 17, 887-896 drate-polyphenol complexation, in Chemistry and Significance
(1991). of Tannins (R. W. Hemingway and J. J. Karchesy, eds.),
PORTER, L. J., Flavans and proanthocyanidins, in The Flavonoids: 307-322, Plenum Press, New York, 1989.
Advances in Research since 1980 (J. B. Harbome, ed.), 21-62, ZUCKER, W. V., Tannins: Does structure detennine function?-An
Chapman & Hall, London, 1988. ecological perspective, Am. Nat., 121, 335-365 (1983).
13
Nonprotein Amino Acids

Introduction usually occur as components of peptides or proteins and are

Nonprotein Amino Acids Derived from Aspartic Acid linked by peptide bonds (Fig. 13.1) or free only in small
Lysine-Derived Nonprotein Amino Acids amounts, although, under unusual circumstances, free pri-
Pipecolic Acid and Related Compounds mary amino acids sometimes accumulate in unusual quan-
L-Mimosine tities. Examples are histidine (1) in ripening bananas, argi-
Homoserine nine (2) in apple trees and some Vida species, and proline
L~Canavanine (3) in Caragana wood. A few primary amino acids such as
Canaline cystine (4) and trans-4-hydroxY-L-proline (5) do not occur in
L-Azetidine-2-carboxylic Acid and Related Amino Acids protein but are synthesized secondarily from primary amino
Nonprotein Amino Acids Derived from Isoleucine acids. The latter of these is found in the cell wall proteins
L-Hypoglycin A and B of higher plants.
L-Homoarginine and Related Compounds from the Genera In addition to these ubiquitous amino acids, an additional
Lathyrus and Vida (Fabaceae) 700 amino acids that are not involved in primary metabolism
L-Homoarginine have been reported. About 300 of these are found in plants
L-Lathyrine (Hunt, 1991). Most nonprotein amino acids occur free, as
Amino Acids from 3-Cyano-L-a1anine and Asparagine their 4-glutamyl (or 'Y-glutamyl) derivatives, or linked to
Nonprotein Amino Acids Developed from Glutamic Acid carbohydrate moieties. There are two main types of nonpro-
Other Nonprotein Amino Acids tein amino acids. A number of nonprotein amino acids are
Sulfur-Containing Amino Acids similar to primary amino acids; others are unrelated to any of
Djenkolic Acid these. Members of the first group may arise in three principal
L-Indospicine
ways. Some nonprotein amino acids arise by simple struc-
A1biziine
taral modifications of protein amino acids; others arise by
5-HydroxY-L-tryptophan
modification of the pathways leading to protein amino acids;
3,4-DihydroxY-L-phenylalanine (L-dopa)
a third group arises by novel routes.
Selenium-Containing Amino Acids
As an example of the first type, tryptophan (6) is con-
Nonprotein Amino Acids in Seeds of the Cucurbitaceae
verted to 5-hydroxY-L-tryptophan (7), which occurs in plants
Compounds Involved in Iron Metabolism in Plants
as its acetyl, 4-glutamyl, and oxalyl derivatives (Fig. 13.2).
Ethylene and Its Precursors
An example of pathway modification (the second type)
Trigonelline, a Hormone Which Controls the Cell Cycle
in Plants is the production of meta-substitoted amino acids such as
Miscellaneous Nonprotein Amino Acids 3(3-carboxyphenyl)a1anine (8) via a modified shikimic acid
Distribution of Nonprotein Amino Acids in Plants pathway (Fig. 13.3) (Bell, 1976).
References An example of the third type (novel pathway) is the pro-
duction of erythro-4-methylglutamic acid (9) from L-Ieucine
(10) in Gleditisia triacanthos (honey locust, Fabaceae) and
INI'RODVCTIOI'I
not from glutamic acid and SAM or from lysine as in many
Approximately 25-30 amino acids are involved in primary other plants (Fig. 13.4).
metabolism (Miflin, 1980; Rosenthal and Bell, 1979). Many Where L-amino acids are concerned, the a-carbon atom
of these are listed in Fig. 13.1. These primary amino acids is usually designated by the prefix D- or L- in accord with

215
216 Nonprotein Amino Acids

co,H
cro,H Co,H co,H

HNO
co,H
I I
I .... H

-<
NB,·C-H NB,-C-H I
NB,C-H NB,C-H
I
CH,

L-oIoDIne
A
~"allne L-leudlle (10)
~
L-........I•• (15) L-p....Une (3)
co,H CO,H
I I

8
NB,~-H NH,.~-H

yu. NH.-~-H
co,H
I
CR,

I
~ ?Iz
SH
CR,
l.-cyItel.e (U) L-..rI.. (11)
L·melhIonIne (16) L-pheJIyIal.nlne
co,H co,H
co,H L-trnotop.... 1
COzH I NB,C·H I

¢
NB,·C-H 1 NH.·C-H
I I
NH:r~-H I co,H CB,
H-C-OH ?liz I
NH,-CH, yu.
I
I
CH, f~0
CR, c:u:C~o
OH
L-threonlne (14) L-aipanoglne (93) glycine L-/yruoiDe L-glu!amloe (Al
co,H CO,H co,H
COzH co,H
I I
I I I NB,C-H NH2"~-H
NH,C-H NH,·C-H
I
NH:r C - H
I
(~ (6JI zl,
[1
CR, CR,
I
co,H I NH

f' ~H, I
C=NH
Co,H NH, I
NHl
(a) L-a&partl. add (22) L-glutamle add (67) L-Iysl•• (23) L-arginine (%) L-bIsIIdlne(l)

CO,H

HO HO
NH,-C-H
I

~CO'H ~CO'H
I
(CH,),
I
I
CH,
CH,
I
NH,
trans-4-hydroxyproline (5) N-metbyl-trans-4-hydroxyproline (72) L-ornithine (71)

CO,H
I CO,H CO,H
CO,H NH,·C-H I I
I
NH,C-H I NH,-C-H NH,-C-H
I CH, I I
CH, I CHz CH:z
I
0---f:..0 CH,
I I
s---s
I
OH

O·••elyl·L..erlne (36) L-homoserine (13) L·eystlne (4)

(b)

Fig. 13.1 (8 & b). Fonnation of the peptide bond and some primary amino acids.
 Nonprotein Amino Acids 217

CJc():,
"'" I I NHz
COZH

----+
HO~COZH
I I NH
NH "'" NH Z

L-tryptophan (6) 5-hydroxy-L-tryptophan (7)

Fig. 13.2. Biogenesis of 5-hydroxY-L-tryptophan.

the structure of D- or L-serine (11) (Rosenthal, 1982). L- (Fowden et aI., 1970; Fried and Sherma, 1982; Hunt, 1991;
Amino acids other than cysteine (12) are (S)-aminoalkanoic Rosenthal, 1982). A number of gas-chromatographic meth-
acids. One major problem is the establishment of chiral cen- ods have been devised for the determination of amino acids
ters in the molecules (Hunt, 1991). The prefixes of carbohy- (Jaeger et aI., 1981). Electrophoresis has also been widely
drate nomenclature, erythro and threo, are still used. It is used. By changing the buffer pH, it is possible to effect
common practice to use the terms cis and trans to refer to separation of different compounds. However, most amino
the positions of hydroxyl or other ring substituents with re- acids are identified and quantified by means of ion-exchange
spect to the carboxyl groups in proline or pipecolic acid resins and automated equipment. High-performance liquid
derivatives. Greek letters are being replaced with numbers, chromatograph (HPLC) with reversed-phase and ion-ex-
but are often seen in the literature. Many workers still prefer change supports has played a major role in simplification of
to give trivial names to novel amino acids (Hunt, 1991; Ro- the analyses of amino acids (Hunt, 1991; Rosenthal, 1991).
senthal, 1982). Various aspects of the purification and characterization of
The toxicity of many nonprotein amino acids to the nonprotein amino acids have been reviewed (Hunt, 1991;
bruchid beetle Callosobruchus maculatus (the southern cow Rosenthal, 1982).
pea weevil) has been evaluated; the larvae normally feed on Most amino acids exist as charged species or zwitterions
the seeds of Vigna unguiculata) (Janzen et aI., 1977). These at physiological pH. No attempt has been made to draw
beetles are adapted to the amino acids accumulated in seeds structures in the ionized form in this chapter.
of the host plant, but they are poisoned by many other non-
protein amino acids at the level of 0.1 % in artificial diets.
The role of nonprotein amino acids as ecological factors in NOl'lPROTEIN AMINO ACIDS DEKIVED FROM
the life history of insects has been reviewed (Bernays, 1983). ASPARTIC ACID
Nonprotein amino acids usually occur free in plants, or
as their 4-glutamyl derivatives. These amino acids generally Homoserine (13), threonine (14), isoleucine (15), methio-
are extracted with ethanol or with alcohol-water mixtures nine (16), and cysteine (12) are all derived from aspartic
(Rosenthal, 1982, 1991). Most amino acids can be detected acid (22). Lysine (23) (Fig. 13.1) is derived via 3-aspar-
by the characteristic reaction with ninhydrin, which produces tylsemia\dehyde and is, in tum, involved in formation of
blue to purple colored chromophores (Rosenthal, 1991). many nonprotein amino acids such as pipecolic acid (17)
Most of the early work with nonprotein amino acids was and mimosine (18) (Figs. 13.5 and 13.6) (Rosenthal, 1982).
done using paper-chromatographic and thin-layer techniques D-Lysine is a precursor of L-pipecolic acid in many plants

chorismic acid isoprephenic acid

isochorismic acid

HOZC~COZH
I 3(3-carboxyphenyl)alanine (8)

"'" NHz

Fig. 13.3. Biogenesis of isoprephenic acid and 3(3-carboxyphenyl)alanine (modified from Rosenthal, 1982; used with permission of the copyright
owner, Academic Press, Orlando, FL).
218 Nonprotein Amino Acids

CH, NH,
CH3
I I
NH2 Gledilsla triacanthos I
HO,CCHCH,CHCO,H
I other plants
lysine
CH,CHCH,CHCO,H F.b.....
ery,hro-4-methylglutamic .cid (9)
L-I.ucin. (10)

NH,

HO,CCH,CH,CHCO,H
I
glutamic acid + methyl

Fig. 13.4. Biogenesis of erythro-4-methylg1utamic acid.

and bacteria. L-Pipecolic acid, in tum, gives rise to L-Iysine amination reaction, cyclization to a I-piperidine-2-carbox-
(Fangmeier and Leistner, 1981). ylic acid (25) and reduction to pipecolic acid or by initial
A number of other nonprotein amino acids are involved conversion to a semialdehyde and conversion to a I_piperi_
in the biosynthesis of lysine (Fig. 13.5). Among these are dine-6-carboxylic acid (24) prior to the formation of pipeco-
2,3-dihydrodipicolinic acid (19), N"-succinyl-2,6-diaminop- lic acid (17) (Rosenthal, 1982). Pipecolic acid is derived
imelic acid (20), and 2,6-diaminopimelic acid (21). from L-Iysine in fungi and D-Iysine in higher plants.
Lysine (23) also serves as a precursor for several major Several other nonprotein amino acids are formed from
types of nonprotein amino acids, (e.g., pipecolic acid (17), pipecolic acid. Among these are a number of compounds
mimosine (18), 4-methyl glutamic acid (9), 4-methyleneglu- illustrated in Fig. 13.7. Fowden (1970) has integrated the
tamic acid (26), and a series of compounds found in the possible metabolic relationships of this group of compounds
families Aceraceae, Hippocastanaceae, and Sapindaceae into a coherent scheme. In this model, rearrangement of exo-
(see below). (cis)-3,4-methanolproline (27) can produce baikiain (28) or
methyleneproline (29) (Rosenthal, 1982).
Ring openings interrelate cis-3- and cis-4-methylated pro-
LYSIl'IE·DERIlIED NOl'lFROTEIN AMINO ACIDS line (30, 31) as well as pipecolic acid (17) into these meta-
bolic pathways. Addition of water to baikiain (28) can lead
Pipecolic Acid and Related Compounds
to trans-4- and/or trans-5-hydroxypipecolic acid (32, 33).
Pipecolic acid (17) can arise from lysine via two distinct 4-Methylglutamic acid (9) can feed into cis-4-methylproline
mechanisms (Fig. 13.6). This synthesis can occur by a trans- (31), 4-methyleneglutamic acid (26) into 4-methylenepro-

aspartic acid (22)

- 3-aspartyl
phosphate
---+- 3-aspartyl
semialdehyde
- o HO,C N CO,H

2,3·dihydropicoJinic
-
o
acid (19)

HO,C N

.6,1'piperidine.2,6.
CO,H- N'.succlnyl·2·amino.6.oxo.
plmelicacld - H,N-CiH
?O,H

(CIH,),
fo,H
?H, -
?H,

- /
dicarboxyJic acid ~O,H H-C - - NH--<:=O
I
H,N-CIH CO,H
lysine (23)

HO/=\ f' (lH,),

H-~-NH,
N'-succinyl.2,6.
diaminopimelic acid (20)
0\J-CH,-CH-CO,H
CO,H

L-mimosine (18) meso·2,6·diaminoplmeJic

acid (21)

Fig. 13.5. Biosynthesis of lysine from aspartic acid in higher plants (modified from Rosenthal, 1982; used with pennission of the copyright owner,
Academic Press. Orlando, FL.).
 Nonprotein Amino Adds 219

<
NH, 2.ox~6~aminohexanoic acid

(C,H')4

H-~-NH, ,11.piperidine-2.carboxylic acid (25)

:,0~
CO,H
2-aminoadipic semialdebyde
lysine (23) pipecolic acid
(17)

~1-piperidine.6.carboxylic acid (24)

Fig. 13.6. Possible biosyntbetic pathways to pipecolic acid (Rosenthal, 1982; modified and used with pemrission of the copyright owner, Academic
Press, Orlando, FL).

line, and 2-aminoadipic acid into pipecolic acid (17). Finally, L-Mimosine
the conversion of cis-2-(carboxycyclopropyl)glycine (34) to L-Mimosine (18) (Fig. 13.5) is a toxic heterocyclic amino
exocyclic proline can be related to the formation of certain acid that has been reported from several legumes. Among
c.-imino acids (Rosenthal, 1982). the plants that contain mimosine are Mimosa pudica and
exo-cis-3,4-Methanoproline (27) is found in Aesculus Leucaena leucocephala. The latter is often cultivated in trop-
parviflora (Hippocastanaceae). Pipecolic acid (17) is found ical countries as a windbreak, a forage crop, and as a shade
in many legume seeds, especially in the genera Acacia and crop for coffee, tea and cacao (Rosenthal, 1991). This native
Prosopis (Rosenthal, 1982). A sulfate of pipecolic acid, of Mexico is capable of fixing nitrogen and is a quite agres-
trans-4-hydroxypipecolic acid-4-sulfate (35), has been re- sive species.
ported from Peltophorum africanum (Fabaceae) (Evans et O-Acetylserine and 3,4-dihydroxypyridine give rise. to
al., 1985). minIosine in the presence of an extract of Leucaena. An L-

tmns-4-hydroxypipecolic cis-3-methylproUne (30) cis-4-methylproline (31) 4-methylglutamic acid (9)

aeid(32) '" , , / h
A
Q NH
++
CO,H
A
~ A
-- ( __
__
NH CO,H
)-.CO,H
NH _ HC
,~
NH
I'
HOD¥ baikiain (28) 4-metbyleneprollne (29) ;-CCH,CHCO,H
exo-3,4-metbanoproline HO,C

/ t
NH C02H (27) 4·methyleneglutamlc acid

f'
(26)

n
trans-5-hydroxypipecolic ISO,"
aeid(33)
NH, ~HO,C(CH,)JCHCO,H r"l
HO'CAJHCO'H ~ __A
NH CO,H
2-aminoadipJcacid ~_._A
NH CO,H
H H

cts-2-(carboxycyclopropyl)glycine (34) pipecolic acid (17) 1rans4·hydroxypipecolic acid-

4-sulrate (35)

Fig. 13.7. Proposed sequence of rearrangements leading to cyclopropanoid and cyclopentanoid nonprotein amino acids (Fowden. 1970; Rosenthal,
1982; modified and used with pennission of the copyright owners, John Wiley and Sons, Ltd, Chichester, and Academic Press, Orlando, FL).
220 Nonprotein Amino Acids

0 0 OR This compound is known from at least 1500 species in 240

IC-O.PO)H I I genera of that subfamily (Bell et aI., 1978). The seeds of
C02K

I
1 C·R ,Hz many of these plants contain 2-4% of L-canavanine and a
I I
r
CR, CR,
number contain 10-15%. In certain legumes, the total pro-
CR,
I
H.C.NHl
--+
I
H.C.NHl
-+
I
H.C.NH1
portion of seed nitrogen allocated to canavanine may repre-
sent 95% of the total amino acid nitrogen of the seeds (Ro-

I
H-C-NHl

COlK
I
COl"
I I
COlK
senthal, 1982).
A proposed metabolic cycle involving canavanine (37) is
CO2" illustrated in Fig. 13.9 (Rosenthal, 1982):
aspartic 3..aspartyl- 3-aspartyl- homoserine (13) The amount of canavanine (37) declines rapidly upon
acid(Zl) phosphate semiaJdebyde gennination of the seed; one function of canavanine is to
Fig. 13.8. Biosynthesis of homoserine from aspartic acid (Rosenthal, serve as a nitrogen storage reserve in the plant. Radioactively
1982; modified and llsed with permission of the copyright owner, labeled canavanine and related metabolites were converted
Academic Press, Orlando. FL). into glutamic acid, glutamine, and alanine in cotyledons of
Canavalia ensiformis (Rosenthal et al., 1988).
mimosine synthase has been isolated from Leueaena /eueo· The potent antimetabolic properties of canavanine result
eepha/a. Some cysteine synthases also cause fonnation of primarily from its ability to function as a highly effective
similar amino acids. One isozyme of such an enzyme from antagonist of arginine (2) metabolism because of its stroc-
Leueaena /eueoeepha/a is capable of producing L-mimosine tural similarity to that acid. This structnral similarity ac-
in the presence of O-acetyl-L-serine (36) (Fig. 13.1) and ap- counts for the ability of arginyl-tRNA synthetase to activate
propriate precnrsors, in this case 3,4·dihydroxypyridine and attach canavanine to the tRNA that nonnally carries
(Jkegami et al., 1990; Rosenthal, 1982). It is probable that arginine to the protein assembly site (Rosenthal, 1991). Ca-
these two enzymes are identical. navanine is incorporated into the proteins of canavanine-
Although considered a promising forage crop, the leaves sensitive organisms, but it is rarely, if ever, encountered in
of Leueaena /eueoeepha/a contain as much as 2% mimosine the proteins of the plants that produce it (Rosenthal, 1988,
dry weight and cause loss of hair and grossly enlarged thy- 1991).
roid glands when used as a food plant for animals. Mimosine The bruchid beetle, Caryedes brasiliensis, feeds only on
inhibits proline hydroxylation and, hence, collagen fonna- the seeds of Dioclea megaearpa, a legume of Costa Rica
tion and hair growth. It has been suggested that the pnre (Rosenthal, 1983). These seeds contain up to 13% of the dry
mimosine can be used to "shear" sheep chemically, but this weight as canavanine (37) (Rosenthal, 1977). Gravid beetles
nonprotein amino acid has proven too toxic for practical use deposit egg clusters on the ovary wall of the fruit, and the
in this regard. newly hatched larvae bore into the pericarp, locate the seeds,
Although it has been suggested that mimosine may serve and consume the cotyledonous tissues (Rosenthal, 1983).
as a tyrosine antimetabolite, the evidence is equivocal (Ro- Although this amino acid is potentially toxic, the insect has
senthal, 1991). an arginyl-tRNA synthetase capable of distingnishing be-
tween canavanine and arginine, and canavanine is not incor-
porated into protein (Rosenthal, 1983). Most of the total
HOJIIOSERIl'IB ninhydrin response (96%) in these seeds is contributed by
canavanine (Rosenthal, 1982, 1983).
Homoserine (13) is an intennediate in the synthesis of threo- The insect uses canavanine as a sonrce of dietary nitrogen
nine (14), isoleucine (IS), methionine (16), and cysteine (12) for the developing larvae (Rosenthal et al., 1982). Arginase
and, thus, found in all plants (Fig. 13.8). Homoserine is accu- activity that hydrolyzes arginine to orinithine also hydro-
mulated and has been studied in Pisum sativum (pea, Faba- lyzes canavanine to canaline (38) and urea. Urea cannot be
ceae). This amino acid is not found in the ungenninated assimiliated directly, rather it is first converted to ammonia.
seed, but during the first week of gruwth, homoserine consti- This insect produces about 30 times more urease that most
tutes 70% of the soluble nitrogen and 12% of the seedling other related insects. Thus, a potentially toxic plant sub-
dry weight (Rosenthal, 1982). stance is converted into a useful resource. The canaline pro-
duced is converted apparently to homoserine (13) and am·
monia and thereby detoxified (Rosenthal, 1982; Rosenthal
and Berge, 1989).
The generalist feeding insect Heliothis vireseens, the to·
L-Canavanine [the L·2-amino-4-(guanidinooxy)butyric acid bacco budwonn, has relatively high resistance to canavanine
structural analog of L-arginine1 (37) is widely distributed in but not to a number of other nonprotein amino acids (Berge
the subfamily Lotoideae (Papilionoideae) of the Fabaceae et al., 1986). In this case, canavanine (37) was shown to be
and, despite some reports to the contrary, is restricted appar- neither excreted nor sequestered, but rather metabolized by
ently to that family (Bell, 1981; Turner and Harborne, 1967). the insect. In Caryedes brasiliensis, Sterneehus tubereu/atus
 Nonprotein Amino Acids 221

NH,
I
c=o
I Q-ureidohomoserine

i
NH

r
ry
aspartic acid

~(rJ'.~'l:ile I ,~ATP'Mg fO,H

OH IH
, J-NH-CH

I I CH,
I
k!H
CH,
I I'
CH I I'
I
H-C-NH,

t-- r'
° CO,H
r' ~
NAD' CHz

H-C-NH,
I
CO,H
INH
H-f- ,
CO,H
CO'Hi'

C=NH
r'
CH,
homoserine (13) canaUne I I
(38) urea NH
I fumaric I
H-C-NH,

°
acid
CO,H
I
CH,
canavaninosuccinic add

I
canavanine
(37) IH,

H-C-NH,

CO,H

Fig. 13.9. Proposed metabolism of canavanine (Rosenthal. 1982; modified and used with pennission of the copyright owner, Academic Press, Orlando,
FL).

(canavanine-using insects), Canavalia ensiformis (a cana- canavanine are considered relatively advanced within the
vanine-storing plant), and Heliothis vireseem (a canavanine- family (Bell, 1981).
resistant insect), canavanine was not incorporated in protein,
whereas in Manduea sexta (a canavanine-sensitive insect)
CANALII'IE
and Glycine max (a canavanine-sensitive plant), canavanine
was incorporated into proteins in place of arginine (2) (Ro- Canaline (38) [the 2-amino-4-(aminooxy)butyric acid ana-
senthal et al., 1987). Because a number of insects can de- log of ornithine and the arginase-mediated hydrolysis prod-
grade canavanine via arginase and others can convert this uct of canavanine (37) is usually found in small amounts in
compound to ammonia via urease, the ability to prevent in- the same legumes that have canavanine (Rosenthal, 1991).
corporation of canavanine into proteins is probably the most Canaline reacts with carbonyl groups and is highly toxic in
important step for utilization of canavanine-containing its own right (Rosenthal, 1991). Canaline exhibits insectici-
plants as hosts (Bleiler et al., 1988). Proteins of vitellogenin dal properties in larvae of the tobacco hornworm, Manduea
from female migratory locusts (Loeusta migratoria migra- sexta, and is a powerful neurotoxin in the adult moth (Rosen-
torioides) that have consumed L-canavanine have disrupted thal et al., 1989). This compound is similar in structure to 4-
tertiary and quaternary structure (Rosenthal et al., 1989). arninobutyric (or 'Y-aminobutyric) acid and is also a powerful
Canavanine has been found in many advanced members inhibitor of pyridoxal phosphate-containing enzymes (Ro-
of the subfamily Papilionoideae, but not in the Mimosoideae senthal, 1982). As mentioned above, the canaline produced
nor in the Caesalpiniodeae (all Fabaceae) (Bell et al .. 1978). is apparently converted to homoserine and ammonia and
This suggests tbat the ability to synthesize this compound thereby detoxified (Rosenthal, 1982; Rosenthal and Berge,
arose after the first two subfamilies diverged from the ances- 1989). Canaline is converted to a novel stable oxime formed
tral stock leading to these plants, but not long after the diver- with canaline and glyoxylic acid in jack beans (Rosenthal
gence of the Papilionoideae. Most of the plants that contain et al., 1989).
222 Nonprotein Amino Acids

OH
I
CH,
I
CH,
I
H'I'NH,
- I
r H,

CH,

H-1-NH 1
--+
t,
CH,

I
I
H,
C=O
--L
H,O

CO,H CO,H
I
CO,H

homoserine 2-oxo-4-aminobutyric
2,4·diaminobutryic acid
acid

o-CO,H
N
- o-CO,H

NH
HOC~NH'
H~CO'H
l-azetine-2-carboxyllc azetidine-l-carboxylic cis-I-amino-3-hydroxycyclobutane-

acid (39) I'carboxylic acid (40)

acid

Fig. 13.10. Proposed biosynthesis of azetidine-2-carboxylic acid (Rosenthal, 1982; modified and used with pennission of the copyright owner,
Academic Press, Orlando, Fl.).

...AmTIDIl'IE·2·CARBOXYLIC ACID amino acids such as 2-amino-4-methylenehex-4-enoic acid

AI'ID RELATED AMINO ACIDS (41) (Fig. 13.11). Labeling studies have shown that uni-
fonnly labeled isoleucine was the best precursor for this
Azetidine-2-carboxylic acid (39) was the fIrst azetidine ring- compound of several tested. However, label from carboxyl-
containing nonprotein amino acid described, but several labeled isoleucine was not incorporated into 2-amino-4-
other compounds with this structural feature are now known. methyIhex-4-enoic acid, suggesting that loss of at least the
Azetidine-2-carboxylic acid is known to occur in a number carboxyl carbon occurs, possibly by decarboxylation, prior
of plants, including: Convallaria majalis (Liliaceae), Poly- to the biosynthesis of 2-amino-4-methylenehex-4-enoic acid
gonatum officinalis (Liliaceae), Busse massaiensis, Delonu (41). Acetate also is incorporated into this acid, suggesting
regia, Parkinsonia aculeata, Peltophorum inerme, Schi- that a C, unit is added to the portion of the molecule derived
zolobium parahybum, and P. africanum (all Fabaceae). This from isoleucine (Fig. 13.11) (Rosenthal, 1982).
unusual compound has been detected in small quantities in Several amino acids of this type are stored in the seeds
the residues from sugar beet processing (Beta vulgaris, Che- of Aesculus californicus-, after for example, 2-amino-4-
nopodiaceae) (Fowden, 1972). cis-I-Amino-3-hydroxy- methyIhex-4-enoic acid (41), 2-antino-4-methyl-6-hydrox-
methylcycIobutane-l-carboxylic acid (40) has been reported yhex-4-enoic acid (42), 2-amino-4-methylhexanoic acid
from seeds of Atelia herbert-smithii (Fabaceae) (Austin et (43), and 2-(methylenecycIopropyl)-3-methylalanine
aI., 1987). (44)-whereas Blighia unijuga (Sapindaceae) stores iso-
Azetidine 2-carboxylic acid (39) is toxic both to plants meric 2-amino-5-methyl-6-hydroxyhex-4-enoic acid (45)
and animals. When introduced into mung bean (Phaseolus and the related compound trans-2-(2-carboxymethylcycIo-
aureus), a plant that does not nonnally have the compound, propyl)glycine (46). Related compounds with ethylenic and
deleterious effects were noted. Azetidine 2-carboxylic acid acetylenic linkages (47-50) are found in Euphoria longan
is incorporated into protein instead of proline. Many of the (Sapindaceae). Compounds 51 and 52 occur in Aesculus par-
Liliaceae have an altered prolyl !RNA synthetase that per- viflora. (Fig. 13.12) (Rosenthal, 1982).
mits them to discriminate against 2-azetidine carboxylic acid
(Rosenthal, 1991).
2-Azetidinecarboxylic acid is synthesized in the plant ...IIYFOOLYCIN A AI'ID B
from homoserine (13) (Fig. 13.10) (Rosenthal, 1982).
Seeds of the sapindaceous tree, Blighia sapida, produce an
1'I0I'IPR0TE1N AMINO ACIDS DERIVED FROM outgrowth which covers the seed, called an aril, that contains
ISOLEUCIl'IE the toxic amino acid hypoglycin A (53) (0.001 % weight).
The 4-glutamyl derivative (hypoglycin B) (54) is also pro-
A number of C7 -amino acids are found in plants of a number duced (Fig. 13.12). This tree, the akee, was introduced from
of genera of the Aceraceae, Hippocastanaceae, and Sapinda- West Africa and is the "national food" of Jamaica today.
ceae. These compounds appear to be based on nonprotein The ripe aril (the least poisonous portion of the plant) is
 Nonprotein Amino Acids 223

isoleucine (15)

CH3CH"
!
transamination
CH3CH,,,
CH,CO,H
CH3CH, I II CH3CH,
NH,
I
°
/CHCCO,H _ CHCCO,H__ 'CHCH,CCO,H- ':::CHCH,CHCO,H
II / I
CH]
I 0 ~ CH3
CH( transamination
OR
CH]
2-amino-4--metbylbexanoic

CO, ~ CH3C~A t~ acid (41)

°II NH
I'
CH CH
3 ~ CH3CH,
OH
I CH3CH ~
CH3CII::"..
~
/CO,H '>:CCHCH CO H _ _ CCH,CCO,H - - + /CCH,CHCO,H
CH3 ------.,/ 1 1 / transamination CH]
CH3 CH3

tiglicacld Z-anlino-4-methylhex-4-enoic
acid (40)

Fig. 13.11. Proposed pathways for the biosynthesis of certain ~ nonprotein amino acids (Fowden and Mazelis, 1971; modified and used with
pennission of the copyright owner, Pergamon Press, Oxford).

commonly eaten and is responsible for the disease known 1991). Osteolathyrism involves bone and mesenchymal tis-
as "vomiting sickness." The problem is most severe in chil- sue aberrations. Consumption of seeds of Lathyrus odoratus
dren and in adults who are malnourished. The immature arils often leads to this malady. Other species, such as Lathyrus
and the seeds are highly toxic. hirsutus and L. pusillus, give rise to both (Rosenthal, 1991).
The aril of the unripened fruit is richer in hypoglycin A Although not often implicated, other genera of plants contain
than the fully matored aril. Both hypoglycin A and B are some of the same amino acids.
found in the unripened aril and the seed (Kean, 1976). These Among the amino acids from Lathyrus are 2,4-diamino-
two compounds also occur in several species of Blighia and butyric acid (56) (a neurologically active factor) (see above),
Billia (Hippocastanaceae) and in members of the genus Acer N3 -oxalyl-2,3-diaminopropionic acid (57) (a potent neuro-
(Aceraceae, the maple family) (Fowden and Pratt, 1973). toxin), 3-cyanoalanine (58) (a neurotoxin), and the 4-gluta-
The most striking manifestation of hypoglycin consump- myI derivative of 3-cyanoalanine. The decarboxylation prod-
tion is hypoglycemia, a condition in which the blood sugar uct of 3-cyanoalanine, 3-aminopropionitrile (59), and its 4-
falls to a very low level. It has been suggested that this effect glutamyl derivative induce many of the osteolathyritic skele-
is produced by inhibition of the oxidation of long-chain fatty tal abnormalities (Rosenthal, 1991) (Fig. 13.13).
acids by hypoglycin A and B. At least 17 species of Vida are able to synthesise the
A lower homolog (55) of hypoglycin A has been isolated arginine derivative, 4-hydroxY-L-arginine (62) ("'(-hydroxy-
from Litchi chinensis (Sapindaceae), Billia, and Acer seeds. L-arginine) (Bell and Tirimana, 1963). This hydroxy amino
In the last two plants, this lower homolog co-occurs with the acid is also found in a number of marine animals. 4-Hy-
corresponding 4-glutamyl peptide. These lower homologs of droxy-L-arginine cyclizes to form a lactone that has been
hypoglyine A and B also have hypoglycemic activity (Fow- isolated from other Vicia species. 4-Hydroxyhomoarginine
den and Pratt, 1973). (63) (",(-hydroxyhomoarginine), a derivative of homoargi-
nine, is found in Lathyrus species along with homoarginine
(Rosenthal, 1982) (Fig. 13.13). erythro-4-Hydroxyhomoar-
....HOMOARGIl'IINE Al'ID RELATeD COJIIPOlJl'lDS ginine (63) has been isolated from Lonchocarpus costari-
FROM THE GEl'IERA LAmYRUS Al'ID VlClA censis (Evans et al., 1985).
(I'ABACEAE)
....Homoarginine
Lathyrus and Vida are closely related genera of the Faba- Homoarginine (L-2-amino-6-guanidinohexanoic acid)
ceae; both are of considerable economic importance. Al- (60) (Fig. 13.13) has been isolated from a large number of
though the seeds of some species such as Vida faba and V.
Lathyrus species. This nonprotein amino acid is toxic when
sativa, are edible, those of many species of both genera are
fed to the larval bruchid beetle Callosobruchus maculatus
poisonous. Consumption of seeds of the genus Lathyrus (Fa-
at the 5% level in artificial diets (Rehr et al., 1973b).
baceae) by man and many of his domestic aJ)imals produces
a syndrome called lathyrism, which may be subdivided into
....Latbyrine
neurolathyrism and osteolathyrism. The most important spe-
cies that cause neurolathyrism are Lathyrus sativus, L. sylv- Lathyrine (61) was first recognized on paper chromato-
estris, L. cicera, L. latifolius, and L. clymenum (Rosenthal, grams because of the unusual vivid red-orange color pro-
224 Nonprotein Amino Acids

CH3 NH, CH3 NH,

I I
H3CCU=CCH,CHCO,H
I
H3CCH,CHCH,CHCO,H
I (43)
2-amino-4-methylhex-4-enoic acid (41)

,H3 iH, H
3 iH
' 7
HOCH,CH=CCH,CHCO,H HOCH,CH,C=CHCH,CHCO,H (45)
2-amino-4-methyl-6-hydroxyhex-4-enoic acid
(42) H

A1:~ VCH'CH(NHR)CO'H

HO,CCH,

tmns-2-(2-carboxymethyl- (53) R = H, L-hypoglycin A

(54) R = OCCH,CH,CH(NH,)CO,H,
cyclopropyl)g1ycine (46)
L-hypogJycin B

VCH(NH,)CO,H

3-(2-metbyJenecyciopropyl) a-(methylene-
(a) 3-methyJalanine (44) cyciopropyJ)gJycine (55)

CH3 NHz CHzOHNHz

I
HC5CCHCHzCHCOzH
I I
HC9::CHCHzCHCOzH
I
(47) (48)

(49)
(50)

-6-
H

H H

trans-2-(2-carboxycyclo- cis-2-(2-carboxycyclo-

(b) propyl)glycine (51) propyl)glycine (52)

Fig. 13.12 (a & b)_ Nonprotein amino acids from plants of the Aceraceae, Hippocastanaceae, and Sapindaceae.
 Nonprotein Amino Acids 225

ro'H
ro
°II
I
C-NH,
I
NH, r
CH,
neurolathyrogen
eHz --+- I
I
CHz ----+
I H-C-NH,
H-C-NH,
H-C-NH, I
I
CO,H
bO,H N'-OxaIYI-2'3-diamin:~;:.nic acid (44)
asparagine (64) 2,3-diamino- fO,H
propionic acid (69)

1
C=O

F,
NH,
I
<EN
I r'
r'
H-C-NH,

I
CO,H
--- r'I --- r'I
H-C-NH,

CO,H
H-C-NH,

CO,H

3-cyanoalanine (58) 2,4-diamino- N'-oxaIyl-2.4-

neurolathyrogen butyric acid (56) dlaminobutyric acid (65)
neurolathyrogen neurolatbyrogen

C:N NHCH3
I I
serine or
r' fH,

r'
cysteine HCNH,

NH,
I
CO,H
3-aminopropionitrile (59) N3.methyl-L-2,3-diaminopropionic acid
(a) osteolatbyrogen (66) osteulathyrogeu

HO,CCH(CH,)4NH-C-NH, H'NyNyyCO'H
I II
NH, NH NV NH,
L-homoarginine (60) L-lathyrine (61)

4-hydroxy-L-arginine (62) H,NC(NH)NH-CH,-CHOH-CH,-CH(NH,)CO,H

(b) 4-hydroxybomoarginine (63) H,NC(NH)NH-CH,-CH,-CHOH-CH,-CH(NH,)CO,H

Fig. 13.13. Biogenesis of 3-cyanoalanine and lathryogenic compounds of the genera Lathyrus and Vida (Fabaceae) (modified from Rosenthal. 1982;
used with pennission of the copyright owner, Academic Press, Orlando. FL).
226 Nonprotein Amino Acids

duced with ninhydrin. In the roots of Lathyrus tingitanus, quickly reproduces the symptoms in monkeys (Spencer et
introduction of labeled L-guanidinohomoarginine leads to a1., 1987). The incidence of this progressive dementia has
the formation of lathyrine. Recent work suggests that lathyr- plummeted with replacement of local foodstuffs with im-
ine is formed from orotate and uracil (Rosenthal, 1982). ported foods.
Tetrahydrolathyrine has been isolated from the seeds of
Lonchocarpus costaricensis. The configuration of the a-car-
bon atom was established by the large positive molar elliptic- NOM"KOTEIl'I AMINO ACIDS DEVELOPED
ity in acid solution (Fellows et a1., 1979). FROM GLUfAJIIIC ACID

The numerous derivatives of glutamic acid (67) and its amide

AMINO ACIDS FROM 3·CYAl'IO·L-ALAI'IIl'IE Al'ID glutamine (68) (Fig. 13.1) constitute the largest group of
ASPAKAGII'IE nonprotein amino acids found in higher plants. Leucine (10),
a derivative of glutamic acid, is also an important precursor
of these compounds. Glutamic acid is formed by transamina-
3-Cyanoalanine (f3-cyanoalanine) (58) is the only nonprotein
tion of 2-oxoglutaric acid from the Krebs cycle; a parallel
amino acid with a cyano group. Normally, cyanide in plants
set of reactions leads to the formation of the oxo derivative
is incorporated into asparagine, but in Vicia sativa, the 4-
of 2-aminoadipic acid, the higher homolog of glutamic acid.
glutamyl derivative of 3-cyanoalanine is produced. The most
4-Methylglutamic acid (9), (2S, 4S)-4-hydroxy-4-meth-
efficient amino acid precursors for 3-cyanoalanine are serine
ylglutamic acid (70) and 4-methyleneglutamic acid (26) ap-
(11) and cysteine (12) (Fig. 13.15) (Rosenthal, 1982).
parently are derived from leucine (Fig. 13.14) (Rosenthal,
3-Cyanoalanine (58) can be decarboxylated to f3-gluta-
1982). All these compounds are found in honey locust,
mylaminopropionitrile (59).
Gleditsia triacanthos (Fabaceae). 4-Methyleneglutamic acid
N'-Oxalyl-L-2,3-diaminopropionic add (57) is derived
(26) is also stored in the peanut, Arachis hypogaea (Faba-
from 2,3-diaminopropionic acid (69), which, in turn, is de-
ceae).
rived from asparagine (64) (Fig. 13.13). The oxalyl deriva-
Ornithine (71) (Fig. 13.1), widely distributed in plants,
tive (57) is a potent neurotoxin found in Lathyrus sativus
can be derived from glutamic acid (orinithine also can be
and certain other legumes. The corresponding N' -methyl de-
derived from arginine). This amino acid serves as an inter-
rivative occurs in certain cycads such as Gycas circinalis
mediate and as a precursor in numerous biosynthetic path-
(Rosenthal, 1991).
ways (for example, see Chapters 29 and 30).
L-2,4-Diaminobutyric (L-a;y-diaminobutyric) acid (56) is
Proline (3) is derived biosynthetically from glutamic acid
derived from 3-cyanoalanine (58). This compound also is
(67). This dicarboxylic acid is activated with ATP and then
found in a number of Lathyrus species. The corresponding
reduced to the corresponding semialdehyde, which sponta-
oxalyl derivative (65) co-occurs with L-2,4-diaminobutyric
neously cyclizes to /lJ-pyrroline-5-carboxylic acid and is
acid (Rosenthal, 1982).
reduced to proline by reduced NAD. Ornithine can be di-
Consumption of seeds of the genus Lathyrus (Fabaceae)
rectly converted to 4-glutamylsemialdehyde, providing an-
by man and his domestic animals produces a syndrome
other route to proline.
called lathyrism. Different amino acids produce the two
4-Hydroxyproline (5) is a major constituent of the cell
forms of this malady: neurolathyrism and osteolathyrism. 3-
walls of plants. This compound is generated by posttransla-
Aminopropionitrile (f3-aminopropionitrile) (59) is the active
tional hydroxylation of peptidyl proline; both plants and ani-
osteolathyritic factor in Lathyrus species. Several factors are
mals require diatomic oxygen for this reaction (Prockop et
responsible for neurolathyrism. 3-Cyanoalanine (58), its 4-
al., 1962). trans-4-Hydroxyproline accounts for as much as
glutamyl derivative, 2,4-diaminobutyric acid (56), and the
20% of the protein amino acids of primary cell walls.
N-oxalyl derivative of 2,3-diaminopropionic (a,f3-diamino-
The imino acid N-methYI-trans-4-hydroxY-L-proline (72)
propionic) acid (57) are all known to be involved. These
(Fig. 13.1) has been isolated from the leaves of two Gopaij-
compounds are found primarily in plants of the genus Lath-
era species (Fabaceae) (Figliuolo et a1., 1987).
yrus. 3-Cyanoalanine (58) and its 4-glutamyl derivative are
also common in Vicia species. Several related series of com-
pounds are found in other members of the Fabaceae.
OTHER NOM"KOTEIl'I AMINO ACIDS
N'-Methyl-L-2,3-diaminopropionic acid (66) (Fig. 13.13)
is found in plants of the genus Gycas, Cycadaceae, a gymno-
Sulfur.Containing Amino Acids
spermous family. Several of these plants are used as human
food sources in Oceania, whereas others are eaten by animals Several sulfur-containing amino acids, other than cyste-
in the South Pacific and in Australia. A form of amyotrophic ine, cystine, and methionine, are found in higher plants. Al-
lateral sclerosis (ALS or Lou Gehrig's disease) used to be lium (LiJiaceae) species often contain sulfur compounds that
endemic among the Chamorros of Guam. An unusual amino are lachrymatory in nature (Fig. 13.17). Among these species
acid N'-methyl-L-amino-2,3-diaminopropionic acid (N~­ are chives, garlic, onion, and leeks. The compounds involved
methylamino-L-a1anine) (66), found in Gycas species, are S-substituted cysteine sulfoxides and their hydrolysis
 Nonprotein Amino Acids 227

CH, OH NH, CH, OH NH,

I I I -- II I
HO,C- C - - CH- CH- CO,H
I
HO,C- CH- CH- CH- CO,H

3-hydroxy-4-metbylglutamic acid 3·hydroxy.4-methyleneglutamic acid

CH, i NH,
I I
CH,-CH-CH,-CH-CO,H
leucine (10)

CH, ~ NH,
I I
° CH, NH,
I I r=== H,N -CII -CH-CH,-CH-CO,H
>
HO,C-CH-CH,--CH- CO,H

4-methylglutamlc add (9) 4·methylglutamine

CH,
~ NH,
I I
HO,C-C -CH,-CH-CO,H
I
OH

4-hydroxy-4-methylglutamlc add (70)

CH,
! NH, ° CH, NH,
II I II II I
HO,C- C - - C H , - CH- CO,H r=== H,N -C -C-CH,-CH-CO,H
>

4-methyle.eglutamlc add (26) 4-methyleneglutamine

Fig. 13.14. Nonprotein amino acids derived from leucine (Rosenthal, 1982; modified and used with pennission of the copyright owner, Academic
Press, Orlando, FL).

products. Co-occurring enzymes convert these amino acid The behavioral responses of many insects to these com-
derivatives into more volatile compounds (Virtanen, 1965). pounds and their derivatives have been reviewed (Stadler,
Alliin (S-allylcysteine sulfoxide) (73), cycioalliin (3-methyl- 1992).
1,4-thiazane-5-carboxylic acid) (74), S-ai1ylcysteine (75), S-
aliylmercaptocysteine (76), S-(2-carboxypropyl)cysteine Djenkolic Acid
(77), S-n-propylcysteine (78), S-(prop-l-enyl)cysteine (79), The sulfur-containing amino acid, djenkolic acid (82), is
and a series of related compounds are found in Allium spe- found in seeds of many species of Pithecellobium, Acacia,
cies, and S-(l,2-dicarboxyethyl)cysteine (80) is found in as- and Mimosa (Fig. 13.16). This compound produces human
paragus, Asparagus ofJicinalis) (Rosenthal, 1982). toxicity in Java when the seeds of P. lobatum are eaten.
Alliin (73) in onions decomposes to yield the lacbryma- Related suifur-containing amino acids are found in these and
tory (tear-producing) compound syn-propanethial S-oxide other mirnosoid legumes.
(81) (Fig. 13.15). 10 the presence of water, this compound
is hydrolyzed to propionaldehyde, sulfuric acid, and H2S ....lndospicine
(Luckner, 1990).
Several species of insects are associated with Allium spe- Indospicine [L-2-amino-6-(amidino)hexanoic acid] (83)
cies. For example, both adults and larvae of Delia antiqua, (Fig. 13.16) has been isolated from Indigofera spicata, a
the onion fly, are attracted to disulfides characteristic of legume that is widely cultivated as a forage crop. The amino
these plants (Stiidler, 1992). Another specialist, the leek acid is quite toxic and causes abortion and hepatotoxicity.
moth, Acrolepiopsis assectella, was found to respond to the Nothing is known of the biosynthetic origin of indospicine
same group of compounds. However, dipropyl sulfmate, the (Hegarty, 1970, 1978).
precursor of the disulfides, is even more stimulatory. A para-
Albiziine
site of the leek moth, Diadromus pulchellus (Ichneumoni-
dae), reacts mainly to the disulfides that emanate from leaves Albiziine (84) is found in the seeds of many species of
damaged by the leek moth (Stiidler, 1992). the genera Acacia and Albizzia (Fabaceae: Mimosoideae)
228 NOlIl'l"Ordll Ami/lo Acids

(Fig. 13.16) (Seneviratne and Fowden, 1968). This amino Griffonia simplicifolia proved to be highly toxic to the larvae
acid has been isolated recently from the genus Dislium (Fa- of southern army worms in artificial diets; 0 normal adults
baceae: Caesalpinioideae). Albiziine often constitutes the were produced by the insects fed the diets with this com-
principal component of the free amino acids of the seeds. pound as compared with control values of about 20 (Rehr
The similarity of the structure of albiziine to glutamine et aI., 1973b; Rosenthal, 1982).
suggests that it could act as an competitive antagonist. Al- Although serotonin is involved in central nervous system
though albiziine has not proven toxic in many types ofbioas- function, this amine cannot cross the blood-brain barrier,
says, in studies with Locusta and Chortoicetes, two grami- whereas 5-hydroxytryptophan can. An excess of sertonin
nivorous feeders, this amino acid was one of the most toxic produces an array of toxic symptoms.
nonproteins tested (Rosenthal, 1982).

5-HydroXY-L-tryptophan 3,4·DihydroXY-L-phenylaianine (L-dopa)

In mammals, conversion of L-tryptophan to 5-hydroxy- 3,4-DihydroxY-L-phenylalanine (L-dopa) (85) (Fig.

tryptophan (7) is the first step in the formation of the biologi- 13.16) is known to occur in a number oflegumes. This amino
cally active amine serotonin or 5-hydroxytryptamine (Fig. acid has been found in seeds of plants from the genera Vicia,
13.16) (Udenfriend et al., 1956). 5-Hydroxytryptophan has Baptisia, Lupinus, Mucuna, and also in Euphorbia lathyrus
been isolated from the legume Mucuna pruriens as well as of the Euphorbiaceae. A massive screening program showed
from cotton and bananas (Rosenthal, 1982). that it occurs above the 0.5% level only in the seeds of
5-Hydroxytryptophan (7) also occurs in large amounts Mucuna species, where it constituted 3-7% of the weight of
(6-10%) in the seeds of Griffonia simplicifolia, a west Afri- the seeds (Daxenbichler et al., 1972). L-Dopa is the principal
can legume. In all tissues except the mature seed, tryptophan nonprotein amino acid in many of these species (Rehr et al.,
serves as a precursor of the 5-hydroxy compound. Seeds of 1973a). The compound may occur free or as a glycoside.

CH,
II alliin_ c) ~ H)=S ~ H)=O
HI/- '\
CH _
I tyase
H.c CH,-CH, 0 CH,- CH,
CH,

~!,.) NH
"
Al~propenylsulfenic acid
(allylsulfenic acid)
syn-propanethioal
S·oxide (81)
propionaJdehyde
+H2S04 +H2S
CH'l:-~ - CO,H CH, CH, o
!
CH, CH,
H
alliin (73)
~ II
CH
II
CH __
II II
CH CH
I I I I
JN~CO'H
lachrymatory precursor
(S~allylcysteine sulfoxide) CH, CH, CH, CH,
I
,e--S--S
I I I
S--S
o diallylsulfide cycloalliin (74)
allicin

NH,
CHz===( ---+-
CH3~
°1/ +NH,
CO,H CO,H

S~a1Iylcysteine (75) H,C:C.CH,.S.CH,.CH(NHz}CO,H

S~allylmercaptocysteine (76) H,C=CH·CH,·S·S·CH,CH(NHz}CO,H

S-(2-carboxypropyl)cysteine (77) HO,~

CH·CH,·S·CH,·CH(NHz}CO,H
CH,/'

S-n-propylcysteine (78) CH,(CH,h·S·CH,·CH(NH,)CO,H

S-(1,2~dicarboxyethyl)cysteine (80) HO,C(CH,),S·CH,.CH(NHz}CO,H

S·(prop·l·enyl)cysteine (79) CH,·CH:CH·S·CH,·CH(NHz}CO,H

Fig. 13.15. Sulfur compounds found in Allium species and fonnation of syn-propanethial S-oxide (Luckner. 1990).
 Nonprotein Amino Acids 229

7
NH,
H
1 ' NH, ,H,
HO,CCHCH,SeCH,CH,CHCO,H
1 ~
~
L-seJenocystathionine (87) 1
tH iH,
NH,
,H,
HO,CCHCH,Se-SeCH,CHCO,H
1 iH, ,H,
H-C-NH, H-,-NH,
L-selenocysteine (86)
I
CO,H CO,H

,H, ,H,
L-albiziine L-glutamine (68)
HO,CCHCH,SCH,CHCO,H
(84)

djenkolic acid (82)

OH

HO'():)):CO'HHofu
I I :::0'1 NH,
::::,... NH, ::::,...
NH CO,H
indospicine (83) 3,4-dihydroxy-

O 0'
5-hydroxy-L-tryptophao (7)
phenylalanine (85)
H CO,H (L-DOPA)
- H
v.CO,H- - --v
H CO,H
(+)-nicotianamine (89) HN

N/~"""""NH~NH,

0 ... -
---COH
H CO,H '
)(~' ~ --v mugineic acid (91)
H COH H CO,H
NH
N' Y . . . NH~OH
OH cucurbitine (88)

H CO,H H CO H H CO,H
~ ~ V' ' --,I avenicacid (90)
HO HN/ ~NH~NH,
Fig. 13.16. Biologically active nonprotein amino acids.

Phenylalanine is not a precursor for L-dopa. Whereas ty- are active selenium accumulators. Some accumulate as much
rosine is incorporated into excised hypocotyls of Vicia faba as 15,000 ppm of selenium, mostly as Se-methylselenocyste-
seedlings, attempts at isolating a tyrosine hydrolase proved ine (86) and selenocystathionine (87) (Fig. 13.16). This phe-
unsuccessful (Griffith and Conn, 1973). nomenon is also known in the fruit of Lecythis ollaria, a
L-Dopa has been used medicinally to treat Parkinson's Central and South American nut that is sometimes eaten. In
disease. The amino acid apparently crosses the blood-brain this case, the compound responsible is selenocystathionine
barrier and is decarboxylated to produce dopamine, a bioge- (Rosenthal, 1991).
netically active amine. Selenium poisoning is a major problem in Australia;
Morinda amplexicaulis and Morinda reticulata (Rubiaceae)
Selenium-Containing Amino Acids are the major species involved. In Morinda species, a selen-
Selenium, normally a minor soil component, is toxic to ium compound represents about 20% of the total nitrogen of
most plants at low levels, but under certain conditions, it the plant.
accumulates in higher plants well in excess of the soil con-
centrations (Shrift, 1969). Of these plants, the best known Nonprotein Amino Acids in Seeds
are about 20 species of the genus Astragalus (Fabaceae). of the Cucurbitaceae
Generally, the compounds formed in Astragalus species are
analogous to common sulfur-containing compounds. Out of A number of nonprotein amino acids are found in seeds
the approximately 1500 species of Astragalus, only a few of members of the Cucurbitaceae. Cucurbitine (88) is active
230 Nonprotein Amino Acids

against Schistosoma japonicum (Fig. 13.16). When mice possible that the compound functions by serving as an iron
were given 400 mg/kg oral cucurbitine for 28 days, the de- chelating agent and is involved in transport of iron to the
velopment of schistosomula was greatly retarded (Borris and sites of chlorophyll synthesis (Ripperger et aI., 1982).
Schaeffer, 1992). Relatively simpleN-substituted asparagine The amino acids avenic acid (90) and mugineic acid {N-
derivatives are formed from asparagine (but not from aspar- [3-(3-hydroxy-3-carboxypropylamino)-2-hydroxy-3-
tic acid) in Ecballium (Cucurbitaceae) seedlings. The N"- carboxypro pyl]-acetidine-2-carboxylic acid} (91) are sider-
ethyl, methyl, and hydroxyethyl derivatives are known to ophores (active scavengers of iron) produced by the roots
occur (Dunhill and Fowden, 1965). The enzymes that lead of grasses in response tu iron depletion. Avenic acid from
to nonprotein amino acids in some cucurbits may be closely oats (Avena sativa, Poaceae) was characterized largely on a
related to those responsible for the synthesis of mimosine basis of its mass and NMR specta; the positive Cotton effect
(Jkegami et aI., 1990). in the circular dichroism (CD) spectrum suggested that both
chiral centers had the S-configuration (Hunt, 1991).
Compounds Involved in Iron Metabolism
in Plants Ethylene and Its Precursors
A number of amino acids and amino acid derivatives such The naturally occurring plant growth regulator ethylene
as ( - )-nicotianamine (89) are involved in iron transport in is derived from the nonprotein amino acid, I-aminocyclopro-
plants (Fig. 13.16) (Winkelmann et al., 1987). This com- pane-I-carboxylic acid (92) (Fig. 13.17) (Yang, 1981). The
pound was identified as the normalizing factor which re- formation of ACC from S-adenosylmethionine is catalyzed
stores chlorophyll synthesis and growth in an auxotroph mu- by ACC-synthase and this is considered to be the rate-con-
tant of Lycopersicon esculenlum. The specific role played trolling step in ethylene biosynthesis (Amrhein et al., 1981).
is not clear, however. ( + )-Nicotianamine, derived from syn- An ethylene-forming enzyme has been observed in plant
thesis, has the same effects in the mutant. The minimum tissues; the reaction is oxygen dependent and activated by
amount that produced a response was 10- 8 mol/plant. It is carbon dioxide (Yang, 1984). As expected, levels of this

Jl'j8denine
CR, CR,
I
1 ATP PP,+P,
+
I
yR' '>- L. yR, R R CR,=CR,
fR, CR, R R +RCN + CO,

R-C-NR,
R-+NH, OR OR [ > < 0 ' ; NR,
!O'R CO,R

Q
L-methionine CO,R
S~adenosylmethionine

RO rl.aminOCYeiopropane

'--~
OR OR
CR, CR,
I I

CR, R 7\ R R
I
fR,
adenine H20 H
OR OR OR OR

R-1- NR,
S-methylthioribose 5 -methylthioadenosine
O,R
L·homoserine

Fig. 13.17. Biosynthesis of ethylene and l-aminocyclopropane-l-carboxylic acid (ACC) and the biosynthesis of ethylene (modified from Adams and
Yang. 1984); used with pennission of the copyright owner, the American Society of Plant Physiology. Rockville. MO).
 Nonprotein Amino Acids 231

CO,H

H,N-+-H

V
i
H CH,
NHC(O)(CH,J,CHCNHz}CO,H
CH, (jHz),
__ CO,H
O=S=NH
I
H H I
CH,
g1abrin (95) trigonelline (94)
(%)

Fig. 13.18. Additional biologically active nonprotein amino acids.

enzyme are low in preclimacteric fruit tissue and young pet- DlSTRlBVl10l'l OF l'I01'lPK0TE1l'I AMIl'IO ACIDS
als of camation flowers but increases rapidly as these tissues Il'IFLAl'ITS
undergo ripening or the senescence process (Yang, 1984).
When ACC is applied to a number of different plant tissues, Nonprotein amino acids are most commonly found in the
ethylene is generated. This suggests that the ethylene-gener- Aceraceae, Agavaceae, Amaryllidaceae, Cucurhitaceae, Fa-
ating enzyme is constitutive in those tissues. Uncouplers of haceae (Leguminosae), Hippocastanaceae, Liliaceae, and the
oxidative phosphorylation are potent inhibitors of ethylene Sapindaceae, but are especially common in the Fabaceae.
formation. Further, ethylene formation is linked to mem- The families Aceraceae (the maple family), Hippocastana-
brane integrity (Yang, 1984). The activity of ethylene-form- ceae (the horse chestnut family), and Sapindaceae are closely
ing enzyme is induced by the presence of ethylene (Manning, related; some phylogenists have placed all three into one
1986). family. The distribution of nonprotein amino acids would
ACC is oxidized by a hydrolase and then fragmented into tend to support this move.
ethylene and cyanoformic acid. Cyanoformic acid is labile
and degraded spontaneously to carbon dioxide (derived from
the carboxyl group of ACC) and hydrogen cyanide (derived
from C-l of ACC). In many plants, hydrogen cyanide is
REFERENCES
converted rapidly into asparagine (93), 3-cyanoalanine (58),
and 'Y-glutamyl-3-cyanoalanine (Adams and Yang, 1977; ADAMS, D. O. & S. F. YANG, Methionine Metabolism in Apple
Yang, 1984). tissue, Plant Physiol, 60, 892-896 (1977).
AMRHeIN, N., D. SCHNEBBECK, H. SKORUPKA, S. P011IOF, and J.
ST6cKIGT, Identification of a major metabolite of the ethylene
TrlgonelUne. a Hormone Which Controls precursor I-aminocyclopropane-I-carboxylic acid in higher
the Ceu Cycle in Plants plants, Naturwissenschaften, 68, 619-620 (1981).
Trigonelline (94), G2 Factor, is commonly found in the AUSTIN, G. N., P. D. BAIRD, H. CHOW, L. E. FELLOWS, G. W. J.
FLEET, R. J. NASH, J. M.I'BACH, R. J. PRYCE, and C. H. STIRTON,
cotyledons of dried seeds and is found in a wide variety of
Isolation fromAtelia herbert-smithii Pittier (Sophoreae, Legum-
plants (Fig. 13.18). This compound is present in the cotyle- inosae) and x-ray structure of cis-I-amino-3-hydroxy-
dons of Pisum sativum and transported to the roots and methylcyclobutane-l-carboxylic acid, an achiral non-protein
shoots after germination. Trigonelline promotes preferential amino acid, Tetrahedron, 43, 1857-1861 (1987).
cell arrest in G2 of the cell cycle (Tramontano et al., 1982). BELL, E. A., "Uncommon" amino acids, FEBS Lett., 64, 29-35
In Pisum sativum, in the absence of trigonelline, root cells (1976).
arrest only in G I . Trigonelline is the first chemically identi- BELL, E. A., Non-protein amino acids in the Leguntinosae, in Ad-
fied hormone that controls the cell cycle in plants or animals vances in Legume Systematics, Vol. 2 (R. M. Polhill and P.
(Tramontano et al., 1982). H. Raven, eds.) 489-499, Royal Botanical Gardens Kew, UK,
1981.
JllisceUaneous l'Ionprotein Amino Acids BELL, E. A. and A. S. L. TnuMANA, Occurrence of -y-hydroxyargi-
nine in plants, Nature, 197, 901-902 (1963).
A neurotoxic principle, glabrin [S-(3-amino-3-carboxy- BELL, E. A., J. A. LACKEY, and R. M. POLHILL, Systematic signifi-
propyl)-S-methyl sulfoxime] (95), found in all parts of cance of canavanine in the Papilionoideae (Faboideae), Bio-
enestis glabra (Connaraceae), possesses an unusual sulfur- chern. Syst. Bcol., 6, 201-212 (1978).
bearing functionality (Jeanodda et al., 1985). BERGE, M. A., G. A. ROSENTHAL, and D. L. DAHLMAN, Tobacco
A nonprotein amino acid (96) has been reported recently budwonn. Heliothis virescens [Noctuidae] resistance to L-cana-
as a component of the defensive secretion of the Colorado vanine. a protective allelochemical. Pesticide Biochem. Phys-
potato beetle (Leptinotarsa decemlineata) (Harborne, 1989). iol., 25, 319-326 (1986).
232 Nonprotein Amino Acids

BERNAYS, E. A., Nitrogen in defence against insects, in Nitrogen in Plant Biochemistry (P. M. Dey and J. B. Harborne, eds.),
as an Ecological Factor (J. A. Lee, S. McNeil, and I. H. Rorison, Academic Press, London I-52, 1991.
eds.), 321-344, Blackwell, Oxford, 1983. IKEGAMI, F., M. MIzuNo, M. KIHARA, and I. MURAKOSHI, Enzy-
BLEILER, J. A., G. A. ROSENTHAL, and D. H. JANZEN, Biochemical matic synthesis of the thyrotoxic amino acid mimosine by cyste-
ecology of canavanine-eating seed predators, Ecology, 69, ine synthase, Phytochemistry, 29,3461-3465 (1990).
427-433 (1988). JAEGER, H., H. U. KLOER, H. DrrsCHUNEIT, and H. FRANK, Glass
BORRls, R. P. and J. M. SCHAEFFER, Antiparasitic agents from plant, capillary gas-liquid chromatography of aroino acids, in Applica-
in Phytochemical Resources for Medicine and Agriculture (H. tions of Glass Capillary Gas Chromatography (W. G. Jennings,
N. Nigg and D. S. Seigler, eds.), 117-158, Academic Press, ed.), Chromatography Science Series Vol. 15,331-364, Marcel
San Diego, CA, 1992. Dekker, Inc., New York, 1981.
DAXENBICHLER, M. E., C. H. VANETIEN, F. R. EARLE, and W. H. JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary
TALLENT, I-Dopa recovery from Mucuna seed, J. Agric. Food compounds to the seed-eating larvae of the hrochid beetle Callo-
Chern., 20, 1046-1048 (1972). sobruchus maculatus, Phytochemistry, 16, 223-227 (1977).
DUNHILL, P. M. and L. FOWDEN. The amino acids of the Cucurbita- JEANODDA, V. L. R., J. VALISOLALAO, E. E. CREPPY, and G. DIR-
ceae, Phytochemistry, 4, 933-944 (1965). HEIMER, Identification of the toxic principle of Cnestis glabra as
EVANS, S. V., L. E. FELLows, D. H. JANZEN, J. CHAMBERS, and R. methionine sulfoximine, Phytochemistry, 24, 854-855 (1985).
C. HIDER, ErytJrro-'1-hydroxyhomo-L-arginine: An aroino acid KEAN, E. A. (ed.), Hypoglycin, Academic Press, New York, 1976.
from seed of Lonchocarpus costaricensis. and its preferential LUCKNER, M., Secondary Metabolism in Microorganisms, Plants,
interaction with borate, Phytochemistry, 24,1289-1292 (1985). and Animals, Springer-Verlag, Berlin, 1990.
FANGMEIER. N. and E. LEISTNER, Conversion of D-Iysine via L- MANNING, K., Ethylene production and l3-cyanoalanine synthase
pipecolic acid in Nicotiana glauca L. plants and cell-suspension activity in carnation flowers, Planta, 168, 61-66 (1986).
cultures, J. Chern. Soc., Perkin I, 1769-1772 (1981). MIFLm, B. J. (ed.), Amino Acids and Derivatives, Vol. 5 of The
FELLOWS, L. E., E. A. BELL, T. S. LEE, and D. H. JANZEN, Tetraby- Biochemistry of Plants (P. K. Stompf and E. E. Conn, eds.),
drolathyrine: A new amino acid from seeds of Lonchocarpus Academic Press, New York, 1980.
costaricensis, Phytochemistry, 18, 1333-1335 (1979). PROCROP, D., A. KAPLAN, and S. UDENFRIEND, Oxygen-18 stodies
FIGLIUOLO, R., S. NAYLOR, J. WANG, and J. H. LANGENHEIM, Unu- on the conversion of proline to hydroxyproline, Biochem. Bio-
sual nonprotein amino acid and its relationship to phenolic and phys. Res. Commun., 9, 162-166 (1962).
nitrogenous compounds in Copaifera, Phytochemistry, 26, REaR, S. S., D. H. JANZEN, and P. P. I'EENY, L-DOPA in legume
3255-3259 (1987). seeds: A chemical barrier to insect attack, Science,I8I, 81-82
FOWDEN, L., The non-protein amino acids of plants, in Progress in (1973a).
Phytochemistry (L. Reinhold and Y. Liwschitz, eds.), 203-266, Rmnt, S. S., E. A. BELL, D. H. JANZEN, and P. P. FEENY,losecticidal
Interscience, London, 1970. aroino acids in legume seeds, Biochem. Syst.,I, 63-67 (1973b).
FOWDEN, L., Amino acid complement of plants, Phytochemistry, RIPPERGER, H., J. FAUST, and G. SCHOLTZ, Synthesis and biological
IJ, 2271-2276 (1972). activity of ( + )-nicotianaroine, Phytochemistry, 21, 1785-1786
FOWDEN, L. and M. MAZELIS, Biosynthesis of 2-amino-4-meth- (1982).
ylhex-2-enoic acid in Aesculus californica: The precursor role ROSENTHAL, G. A., Nitrogen allocation for L-canavanine synthesis
of isoleucine, Phytochemistry, 10, 359-365 (1971). and its relationship to chemical defense of the seed, Biochem.
FOWDEN, L. and H. M. PRATT, Cyclopropylaroino acids of the genus Syst. Ecol., 5, 219-220 (1977).
Acer, Phytochemistry, 12, 1677-1681 (1973). ROSENTHAL, G. A., Plant Nonprotein Amino and Imino Acids, Aca-
FowDEN, L., J. W. ANDERSON, and A. SMlTIf, A comparative study demic Press, New York, 1982.
of the aroino acids and phenylalanyl-tRNA synthetases of ROSENTHAL, G. A., A seed-eating beetle's adaptations to a poison-
Aesculus spp., Phytochemistry, 9, 2349-2357 (1970). ous seed, Sci. Am., 249, 164-171 (1983).
FRffiD, B. andJ. SHERMA, Thin Layer Chromatography, Chromatog- ROSENTHAL, G. A., The protective action of a higher plant toxic
raphy Science Series Vol. 17,230-245, Marcel Dekker, Inc., product, BioScience, 38,104-109 (1988).
New York, 1982.
ROSENTHAL, G. A., Nonprotein amino acids as protective allelo-
GRIFFITH, T. and E. E. CONN, Biosynthesis of3,4-dihydroxyphenyl- chemicals, in Herbivores: Their Interactions with Secondary
alanine in Vicia sativa, Phytochemistry,n, 1651-1656 (1973). Plant Metabolites (G. A. Rosenthal and M. R. Berenbaum, eds.),
HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod. 1-34, Academic Press, San Diego, CA, 1991.
Rep., 7, 85-109 (1989). ROSENTHAL, G. A. and E. A. BELL. Naturally occurring toxic non-
HEGARTY, M. P., Indospicine, a hepatotoxic amino acid from Indi- protein amino acids, in Herbivores: Their Interactions with Sec-
go/era spicata: Isolation, structure, and biological studies. Aust. ondary Plant Metabolites (G. A. Rosenthal and D. H. Janzen,
J. BioI. Sci., 23,831-842 (1970). eds.), 353-385, Academic Press, New York, 1979.
HEGARTY, M. P.. Toxic amino acids of plant origin, in Effects of RosENTIIAL, G. A. and M. A. BERGE, Catabolism of L-canavanine
Poisonous Plants on Livestock (R. F. Keeler. K. R. van Kampen. and L-canaline in the jack bean, Canavalia ensiformis (L.) DC.
and L. F. Jaroes, eds.), 575-585, Academic Press, New York, (Leguminosae), J. Agric. Food Chern., 37, 591-595 (1989).
1978. ROSENIHAL, G. A., M. A. BERGE, J. A. BLEll.ER, and T. P. RUDD,
HUNT, S., Non-protein amino acids, in Amino Acids, Proteins, and Aberrant, canavalyl protein formation and the ability to tolerate
Nucleic Acids (L. J. Rogers, ed.), Vol. 5 of Modern Methods or utilize L-canavanine, Experientia, 43, 558-561 (1987).
 Nonprotein Amino Adds 233

ROSENTIiAL, G, A.. M. A. BERGE, A. J. OZINSKAS, and C. G. TRAMONTANO, W. A., C. M. HARTNETT, D. G. LYNN, and L. S.
HUGHES, Ability of L-canavanine to support nitrogen metabo- EVANS, Relationship between trigonelline concentration and
lism in the jack bean, Canavalia ensiformis (L.) DC., J. Agric. promotion of cell arrest in G2 in cultured roots of Pisum sati-
Food Chern., 36, 1159-1163 (1988). vum, Phytochemistry, 21, 1201-1206 (1982).
ROSENTHAL, G. A., C. G. HUGHES, and D. H. JANZEN, L-Canavanine, TuRNER, B. L. and J. B. HARBORNE, Distribution of canavanine in

a dietary nitrogen source for the seed predator Caryedes bra- the plant kingdom, Phytochemistry, 6, 863-866 (1967).
siliensis (Bruchidae), Science, 217, 353-355 (1982). UDENFRIEND, S., E. TITus, H. WEISSBACH. and R. E. PETERSON,

ROSENTHAL, G. A., J. RErCHHART, and J. A. HOFFMANN, L-Canavan- Biogenesis and metabolism of 5-hydroxyindole compounds, J.
ine incorporation into vitellogenin and macromolecular confor- BioI. Chern., 219, 335-344 (1956).
mation,1. BioI. Chern., 264, 13693-13696 (1989). VIRTANEN, A. l., Studies on organic sulphur compounds and other
labile substances in plants, Phytochemistry, 4, 207-228 (1965).
SENEVIRATNE, A. S. and L. FOWDEN, The amino acids of the genus
WINKELMANN, G., D. VAN DBR HELM, and J. B. NBILANDS, Iron
Acacia, Phytochemistry, 7, 1039-1045 (1968).
Transport in Microbes, Plants, and Animals, VCH VerIagsges.,
SHRlFI', A., Aspects of selenium metabolism in higher plants, Annu. Weinheim, 1987.
Rev. Plant Physiol., 20, 475-494 (1969).
YANG, S. F.. Biosynthesis of ethylene and its regulation, in Recent
SPENCER, P. S., P. B. NUNN, J. HUGON, A. C. LUDOLPH, S. M. Ross, Advances in the Biochemistry of Fruits and Vegetables (1.
D. N. RoY, and R. C. ROBERTSON, Guam amylotrophic lateral Friend and M. J. C. Rhodes, eds.), Annual Proceedings of the
sclerosis-ParkinsonisM-Dementia linked to a plant excitant Phytochemistry Society of Europe No. 19),89-106, Academic
neurotoxin, Science, 237, 517-522 (1987). Press, London, 1981.
STADLER, E., Behavioural responses of insects to plant secondary YANG. S. F., The fonnation of ethylene from l-amioocyclopropane-
compounds, in Herbivores: Their Interactions with Secondary I-carboxylic acid, in Ethylene. Biochemical, Physiological and
Plant Metabolites, Vol. 2 (G. A. Rosenthal and M. R. Beren- Applied Aspects (Y. Fuchs and E. Chalutz, eds.), 1-10, Nijhoff
bawn, eds.), 45-88, Academic Press, San Diego, CA, 1992. Junk, The Hague, 1984.
14
Peptides

Introduction Many nonprotein amino acids occur in plants as their 'Y-

Peptides glutamyl derivatives and are discussed in Chapter 13.
Peptide Antibiotics
Antimetabolites and Antitwnor Peptides
Peptide Antibiotics
Peptides from Fungi
Phytotoxins Produced by Bacteria and Fungi A number of peptides, especially those from microorga-
Opines nisms, have antibiotic properties. These compounds often
Peptides from Blue-Green Algae have molecular weights in the range 300-2000, contain non-
Siderophores protein and D-amino acids, are cyclic, and tend to occur in
Peptides from Higher Plants families in which individual amino acids are replaced by
Glutathione others. For example, polymyxin B (1) (Fig. 14.1), from the
'Y-Glutamyl Peptides
bacterium Bacillus polymyxa, contains 10 amino acid units.
Phytochelatins
Among these are six units of n,'Y-diaminobutyric acid and
Proteins
one of D-phenylalanine. Polymyxin D has D-leucine instead
Phytohemoglobins
of D-phenylalanine, and leucine instead of threonine. Inter-
Lectins or Hemagglutinins
estingly, L-amino acids are better incorporated than D-amino
Biological Activity of Lectins
acids, suggesting that the L-form is converted to D-amino
Proteinase Inhibitors
acids in the producing organism. Chloramphenicol, puromy-
Sweeteners
cin, and tetracycline inhibit protein synthesis when added to
References
cultures of Bacillus polymyxa, but do not inhibit polymyxin
biosynthesis (Berdy, 1980-1985; Gottlieb and Shaw,
1967-1983). Gramicidin S (2), from Bacillus brevis, con-
Il'ITRODVCTIOI'l tains only five different amino acids (Sneader, 1985).
Penicillins and cephalosporins contain a [I-lactam ring,
Peptides, molecules comprised of two or more amino acid which is fused with a thiazolidine ring. These types of com-
units joined by peptide linkages, are widespread and occur pounds are synthesized by both prokaryotic and eukaryotic
both free and as components of proteins in bacteria, fungi, microorganisms, including Streptomyces, Penicillium, As-
animals, and plants. Some peptides arise from degradation pergillus, and Cephalosporium species (Luckner, 1990). Im-
of proteins, whereas others are formed from activated amino portant precursors are L-2-aminoadipic acid, L-cysteine, and
acids (Luckner, 1990). Because peptides were difficult to L-valine. An important intermediate is 6-(L-aminoadipyl)-L-
isolate and study, those that were toxic or had some impor- cysteinyl-D-valine (Luckner, 1990). The valine unit with a D-
tant physiological property tended to be studied preferen- configuration found in one portion of the molecule is labeled
tially. Today, new analytical and separation techniques have when radioactively labeled L-valine is added to the culture.
greatly simplified work with this group of compounds. Re- The most important antibiotic of this group, penicillin G (3)
cent studies in molecular biology suggest that peptides are from Penicillium spp., is derived from similar precursors.
important in virtually all aspects of cell development. Penicillins and cephalosporins inhibit cell wall formation

234
 Peptides 235

leu-Dab

/
n.phe
"- nab

IDab thr
I
""--nab / '
I
nab
I
thr
I
nab.0C(cH,J4~H1H,

CH3 CH3

polymyxin B, (1) actinomycin D

Dab = L-2,4-diaminobutyric acid sar = sarcosine
meval = N-methylvaline

()YNHtQ<H
~ H'N~N)j~S
N N,9 0,/
o 0 ~
CO,H CO,H 0
benzyl peniclllln cephalosporin C (2)
(penicillin G)

HO

I ~H3 o~'(~T
MeVai "-

cyclosporin A (5)

(a) MeLeu - MeLeu-D-Ala-Ala -MeLeu-Vat

Fig. 14.1 (8-C). Some polypeptide antibiotics.

in bacteria. These antibiotics are inactivated by bacterial 13- of this drug is that it does not cause bone marrow depression
lactamases and stomach acid (Luckner, 1990). (Sneader, 1985). Its main clinical value is in the treatment
The history of the development of penicillin and a series of squamous cell carcinoma and it is administered to treat
of antibiotic substances from bacteria of the genus Cepha· head and neck tumors, non-Hodgkin's lymphoma, and testic-
losporium [e.g., cephalosporin C (4)] has been reviewed ular teratomas (Sneader, 1985).
(Sneader, 1985).
I"eptides from Fungi
AntimetaboUtes and Antitumor I"eptides Most members of the genus Amanita are poisonous, but
several species that produce cyclic peptides are mong the
A number of peptides of animal origin were shown to most poisonous fungi in the world (Bresinsky and Besl,
playa role in cancer therapy as antimetabolites. Later, sev- 1990; Litten, 1975). Amanita species are common in both
eral peptide antibiotics were demonstrated to have similar Europe and the United States. Most important among these
properties. One of these, cyclosporin A (5), produced by are Amanita phalloides, A. verna, A. virosa, and A. panth·
certain fungi imperfecti, was shown to prevent attack on host erina. Furthermore, most fatalities from eating poisonous
tissues by transplanted cells (Sneader, 1985). Another series mushrooms are attributable to this genus (about 90% in Eu·
of peptides from Streptomyces verticillus, of which bleomy- rope) (Bresinsky and Besl, 1990). Similar peptides are found
cin A2 (Fig. 14.2) is the main component used clinically, in a number of Galerina species and in some Lepiota species
has pronounced antitumor activity. The most striking feature (Bresinsky and Besl, 1990).
236 Peptides

(b) bacitracin A

gramicidin S (3)

Fig 14.1 (continued)

 Peptides 237

C I ,,~
=t
, ~, x~~r. ~,u---r
H
,

.. J-...NH~CONH' 0 /S+
NH
0

I I
l' T
AJl.#o
N 9' N CH':t.

HN- 0 H
NH
NH N I S

ooild lj bleomycin Az

OH ~OH
o OH

OH 0 NH,

Fig. 14.2. Bleomycin A2.

When cooked, these mushrooms apparently taste good. which leads to the rapid conversion of monomeric G-actin
The estimated lethal dose of amatoxins for man (depending to polymeric F-actin and inhibition of the reverse (depolym-
on body weight) is only 5-7 mg, the amount found in ap- erization) reaction. Phalloidin (9) administered interperito-
proximately 50 g of fresh fungus (less than a single fruit- neally has an almost immediate effect, whereas amatoxins
body of Amanita phalloides (Bresinsky and Besl, 1990). do not cause death in less than 15 h regardless of the dose.
Onset of poisoning typically requires 8-12 h (6-24 h), sev- Phalloidins are not absorbed readily by the gastrointestinal
eral phases follow, and death often ensues within 6-8 days. tract (Bresinsky and Besl, 1990).
The liver is usually the most affected organ (Bresinsky and A third group of peptides, the virotoxins, appear to be
Besl, 1990; Kingsbury, 1964). There is no entirely effective similar in action to the phallotoxins (Bresinsky and Besl,
treatment, although removal of toxins from the digestive 1990).
tract, chemotherapy (in Europe, thioctic acid often is used),
and symptomatic treatment are used. In the United States, Pbytotoxius Produced by Bacteria or Fungi
liver transplants have been used in some instances. Mortality
is about 50-90% compared to about 10% for a typical small- As discussed previously (see Chapter 5), pathogenic bac-
pox epidemic. Those who contemplate eating wild-collected teria and fungi synthesize a number of compounds that help
mushrooms should read accounts of poisoning (Bresinsky to break down host tissues and to weaken the host plant.
and Besl, 1990; Kingsbury, 1964). Similar phytotoxins, especially those produced by bacteria,
Two major types of cyclic peptides have been isolated are peptides typically with molecular weights less than 600.
from Amanita species. Phallotoxins, such as phalloin (6), However, in contrast to the compounds of fungi, most bacte-
have seven amino acids in the ring, whereas, amatoxins [e.g., rial toxins exhibit an overall lack of specificity (Mitchell,
,,-amanitin (7)], have eight (Fig. 14.3). Certain portions of 1981). In some cases, these phytotoxins are produced in con-
the molecules are crucial for toxicity; for example, in the junction with phytotoxins frum other biosynthetic groups
amatoxin amanullin (8), elimination of a single hydroxyl of compounds. For example, in addition to the polyketide-
group from one amino acid eliminates activity. However, derived compounds involved in the attack on elm trees by
generalizations about the effects of structure changes on tox- Ceratocystis uimi, phytotoxic glycoproteins are also released
icity cannot yet be made reliably. (Harborne, 1988; Wood et aI., 1972).
The toxicity of ,,-amanitin is 10-20 times greater than Attack of the fungus Fusarium oxysporum on the tomato
phallotoxins, despite its slower action. ,,-Amanitin (7) inhib- appears to involve both fusarlc acid (10) (a pyridine deriva-
its RNA-polymerase II in the cell nucleus, resulting in a tive) and a small peptide, Iycomarasmin (11) (Fig. 14.4)
decrease in RNA content. The transcription of DNA by (Ballio, 1981; Harborne, 1988). The former compound is
mRNA is completely inhibited by an amanitin concentration linked to chelation of metal ions, whereas the peptide is
as low as 10- 8 M (Bresinsky and Besl, 1990). involved in water-permeability effects (Ballio, 1981).
Phallotoxins also attack liver cells. These peptides alter Tabtoxin or wildfire toxin (12), produced by Pseudomo-
membranes because they have affinity for actin filaments, nas syringae pv. tabaci and related bacteria, on tobacco pro-
238 Peptides

duces light-dependent chlorosis in the plants (Ballio, 1981; of S-adenosylmethionine to I -aminocyclopropane- I -carbox-
Mitchell, 1981). The peptide responsible possesses a i3-lac- ylic acid (Ballio, 1981).
tam and is highly unstable. The active peptide is converted Phaseolotoxin (17), a tripeptide, has been isolated from
to an inactive isomer, isotabtoxin (13), on standing (Ballio, cultures of Pseudomonas syringae pv. phaseolicola. This
1981; Mitchell, 1981). A related compound, (2-serine)tab- toxin and related compounds which co-occur with it, irre-
toxin (14), is produced along with tabtoxin in many in- versibly inhibit ornithine carbamoyltransferase and produce
stances. Other types of Pseudomonas syringae produce simi- chlorosis in the host (Ballio, 1981).
lar compounds such as coronatine (15). This peptide is The toxins of many members of the fungal genus Al-
responsible for chlorosis in a number of grasses (Mitchell, ternaria produce phytotoxins; of these, many [such as AM-
1981). toxin I (18)] are peptides. The fungal pathogen Alternaria
A similar peptide toxin, rhizobitoxin (16), is produced by mali on apples produce necrotic lesions on the leaves, shoots,
the root-nodulating organism Rhizobium japonicum. Rhi- and fruits of susceptible cultivars (Natori et aI., 1981). Al-
zobitoxin causes chlorosis in the developing leaflets of ternaria kikuchiana, which causes black spot disease on the
plants, such as soybean (Glycine max), which have nodules Japanese pear (Pyrus serotina) produces phytotoxins which
colonized with these strains of Rhizobiumjaponicum (Mitch- are based on N-acetylphenylalanine and a C IO fatty acid
ell, 198 I). This compound is a irreversible inhibitor of ethyl- (Harborne, 1986).
ene production from methionine, as it blocks the conversion Enniatins are peptides produced by Fusarium species. En-

alanin~ef ~ 'i~0
H" N~N
I! 0 OH

"" ~,l~" ~ "~"

H>Cl,-~o S
o
N
I H
tryptophan
0
hydroxyproline HNI ~k "~H
~ alanine

OH

!<
threonine
phalloidin (8)
(a phallotoxin)

~IH I! 0II i

>Ct
alanine 0
H" N~N
Nii ; H

~YPtOPhan
H, 0 '-"',
N /
HO ' 0 h ~ #

r
H NOS
~ I
hydroxyproline ~N-;CH
0
k
H 0
"-"'H alanine

I threonine
H OH

phalloin (6)

(a) (a phallotoxin)

Fig. 14.3 (a & b). Phallotoxins and amatoxins.

 rr
Peptide.~ 239

hydroxylated

~ --LJ--
OH OH tryptOPhan

""d:EN"' )cQ~'
a-amanitin (7) H 0 H glycine
(an amatoxin) I I °

H ° HI O">s, H 0 0 ~ isoleucine

r
hydroxyproline H ~
N", ,.~ ~ N
">"'H --"'N 'H
asparagine 0 ~ glycine
o NH2 cysteine
hydroxylated
tryptophan

~ i ~ if 'j

m ':."
glycine 0

H,. N-r\N~
~\,l:
""><-t. 0

O~ A _~ isoleucine

H ~N i I H7
r II
0 0
hydroxyproline ...., N............ I.~ H N,
""H -"'N~ H

o cysteine ~ glycine
asparagine 0 NHz amanullin (14)
(b)

Fig. 14.3. (continued)

niatin (19) (10-80 fLg/ml) reduces growth of wheat seed- selective advantage from opine synthesis by transformed
lings. Root elongation is inhibited more than leaf develop- plant cells (Tremblay et aI., 1987).
ment (Burmeister and Plattner, 1987).
Peptides from Blue-Green Algae
Opines A number of blue-green algae are known to produce
highly toxic compounds. When individuals of these organ-
Infection of susceptible plants with virulent cells of Agro-
isms occur in large numbers, as in algal blooms, they may
bacterium results in the development of crown gall tumors
produce severe poisoning problems in animals. The agents
and the synthesis of peptide metabolites called opines (Chil-
responsible belong to many diverse groups of compounds
ton et aJ., 1985) (Fig. 14.5). These compounds involve con-
(Moore, 1977, 1981); the toxin from Microcystis aeruginosa
densation of 2-oxo acids and basic amino acids such as L-
is a cyclic peptide. This compound is heat stable, dialyzes
arginine, L-histidine, and L-lysine (Luckner, 1990). The
slowly, and has an LDso intetperitoneally of 0.47 mg/kg in
genes of opine synthesis are carried by bacterial Ti or Ri
mice. Poisoning is rapid; symptoms appear in 15-45 min.
plasmids and are transferred to the plant cells in which they
One peptide from this organism, cyanoginosin-LR (20), has
are then expressed. The particular opines that are found in
been studied in some detail (Botes et aJ., 1984) (Fig. 14.6).
a given crown gall tumor correspond to those that can be
metabolized by the inciting bacteria. Further, opine synthesis
Siderophores
is a useful criterion for transfer of genetic material because
opines are unique products of transformed cells and are not A number of fungi, blue-green algae, and bacteria release
produced by free-living bacteria or nontransformed plant compounds into the environment that are highly effective
cells (Sederoff et aJ., 1986). Agrobacteria appear to gain a in chelating iron (also see Chapter 13). These organisms
240 Peptides

~(CH'hCH'

HO,C N

fusaric acid (10) coronatine (13)

CO,H
I (CH,hCHCH,", /
CO-NCH,
\
/CH2~H CH CHCH,
HO,C-CH- NH
I
NH
I kH bo
HOzC-CHz CHzCONHz
fO kH
Iycomarasmin (11) C,H,CH =C ):H'
\CH,-CO
o
II tentotoxin
H,N·1H.C.NH.~H.CO,H Alternaria tenuis, cotton chlorosis
CH, CH.OH
I I
o ~H, NH,
~C--C.OH
I I
NH-CH,

tabtoxin (12)

D-cys
/'
L-Ieu "- D-cys
\
D-Ieu _
/
L-val

malformin
(a) Aspergillus niger

Fig. 14.4 (a-b). Peptide phytotoxins.

apparently deplete iron from the surrounding medium to the ber of lectins or hemagglutinins are composed of peptide
extent that competing organisms cannot survive. Several of units, but are discussed under Proteins below.
the compounds involved are peptides (Fig. 14.7) (Aaronson, Several peptides can be absorbed intact by plants. The
1981; Winkelmann et al., 1987). three peptides DL·ala·DL·asp, DL·ala·DL·met, and DL·ala·
DL·leu are not hydrolyzed by the pitcher liquid of Ibe insec·
Peptides from Higher Plants tivorous plant Sarracenia, but are taken up intact into leaves.
The mechanism of absorption is unknown (Higgins and
Peptides in higher plants can be divided into two major Payne, 1982). However, during the germination of barley
categories. Some have unique structures and functions, grains, peptides produced by breakdown of storage proteins
whereas others are involved in the synlbesis or degradation are actively transported by the scutellum (Higgins and
of proteins (Higgins and Payne, 1982). In this treatment, Payne, 1982).
emphasis will be given the former type. There are limited
data concerning peptides of either type from plants. A series Glutathione
of peptides known as phytochelatins has been shown to che·
late heavy metals. A number of plants, including those of Glutathione ('Y·glu·cys·gly) can occur in a reduced or an
certain members of the Celastraceae, Euphorbiaceae, and oxidized form. The two are readily interconverted by the
Rharnnaceae, contain peptide alkaloids which are discussed enzyme glutathione reductase or by numerous redox re-
under Peptide and Macrolide alkaloids (Chapter 37). Anum· agents (Higgins and Payne, 1982). Both the peptide and the
 Peptide.s 241

CD3 CD3
"'-./
CH
O~ I
~C---C-O ",0
/ '-.... /,?
HN C

CH30O-CH'-CH'-CH''''''''"~H Hl""'CH3

- -:::?C, NH
0" /
HN _____ C..---c~
II ~O
AM-toxin I (19) CH,

~""OH
o CH
I'
CH, CH,OH
I
H,N-CH-CO-NH-CH-CO,H
I
serine tabtoxin (17) isolabloxin (16)

rhizobitoxin (14)

o
II ~H
HN-P(OH)-O~O,NH, NH-C-NH
I
(CH,h
I 2

I 1 3
CH (CH')4
I
H,N--CH---Co- NH-CH-CO --NH-CO,H

phaseolotoxin (15)

(b) (NO-phosphosulfamyl)ornithylanylhomoarginine

Fig. 14.4. (continued)

enzyme appear to be ubiquitous in plants and animals. Al- Payne, 1982). Individual compounds of this type are dis-
though several possible functions in animals have been pro- cussed in Chapter 13.
posed, the role is not as clear in plants. Glutathione is a key
compound in the ,,{-glutamyl cycle and may be involved in Phytochelatins
the transport of amino acids and peptides. In the chloroplast,
Many plants respond to the presence of heavy metals
glutathione may be involved in maintaining reducing condi-
by the production of peptides that are rich in cysteine. The
tions and the stability of enzymes containing sulfhydryl
presence of "{-glutamyllinkages indicates that these peptides
groups (Higgins and Payne, 1982).
are not primary gene products. Glutathione or "{-glutamyl-
cysteine have been suggested as precursors (Gekeler et aI.,
y-Olutamyl Peptides
1988). Many have the structure ,,{-glutamic acid-(cysteine)n-
Although ,,{-glutamyl peptides are common in plants, they glycine with n = 2-11. Those from several legumes have
are not found in animal cells. The majority are dipeptides, the general structure (,,{-glutarnylcysteine)n-f\-alanine with n
although a number oftripeptides are found. Significant quan- = 2-7 (Grill et aI., 1986). Phytochelatins have been found
tities of "{-glutarnyl peptides occur, especially in storage or- in several plants, including Anethum graveolens, Berberis
gans of plants. For example, 34% of the nonprotein amino stolonifera, Catharanthus raseus, Dioscarea camposita, Fu-
acid nitrogen of kidney bean seeds (Phaseolus vulgaris) is maria parviflora, Galium mo/lugo, Glycine max, Malva sylv-
present as "{-glutamyl-S-methyl-L-cysteine (Higgins and estris, Phaseolus aureus, Phaseolus multiflorus, Phaseolus
242 Pep/ides

D,L-succinamopine L,L-succinamopine L,L-succinamopine

ladam

H. n
~ __ ? O NH yCOH
' HO'VyCO,H
HO,c~ N A ~!'IH ~ H"NH
HOC V H,N N~ HN NH _ H,[
2 ~"""'C02H C02H 2 ~ ~"C02H

L,L-succinopine octopine nopaline

ladam

HO,C~CO'H H02C~C02H
o H 'NH
H , H --NH

H2N~C02H AX CO,H

succinamopine leucinopine

Fig. 14.5. Representative opines.

vulgaris, Rauvolfia serpentina, Rhazya stricta, Rosa canina,

Silene cucubalus. Solanum marginatum, and Thalictrum
dipterocarpum (Grill et aI., 1985, 1986).
A similar cadmium-binding complex of peptides is pro-
duced by the alga Chlorella fusea as well as other Phyto-
phyta (Gekeleret al., 1988). Phytochelatins have been identi-
fied in the roots of heavy-metal-sensitive Aeer
pseudoplatanus and resistant Silene cueubalus plants grown
in zinc-rich soil, whereas plants grown in the absence of this
metal lacked these peptides. Metal-binding phytochelatins
appear to be specifically induced in plants in heavy-metal-
enriched ecosystems (Grill et aI., 1988).

PROTEIl'IS

Proteins are found in all organisms, play many roles, and

vary widely in structure and properties. The enzyme ribulose
1,5-bisphosphate carboxylase (EC 4.1.1.39) is probably the
world's most abundant protein, as it is the major protein
present (up to 50%) in the leaves of plants (Ramshaw, 1982).
However, in the following discussion, only those proteins
with toxic or inhibitory properties to animals or fungi or
other noteworthy activity will be discussed. Several groups
of proteins from plants are known to be toxic to animals.
The most important group of these is called lectins or hemag-
glutinins. Many legume seeds also contain protease (or tryp-
cyanoginosin-LR (20) sin) inhibitors (Weder, 1981). Similar compounds are found
in plants of the Solanaceae (Ramshaw, 1982).
Fig. 14.6. Cyanoginosin-LR.
Phytohemoglobins
Plant hemoglobin is known to occur in eight families of
dicotyledonous plants. These farailies, the Betulaceae, Casu-
 Peptides 243

aerobactin [Aerobacter (Klebsiella) aerogenesJ

HH Hn HH
H,N(CH,J,N.C(CH,J,CONH(CH,J,N.C(CH,J,CONH(CH,J,N.CCHJ

desferal (Streptomyces pilosus)

IT fH iO,H HITI
CHJC.N(CH,JJNHIiCH'iCH'IiNH(CH,JJN.CCHJ

° OH °
(a) schizokinen (Bacillus megaterium) enterochelin (Escherichia coli) (b)

(c) ferrichrome (Ustilago sphaerogena)

Fig. 14.7 (a-c). Structures of several siderophores.

244 Peptides

arinaceae, Coriariaceae, Datiscaceae, Eleagnaceae, Faba- be highly toxic (Anon., 1987; Griffiths et al., 1987). Protein
ceae, Myricaceae, Rhamnaceae, Rosaceae, and Ulmaceae extracts of the seeds of Ricinus communis, castor bean (Eu-
(Landsmann et al., 1986), are not closely related; most of phorbiaceae), contain proteins that are antigenic and aggluti-
the plants have a symbiotic relationship with nitrogen-fixing nate defibrinated blood and red blood cells in vitro. As few
bacteria or actinomycetes. The plant gene has a 4-exon-3- as two to four seeds of Ricinus communis can be deadly
intron structure, while in vertebrates exons 2 and 3 are fused, and eight are almost always lethal (Ishiguro et al., 1971;
forming a long internal exon (Kubitzki et al., 1991). In order Kingsbury, 1964). Ricin is a dimeric protein with two differ-
to account for the peculiar distribution of plant hemoglobin, ent peptide chains (63,000 MW). The chains have been puri-
either the gene must be widespread, but only occasionally fied and the amino acid sequence of both chains reported.
expressed, or horizontal transfer of genetic material must The A chain is capable of inactivating the 60 S subunit of
have occurred-possibly from vertetrates (Kubitzki et aI., the ribosomes of eukaryotic cells resulting in inhibition of
1991). At present, neither possibility can be confirmed. protein synthesis. The B chain binds galactose and saccha-
rides containing nonreducing j3-galactosyl units and permits
Lectins or HemaggIutinins entry into the cell by the toxic subunit (Liener et al., 1986).
A number of other agglutinins are found in seeds of this
Lectins or hemagglutinins are well known among the nat-
plant.
urally occurring plant proteins with toxic effects in animals
The seeds of Aleurites [ordii, the tung nut (Euphor-
(Liener, 1991; Lis and Sharon, 1981). This is a somewbat
biaceae), formerly cultivated in the south central United
heterogeneous collection of proteins that agglutinate cells
States, and presently cultivated in Argentina and China pro-
and exhibit antibody-like sugar-binding activity (Liener et
duce toxic proteins. There are many references to human
al., 1986; Ramshaw, 1982). Agglutination is inhibited by
poisoning by this plant, as the seed is large and, to many,
sugars, oligosaccharides, glycosides, and glycoproteins. Pre-
appears edible. Press cake from the manufacture of tung
cipitation occurs in the presence of glycoproteins and poly-
oil also is toxic and difficult to detoxify. Cocarcinogenic
saccharides (Liener et al., 1986). Lectins are necessarily
diterpenes have been recently reported from this plant and
multivalent; that is, they bind at least two sugar molecules
are undoubtedly responsible for part of the toxic activity
in order to cause precipitation. The sugar specificity of lec-
(Beutler et al., 1989). Plants of Jatropha curcas and J. mul-
tins is usually dermed in terms of the monosaccharide(s}
tiftda, also from the Euphorbiaceae, contain toxic proteins,
that inhibit lectin-induced agglutination (Liener et al., 1986).
sometimes called •• curcin. " Both species are planted widely
Lectins often constitute 6-11 % of the plant's total protein
in the tropics.
(Liener, 1991).
Several members of the Loranthaceae (the mistletoe fam-
These proteins occur in the seeds of many plants but are
ily) have long been used as medicinal plants. Viscum album,
especially common in the Fabaceae (Leguminosae) (more
a European species, was used in this manner and there is
than 600 species) and the Euphorbiaceae (lectins also occur
an extensive folklore associated with it. The toxic fraction
in other organisms such as fungi, bacteria, and animals). In
(sometimes called viscotoxin) is a mixture of several related
legnmes, lectins are usually found in the cotyledons (Liener,
peptides which affect heart muscle (Ramshaw, 1982). One
1991). Well-known plant lectins are concanavalin A, from
of these, viscotoxin B, has 46 amino acids, a 9170 MW, and
jack beans (Canavalia ensiformis), favin, from the broad
an LD,o (mouse) of 0.78 mg/kg. Viscotoxin A has 46 amino
bean (Vicia Jabal, and phasin, from the kidney bean
acids (Olson and Samuelsson, 1972; Samuelsson and Pet-
(P haseolus vulgaris) (Ramshaw, 1982). The toxicity of lec-
tersson, 1971; Sandberg and Samuelsson, 1961). Although
tins differs considerably (Liener, 1991; Liener et al., 1986).
the froits of the southeastern American species, Phoraden-
Data for lectins are given in Liener et al. (l986) and Lis and
dron villosum and P. flavescens (mistletoe), are also poison-
Sharon (1981).
ous, they contain j3-phenethylamine and tyramine (see Chap-
The brilliant red black seeds of Abrus precatorius (preca-
ter 28).
tory bean) contain abrin, an extremely toxic protein (Ghosal
and Dutta, 1971). Seeds of this plant are commonly used in
Biological Activity of Lectins
the tropics to make necklaces and other jewelry. One seed,
if thoroughly masticated, is sufficient to cause death in a The interactions of Rhizobium bacteria and plants of the
child (Kingsbury, 1964). The lectins of this plant primarily Fabaceae are highly specific. It is believed that lectins play
bind galactose and are similar in properties to those of ricin a major role in this specificity (Etzler, 1986, Liener, 1991;
(see below). The lectin has a molecular weight of 65,000 Sharon and Lis, 1989). Several models have been proposed
and is composed of dissimilar peptides. to explain this interaction. The lectin appears to act as a
Robinia pseudoacacia, the black locust (Fabaceae), is a "glue" to hold the bacterium in place (Riidiger, 1984). It
common cultivar in the United States and in Europe and appears that binding of host lectins may send a signal to the
is commonly escaped from cultivation. The bark contains bacterium that turns on a gene, which leads to the synthesis
several toxic proteins (Kingsbury, 1964). The seeds are also of a specific polysaccharide on the bacterial cell wall that
known to contain a distinct lectin (Liener et aI., 1986). serves as a receptor site for the lectin (Liener, 1991).
Seeds of members of the Euphorbiaceae are also known to A mixture of lectins of Phytolacca americana, pokeweed
 Peptides 245

(Phytolaccaceae), is known to be highly mitogenic (pro- pound of serendipity froit, monellin, is 2500 times sweeter
motes mitotic cell divisions). Individual components of the than sucrose and has a molecular weight of 11 ,000. The
mixture have been isolated and their binding properties de- proteins from miracle froit cause sour substances to taste
termined (Liener et aI., 1986). sweet but have no inherent taste. These compounds are dena-
Another role of lectins is in the interactions of the plant tured by heating, limiting their usefulness as sweetening
against insects and microbial pathogens (Etzler, 1986, agents (Inglett, 1984). Thaumatin, a protein of 207 amino
Liener, 1991; Sharon and Lis, 1989). The lectin from acid residues, occurs in the West African shrub Thaumato-
PfUlseolus vulgaris has a lethal effect on the larvae of a coccus danielli (Marantaceae) and is about 5000 times swee-
bruchid beetle, presumably because of the binding of the ter than sucrose. It is used as a food additive in some coun-
lectin to epithelial cells lining the midgut of this insect. Such tries, but high cost has limited its wider use. The genes
binding inhibits uptake of nutrients (Liener, 1991). Soybean encoding thaumatin have been cloned and expressed in Esch-
lectin inhibits larval growth of Manduca sexta (Liener, erichia coli, but yields were disappointingly low. Expression
1991). A protein from the seeds of a wild variant of Phaseo- in SaccfUlromyces cerevisiae was better (McPherson and
Ius vulgaris is toxic to a bean weevil (Zabrotes subfascia- Parish, 1987).
tus), which is a common bean pest. Transfer of the allele
for the production of this lectin to nonresistant bean cultivars
and back-crossing resulted in plants with high levels of insect REFERENCES
resistance (Liener, 1991).
Lectins from wheat germ, potato, and soybean have been ANON" Castor oil shares its poison with dysentery bacterium, New
shown to inhibit fungal growth (Liener, 1991). Scientist, 115, 22 (20 August 1987).
The effect of lectins in the diets of humans has been AARONSON. S., Molecules excreted by microorganisms. Vois. 1 and
reviewed (Liener, 1991; Liener et al., 1986). Clearly, un- 2 of Chemical Communication at the Microbial Level, Vol. I,
cooked legume seeds should not be eaten. The effects of 1-27, CRC Press, Boca Raton, FL, 1981.
cooking on lectins is not clear, but uncooked seeds contain- BALUO, A., Structure-activity relationships, in Toxins in Plant Dis-
ing these compounds should be considered toxic and inedible ease (R. D. Durbin, ed.), 395-441, Academic Press, New York,
under any conditions. 1981.
BERny, J., Handbook of Antibiotic Compounds, Vols. 1-12, CRC
Proteinase InbIbitors Press, Boca Raton, FL, 1980-1985.
BEUTLER, J. A., A. B. ALVARADO, T. G. MCCLOUD, and G. M.
A number of proteinase inhibitors, often called trypsin
CRAGG, Distribution of phorbe! ester bioactivity in the Euphor-
inhibitors, are known to occur in seeds of legumes (Ryan, biaceae, Phytother. Res .. 3, 188-192 (1989).
1981; Weder, 1981). These polypeptides often inhibit both
BOTES, D. P., A. A. TUINMAN, P. L. WESSELS, C. C. VlLJOEN, H.
trypsin and chymotrypsin and block digestion of many leg- KRUGER, D. H. WILLIAMS, S. SANTIKARN, R. J. SMITH, and S.
ume seeds. Cooking deactivates most of these compounds. J. HAMMOND, The structure of cyanoginosin-LR, a cyclic hepta-
peptide from the cyanobacterium Microcystis aeruginosa, J.
Sweeteners Chern. Soc. Perkin I, 2311-2318 (1984).
BRESINSKY, A. and H. BESL, A Colour Atlas of Poisonous Fungi.
A number of peptides and proteins modify taste in hu- Wolfe, London, 1990.
mans and presumably in other animals (Ramshaw, 1982).
BURMEISTER, H. R. and R. D. PLATINER. Enniatin production by
Aspartame (21) (Fig. 14.8), a purely synthetic dipeptide, is
Fusarium tricinctum and its effect on genninating wheat seeds,
widely used as a sweetening agent in many countries. Other Phytopathology, 77, 1483-11487 (1987).
sweet-tasting compounds, such as those of serendipity ber-
CHILTON, W. S .• E. HOOD, and M. CHILTON, Absolute stereochemis-
ries (Dioscoreophyllum cumminsii, Menispermaceae), mira- try of leucinopine, a crown gall opine, Phytochemistry, 24,
cle froit (Richardella dulcifera, formerly known as Synsepa- 211-224 (1985).
lum dulcificum, Sapotaceae), and katemfe (TfUlumatococcus E1ZLER. M. E., Distribution and function of plant leetins, in The
danielli, Marantaceae) are proteinaceous. The sweet com- Lectins. Properties, Functions, and Applications in Biology and
Medicine (I. E. Liener, N. Sharon, and I. 1. Goldstein. eds.),
371-435. Academic Press, New York. 1986.
fH,CO,H
GEKELER, W., E. GRILL, E. WINNACKER, and M. H. ZEN!<, Algae
sequester heavy metals via synthesis of phytochelatin com-
H,NCHCONHCHCO,CH, plexes, Arch. Microbiol., 150. 197-202 (1988).

I
CH,C,H,
GHOSAL, S. and S. K. DUTIA, Alkaloids of Abrus precatorius, Phy-
tochemistry, 10, 195-198 (1971).
GOTILIEB, D. and P. D. SHAW (eds.). Antibiotics, Vols. 1-6,
Springer-Verlag, New York and Berlin, 1967-1983.
aspartame (21)
GRIFFITHS, G., A. LEITE, and M. GREEN. Proteins that play Jekyll
Fig. 14.8. A'partame. and Hyde, New Scientist, 115, 59-61 (16 July 1987).
246 Peptides

GRILL, E., W. GEKELER, E. WlNNACKER, and M. H. ZENK, Homo- MITcHELL, R. E., Structure: Bacterial, in Toxins in Plant Disease (R.
phytoche1atins are heavy metal-binding peptides of homo-gluta- D, Durbin, ed.), 259-293, Academic Press, New York, 1981.
thione containing Fabales, Federation of European Biochemical MOORE, R. E., Toxins from blue~green algae, BioScience, 27,
Societies (FEBS) Lett, 205, 47-50 (1986), 797-802 (1977).
GRILL, E., E. WINNACKER, and M. H. ZENK, Phytochelatins: The MOORE, R. E., Constituents of blue-green algae, in Marine Natural
principal heavy-metal complexing peptides of higher plants, Prod., Vol. 4 (P. Scheuer, ed,), I-52, Academic Press, London,
Science, 230, 674-676 (1985), 1981.
GRILL, E., E. WINNACKER, and M. H. ZENK, Occurrence of heavy NATORI, S., N. IKEKAWA andM. SUZUKI, Advances in Natural Prod-
metal binding phytochelatins in plants growing in a mining re- ucts Chemistty, Wiley, New York, 1981.
fuse area, Experientia, 44, 539-540 (1988), OLSON, 1. and G. SAMUELSSON, The amino acid sequence of visco-
HARBORNE, 1. B., Recent advances in chemical ecology, Nat. Prod. toxin A2 from the European mistletoe (Viscum album), Acta
Rep,,4, 323-344 (1986), Chern, Scand, 26, 585-595 (1972).
HARBORNE, J. B., Introduction to Ecological Biochemistry, 2nd ed., RAMSHAW, 1. A. M., Structures of plant proteins, in Nucleic Acids
Academic Press, London, 1982; 3rd edition, 1988. and Proteins in Plants I (D. Boulter and B. Parthier, eds.), Ency~
HIGGINs, C. F. and 1. W. PAYNE, Plant peptides, in Nucleic Acids clopedia of Plant Physiology, N,S., Vol. 14A, 229-290,
and Proteins in Plants I (D, Boulter and B, Parthier, eds,), Ency- Springer-Verlag, Berlin, 1982,
clopedia of Plant Physiology, N, S" VoL 14A, 438-458, RUDIGER, H., On the physiological role of plant lectins, BioScience,
Springer-Verlag, Berlin, 1982. 34, 95-99 (1984).
INGLETI, G. E., Sweeteners: An overall perspective, in Aspartame, RYAN, c. A., Proteinase inhibitors, in Proteins and Nucleic Acids
Physiology and Biochemistry (L. D, Stegink and L. 1. Filer, Jr., (A, Marcus, ed.), Vol. 6 of The Biochemistty of Plants (P, K.
eds.), 11-25, Marcel Dekker, Inc" New York, 1984. Stumpf and E, E. Conn, eds.), 351-370, Academic Press, New
York, 1981.
ISHIGURo, M., F. GUNKI, and F. MASARU, Biochemical studies on
ricin IV, Agric, BioL Chern" 35, 729-733 (1971), SAMUELSSON, G. and B. M. PETIERSSON, The amino acid sequence
of viscotoxin B from the European mistletoe (Viscum album
KINGSBURY, 1. M., Poisonous Plants of the U.S. and Canada, 91,
L., Loranthaceae), Eur. 1. Biochem" 21,86-89 (1971),
Prentice-Hall, Englewood Cliffs, NJ, 1964.
SANDBERG, F. and G. SAMUELSSON, Toxic peptides from the Loran-
KUBI1ZKI, K., P. v. SENGBUSCH, and R. POPPENDIECK, Parallelism, thaceae, Symposium on Phytochemistty, 54, (H. R. Arthur, ed,),
its evolutionary origin and systematic significance, Aliso, 13,
Hong Kong University Press, Hong Kong, 1961.
191-206 (1991),
SEDEROFF, R., A. STOMP, W. S. CHILTON, and L. W. MOORE, Gene
LANDSMANN, 1., E. S. DENNIS, T. 1. V. HIGGINS, C. A. APPELBY, transfer into loblolly pine by Agrobacterium tumefaciens,
A. A. KORTI, and W. 1. PEAcOCK, Common evolutionary origin Bio/Technology, 4, 647-648 (1986).
of legume and non-legume plant hemoglobins, Nature, 324,
SHARON, N. and H. LIS, Lectins as cell recognition molecules, Sci-
166-168 (1986).
ence, 246, 227-246 (1989).
LIENER, 1. E., LECTINS, in Herbivores: Their Interactions with Sec-
SNEADER, W., Drug Discovery: The Evolution of Modem Medi-
ondary Plant Metabolites (G, A. Rosenthal and M. R. Beren-
cines, John Wiley & Sons, Ltd., Chichester, 1985.
baum, eds,), 327-353, Academic Press, San Diego, CA, 1991.
TREMBLAY, G., R. GAGLIARDO, W. S. CHILTON, andP. DION, Diver-
LIENER,1. E., N. SHARON, and 1. 1. GOLDSTEIN (eds.), The Lectins, sity among opine-utilizing bacteria: Identification of Coryne-
Academic Press, Orlando, FL, 1986. form isolates, AppL Env, Microbiol., 53, 1519-1524 (1987),
LIs, H. and N. SHARON, Lectins in higher plants, in Proteins and WEDER, 1. K. P., Protease inhibitors in the Leguminosae, in Ad-
Nucleic Acids (A. Marcus, ed,), VoL 6 of The Biochemistty of vances in Legume Systematics (R. M. Polhill and P. H. Raven,
Plants (P. K. Stumpf and E, E. Conn, eds.), 371-447, Academic eds.), 533-560, Royal Botanical Gardens, Kew, UK, 1981.
Press, New York, 1981.
WIELAND, T., Poisonous principles of mushrooms of the genus
LrITEN, W., The most poisonous mushrooms, Sci. Am., 232, Amanita, Science, 159, 946-952 (1968).
90-101 (1975), WINKELMANN, G., D. VAN DER HELM, and 1. B. NEILANDS, Iron
LUCKNER, M., Secondary Metabolism in Microorganisms, Plants, Transport in Microbes, Plants and Animals, VCR Verlagsge-
and Animals, Springer-Verlag, Berlin, 1990. selL, Weinheim, 1987.
McPHERSON, M. J. and J. H. PARISH, Applications of recombinant WOOD, R. K. S., A. BALLIO, and A. GRANm, Phytotoxins in Plant
DNA in biotechnology, Nat. Prod. Rep., 4, 205-224 (1987). Diseases, Academic Press, London, 1972.
15
Carbohydrates

Introduction other compounds as glycosides. Whereas a few sugars are

Simple Sugars primary metabolites and are Ubiquitous in plants (Fig. 15.1),
Simple Sugars-Secondary Metabolites many others are limited in distribution. Some sugars are
Nomenclature potent inhibitors of metabolism (Engel, 1992).
2-Deoxyhexoses Monosacchatides are polyhydroxy aldehydes (aldoses) or
6-Deoxyhexoses polyhydroxy ketones (ketoses). These monosacchatides usu-
Methoxy Sugars ally contain several chiral centers; those which have the same
Branched-Chain Sugars configuration as o-glyceraldehyde at the chiral C-atom
Sugar Carboxylic Acids farthrest from the carbonyl group belong to the o-series (0-
Amino Sugars glucose, o-fructose, etc.), whereas their counterparts belong
Biological Activity of Sugars to the L-series (L-rharnnose, L-fucose, etc.) (Engel, 1992;
Disacchatides and oligosacchatides Luckner, 1990). Monosacchatides exist as cyclic hemiace-
Sucrose tals (pyranoses and furanoses) rather than as open chain
OJigosacchatides forms by which they are frequently represented. The py-
Fructans ranose and furanose forms involve an additional, anomeric
Polysacchatides: Reserve and Structural Forms chiral center, represented by at- or 13-. The anomeric forms
Energy Storage Compounds Derived from Sugars of monosacchatides equilibrate in solution (Luckner, 1990).
Structural Carbohydrates The six-membered forms are best represented by chair con-
Callose formations; in general, several conformers of a sugar may
Pectins exist and can be interconverted.
Plant Gums There are several ways in which sugar structures are rep-
Algal Polysacchatides
resented. In some instances in this book, sugars are repre-
Chitin
sented in a chair-boat type of structure. However, many of
Biological Activity of Oligosacchatides and
the following figures include Fischer projections [in which
Polysacchatides
the aldehyde or keto group is at the top, and the penultimate
Sugar Alcohols
(from the bottom) OH group is on the right in o-sugars (R),
Polyols
the same as the relationship at the chiral carbon of ( + )-0-
Cyclitols
glyceraldehyde, and on the left in L-SUgars (S)], Haworth
L-Ascorbic Acid
Glycosides in Plants perspective formulas (El Khadem, 1988), and other repre-
Indigo-producing Glycosides sentations of sugars.
Lactone-forming Glycosides Solutions of aldoses at equilibrium contain mixtures of
Sugar Esters at least two (at and 13) furanoses and two (at and 13) pyranoses.
References Typically, the two pyranose forms predominate at equilib-
rium. The anomeric carbon is chiral (C-I of aldoses and C-
2 of ketoses). In the o-series, if the OH on the anomeric
Il'ITRODUCTION
carbon is to the right in a Fischer projection or below the
A large number of sugars or sacchatides are found in nature. plane of a Haworth perspective formula, the isomer is at and
These may occur singly (monosacchatides) or as dimeric, it is 13 if the OH is to the left or above. Conversely, in the
trimeric, oligomeric, or larger aggregates or combined with L-series, the at anomer has the OH at carbon 1 to the left or

247
248 Carbohydrates

above and the 13 anomer has this OH to the right or below ars that possess a hemiacetal linkage may be easily oxidized
(El Khadem, 1988). and are known as reducing sugars. All monosaccharides are
reducing, whereas many complex saccharides are nonre-
ducing.
Many other sugars are involved in primary metabolic
SJlIWLE SUGARS
pathways such as the oxidative and reductive pentose cycle,
glycolysis, and photosynthesis.
o-Glucose (1), o-galactose (2), o-glucuronic acid (3), D-ga- Glucose, xylose, mannose, and arabinose often are associ-
lacturonic acid (4), o-xylose (5), and L-arabinose (6) consti- ated with wall materials in plants (Fig. 15.1).
tute the major sugars of carbohydrates of higher plants (El The Calvin cycle, associated with the photosynthetic as-
Khadem, 1988). similation of CO2 , represents the principal source of phos-
Monosaccharides are divided into two major groups de- phorylated sugars in plants. o-Fructose 6·phosphate acts as
pending on whether their acyclic forms possess an aldehyde the precursor of many of the monosaccharide moieties in
or a keto group, that is, whether they are aldoses or ketoses. plants (Feingold and Avigad, 1980). Kinases capable of
Monosaccharides can also be grouped by the size of the ring phosphorylating o·glucose, o-fructose (8), o·mannose, and
that they form, for example, furanoses or pyranoses. Both o-glucosamine (9) at C-6 and L-arabinose, D·galactose, and
aldoses and ketoses may be designated as to size, for exam- D-galacturonate at Col have been demonstrated in a large
ple, as aldopentoses or 2-hexuloses (El Khadem, 1988). Sug- number of plants (Feingold and Avigad, 1980).

CHO CHO
I I
CHO H-C-OH HO·C-H
I I I
H-C-OH H-C-OH H-C-OH
I I I
CH,OH CH,OH CH,OH

D-glyceraldebyde D-erythrose D-threose

CHO CHO CHO

CHO
I I I I
H-C-OH HO-C-H H-C-OH HQ-C-H
I I I I
H-C-OH H-C-OH HO·C-H HO-C-H
I I I I
H-C-OH H-C-OH H-C-OH H-C-OH
I I
I I CH,OH
CH,OH CH,OH CH,OH

D-ribose D-arabinose D-xylose D-Iyxose

CHO CHO CHO CHO

I I I I
H-C-OH HO'C-H H-C-OH HQ-C-H
I I I I
H-C-OH H-C-OH HO·C-H HO-C-H
I I I I
H-C-OH H-C-OH H-C-OH H-C-OH
I I I I
H-C-OH H-C-OH H-C-OH H-C-OH
I I I I
CH,.OH CH,.OH CH,.OH CH,·OH
D-allose D-altrose D-gJucose D-mannose
CHO CHO CHO CHO
I I I I
H-C-OH HO-C-H H-C-OH HO-C-H
I I I I
H-C-OH H-C-OH HO-C-H HO-C-H
I I I I
HO-C-H HO-C-H HO-C-H HO-C-H
I I I I
H-C-OH H-C-OH H-C-OH H-C-OH
I I I I
CH,·OH CH,·OH CH,·OH CH,.OH

(.) D-gulose D-idose D-galactose D-talose

Fig. 15.1 (a & c). Structures of some important primary monosaccharides.

 Carbohydrates 249

~
OH o
OHPH ~OH
HO 0
~OH
0
HO
HO ~:H~H 0 HO
HO H HO
HO
NH, H
OH OH OH

a-D-glucopyranose (1) a-D-galactopyranose (2) o:-D-mannopyranose (7) a-D-glucosamine (9)

CO,H 0 O~'=OlH HO 0 HOCHDo",--PH

~HHO
HO OHO
OH H ~CHlOH.I . ,
OHCH,OH
OH OH HO OH OHH

a-glucuronic acid. (3) ,,-gaIac!1lronic acid (4) ,,-D-fructopyranose (8) P-D-fructofuranose

"x::.. ';;-t.,0
HOCHYu°~OH
H
OH 0 H 0 CH,OH

H~H'H HOCH, HH<lu~HHDoH

OH H H OH HO OHOH
poD-ribose (10) ,,-D-xy)ose (5) a-D-ribulose

~HlOH

t---r
HX:°'6H<CHl OH C=O

H OH
HO-~-H
I
H-{;-OH
OH H
I
CH,OH
a-D-xylulose D-xylulose (open chain)
(b)

Fig. 15.1. (continued)

Many sugars occur in combined forms with nucleotides Techniques for the analysis of carbohydrates have been
(Feingold and Avigad, 1980). The amount of nucleotide sug- reviewed (Churms, 1990; EI Khadem, 1988; Sturgeon,
ars is usually 10-25% of the pool of soluble nucleotides 1990).
extracted from plant tissues, but extraction and chromato-
graphic separation may lead to degradation of these nucleo-
tides. UDP-G1ucose usually predominates and is ubiquitous SIMPLE SUGARS-SECONDAKY IllETABOLITES
in plants, but in seeds in which starch synthesis is intense, the
major nucleotide sugar is adenosine 5'-(a-D-glucopyranosyl Although most of the sugars discussed above are distributed
pyrophosphate) (ADP-glucose) (Feingold and Avigad, widely, a number of others are limited in distribution and
1980). Techniques for analysis of nucleotide sugars have may be considered to be plant secondary compounds. A
been reviewed (Feingold and Barber, 1990). number of unusual sugars are found in glycosides, especially
Several epimeric pairs of monosaccharides are produced in cardiac glycosides and glycosidic Solanum alkaloids.
by epimerization of nucleotide sugars (Luckner, 1990). For Other uncommon sugars are found in antibiotics produced
example, UDP-D-glucose-4-epimerase converts UDP-D-glu- by bacteria and fungi.
cose to UDP-D-galactose. Enzymes that produce the other
epimers are also known. A C-2 epimerase may be involved Nomenclature
in the interconversion of D-glucose and o-mannose (Karr,
1976). The trivial names of the common sugars, including a1-
The reactions of monosaccharides and oligo saccharides doses with up to six carbon atoms and ketohexoses, are still
have been summarized (Dey, 1990; Gander, 1976). used (Fig. 15.1). These include the D- and L-forms of glycer-
Combined monosaccharides are widespread and occur in aldehyde, erythrose, threose, ribose, arabinose, xylose, lyx-
all plants; free monosaccharides also are widespread, but ose, allose, altrose, glucose, mannose, gulose, idose, galac-
usually are not accumulated. tose, and talose (El Khadem, 1988). The prefixes derived
250 Carbohydrates

from these names are used to designate the configuration of dehydrogenases in analogy to the biosynthesis of 2-deoxy-
a group of consecutive chiral carbon atoms to which other n-ribose (11) [which should more correctly be named 2-
groups are attached. The italicized prefix derived from the deoxy-n-erythro-pentose (El Kbadem, 1988)] from n-ribose
name of the aldose with the same configuration is preceded (10) (Luckner, 1990). (See Fig. 15.2.) Both n-ribose and 2-
by the appropriate n or L symbol and followed by the root deoxy-n-ribose are components of nucleic acids.
denoting the total number of carbon atoms in the chain. Keto- Among the 2-deoxy sugars found as the carbohydrate
ses in which the carbonyl group contains C-2 are named by portion of cardiac glycosides and other glycosides in plants
prefixing one of these prefixes to a root denoting the total are n-cymarose (2,6-dideoxy-3-0-methyl-n-ribo-hexose)
number of carbon atoms in the chain and suffixing "ulose" (12), n-sarmentose (2,6-dideoxy-3-0-methyl-n-xylo-hex-
to make, for example, n-arabino-hexulose (n-fructose) ose) (13), L-oleandrose (2,6-dideoxy-3-0-methyl-L-arabino-
[however, the trivial names of the four n-hexuloses (n-fruc- hexose) (14), n-diginose (2,6-dideoxy-3-0-methyl-n-lyxo-
tose, n-psicose, n-sorbose, and n-tagatose) are accepted]. If hexose) (15), n-digitoxose (2,6-dideoxy-n-ribo-hexose)
the carbonyl group is at a higher numbered position, the (16), and n-boivinose (2,6-dideoxy-n-xylo-hexose) (17)
appropriate number is used, for example, n-arabino-3-hexu- (Courtois and Percheron, 1970).
lose (El Kbadem, 1988). Replacement of a hydroxy group
with a hydrogen yields deoxy sugars such as 6-deoxy-n- 6·Deoxyhex08es
galactose and 6-deoxY-L-mannose (which have the accepted
names of n-fucose and L-rhamnose). The names of deoxy A number of 6-deoxyhexoses also are widespread in
sugars may be used to name a product in which the hydroxyl plants, but these compounds are especially common in car-
group of a sugar has been replaced by an amino group. For diac glycosides (Courtois and Percheron, 1970). These sug-
example, n-glucosamine is named 2-amino-2-deoxy-n-glu- ars are formed from the corresponding hydroxylated mono-
cose. For more complex sugars, El Kbadem (1988) should saccharides (Luckner, 1990). Among the 6-deoxyhexoses
be consulted. are L-acofriose (6-deoxy-3-0-methyl-L-mannose) (18), L-
acovenose (6-deoxy-3-0-methyl-L-talose) (19), n-allometh-
2·Deoxybexoses ylose (6-deoxy-n-allose) (20), L-altromethylose (6-deoxY-L-
2-Deoxy sugars may be formed by direct reduction at altrose) (21), n-antiarose (6-deoxy-n-gulose) (22), n-boivi-
the level of nucleotides by pyridine nucleotide-dependent nose (2,6-dideoxy-n-xylo-hexose) (17), n-cymarose (2,6-di-

,
CH,OH
, CH,OH
,
CH,OH
,
c=o ,
c=o
,
CH,OH
c=o ,
I
c=o
HOC-H
,
H-C-OH

I
CH,OH
,
H-C-OH H-C-OH
, ,
H-C-OH
CH,OH CH,OH CH,OH

dihydroxyacetone D~erythrulose D-xylulose D-ribulose

,
CH,OH ,
CH,OH
, CH,OH
,
CH,OH

,
c=o ,
c=o ,
c=o ,
c=o
,
HOC-H ,
H-C-OH
,
HOC-H
,
H-C-OH

,
HOC-H HOC-H
, ,
H-C-OH
,
H-C-OH

,
H-C-OH ,
H-C-OH
,
H-C-OH
,
H-C-OH
CH,-OH CH,-OH CH,.OH CH,-OH

D~tagatose D-sorbose D-fructose D-psicose

CHO
, ,
CHO ,
CHO
,
CHO
HOC-H
, ,
H-C-OH HO·C-H
, ,
H-C-OH

,
H-C-OH
,
HOC-H ,
H-C-OH HOC-H
,
,
HOC-H
,
H-C-OH ,
H-C-OH HOC-H
,
,
HOC-H
,
H-C-OH ,
HOC-H
CH,-OH
,
H-C-OH
CH,-OH
CH,-OH CH,.OH

(c) L-glucose D-glucose L~galactose D-galactose

Fig. 15.1. (continued)
 Carbohydrates 251

NH2 reduced thioredoxin NU 2

O' O' (xN) ( OXidtz:~:~or~:OXin O' (x>

I INN ~ I I N

I'
O'-P-O-P-O~
tI
00 HHHH
ribonucleotide
reductase
O'-P-O-P-O~
II II
00 HHHH

OH OH OH H

adenosine diphosphate (ADP) 2'-deoxyadeoosine diphosphate (dADP)

H HHH HO~O
HOCHX-;;0';;j<0H HOCHx-;;0:t<0H
CH, OH
H H H OCH,H H

OH OH OH H CH, H
2,6-dideoxy-3-C-methyl-O-methyl-
D-ribose (10) 2-deoxy-D-ribose (11) L-ribo-hexose or L-c1adinose

CHO CHO CHO CHO

I I I I
CH, CH, CH, CH,
I I I I
H-C-OH H-C-OCH, H-C-OCOCH, CH,OC -H
I I I I
H-C-OH H-C-OH H-C-OH H-C-OH
I I I I
H-C-OH H-C-OH H-C-OH H-C-OH
I I I I
CH, CH, CH, CH,

D-digitoxose (16) D-cymarose (12) D-3-acetyl digitoxose D-sarmentose (13)

CHO CHO
I
CH,
,
CHO
,
CHO CHO
I I
CH, CH, CH, CH,
I
H-C-OCH,
I I I
I
H-~-OCH3
,
HO-C-H CH,-C-OCH,
I
HO·C-H
I
HOC-H
I
H-C-OH
, ,
H-C-OH H-C-OH
I
HO-C-H
HOC-H
I
H-C-OH
, ,
H-C-OH
H-C-OH
I
HO·C-H
I

CH, CH, CH, I

CH, CH,
L-oleandrose (14) D-diginose (15) D-boivinose (17) D-cladinose L-mycarose

Fig. 15.2. 2-Deoxyhexoses.

deoxy-3-0-methyl-D-ribo-hexose) (12), D-diginose (2,6-di- by the action of S-adenosyl-L-methionine-dependent methyl-

deoxy-3-0-methyl-D-lyxo-hexose) (15), D-digitalose (6- transferases (Luckner, 1990). Among the methoxy sugars
deoxy-3-0-methyl-D-galactose) (23), D-digitoxose (2,6-di- are o-cymarose (2,6-dideoxy-3-0-methyl-o-ribo-hexose)
deoxY-D-ribo-hexose) (16), D-fucose (6-deoxY-L-galactose) (12), D-sarmentose (2,6-dideoxy-3-0-methyl-D-xylo-hex-
(25), L-fucose (6-deoxY-L-galactose) (24), L-oleandrose (2,6- ose) (13), L-oleandrose (2,6-dideoxy- 3-0-methyl-L-arabino-
dideoxy-3-0-methyl-L-arabino-hexose) (26), D-quinovose hexose) (14), D-diginose (2,6-dideoxy-3-0-methyl-o-lyxo-
(6-deoxY-D-glucose) (27), L-rhamnose (6-deoxY-L-mannose) hexose) (15), L-acofriose (6-deoxy-3-0-methyl-L-mannose)
(28), D-sarmentose (2,6-dideoxy-3-0-methyl-o-xylo-hex- (18), L-acovenose (6-deoxy-3-0-methyl-L-talose) (19), D-
ose) (13), L-talomethylose (6-deoxY-L-talose) (29), D-theve- digitalose (6-deoxy-3-0-methyl-D-galactose) (23), D-theve-
tose (6-deoxy-4-0-methyl-D-glucose), L-thevetose (6- tose (6-deoxy-4-0-methyl-D-glucose), and L-thevetose (6-
deoxy-4-0-methyl-L-glucose) (30) (Fig. 15.3) (Courtois and deoxy-4-0-methyl-L-glucose) (30) (Courtois and Percheron,
Percheron, 1970). D-Fucose (25) is a component of glycolip- 1970).
ids found in seeds of the Convolvulaceae.
Branched·Chain Sugars
lIfethoxy Sugars
The branched-chain sugar, apiose (31), was first isolated
Methoxy sugars arise by methoxylation of other sugars from PetroseUnum hortense (parsley, Apiaceae) about 1900.
(Fig. 15.2 and 15.3). For example, o-cymarose (12) arises A similar sugar, hamamelose (32), was later isolated from
252 Carbohydrates

the bark of Hamamelis virginiana (witch hazel, Hamameli- D-Hamamelose (32) arises by rearrangement of fructose-
daceae). Although considered to be rare secondary metabo- I ,6-bis(phosphate).
lites for many years, these sugars are now known to occur in D-Hamamelose (32) and the corresponding polyol, D-ha-
several species of plants (Beck and Hopf, 1990; Grisebach, mamelitol, are very widely distributed among plants. The
1980). digalloyl ester of hamamelose has been isolated from witch
D-Apiose (31) is synthesized from D-glucuronic acid me- hazel, Castanea sativa (chestnut, Fagaceae), and red oak
tabolism in a reaction that involves loss of the carboxyl (Quercus rubra, Fagaceae). Free hamamelose also occurs
group and C-3 of D-glucuronic acid. The enzyme which re- in these plants. The overall pattern of distribution has been
quires NAD+ has been isolated (Grisebach, 1980). reviewed (Grisebach, 1980).
D-Apiose (31) is widely distributed in the Apiaceae (Um- Techniques for the analysis of branched-chain carbohy-
belliferae). This sugar generally occurs in the D-erythro-fu- drates have been reviewed (Beck and Hopf, 1990; Sturgeon,
ranose form as a component of glycosides such as apiin (33) 1990).
(Fig. 15.4). Apiose also occurs as a component of polysac-
charides, especially in aquatic monocotyledonous plants Sugar Carboxylic Acids
(Grisebach, 1980). Relatively simple, specific methods for
screening for this sugar as a component of glycosides are Aldose-derived carboxylic acids are synthesized in bacte-
available (Grisebach, 1980). ria, fungi, plants and animals (Fig. 15.5) (Luckner, 1990).

CHo. CHo. CHo. CHo.

I r r I
H-C-o.H HQ.C-H H-C-o.H CH,
I I I r
H-C-o.H H-C-o.H CH,o.C-H H-C-o.CH,
I r r r
HQ.C-H H-C-o.H H-C-o.H HQ.C-o.H
I I r r
Ho.C-H HQ.C-H H-C-o.H Ho.'C-H
I r I r
CH, CH, CH, CH,
D-thevetose (30) L-oleandrose (26)
L-rhamnose (28) L-fucose (24) 6-deoxy-3-0-D-glucose 2,6-dideoxy-3-0-methyl-
arabino-hexose
CHo. CHo. CHo. CHo.
I r r r
H-C-o.Co.CH, H-C-o.H H-C-o.H H-C-o.H
r r r I
CH,o.C-H Ho-C -H H-C-o.CH, H-C-o.CH,
I r r r
Ho.·C-H H-C-o.H Ho.·C-H H-C-o.H
I I r r
Ho.·C-H H-C-o.H HQ.C-H HQ.C-H
I I r r
CH, CH, CH, CH,
2-0-acetyl-L-vallarose D-quinovose (27) L-acofriose (18) L-acovenose (19)
6-deoxy-D-glucose 6-deoxy-3-0-methyl-L- 6-deoxy-3-0-methyl-L-
mannose talose
CHo. CHo. CHo.
CHo. CHo. I
I I r
I H-C-o.H
H-C-o.H H-C-o.H H-C-o.H H-C-o.H
I I I r I
H-C-o.H HQ.C-H H-C-o.H CH,Q.C-H H-C-o.H
I I I r I
H-C-o.H HQ.C-H Ho.C-H Ho.C-H H-C-o.H
I I I r I
H-C-o.H HQ.C-H H-C-o.H H-C-o.H Ho-C-H
I I r r I
CH, CH, CH, CH, CH,

D-aJlomethyJose (20) L-altromethyJose (21) D-antiarose (22) D-digitalose (23) L-talomethylose (29)
6-deoxy-D-allose 6-deoxy-L-altrose 6-deoxy-D-gulose 6-deoxy-3-0-methyl- 6-droxy-L-lalose
D-galactose

~~o.\. .o.H
Ho. ~o." H Ho.~o.H
~~ o.H H
~o.H
o.H H

a-L-rhamnopyranose (28) a·D-fucose (25) (t,-L·fucose (24)

Fig. 15.3. 6-Deoxyhexoses.

 Carbohydrates 253

CHO
CHO I
H~OH
""-- H - OH
I
C-
HOH'C~OOH
.L..-
HOH,CC-OH
I
CH,OHH ..." I H HOH,C "7 H-C-OH
H OH HOH,c-C-OH H H H-~-OH
OH OH
I
CH,OH OH OH
I
CH,OH

D-.piose (31) D-hamamelose (32)

H~O
H 0 H

CHO
H H,OH
~~~-
CH, H ~ ~O-UDP
OH OH OH OH

streptose UDP-D-apiose
OH

HO~OH
HO
0 0

H 0 0

~~;,;;'l H
H~~H
OH OH
apiin (33)

Fig. 15.4. Branched-chain sugars.

Several types of acids may be distinguished; some form lac- mans. The sugars responsible usually are glucose (I), fruc-
tones readily. D-Glucuronic acid, a common component of tose (8), or sucrose (37) (Fig. IS.9). However, many sugars
glycosides, can be derived from D-glucose 6-phosphate di- have diverse taste properties to humans and, presumably,
rectly or via myo-inositol (Gander, 1976). UDP-D-Xylose other mammals. Some have sweet tastes, whereas others are
and UDP-L-arabinose are formed by decarboxylation of bitter. Although, specific functions for most sugars have not
UDP-D-glucuronic acid and UDP-D-galacturonic acid, re- been elucidated, several have pronounced physiological ac-
spectively (Luckner, 1990). Ribulose S-phosphate (34) is tivity.
formed by the action of a NADP-requiring dehydrogenase A number of nitrogenous molecules with piperine rings
on 6-phosphogluconic acid (35). act as sugar mimics (Fig. IS.8) (Fellows, 1987; Fellows et
al., 1989). These compounds have been isolated from mi-
Amino Sugars crobes of the genera Streptomyces and Bacillus, and from
A number of unusual sugars are found in antibiotics the fungal genera Nectria, Rhizoctonia, and Metarrhizium.
(Hanessian and Haskell, 1970; Sueader, 1985). Many of the Similar nitrogenous compounds also are known from M orus
sugars contain amine, methoxyl, and methyl groups (Fig. (Moraceae), Omphalea (Euphorbiaceae), Derris, Loncho-
IS.6). Some antibiotics, mostly of bacterial or fungal origin, carpus, Swainsona, Astragalus, Oxytropis, Castanosper-
such as steptomycin (36), consist primarily of sugar units mum, Alexa, Angylocalyx, Xanthocercis (Fabaceae), and a
(Fig.IS.7). fern Arachnoides standishii (Fellows et al., 1989). The
Amino sugars are formed from the corresponding ketoses. larvae and adults of day-flying and brightly colored urania
Ammonia or glutamine usually act as the amino donors moths that feed on Derris species in the larval stages con-
(Luckner, 1990). tain 2R,SR-dihydroxymethyl-3R,4R-dihydroxypyrrolidine
N-Acetyl-D-glucosamine is the precursor of chitin in (DMDP) (Fellows et aI., 1989).
fungi and animals. This sugar is also found in the cell walls The glucose mimic, desoxynojirimycin (38) (from Morus,
of bacteria (Luckner, 1990). mulberry, Moraceae), inhibits a-glucosidases in the mamma-
lian gut. A mannose analog, desoxymannojirimycin (39), in-
Biological Activity of Sugars hibits mannosidases. Another alkaloid from Derris (Faba-
Many sugars and sugar-like molecules are accumulated ceae) inhibits the action of invertase but does not prevent
in plants. Most plant materials that contain S% or more mo- hydrolysis of this disaccharide in the mammalian gut. In artifi-
nosaccharides or disaccharides taste noticeably sweet to hu- cial diets with Locusta migratoria, 0.001 % of DMDP in the
254 Carbohydrates

CHo. Co.,H Co.2H

CHo. I
I I I
H-C-o.H H-C-o.H H-C-o.H H-C-o.H
I I I I
HOC-H Ho.-C-H HOC-H HOC-H
I I I I
H-C-o.H Ho.-C-H H-C-o.H H-C-o.H
I I I I
H-C-o.H H-C-o.H H-C-o.H H-C-o.H
I I I I
Co.2H Co.,H CH,.o.H Co.,H
D-galacturonic acid (4) D-glucaric acid
D-glucuronic acid (3) D-gluconic acid
CHo. CHo.
Co.2H
I CH,o.H I I

-
H-C-o.H I H-C-o.H HOC-H
I C=o. I I
Ho.·C-H I H-C-o.H Ho.-C-H
I H-C-o.H I I
H-C-o.H I Ho.-C-H H-C-o.H
I H-C-o.H 0.- I I
H-C-o.H 0.- H-C-o.H
I I I I H-C-o.H
CH,-o.-P-o.- CH,-o.-P-o.- I I
II Co.,H Co.,H
"
0. 0.
6-phosphogluconic acid (35) D-guluronic acid (52)
ribulose-5-phosphate (34) D-mannuronic acid (51)
OP NAD+-enzyme H+ ~

Ho.~<-
Ho.~ ~~ H_Ho.
~o.ft 0. _
Ho.~o. Ho.
o.P
o.H_
H 0. Ho. ~H o.H
0., o.H]( H
H c...
0 NADH-enzyme
D-glucose 6-phosphate H+

Ho.~P Co.,H 0.
Ho. o.HHo.~H
Ho. o.H __
Ho.~o.,~
Ho.~ H
o.Ho.H -- 0.
H o.Ho.H o.H

myo-inositol D-glucuronic acid

Fig. 15.5. Carboxylic acids derived from sugars (in part modified from Goodwin and Mercer, 1983; used with pennission of the copyright owner,
Pergamon Press, Oxford).

diet deters locust nymphs from feeding, although force-fed whereas others appear to be esoteric secondary metabolites
nymphs showed no signs of poisoning at this level (Fellows, (Pazur, 1970). a,a-Trebalose (a-n-glucopyranosyl a-n-glu-
1987; Fellows et al., 1986). Effects on other insects varied. copyranoside) (42) is common in fungi and insects, but ap-
A more complex alkaloid, swainsonine (40) (from Swain- parently rare in plants (Fig. 15.9). However, trehalose is
sona species, Fabaceae), also inhibits a-mannosidase activ- found in algae, fungi, and ferns (Kandler and Hopf, 1980).
ity. A similar complex alkaloid, castanospennine (41) (from Lactose (4-0-J3-n-galactopyranosyl-n-glucopyranose) is
Castanospermum austraie, Fabaceae) inhibits J3-glucosi- common in mammary secretions of animals. Maltose (4-
dases [but not a-glucosidases as in (38)] (Fellows, 1987; O-a-n-glucopyranosyl-n-glucopyranose) (43) isa structural
Fellows et aI., 1992). component of two important n-glucans, starch and glycogen
Castanospennine, deoxynojirimycin (DN), and DMDP (Greenwood, 1970; Pazur, 1970). In a similar manner, cello-
reduce the infectivity of the HIVl and the Moloney murine biose (44) is a part of the repeating units of cellulose and
leukemia viruses (Fellows et al., 1989). These compounds certain other wall polysaccharides. Cellobiose (4-0-J3-n-glu-
also have antitumor activity. copyranosyl-n-glucopyranose) also occurs as the carbohy-
drate constituent of many plant glycosides (cellobiosides)
DlSACCHARIDES AI'ID OLiGOSACCHARIDES (Pazur, 1970). Gentiobiose (6-0-J3-n-glucopyranosyl-n-glu-
copyranose) (45) and primeverose (6-0-J3-n-xylosyl-n-glu-
Disaccharides and oligosaccharides are ubquitous in plants cose) (47) are components of many glycosides; the best
and fungi. Several of these, [e.g., sucrose (37)] are among known of these are amygdalin, vicianin, and crocin. Lami-
the most important energy transfer compounds in plants, narabiose (3-0-J3-n-glucopyranosyl-n-glucopyranose) (46)
 CHO CHO CHO
CHO
I I I I
H-C-NH, H-~-NHCOCH, H,N-C-H H-C-OH
I I CH" I
HGC-H H()'C-H HO-C-H ;N-C-H
I I I CH, I
H-C-OH H-C-OH H-C-OH H-C-OH
I I I I
H-C-OH H-C-OH H-C-OH H-C-OH
I I I I
CH,-OH CHrOH CHrOH CH,

n-glucosamine (9) N-acetyl-D-glucosamine (53) D-mannosamine D-mycaminose

-r:Jf""-......O~OH
H,N~H
H

kosugamine desosamine tolyposamine

2,4.diamino-2,3,4,6- 3-dimethylamino-3,4,6-trldeoxy- 4-amino-2,3,4,6-tetradeoxy·
tetradeoxy-D-arabino-hexose 8-D-xylo-bexose L-erythro-hexose

2-deoxy-2-methylamino- neosamine B 6-amino-6-deoxy· angolosamine

L-glucose 2,6-diamino-2,6-dideoxy- D-glucose 3-dimetbylamino-2,3,6-
L-idose trideoxy-D-xylo-hexose

neosamine C perosamine rhodosamine

kanosamine 2-dimethylamino-2,3,6-
3-amino-3-deoxy· 2,6-diamino-2,6- 4-amino-4,6-dideoxy-
D-glucose dideoxy-D-glucose D-mannose trideoxy-L-lyxo-hexose
(a)

HO~O, OH
HO~
H

digitoxose

digitoxose-digitoxose-digitoxose- 0

~ HO~OH
OH
o

~~
o HO 0 OH
H,N 0
00 H HO
HO H
H,N OH H NH,
OH NH, OH H
OH

kanosamine daunosamine perosamine D-gulosamine

(b)

Fig. 15.6 (a & b). Hexoses from antibiotics.

255
256 Carhohydrates

also occurs as a component of glycosides. These disaccha- 1976; Akazawa and Okamoto, 1980; Hawker, 1985; Lucas
rides are rare as free sugars (Pazur, 1970). The structures, and Madore, 1988). Invertase cleaves sucrose to yield D-
properties, and distribution of a number of other disaccha- glucose and D-fructose. Both enzymes are common in plant
rides have been reviewed (Dey and Dixon, 1985; Pazur, tissues (Feingold and Avigad, 1980).
1970). Techniques for analysis of disaccharides have been
reviewed (Avigad, 1990; El Khadem, 1988). Oligosaccharides
Oligosaccharides, compounds composed of several sugar
Sucrose
units (usually 2-10), are also widespread in plants (Dey,
A major part of the CO2 assimilated in plants during the 1990; Pazur, 1970). At least SOO oligosaccharides are known
operation of the photosynthetic carbon reduction cycle ap- (Kandler and Hopf, 1980); most are comprised of a small
pears in sucrose (j3-D-fructofuranosyl a-D-glucopyranoside) number of monosaccharide units (Pazur, 1970). Other oligo-
(37) (Fig. IS.9). In many plants, sucrose is the principal saccharides are formed as breakdown products of polysac-
or only common sugar to be translocated (Akazawa, 1976; charides. Raffinose [j3-D-fructofuranosyl O-a-D-galactopy-
Akazawa and Okamoto, 1980; Hawker, 1985; Lucas and ranosyl-(I - 6)-a-D-glucopyranoside] (49) and stachyose
Madore, 1988). Thus, sucrose is found in all plants and has [O-a-D-galactopyranosyl-(l - 6)-O-a-D-galactopyranosyl-
been reported from seeds, leaves, fruits, flowers, and roots (I - 6)-O-a-D-glucopyranosyl-(1 - 2)-j3-D-fructofurano-
(Pazur, 1970). Honey consists largely of sucrose and its hy- side] (48) are particularly common (Fig. IS.IO) (Goodwin
drolysis products glucose and fructose. Most commercial and Mercer, 1983; Kandler and Hopf, 1980); raffinose is
sucrose is isolated from sugar cane and sugar beets. Sucrose distributed almost as widely as sucrose, but is usually present
synthetase catalyzes the formation of sucrose from UDP- at only low concentrations (Dey, 1985; Pazur, 1970). How-
glucose and D-fructose. This reaction is also reversible and ever, raffinose is accumulated in cereal grains, sugar beets,
the reversal of the sucrose synthetase reaction appears to cottonseed, soybeans, and many other legumes. Stachyose
be the principal mechanism of sucrose cleavage (Akazawa, (48), usually associated with raffinose and sucrose, is found

NH,
2'6_diaminO_D_gluc~oe
0
HO
HO H HO
OH
NH
HNyNH
deoxystreptamine

"8.
H,N~NH,.o
HO~ H,N 0 °
HOH'Q<0
° streptose
H H H
D-ribose H
OH 0
OH 0
HO~
HO~~H HO0

HO
NHCH,

NH2 2,6-diamino-D-glucose 2-methylamino-L-gJucose

neomycin C streptomycin (36)

HO
~
NH'
00
0

OH
H
H~NH' OH
0
0
00
6·amino·D-glucose o~ 3-amino·D-glucose
HO~ NH,
H,N
deoxystreptamine

kanamycin A

Fig. 15.7. Carbohydrate antibiotics.

 Carbohydrates 257

HO ~°
HO
OHHO

OH
H HO ~
HO
OHHO
NH
HO 4
HO
0H NH

OH
OH
(X~D-mannose (7) desoxymannonojirimycln nojirimycin
DMJ (a maun..e mimic) (39)

r
~
OH
i----Y
NH HOCHQ0"---- HOCHl NH H

HO
H oH\ k:--~
HO HO CHlOH HO CHlOH
OH
OH H OH H
desoxynojirlmycin
poD-fructose (8) DMDP (4) (a fructose mimic)
DNJ (a glucose mimic) (38)

HO
HOCHVuNH~
H

W
HO'N:

" N c : r OH OH H

castanospermine (41) DABI (from Angylocalyx) swalnsonine (40)

Fig. 15.8. Nitrogenous compounds that mimic sugars.

in many genera of the Fabaceae and is a dominant constituent Iyfructosans. Inulin and other related fructosans with 13-1,2-
of many plants in the Lamiaceae (Labiatae), the mint family linkages are found in the roots and tubers of many members
(Gander, 1976; Pazur, 1970). Melizitose [O-a-D-glucopyra- of the Asteraceae and Campanulaceae (Akazawa, 1976;
nosyl-(1-3)-13-D-fructofuranosyl a-D-glucopyranoside] Pontis and del Campillo, 1985). Inulin is rich in the tubers of
(50) is present in the sweet exudations of many plants such Helianthus tuberosus, the Jerusalem artichoke (Asteraceae).
as the "honeydew" of limes and poplars and the manna of This polyfructosan makes up more than 50% of the dry
Douglas fIr, Virginia pine, and larch (Pazur, 1970). Honey weight of the tubers (Akazawa, 1976).
sometimes contains this trisaccharide, which cannot serve Methods for the analysis of fructans have been reviewed
as food for bees, probably because they lack the enzymes (pontis, 1990).
to degrade it (Pazur, 1970). The properties and distribution
of a number of other oligosaccharides have been reviewed
(Kandler and Hopf, 1980; Pazur, 1970). POLYSACCHARIDES: RESERVE AI'ID
STRUCTURAL FORMS
Fructans
Energy Storage Compounds Derived
Fructans (or fructosans) are related both structurally and
from Sugars
metabolically to sucrose (Pollock and Chatterton, 1988).
Under some conditions, fructans can be the major site of Sugars often occur as part of large polymeric units.
storage of fIxed carbon. Fructans are most important in the Among these are cellulose and starches (Eastwood and Ed-
Agavaceae, Asteraceae, Boraginaceae, Campanulaceae, wards, 1991; Greenwood, 1970; Manners, 1985; Preiss and
Goodeniaceae, Haemadoraceae, Irldaceae. Liliaceae, Levi, 1980; Whistler et aI., 1984). Starch based on glucose
Poaceae, and Stylidiaceae (Pollock and Chatterton, 1988; occurs in green algae and most other plants. Starch is an
Pontis and del Campillo, 1985). In some members of the "end product" of the phytosynthetic reduction of CO2 , This
Asteraceae, Liliaceae, and Poaceae, more than 50% dry assimilation starch is a transitory reserve carbohydrate that
weight of the tissues may be comprised of fructans (Pontis, disappears in the dark and is transported as sucrose through
1990). In the Asteraceae, these compounds are usually accu- the phloem to other tissues (Akazawa, 1976).
mulated in underground storage structures, whereas in the Starch consists of two chemically distinct polysaccharide
Poaceae, they are found throughout the plant. fractions, amylose (soluble in hot water) and amylopectin
In some plants, photosynthetic products are stored as po- (insoluble in hot water). Amylose is a linear polymer of a-
 Carbohydrates 259

H~O,
)~
H H~H ____ O, H

~
OHO
[O~

H~-----O'
HO
HO H

~
H
o 0 OH
O~
~
HO OH 0
o HO H o
OH HO

HO~H HOCHVO~
HO 0 H

HOCHVu°~
Ii W CH,OH
W
HOCH'K::0';;/,.

H W CH,OH
OH H
Ii
OH H
CH,OH

stachyose (48) OH H raffinose (49) melizitose (50)

Fig. 15.10. SbUctures of some common oligosaccharides.

I ,4-linked glucose with a helical conformation. The molecu- in the cell wall and is packed in an ordered manner to form
lar weight ranges from 10,000 to 100,000. In amylopectins, compact aggregates called microfibrils (Baeic et ai., 1988).
many chain segments with 20-25 glucose units are linked The glucosyl donor in a number of cases appears to be GDP-
to the main chain by a-I,6 bonds. These molecules are only glucose (guanosine diphosphate-o-glucose), but there is
partially attacked by 13-amylases and phosphorylases. The some controversy about the role of this compound as the
hydrolysis products are called dextrins. So-called de- precursor (Karr, 1976). Cellulose is the most abundant or-
branching enzymes break down the a-I,6-linkages. ganic compound.
Starch synthetase catalyzes the stepwise transfer of the Methods for the analysis of cellulose have been reviewed
glucose moiety from the nucleotide glucose molecule to the (Franz and Blaschek, 1990).
nonreducing end of the accepting a-glucan molecule Cellulose microfibrils are usually embedded in a continu-
(primer) by forming the linear a-I ,4-glucan amylose (Aka- ous phase of lignin, pectin, and hemicellulose; hemicellulose
zawa, 1976; Preiss, 1988). The formation of a-I,6 bonds in usually predominates (Whistler and Richards, 1970). Most
the glycogen molecule is known to be catalyzed by "branch- of the hemicellulose is found mixed with cellulose in both
ing enzyme" or amylo-(I,4->I,6)-translucosylase (Aka- the primary and secondary cell walls. Hemicellulose is now
zawa, 1976; Manners, 1985; Preiss, 1988). known not be a precursor to cellulose. In general, hemicellu-
Alternatively, phosphorylases can catalyze the reversible loses are the cell wall polysaccharides other than cellulose
phosphorylation of a-glucan molecules and lead to the for- and pectin. These polymers are classified on the basis of the
mation of starch. Starch is broken down in plants by a- type of sugar residues present. For example, o-xylan is made
amylases. Methods for the analysis of starch have been re- up of o-xylose, o-mannan of o-mannose, and o-galactan of
viewed (Morrison and Karkalas, 1990).
o-galactose units (Whistler and Richards, 1970). In practice,
few hemicelluloses contain ouly one sugar, most contain
structural Carbohydrates two to four. Furthermore most hemicelluloses have branched
structures. The major types of hemicellulose and their distri-
Young plant cell walls are primarily made up of polysac-
bution have been reviewed (Whistler and Richards, 1970).
charides and protein. Cells become more highly lignified as
Methods for the analysis of mannans have been reviewed
they become older. The degree of modification is linked to
(Matheson, 1990).
the function of the particular cell involved. Although the
composition of polysaccharides may vary in different plant
species, composition in a particular plant species is usually Callose
constant from plant to plant. The amount and type of g1yco-
proteins present in the cell wall are usually characteristic for Callose is a morphologically distinctive polysaccharide
different plants. The cell wall composition changes during that occurs in granular form deposited around sieve plates
growth and development (Karr, 1976). and on the side of sieve-tube pores. Callose is a 13-1 ,3-linked
Cellulose, a 13-1 ,4-glucan, is synthesized by many organ- o-glucan with no detectable branching (Aspinall, 1980). In
isms from bacteria to higher plants (Aspinall, 1980; Colvin, higher plants, callose is deposited as a response to wounding
1980; Ward and Seib, 1970). This polymeric material occurs or infection by microorganisms (Baeic et ai., 1988).
260 Carbohydrates

Pectins strong acids and exist as salts (Percival, 1979). Those of

Pectin, a polymer rich in ,,-I ,4-linked o-galacturonic acid Cladophora and Codium are often arabinogalactans [with 0-
residues, is important in plants as the material that glues galactose (2) and L-arabinose (6)]. Many species synthesize
plant cell walls together (Aspinall, 1970, 1980). Although comparatively simple xylans and mannans (percival, 1970,
o-galacturonic acid (4) is the main constituent of pectic sub- 1979).
Many red algae (Rhodophyceae) contain complex carbo-
stances, various proportions of other sugars are found, in-
hydrate molecules. These algae produce o-glucans, known
cluding o-galactose (2), L-arabinose (6), o-xylose (5), and
as f10ridean starch, as their food reserve material (Percival,
L-rhamnose (28). In most instances, methylated sugars also
1970, 1979). This starch differs in that no amylose is present,
are present. An enzyme that converts UDP-o-galacturonic
but it resembles amylopectins in many of its properties.
acid to pectin has been isolated from mung beans (Karr,
There is little evidence for starch in other red algae [e.g.,
1976). Extensive methyl esterification takes place after the
Rhodymenia palmata (dulse)]. In this alga, the major poly-
polygalacturonic acid molecule is assembled.
saccharide is a water-soluble xylan made up of random 1,3-
Many of the chemical properties and economic uses of
and 1,4-linked units. Other 1,3-, 1,4-, and mixed 1,3- and
pectins have been reviewed (Aspinall, 1970, 1980; Fishman
1,4-xylans have been isolated from a number of red algae
and Jen, 1986). Most commercial pectins come from apple
(Percival, 1970, 1979).
pomace, citrus waste, and sugar beet pulp.
The major polysaccharides of most Rhodophyceae are
Techniques for the analysis of pectins have been reviewed
galactans consisting of 1,3-linked galactose units. Agar and
(O'Neill et al., 1990).
carrageenan are among these. Agars contain 0- and L-galac-
Plant Qums tose units, whereas carrageenans contain only o-galactose
(2) (percival, 1979). In each, some of the units are sulfated
A number of gum exudates from plants have a composi-
and others OCCut as 3,6-anhydrogalactose. A carbohydrate
tion similar to pectins. These gums may form spontaneously from Porphyra is known to have glucomannans with o-man-
at sites of injury to the plant, or may be induced by deliberate nose (7) in a (3-1,4-linkage.
incisions; the actual site of formation of gums is not well In the Phaeophyceae (brown algae), the food reserves are
established (Joseleau and Ullmann, 1990). The most impor- laminarin, a water-soluble (3-o-(l-3)-linked o-g1ucan and a
tant gum is gum arabic (or gum acacia) from Acacia senegal sugar alcohol, mannitol. Laminarin contains 20-25 glucose
and A. catechu (Fabacehe). The polysaccharides all contain units (Percival, 1970, 1979).
highly branched structures, usually based on o-g1ucutonic The major polysaccharide of brown algae is alginic acid.
(3) and/or o-galacturonic acids (4) (Aspinall, 1970). Many This polymer consists of unbranched chains of contiguous
gums contain L-arabinofuranose (6) residues attached to the (3-I,4-linked o-mannutonic acid (51) and blocks of contig-
periphery of the molecules. Gum arabic contains L-arabinose uous ,,-1,4-linked L-guluronic acid (52) (Fig. 15.5) (Percival,
(6), o-galactose (2), L-rhamnose (28), and o-glucutonic acid 1970, 1979). Alginate is present as the salt of different met-
(3) in the ratio of3:3: I: I (Aspinall, 1970). Carob and guar als, usually sodium and calcium. Calcium ions are important
gum (from the seeds of Ceratonia siliqua and Cyamopsis in determination of gel strength.
tetragonolobus, respectively) (Fabaceae) are characterized A sulfated polysaccharide from brown algae is called
by the presence of o-galacto-o-mannans (Reid, 1985). "fucan." This is a complex highly branched mixture, con-
Techniques for the analysis of plant gums have been re- taining fucose and other sugars and sugar acids (Percival,
viewed (Stephen et al., 1990). 1970, 1979).
Algal Polysaccharides Teclmiques for the analysis of algal polysaccharides have
been reviewed (Percival and McDowell, 1990).
Algae belong to three major groups: the Chlorophyceae
(green algae), the Phaeophyceae (brown algae), and the Rho- Chitin
dophyceae (ted algae). Polysaccharides are common in rep- Chitin is a ubiquitous component of fungal cell walls,
resentatives of each group (Percival, 1970; Percival and Mc- being absent only from the Oomycetes and Trichomycetes
Dowell, 1981, 1985). These polysaccharides often are (Cabib, 1981). Chitin also makes up much of the wall mate-
different from those of land plants, but some algae do synthe- rial of insects and crustaceans. This polysaccharide is com-
size cellulose and starch. Evidence has been advanced that posed of (l-4)-(3-linked N-acetylglucosamine (53) (Fig.
these polysaccharides are involved in mechanisms by which 15.6) units (Cabib, 1981).
the algae selectively absorb potassium and calcium from Teclmiques for the analysis of chitin have been reviewed
seawater (Percival, 1970). For convenience, these com- (Lezika and Quesada-Allue, 1990).
pounds are divided into three groups: structural carbohy-
drates, food reserves, and sulfated polysaccharides. BiolOgical Activity of Oligosaccharides and
Many green algae (Chlorophyceae) contain starch which Polysaccharides
can be fractionated into amylose and amylopectins. The These compounds probably are most important in the
major polysaccharides, however, are polysaccharides in diets of animals as sources of energy, but many other roles
wbich at least some of the hydroxyl groups of the sugars also are known. As is true for simple saccharides, many
are substituted by half ester groups. These compounds are oligosaccharides possess sweet tastes (Pazut, 1970). Poly-
 Carbohydrates 261

saccharides exhibit antitumor, immunological, anti-inflam- tions between pathogenic fungi and their host plants indicate
matory, anticoagulant, hypoglycemic, and antiviral activities that host cell wall fragments, often consisting of oligomeric
(Srivastava and Kulshreshtha, 1989). saccharides released by fungal enzymes, can elicit cyto-
Humans, when awake, release 20-50 ml of flatus per hour plasmic changes. One well-studied mechanism of defense
under nonnal conditions; the dry bean (Phaseolus vulgaris) is involves the accumulation of phytoalexins at the site of in-
the champion gas producer among common foods. Stachyose fection (Kogel and Beillman, 1992; Kosuge, 1981; Ralton et
(48), raffinose (49)(Fig. 15.10) (Dey, 1985), and sucrose (37) aI., 1987). The presence of compounds produced by several
are the major molecules that are fennented by Clostridium mechanisms, but usually by degradation of wall materials of
perJringens to a mixture of carbon dioxide, hydrogen, and the host or of the pathogen, elicit fonnation of phytoalexins
other compounds (Murphy, 1968; Rockland, 1968). (Ebel, 1986; Eilert, 1987; Kogel and Beillmann, 1992).
Oligomers of N-acylated, N-acetylglucosamine (53) from A glucan from yeast extract containing 3-, 6-, and 3,6-
Rhizobium meliloti (nod factors) are involved in depolariza- linked glucosyl residues was shown to elicit fonnation of
tion of root hair cells in alfalfa (Medicago sativa) at nanomo- glyceollin in soybeans (Glycine max, Fabaceae). The yeast-
lar concentrations (Ehrhardt et aI., 1992). Other chemical derived elicitor stimulates accumulation of this isoflavone
messengers are known to be involved in establishment of when as little as 15 ng of the elicitor is applied to soybean
nodulation in legnmes (see Chapter 11). cotyledons (Hahn and Albersheirn, 1978).
The structure of cell wall carbohydrates is important in A heat-labile endopolygalacturonase from culture fil-
recognition of surfaces in plant-microbe interactions involv- trates of Rhizopus stolonifer elicited accumulation of the
ing pathogens, mycorrhizal fungi, nitrogen-fixing bacteria, phytoalexin casbene in castor bean (Ricinus communis).
and in other interactions. Cell recognition is the initial event This material releases heat-stable water-soluble cell wall
of cell-cell communication which elicits a defined biochem- fragments from the host plant. These wall fragments contain
ical, physiological, or morphological response (Ralton et aI., a high fraction of galacturonic acid or its methyl ester, indi-
1987). Many of these interactions involve proteins, often cating their derivation from pectic polysaccharides (Ralton
those of the cell membrane, whereas others involve carbohy- et aI., 1987).
drates. Lectins bind to specific sugar moieties on plant sur- Solubilized polysaccharide fractions from Glycine max
faces (see Chapter 14) (Kauss, 1981; Schmidt and Bohlool, (soy bean) cell walls elicit fonnation of phytoalexins in that
1981; Sharon and Lis, 1990). Cell Walls often serve as plant (Hahn et aI., 1981). These fragments are rich in galact-
sources of secondary signal molecules. In instances where uronic acid (4) and appear to come from the pectic materials
the primary signal from one cell is an enzyme such as a of the plant. A ~-glucosyl hydrolase from soy bean cell walls
hydrolase or lyase, it may catalyze the release of fragments has the ability to degrade the elicitor (Cline and Albersheim,
from the cell wall of the target cell. These wall .fragments 1981). Fractions from the soybean pathogen Phytophthora
may become involved in the recognition process by interact- megasperma var. sojae also serve as elicitors. The activity
ing directly with a plasma membrane receptor (Ralton et aI., is associated with the glucan fraction (Ayers et aI., 1976).
1987). Wall-bound lectins and agglutinins may bind these Only one of a series of eight hepta-~-glucoside alditols iso-
secondary extracellular signals. lated was capable of eliciting phytoalexin production (Fig.
Many pathogens produce pectinases in the process of pen- 15.11) (Ralton et aI., 1987).
etrating tissues of the host plants (Collmer, 1987). Interac- Cell suspension cultures of parsley, PetroseUnum

~
OH 0

HO

Ho~HOO 0 H o0h
O

HO OH 0 O~O
H HO 0

O
HOOH OHk

HO~HOo 0 0

HO OH OH~
0
H HO
HO
OH H,OH

Fig. 15.11. An active hepta-(3-glucoside alditol from an acid hydrolysate of Phthophthora megasperma cell walls.
262 Carbohydrates

crispum, accumulate coumarin phytoalexins when treated zation of the matrix polysaccharides provide cues that help
with either a purified ,,-1,4-D-endopolygalacturonic acid aphids to locate and or ingest materials from the phloem.
lyase from Erwinia carotovora or oligosaccharides solubi- Some of these breakdown products also may act as elicitors
lized from parsley cell walls by endopolygalacturonic acid of plant hypersensitive reactions and phytoalexins (Camp-
lyase (Davis and Hahlbrock, 1987). The plant cell wall elici- bell and Dreyer, 1990; Dreyer and Campbell, 1987).
tor was found to act synergistically with the fungal glucan Other oligosaccharides, such as streptomycin (36) (Fig.
elicitor in the induction of coumarin phytoalexins. 15.7) from Streptomyces griseus isolated from soil samples,
Aphids have flexible, stylet-like mouthparts adapted for have potent antibiotic properties (Sneader, 1985). Strepto-
probing of plant tissues. By this means, the aphid must use mycin was originally of great interest because of its activity
chemical cues from the plant to determine if the plant is a against Mycobacterium tuberculosis (Sneader, 1985).
suitable host (host plant quality), where the aphid is on the
plant, the location of the stylet within the plant tissues, and
the direction to probe to locate the plant phloem (Campbell SUGAR ALCOHOLS
and Dreyer, 1990; Dreyer and Campbell, 1987). Aphids
avoid many of the toxic compounds stored in plant cells by
Polyols
probing between the cell walls. These insects are able to
penetrate the intercellular spaces by producing a variety of Although some representatives of this group are wide-
digestive enzymes which depolymerize the pectins and spread in plants, only about 35 structures have appeared in
hemicelluloses forming the intercellular-cell wall matrix. the literature (Fig. 15.12). These compounds are derived by
Aphid-host plant compatibility is associated to the extent the reduction of aldoses or ketoses with pyridine nucleotide-
that these enzymes depolymerize their respective substrates, dependent dehydrogenases (Luckner, 1990). Polyols corre-
and a reduced rate of depolymerization is often associated sponding to C2 , C" C4 , Cs , C6 , and C7 are all known (Plou-
with host plant resistance. Differences in methoxylation, vier, 1963). Esters of ethylene glycol probably occurin small
acetylation, or neutral sugar composition in the matrix poly- amounts in many plants, and esters of glycerol (i.e., triglycer-
saccharides are often involved in this reduced depolymeriza- ides) are found in all plants (see Chapter 2). Compounds
tion. Further, specific breakdown products from depolymeri- with four and five carbons are common in many fungi. Sorbi-

CH,OH CH,OH CH,OH CH,OH

I I I
I H-C-OH H-C-OH
H-C-OH HO-C-H
I I
I I HO-C-H HOC-H
H-C-OH H-C-OH I
I
I I HO-C-H H-C-OH
CH,OH CH,OH I I
CH,-OH CH,OH
meso-erythritol (56) D·threitol L-arabinitol (57) meso-xylitol (58)

CH,OH CH,OH CH,OH

CHzOH
I I I I
H-C-OH HO-C-H H-C-OH
H-C-OH
I I I I
HO-C-H HOC-H
H-C-OH HO-C-H
I I
I I H-C-OH
H-C-OH HOC-H H-C-OH
I I
I I H-C-OH
H-C-OH H-C-OH H-C-OH
I I
I I CH,.OH
CH,.OH CH,.OH CH,.OH

meso-allitol (59) meso-galactitol (60) D-mannitol (55) D-sorbitol (54)

CH,OH CH,OH CH'-:=l

CH,~
HO-? -H I
I
H-C-OH
I
H-C-OH
I
I
H-?-OH I
HQ-C-H 0 HO-C-H HOC-H HO-C-H 0

H-?-OH I
I
H-C-OH
I
H-C-OH
I

I
H-?-OH I
H-C HO·C-H H-C-OH H-C
I I I I
CH,.OH CHrDH CH,-OH CH,.OH

1,5-anhydro-D.mannito) (61) L·iditol D-glucitol 1,5-anhydro·D-glucitol

Fig. 15.12. Polyols.

 Carbohydrate.~ 263

OH OH HO OH HO

~ OH
OH OH
+-
~ OH
OH +- ~o, OH OH
OH OH OH
L-chiro-inositol (71) myo-inositol (62) D-chiro-inositol (72)

I / \ \
CH,O HO ~OHCHoHO CH.O HO CH'~O
HO

~OH;)H 3 OH3~O --:H OH

~
OH
OH
OH OH
OHOH
OH
OH
D-(-)-bornesitol (65) sequoyitol (68) D-pinitol (63)

~
OH ~ ~
~H~
O~H
HO OCH,

OH
OH

scyllo-inositol (69)
OH

D-(+)-ononitol (67)
OH

L-(+)-bornesitol (66)

~OHJH
~ OH HO~OH
1 / D-I-O-methyl-muco-jnositol(70)
HO~OH
(a) myo-inositol (62) OH OH

H~
HO

HO~OH HO~
Yl
HO OH
OH OH OH
OH HO OH

myo-inositol (62) D-quercitol (73) D-pinitol (63)

'~~O"
HO OH
"O~o~" 03P~1
,)3~:"" OH OH

~PO;'
(b) sequoyitol (68) quebrachitol (64) phytic acid (74)

Fig. 15.13 (a & b). Cyclitols (modified from Loewus and Loewus, 1980; used with pennission of the copyright owner, Academic Press, Orlando, FL).
264 Carbohydrates

tol (54), a C6 polyol, is common in the Rosaceae, especially higher plants (Loewus and Loewus, 1980). Nine of these
in the Spiraeoideae, Maloideae, and the Amygdaloideae, but compounds occur commonly: 0-( +)- (63) and L-( - )-pinitol
seems to be lacking in other subfamilies. As much as 13.6% (lo- and IL-3-0-methyl-chiro-inositol), L-( - )-quebrachitol
of one red alga is composed of sorbitol. Mannitol (55), an- (lL-2-0-methyl-chiro-inositol) (64), 0-( - )- (65) and L-( + )-
other C6 polyol, is found in at least 50 families and is proba- bornesitol (66) (10- and IL-I-O-methyl-myo-inositol), 0-
bly the most widely distributed polyol. Hamamelitol, the (+ )-ononitol (l0-4-0-methyl-myo-inositol) (67), sequoyitol
polyol corresponding to hamamelose, is accumulated in (5-0-methyl-myo-inositol) (68), O-methyl-scyllo-inositol
plants of the genus Primula (section Auricula) (Primulaceae) (69), and 10-I-O-methyl-muco-inositol (70).
(Grisebach, 1980). . Epimerization of myo-inositol (62) at C-I, C-2, or C-3
Many polyols [such as meso-erythritol (56), 0- and L- produces L-chiro- (71), scyllo-, (69), or o-chiro-inositol (72),
arabinitol (57), meso-xylitol (58), meso-allitol (59), meso- respectively (Fig. 15.13). Methylation at C-3, C-I, C-5, or
galactitol (60), o-mannitol (55), 1,5-anbydro-o-mannitol C-6 in the absence of epimerization gives 0-( - )-bornesitol
(61), and o-sorbitol (54)] taste sweet (Beck and Hopf, 1990). (65), L-( + )-bornesitol (66), sequoyitol (68), or 0-( + )-onon-
itol (67), respectively. All of these conversions occur in
CycHtols plants (Loewus and Loewus, 1980). Methyltransferases that
convert inositol to 0- (65) or L-bornesitol (66) have been
A number of cyclic polyols also are known to occur (Fig. characterized. An oxidoreductase that converts sequoyitol
15.13). myo-Inositol (62), widespread in animals, is a growth (68) to o-pinitol (63) also has been studied. o-Pinitol (63)
factor in yeasts and a group B vitamin. This compound, one is converted to o-I-O-methyl-muco-inositol (70) (presum-
of nine possible stereoisomers, is widespread in plants (six ably by epimerization). A number of other epimerization
other isomers also are naturally occurring). When plant cells reactions are known to occur (Loewus and Loewus, 1980).
are incubated with radiolabeled myo-inositol, a large portion o-Quercitol (73) is common in members of the genus
of the label appears in the pentose and uronic acid residues Quercus, oaks, of the family Fagaceae. This cyclitol deriva-
of the cell wall (Karr, 1976; Loewus and Loewus, 1980). tive is known from at least nine other families. o-Pinitol (63)
myo-Inositol serves as a precursor for o-glucuronic acid (3). is known from six families of gymnosperms, in which it is
Furthermore, when glucose and other monosaccharides are widespread, but it is also known from a number of angio-
incorporated into cell walls, myo-inositol appears to be an sperms.
intermediate (Karr, 1976; Loewus and Loewus, 1980). Ana- The hexakis-o-phosphate of myo-inositol, phytic acid
lytical techniques for the study of cyclitols and sugar alco- (74), occurs in most plants. It is recovered from most plants
hols (polyols) have been reviewed (Beck and Hopf, 1990; as phytin, the calcium-magnesium salt. As much as 90% of
Loewus, 1990). the organic phosphate of seeds may be stored in this form
The corresponding methyl ethers often are found in (Loewus and Loewus, 1980). Three inositol dimethyl ethers

H
HO i OH OH OH

?-+----r
~o~ H~oAo H.
0H

OH OH HO H

L-ascorbic acid (75) 3,6-anhydro-L-xylo-hexulono-

1,4·Jactone hydrate (76)
(dehydroascorbic acid)

C02H
o I
C02H CH20H \ C02H

H-t-OH H-~-OH Ho-c'l Pelargonium crispum

oxalic acid

HO-C-H
I -
HO-C-H
I+-O
" I
~02H I 80- C ---.J geranium CO,"
I

L-(+)-tartaric acid (77) C0 H2

H- b H-C-OH
I

I i:. _ H HO-C-H
L-threonic acid HO _ I
carbohydrate metabolism I C02H
---...... CH2-OH
[C 3 -fragment] L-(+)·lartaric acid (77)
L·ascorbic acid (75)

Fig. 15.14. Oxidation of L-ascorbic acid (Loewus, 1980; modified and used with pennission of the copyright owner, Academic Press, Orlando, FL.).
 Carbohydrates 265

OH "o--R
HO~o O)H
OH 0 0
C:N
I I
(, /I /I A
UDPG(35) ~O-!;O-r:OON 0

!
H H
H H
glycosyl transferase OH OH

OH

OH ~N
HO~OR HO-~-OJ-o I NA.O
HO OH + I
OH I
OH ~
H H
H H
OH OH
(a)
a p-glycoside UDP

(b)
UDP-glucose dburrin

Fig. 15.lS (a & b). Synthesis of glycoside, by action of UDP-gluco,e.

are known: dambonitol (1,3-di-O-methyl-myo-inositol), 0- contain little, if any, ascorbic acid, upon germination there
(- )-liriodentritol (l-o-I,4-di-O-methyl-myo-inositol), and is a rapid synthesis from carbohydrate reserves. Walnut hulls
(+ )-pinpollitol (lo-di-O-methyl-chiro-inositol). In view of (Juglans regia), rose hips (Rosa spp.), and the West Indian
the amounts of these compounds encountered in plants, a cherry (Malpighia glabra, Malpighiaceae) all contain more
storage function is suggested (Loewus and Loewus, 1980). than 1% dry weight L-ascorbic acid (Loewus, 1980).
Several C-methyl-branched inositols are known from Labeling studies indicate that in the conversion of o-glu-
green, red, and brown algae (Plouvier, 1963). The biosyn- cose to L-ascorbic acid in plants, the carbon chain is con-
thesis of these compounds does not proceed by C-methyla- served. Although the steps of biosynthesis are not clear, they
tion of inositols, but by cyclization of a I-deoxyheptulose involve oxidation at C-l, an internal oxidation of C-2 or C-
(Grisebach, 1980). 3, and epimerization at C-5 of glucose. One possible se-
Pinitol (63) is a feeding stimulant for the larvae of the quence is as follows: o-glucose is converted to o-glucosone,
butterfly, Eurema hecabe mandarina and also inhibits H eli- which compound to L-sorbosone, which compound to L-
coverpis zea larval growth in soybeans (Ley et aI., 1987). ascorbic acid (Loewus, 1988).
Ascorbic acid is oxidized by plants with ascorbate oxi-
dase tu yield dehydroascorbic acid (3,6-anhydro-L-xylo-hex-
L-ASCORBIC ACID ulono-l,4-lactone hydrate) (76) (Loewus, 1980, 1988). In
plants, ascorbic acid is rarely accumulated, but is usually
L-Ascorbic acid (75), vitamin C, is an important component converted into tartaric (77) and oxalic (78) acids (Fig. 15.14).
of many fruits and is important in human nutrition (Sneader, Ascorbic acid is involved in removal of active oxygen
1985). Methods for the determination of L-ascorbic acid have during photosynthesis, with activation and deactivation of
been reviewed (Loewus, 1980, 1988). L-Ascorbic acid is enzymes, mobilization of ferrous ion, growth regulation, and
produced in actively growing plants. Although dry seeds the oxidation of organic compounds. One well-known exam-
266 Carbohydrale.s

HO 4
HO
0H

°
OH
H
O+H
CH,OH

CH,OCOCH'OH
HO
HO
~OH
°
OH
O-CH,

H+OH

~
[H'OH liIiosideC CH,OH
liIioside A HOHO 0 H

OH CHOH
H '
liIiosideB OH
OH

~
liIiOSideE
HO
HO ~ OH
HO
O-+CH,
H
HO

HO OH O+CH'
HO H
CH,OH CH,OCOCH,
liIiosideD
H
° CH,OCOCH,
HO
OH
oOH

~t"~~"
(104)

HO
]r ~
oH
OHH
H
H ~OH
OH

stachysoside A

HO

Fig. 15.16. Some unusual types of glycosides.

pie involves the activation of the glucosidase (myrosinase) algae. Diglycosides, especially those involving gentiobiose
that hydrolyses the glucosinolate sinigrin to allyl isothiocya- [O-J3-o-glucopyranosyl-(l-6)-J3-glucose] (45), sophorose
nate (Loewus, 1980, 1988). (79), vicianose [O-a-L-arabinopyranosYl-(l-6)-J3-o-glu
cose] (81), rutinose (80), and primeverose (47) (Fig. 15.9)
are widespread (Courtois and Percheron, 1970; Dey and
GLYCOSIDES Il'I PLANTS Dixon, 1985). Glycosides involving oxygen, sulfur, nitro-
gen, and other atoms are known. In plants, most glycosides
Many sugar molecules in plants occur as glycosides. Glyco- are accompanied by enzymes capable of hydrolyzing them
sides have modified solubility properties, toxicity and trans- (Nisizawa and Hashimoto, 1970).
port possibilities. Glucosides and galactosides are the most In general, these compounds are derived via UDP-glucose
common glycosides. Mannosides are known only from (UDPG) or a similar sugar nucleotide (Fig. 15.15). In gen-

~O.gIUCOSYI OH
° °
0-('~ 0-~
~N} \ ~N} ~N~N~
glucose
indican (82) indoxyl (83) indigo (84)

Fig. 15.17. Hydrolysis and oxidation of indican to produce indigo (modified from Courtois and Percheron. 1970; used with pennission of the copyright
owner, Academic Press, Orlando, FL).
 Carbohydrates 267

eral, inversion is observed when the glycoside is formed roots may be the main site of biosynthesis. On hydrolysis,
(Dey and Dixon, 1985). indican yields o-glucose and indoxyl (83), which, on oxida-
Glycosides are treated in the chapters that deal with the tion in air, yields the insoluble dye indigo (84) (Fig. 15.17).
aglycone portion of the molecule, but a few types, mostly
those of unclear origin, are described below. Other unusual Lactone.Porming Glycosides
types include compounds such as glycerol glucosides from
A number of low-molecular-weight glycosides that pro-
Ulium longiflorum (Fig. 15.16) (Kaneda, 1990). Phenethyl
duce 5- or 6-membered lactones have been encountered in
alcohol glycosides are widely distributed among plants and
several plant families, but are best known from members of
have various types of biological activity. These glycosides
the Ranunculaceae and Liliaceae. These compounds often
have been reported to be antibacterial, antifeedant, cytotoxic,
occur at concentrations of 0.1 % or more dry weight in the
and enzyme-inhibitory activity against AMP phosphodies- plants and are biologically active, having pronounced fungi-
terase and 5-lipoxygenase (Nishimura et al., 1991). toxic andlor allergenic properties (Hegnauer, 1973). These
lactones often produce intense irritation on human skin.
Indigo-Producing Glycosides
Nothing is known about the biosynthesis of this group of
Indican (3-hydroxyindole j3-o-glucoside) (82) occurs compounds in plants. Many of these compounds can be de-
mainly in the genera Indigo/era (Fabaceae),!satis (Brassica- tected by Kedde' s reagent, antimony (III) chloride, and other
ceae), and Polygonum (Polygonaceae). When L_[5_3 Hjtryp_ reagents for cardiac glycosides (which possess butenolide
tophan was adrnirtistered to young plants of !satis tinetoria or other lactone rings) (Fung et al., 1988).
(woad) and Polygonum tinetorium, radioactively labeled in- The biting taste and blistering properties of many species
dican was isolated (Maier et al., 1990). In the woad plant, of the Ranunculaceae are well known. These properties are

rr OH

--
o:r-CH,-CH,-C-~H,

? ? 2HO (,0. O~o

~
I (-
gluiYI gluCjyl
o 0
I I )-ranuDculin (87)
CH,-C-CHrCH,-C=O
II
o
ranunculoside (86)
Ranunculaceae
anemonin (88)

OCH,

O£:\OH

n
nartheside (93), Liliaceae
o o

P~'" -
parasorboside (95) Rosaceae
OH H
parasorbic acid

c:f c\o
OH
~OH

0" -
o 0 mevaloside (94)
(a) Mespilus germanica, Rosaceae anhydromevalonic acid lactone

Fig. 15.18 (8 & c). Lactone-fanning glycosides and their hydrolysis products.
268 Carbohydrates

cSo o
a-methylene-y-
butyrolactone (91)
HO 0

HO~OH
tuliposide A (89)
a-methylene-r-hydroxy-
butyric acid

HO,
~ If
HO
A AO~OH
OH
0

tuliposide B (90)
OH

-
--............
HO~O
o
p-hydroxy-a-methylene-
y-butyrolactone (92)

HO 0

HO~OH
a-methylene-p,y-dihydroxy
(bJ butyric acid
Fig. 1S.18. (continued)

c," ,,,"~ru,, ~
O~C02H (~~O
o 0
0 0

Holygarna lactone (98) biglandulic acid (97) massoialactone (101)

Anacardiaceae Euphorbiaceae
Lauraceae

CH3(CH')~O
OAc
o
OAc OAc
hyptolide (99) argentilactone (100)
Lamiaceae Aristilochia sp.

C:~' o
(c) siphonidin (102) isosiphonidin (103)

Fig. 1S.18. (continued)

 ('arlm"'ydrltl(,.~ 269

caused by protoanemonin (85), which is formed by a com- saceae [parasorboside (95) in Sorbus), mevaloside (94) in
plex set of reactions. The precursor ranunculoside (86) is Mespillus germanica (Rosaceae) (Davies-Coleman and Riv-
converted to ranunculin (87), hence to protoanemonin, and ett, 1989; Tschesche et al., 1969), Holygama lactone in the
subsequently by dimerization to anemonin (88) (Hegnauer, Anacardiaceae (98), biglandulic acid (97) in the Euphor-
1984; Nahrstedt, 1979). Protoanemonin, which arises by biaceae, hyptolide (Hyptis pectinata) (99) in the Lamiaceae,
cleavage of ranunculin by glucosidases found in the plant, argentilactone (100) (Aristolochia argentina) in the Aristo-
also has been isolated by steam distillation of plant material. lochiaceae, and massoia lactone (101) (Cryptocarya mas-
Protoanemonin spontaneously dimerizes to form anemonin soia) in the Lauraceae (Davies-Coleman and Rivett, 1989;
(Hegnauer, 1973). Both anemonin and protoanemonin have Hegnauer, 1984; Nahrstedt, 1979).
antifungal properties (Misra and Dixit, 1980). Another butenolide, siphonidin (102) and its glucoside,
Protoanemonin (85) yielding compounds are known to siphonoside, have been isolated from the leaves of Siphono-
occur in the genera Anemone, Ceratophalus, Clematis, don australe (Celastraceae) (Wagner and F1itsch, 1981;
Clematopsis, Helleborus, Hepatica, Knowltonia, Myosurus, Wagner et al., 1981). A new butenolide, isosiphonodin [3-
Pulsatilla, Ranunculus, and Trautvetteria (Hegnauer, 1973; hydroxymethyl-2(SH)-furanonel (103) and a small amount
Ruijgrok, 1966). The results of numerous tests indicate that of siphonidin (102) were isolated from fifth-instar larvae of
these glycosides do not occur in Aconitum, Actaea, Adonis, Yponomeuta cagnagellus. The larvae of this moth feed on
Anemonella, Anemonopsis, Aquilegia, Callianthemum, Cal- the foliage of the spindle-tree, Euonymus europaeus (Celas-
tha, Cimifuga, Consolida, Coptis, Delphinium, Erantis, Hy- traceae) (Fung et al., 1988).
drastis, lsopyrum, Leptopyrum, Nigella, Semiaquilegia,
Thalictrum, Trollius, and Zanthorhiza (Hegnauer, 1973). Sugar Esters
Tuliposide A (89) is the ester of glucose with a-methyl-
The exudate of the glandular trichomes of Solanum ber-
ene-'Y-hydroxybutyric acid. This compound often is accom-
thaultii are effective against aphids and similar insect pests.
panied by the l3-hydroxyderivative, tuliposide B (90) (Fig.
One part of the defense involves the immobilization of the
IS.18) (Nahrstedt, 1979). The hydroxyacids released by en-
insects in the exudate which hardens (Harbome, 1989). This
zymic or spontaneous hydrolysis of tuliposides lactonize to
exudate proved to be a mixture of sucrose esters, of which
form a-methylene-'Y-butyrolactones (91,92) (Slob et al.,
3,4-di-O-isobutyryl-6-0-caprylsucrose is a major compo-
1975). The enzymes necessary to hydrolyze tuliposides often
nent. These compounds also appear to provide resistance to
co-occur with the glycosides (Slob et al., 1975). These com-
potato blight (Harbome, 1989). A similar series of com-
pounds have strong fungitoxic and allergenic properties. Tu-
pounds is found in the trichomes of other solanaceous plants
liposide A is found in the bulbs, stems, leaves, and flowers
of the genera Datura, Lycopersicon, Nicotiana, and Petunia.
of cultivated tulips (Slob et al., 1975). The NMR spectra of
A series of esters of sucrose with E-p-coumaric acid (such
tuliposides have been reviewed (Tschesche et al., 1969).
as 104) and acetate from Prunus maximowiczii proved to be
TuJiposides are widespread in the genera Tulipa, Ery- responsible for the very bitter taste of the fruits (Shimazaki
thronium, Gagea, Bomarea, and Alstroemeria (Liliaceae) et al., 1991) (Fig. IS.18).
(Hegnauer, 1973; Siobetal., 1975). The genus Alstroemeria
is sometimes placed in the family Alstroemeriaceae, but re-
sembles the Liliaceae in the content of tuliposides. Tulipo-
REFERENCES
side A (89) occurs in large amounts in tulips (Slob et al.,
1975).
Tuliposides appear to play a role in resistance of tulip AKAZAWA. T., POLYSACCHARIDES, in Plant Biochemistry, 3rd ed.
(1. Bonner and J. E. Varner, eds.), 381-403, Academic Press,
cultivars to various fungi (Tschesche et al., 1969). These
New York, 1976.
compounds serve as preformed defensive substances. Tulips
AKAzAWA. T. and K. OKAMOTO, Biosynthesis and metabolism of
often are attacked by Fusarium species, especially at the end
sucrose, in Carbohydrates: Structure and Function (J. Preiss,
of the growing season (Nahrstedt, 1979). These glycosides ed.), Vol. 3 of The Biochemistry of Plants (P. K. Stumpf and
apparently provide most of the resistance to Botrytis tulipae E. E. Conn, eds.), 199-220, Academic Press, New York, 1980.
in resistant lines. However, it is not the intact glycosides AsPINALL, G. 0., Pectins, plant gums, and other plant polysaccha-
themselves but their hydrolysis products that are antifungal. rides, in The Carbohydrates, 2nd Ed., Vol. 2B (R. Pigman and
The greatest amount of tuliposides in tulips occurs in the D. Horton, eds.), 515-536, Academic Press, New York, 1970.
stigmatic branches of the pistil-as much as 32% of the dry AsPINALL, G. 0., Chemistry of cell waJl polysaccharides, in The
weight or 32,000 ppm based on fresh weight (Nahrstedt, Biochemistry of Plants (P. K. Stumpf and E. E. Conn, eds.),
1979). As little as 100 ppm is active in vitro. 473-500, Academic Press, New York, 1980.
Nartheside (a-methoxY-'Y-hydroxy-aa -butenolide) (93), AVIGAD, G., Disaccharides, in Carbohydrates (P. M. Dey, ed.),
a glucoside that yields a-methoxy-aa-butenolide on hydrol- Vol. 2 of Methods in Plant Biochemistry (P. M. Dey and J. B.
ysis, occurs in Narthecium ossifragum and probably other Harbome, eds.), 111-188, Academic Press, London, 1990.
plants of this genus (Liliaceae) (Hegnauer, 1973). AVERS, A. R., J. EBEL, B. VALENT, and P. ALBERSHElM, Host-patho-
Other unsaturated lactone glycosides are found in the Ro- gen interactions. X. Fractionation and biological activity of an
270 Carbohydrates

elicitor isolated from the mycelial walls of Phytophthora meg- of alfalfa root hair membrane potential by Rhizobium meliloti
asperma var. sojae, Plant Physiol., 57, 760-765 (1976). Nod factors, Science, 256,998-1000 (1992).
BACIC, A., P. J. HARRIS, and B. A. STONE, Structure and function EILERT, D., Elicitation: Methodology and aspects of application, in
of plant cell walls, in The Biochemistry of Plants (P. K. Stumpf Cell Culture and Somatic Cell Genetics of Plants, Vol. 4 (F.
and E. E. Conn, eds.), 297-371, Academic Press, New York, Constabel and I. K. Vasil, eds.), Academic Press, San Diego,
1988. CA,1987.
BECK, E. and H. HoPF, Branched chain sugars and sugar alcohols, EL KHADEM, H. S., Carbohydrate Chemistry, Monosaccharides and
in Carbohydrates, (P. M. Dey, ed.), Vol. 2 of Methods in Plant Their Oligomers, Academic Press, Orlando, FL, 1988.
Biochemistry (P. M. Dey and J. B. Harborne, eds.), 235-289, ENGEL, M., A projection on Fischer, Chern. Br., 1106-1109 (1992).
Academic Press, London, 1990.
FEINGOLD, D. S. and G. AVIGAD, Sugar nucleotide transformations
CABm, E., Chitin: Structure, metabolism, and regulation of biosyn- in plants, in Carbohydrates: Structure and Function (J. Preiss,
thesis, in Plant Carbohydrates II. Extracellular Carbohydrates ed.) Vol. 3 of The Biochemistry of Plants (P. K. Stumpf and
(W. Tanner and F. A. Loewus, eds.), 395-415, Springer-Verlag, E. E. Conn, eds.), 101-170, Academic Press, New York, 1980.
Berlin, 1981.
FEINGOLD, D. S. and G. A. BARBER, Nucleotide sugars, in Carbohy-
CAMPBELL, B. C. and D. L. DREYER, The role of plant matrix poly- drates (P. M. Dey, ed.), Vol. 2 of Methods in Plant Biochemistry
saccharides in aphid-plant interactions, in Aphid-Plant Geno- (P. M. Dey and J. B. Harborne, eds.), 39-78, Academic Press,
type Interactions (R. D. Eikenbary and R. K. Campbell, eds.), London, 1990.
149-170, Elsevier, Amsterdam, 1990).
FELLOWS, L. E., Sugar-shaped bullets from plants, Chern. Br.
CHURMS, S. C., CRC Handbook of Chromatography: Carbohy- 842-844 (1987).
drates, Vol. 2, CRC Press, Boca Raton, FL, 1990.
FELLows, L. E., G. C. KITE, R. J. NASH, M. S. J. SrMMONDS, and A.
CLINE, K. and P. ALBERSHEIM, Host pathogen interactions. xvn. M. SCOFIELD, Distribution and biological activity of alkaloidal
Hydrolysis of biologically active fungal glucans by enzymes
glycosidase inhibitors from plants, in Nitrogen Metabolism of
isolated from soybean cells, Plant Physiol., 68, 221-228 (1981).
Plants, K. Mengel and D. J. Pilbeam, eds., Proc. Phytochem.
COLLMER, A., Pectin enzymes and bacterial invasion of plants, in Soc. Europe, 33, 271-282, Clarendon Press, Oxford, 1982.
Plant-Microbe Interactions. Molecular and Genetic Perspec-
FELLOWS, L. E., S. V. EVANS, R. J. NASH, and E. A. BELL, Polyhy-
tives, Vol. 2 (T. Kosuge and E. W. Nester, eds.), 253-284,
droxy plant alkaloids as glycosidase inhibitors and their possible
Macmillan, New York, 1987.
ecological role, in Natural Resistance of Plants to Pests M. B.
COLVIN, 1. R., The biosynthesis of cellulose, in The Biochemistry of Green and P. A. Hedin, eds.), ACS Symposium Series, Ameri-
Plants (P. K. Stumpf and E. E. Conn, eds.), 543-570, Academic can Chemical Society, Washington, DC, 1986.
Press, New York, 1980.
FELLOWS, L. E., G. C. KITE, R. J. NASH, M. S. J. SIMMONDS, and
COURTOIS, E. and F. PERCHERON, Complex glycosides, in The Car- A. M. SCOFIELD. Castanospermine, swainsonine, and related
bohydrates, Vol. 2A, 2nd ed. (R. Pigman and D. Horton, eds.), polyhydroxyalkaloids: Structure, distribution, and biological ac-
213-240, Academic Press, New York, 1970. tivity, in Plant Nitrogen Metabolism (J. E. Poulton, I. T. Romeo,
DAVffiS-COLEMAN, M. T. and D. E. A. RIVEIT, Naturally occuning and E. E. Conn, eds.), Recent Advances in Phytochemistry Vol.
6-substituted 5,6-dihydro-a-pyrones, Fortschr. Chern. Org. Na- 23,395-427, Plenum Press, New York, 1989.
turst., 55, 1-35 (1989). FISHMAN, M. L. and J. J. JEN (eds.), Chemistry and Function of
DAVIS, K. R. and K. HAHLBROCK, Induction of defense responses Pectins, ACS Symposium Series 310, American Chemical Soci-
in cultured parsley cells by plant cell wall fragments, Plant ety, Washington, DC, 1986.
Physiol.,85, 1286-1290 (1987). FRANZ, G. and W. BLASCHEK, Cellulose, in Carbohydrates (P. M.
DEy, P. M., D-Galactose-containing oligosaccharides, in Biochem- Dey, ed.) Vol. 2 of Methods in Plant Biochemistry (P. M. Dey
istry of Storage Carbohydrates in Green Plants (P. M. Dey and and J. B. Harborne, eds.), 291-322, Academic Press, London,
R. A. Dixon, eds.), 53-129, Academic Press, London, 1985. 1990.
DEY, P. M., Oligosaccharides, in Carbohydrates (P. M. Dey, ed.), FuNG, S. Y., W. M. HERREBOUT, R. VERPOORTE, and F. C. FISCHER,
Vol. 2 of Methods in Plant Biochemistry (P. M. Dey and J. B. Butenolides in small ermine moths, Yponomeura spp. (Lepidop-
Harborne, eds.), 189-218, Academic Press, London, 1990. tera: Yponomeuridae), and spindle-tree, Euonymus europaeus
DEY, P. M. and R. A. DIXON, Glycosides, in Biochemistry of Stor- (Celastraceae), J. Chern. Bcol., 14,1099-1111 (1988).
age Carbohydrates in Green Plants (P. M. Dey and R. A. Dixon, GANDER, I. E., Mono- and oligosaccharides, in Plant Biochemistry,
eds.), 131-148, Academic Press, London, 1985. 3rd ed. O. Bonner and J. E. Vamer, eds.), 337-380, Academic
DREYER, D. L. and B. C. CAMPBELL, Chemical basis of host-plant Press, New York, 1976.
resistance to aphids, Plant Cell Environ., 10, 353-361 (1987). GOODWIN, T. W. and E. I. MERCER, Plant Biochemistry, 2nd ed.,
EASTWOOD, M. A. and C. A. EDWARDS, Fibrous polysaccharides, Pergamon Press, Oxford, 1983.
in Toxic Substances in Crop Plants (J. P. F. D'Mello, C. M. GREENWOOD, C. T., Starch and glycogen, in The Carbohydrates,
Duffus, and 1. H. Duffus, eds.), 258-284, Royal Society of Vol. 2B, 2nd ed. (R. Pigman and D. Horton, eds.), 471-513,
Chemistry, Cambridge, 1991. Academic Press, New York, 1970.
EBEL, I., Phytoalexin synthesis: The biochemical analysis of the GRISEBACH, H., Branched-chain sugars: Occurrence and biosyn-
induction process. Annu. Rev. Phytopathol., 24, 235-264 thesis, in Carbohydrates: Structure and Function O. Preiss, ed.),
(1986). Vol. 3 of The Biochemistry of Plants (P. K. Stumpf and E. E.
EHRHARDT, D. W., E. M. ATKINSON, and S. R. loNG, Depolarization Conn, eds.), 171-197, Academic Press, New York, 1980.
 Cllrhohydrate.f 271

HAHN. M. G. and P. ALBERSHEIM. Host-pathogen interactions. LOEwus, F. A., Cyclitols, in Carbohydrates (P. M. Dey, ed.), Vol.
XIV. Isolation and partial characterization of an elicitor from 2 of Methods in Plant Biochemistry (P. M. Dey and J. B. Har-
yeast extract. Plant Physiol. 62. I07-1I1 (1978). borne, eds.), 219-233, Academic Press, London, 1990.
HAHN. M. G.. A. G. DARVlll. and P. AlBERSHEIM, Host-pathogen LoEWUS, F. A. and M. W. LoEWUS, myo-Inositol: Biosynthesis and
interactions. XIX. The endogenous elicitor, a fragment of a plant metabolism, in Carbohydrates: Structure and Function (J. Preiss,
cell wall polysaccharide that elicits phytoalexin accumulation ed.), Vol. 3 of The Biochemistry of Plants (P. K. Stumpf and
in soybeans. Plant Physiol .. 68. 1161-1169 (1981). E. E. Conn, eds.), 43-76, Academic Press, New York, 1980.
HANESSIAN, S. and T. H. HASKEll, Antibiotics containing sugars, LUCAS, W. 1. and M. A. MADORE, Recent advances in sugar trans-
in The Carbohydrates. Vol. 2A. 2nd ed. (R. Pigman and D. port, in Carbohydrates 0. Preiss, ed.), Vol. 3 of The Biochemis-
Horton. eds.). 139-211. Academic Press. New York. 1970. try of Plants (P. K. Stumpf and E. E. Conn, eds.), 35-84, Aca-
HARBoRNE. J. B., Recent advances in chemical ecology, Nat. Prod. demic Press, New York, 1988.
Rep., 7. 85-109 (1989). LUCKNER, M., Secondary Metabolism in Microorganisms, Plants,
HAWKER, J. S., Sucrose, in Biochemistry of Storage Carbohydrates and Animals, Springer-Verlag, Berlin, 1990.
in Green Plants (P. M. Dey and R. A. Dixon, eds.), I-52, Aca- MAlER, W., B. SCHUMANN, and D. GROGER, Biosynthesis of indoxyl
demic Press, London, 1985. derivatives in /saris tinctoria and Polygonum tinctorium, PhytoR
HEGNAUER, R., Chemotaxonomie der Pflanzen, Vol. 6,17-20, 285, chemistry, 29, 817-819 (1990).
328, Birkhauser Verlag, Basel, 1973. MANNERS, D. J., Starch, in Biochemistry of Storage Carbohydrates
HEGNAUER, R., Bedeutung der Chemotaxonomie fiir die Pbanna- in Green Plants (P. M. Dey and R. A. Dixon, eds.), 149-203,
zeutische Biologie, in Biogene Arzneistoffe (F. Czygan, ed.), Academic Press, London, 1985.
157-175, Vieweg und Sohn, Braunschweig, 1984. MATHESON, N. K., Marmose-based polysaccharides, in Carbohy-
JOSELEAU, J. and G. ULLMANN, Biochemical evidence for the site drates (P. M. Dey, ed.), Vol. 2 of Methods in Plant Biochemistry
of fonnation of gum arabic in Acacia senegal, Phytochemistry, (P. M. Dey andJ. B. Harborne, eds.), 371-413, Academic Press,
29, 3401-3405 (1990). London, 1990.
KANDLER, O. and H. HOPF, Occurrence, metabolism, and function MIsRA, S. B., and S. N. DIXIT, Antifungal principle of Ranuneulus
of oligosaccharides, in Carbohydrates: Structure and Function, seeleratus, Bcon. Bot., 34, 363-367 (1980).
(1. Preiss, ed.), Vol. 3 of The Biochemistry of Plants (P. K. MORRISON, W. R. and J. KARRALAS, Starch, in Carbohydrates (P.
Stumpf and E. E. Conn, eds.), 221-270, Academic Press, New M. Dey, ed.), Vol. 2 of Methods in Plant Biochemistry (P. M.
York,1980. Dey and J. B. Harborne, eds.), 323-352, Academic Press, Lon-
KANEDA, M., Lilioside A from Ulium longiflorum: Synthesis and don, 1990.
absolute configuration, Phytochemistry, 29, 3559-3564 (1990).
MuRPHY, E. L., Whai do we know about flatulence in 1968? Ninth
KARR, A. L., Cell wall biogenesis, in Plant Biochemistry, 3rd ed. Dry Bean Res. Conf., 1968, pp. 86-92.
(J. Bonner and J. E. Varner, eds.), 405-426, Academic Press,
NAHRSTEDT, A., Chemische Waffen bei hOheren Pflanzen, Pharma-
New York, 1976.
zie in unserer Zeit, 8, 129-138 (1979).
KAuss, H., Lectins and their physiological role in slime molds
NtslDMURA, H., H. SASAKI, N. iNAGAKI, M. CHlN, and H. MITsu-
and in higher plants, in Plant Carbohydrates n. Extracellular
HASHI, Nine phenethyl alcohol glycosides from Staehys siebol-
Carbohydrates CW. Tarmer and F. A. Loewus, eds.), 627-657,
dU, Phytochemistry, 3D, 965-969 (1991).
Springer-Verlag, Berlin, 1981.
NlSlZAwA, K. and Y. HASHIMOTO, Glycoside hydrolases and glyco-
KooEL, K. and B. BEIllMANN, Isolation and characterization of elici-
syl transferases, in The Carbohydrates, Vol. 2A, 2nd ed. (R.
tors, in Plant Toxin Analysis, Vol. 13 of Modem Methods in
Pigman and D. Horton, eds.), 241-300, Academic Press, New
Plant Analysis (H. F. Linskens and J. F. Jackson, eds.), Springer-
York, 1970.
Verlag, Berlin, 1992.
O'NEILL, M., P. ALBERSHBlM, and A. DARVlLL, The pectic polysac-
KOSUGE, T., Carbohydrates in plant-pathogen interaction, in Plant
Carbohydrates n. Extracellular Carbohydrates (W. Tarmer and charides of primary cell walls, in Carbohydrates (P. M. Dey,
F. A. Loewus, eds.), 584-623, Springer-Verlag, Berlin, 1981. ed.), Vol. 2 of Methods in Plant Biochemistry (P. M. Dey and
1. B. Harborne, eds.), 41 5-441, Academic Press, London, 1990.
LEY, S. V., F. STERNFELD, and S. TAYLOR, Microbial oxidation in
synthesis: A six step preparation of ( + )-pinitol from benzene, PAZUR, J. H., OIigosaccharides, in The Carbohydrates, Vol. 2A,
Tetrahedron Lett., 28, 225-226 (1987). 2nd ed. (R. Pigman and D. Horton, eds.), 69-137, Academic
Press, New York, 1970.
LEZlKA, R. P. and L. QUESADA-ALLUE, Chitin, in Carbohydrates
(P. M. Dey, ed.), Vol. 2 of Methods in Plant Biochemistry (P. PBRClVAL, E., Algal polysaccharides, in The Carbohydrates, Vol.
M. Dey and J. B. Harborne, eds.), 443-481, Academic Press, 2B, 2nd ed. (R. Pigman and D. Horton, eds.), 537-568, Aca-
London, 1990. demic Press, New York, 1970.
Loewus, F. A., L-Ascorbic acid: Metabolism, biosynthesis, func- PERCIVAL, E., The polysaccharides of green, red, and brown seawR
tion, in Carbohydrates: Structure and Function (1. Preiss, ed.), eeds: Their basic structure, biosynthesis aid function. Br. PhyR
Vol. 3 of The Biochemistry of Plants (P. K. Stumpf and E. E. col. 1.,14,103-117 (1979).
Conn, eds.), 77-99, Academic Press, New York, 1980. PERClVAL, E. and R. H. McDoWELL, Algal walls-Composition
LOEWUS, F. A., Ascorbic acid and its metabolic products, in Carbo- and biosynthesis, in Plant Carbohydrates n. Extracellular Car-
hydrates (J. Preiss, ed.), Vol. 3 of The Biochemistry of Plants bohydrates (W. Tarmer and F. A. Loewus, eds.), 277-316,
(p. K. Stumpf and E. E. Conn, eds.), 85-107, Academic Press, Springer-Verlag, Berlin, 1981.
New York, 1988. PERcrvAL, E. and R. H. McDOWELL, Algal polysaccharides, in Bio-
272 Carbohydrates

chemistry of Storage Carbohydrates in Green Plants (P. M. Dey SCHMIDT. E. L. and B. B. BOHLOOL, The role oflectins in symbiotic
and R. A. Dixon, eds.), 305-348, Academic Press, London, plant-microbe interactions, in Plant Carbohydrates II. Extracel-
1985. lular Carbohydrates (W. Tanner and F. A. Loewus, eds.),
PERCIVAL, E. and R. H. McDoWELL, in Algal polysaccharides, in 658-677, Springer-Verlag, Berlin, 1981.
Carbohydrates (P. M. Dey, ed.), Vol. 2 of Methods in Plant SHARON, N. and H. LIS, Carbohydrate-protein interactions, Chern.
Biochemistry (P. M. Dey and 1. B. Harbome, eds.), 523-547, Br., 679-682 (1990).
Academic Press, London, 1990. SHIMAZAKJ, N., Y. MIMARI, and Y. SASHlDA, Prunasin and acety-
PLoUVlER, V., Sur la recherche des itols a chaine droite et des lated phenylpropanoic acid sucrose esters, bitter principles from
cyclitols chez les vegetaux. Relations entre leur repartition et the fruits of Prunus jamasakura and P. maximowiiczii. Phyto-
la classification systematique, Bull. Soc. Chim. Bioi., 45, chemistry,30, 1475-1480 (1991).
1079-1118 (1963).
SWB, A., B. JEKEL, B. DE JONG, and E. SCBLATrMANN, On the
POLLOCK, C. J. and N. J. CHATl'ERTON, Fructans, in Carbohydrates occurrence of tuliposides in the Liliiflorae, Phytochemistry, 14,
O. Preiss, ed.), Vol. 3 of The Biochemistry of Plants (P. K. 1997-2005 (1975).
Stumpf and E. E. Conn, eds.), 109-140, Academic Press, New
SNEADER, W., Drug Discovery: The Evolution of Modem Medi-
York,1988.
cines, John Wiley & Sons, Ltd., Chichester, 1985.
PONTIS, H. G.. Fructans in Carbohydrates (p. M. Dey, ed.), Vol. 2
of Methods in Plant Biochemistry (P. M. Dey and J. B. Har- SRlVASTAVA. R. and D. K. KULSHRESHTHA, Bioactive polysaccha-
borne, eds.), 353-369, Academic Press, London, 1990. rides from plants, Phytochemistry, 28, 2877-2883 (1989).
PONTIS, H. G. and E. DEL CAMPILLO, Fructans, in Biochemistry of STEPHEN, A. M., S. C. CHURMS, and D. C. VOGT, Exudate gums,
Storage Carbohydrates in Green Plants (P. M. Dey and R. A. in Carbohydrates (P. M. Dey, ed.), Vol. 2 of Methods in Plant
Dixon, eds.), 205-227, Academic Press, London, 1985. Biochemistry (P. M. Dey and J. B. Harbome, eds.), 483-522,
i'REIss, J., Biosynthesis of starch and its regulation, in Carbohy- Academic Press, London, 1990.
drates (J. Preiss, ed.), Vol. 3 of The Biochemistry of Plants (P. STURGEON, R. J., Monosaccharides, in Carbohydrates (p. M. Dey,
K. Stumpf and E. E. Conn, eds.), 181-254, Academic Press, ed.), Vol. 2 of Methods in Plant Biochemistry (p. M. Dey and
New York, 1988. J. B. Harbome, eds.), 1-37, Academic Press, London, 1990.
PREISS, J. and C. LEVI, Starch biosynthesis and degradation, in TscHESCBE, R., F. KAMMERER, and G. WULFF, Dber die Struktur
Carbohydrates: Structure and Function (J. Preiss, ed.), Vol. 3 der antibiotisch aktiven Substanzen der Tulpe (Tulipa gesneri-
of The Biochemistry of Plants (p. K. Stumpf and E. E. Conn, ana L.), Chem. Ber., 102, 2057-2071 (1969).
eds.), 371-423, Academic Press, New York, 1980. WAGNER. H. and K. FUTSCH, Neue monocyclische Lactone aus
RALTON,1. E., M. G. SMART, and A. E. CLARKE, Recognition and Siphonodon australe, Plant. Medica, 43, 245-248 (1977).
infection processes in plant pathogen interactions, in Plant-Mi-
WAGNER, H., K. PurSCH, and K. JURCIC, Cytotoxizat von Siphono-
crobe Interactions, Molecular and Genetic Perspectives, Vol. 2
don und aliphatischen Estern des Siphonodins, Planta Medica,
(T. Kosuge and E. W. Nester, eds.), Macmillan, New York,
43, 249-251 (1981).
1987.
REID, J. S. G .. Galactomannans, in Biochemistry of Storage Carbo- WARD JR., K. and P. A. SEIB, Cellulose, lichenan, and chitin, in
hydrates in Green Plants (P. M. Dey and R. A. Dixon, eds.), The Carbohydrates, Vol. 2A, 2nd ed. (R. Pigman and D. Horton,
265-288, Academic Press, London, 1985. eds.), 413-445, Academic Press, New York, 1970.
ROCKLAND, L. B., The search for a convenient assay method for WHISTLER, R. L., J. N. BEMn.LER, and E. F. PASCHALL, Starch:
the flatulence factor in dry beans, Ninth Dry Bean Res. Conf., Chemistry and Technology, 2nd ed., Academic Press, Orlando,
1968, pp. 78-86. FL,1984.
RUIJGROK. H. W. L., Distribution of ranunculin and cyanogenetic WHISTLER, R. L. and E. L. RICHARDS, Hemicelluloses, in The Car-
compounds in the Ranunculaceae, in Comparative Phytochem- bohydrates, Vol. 2A, 2nd ed.. (R. Pigman and D. Horton, eds.),
istry (T. Swain, ed.), 175-186, Academic Press, London, 1966. 447-469, Academic Press, New York, 1970.
16
Cyanogenic Glycosides and Cyanolipids

Introduction ric acid) and the Feigl-Anger methods (Feigl and Anger,
Biosynthesis 1966; Seigler, 1991; Tantisewie et aI., 1969).
Major Structural Types of Cyanogens The actual cyanogens have been isolated and studied from
Cyanogens Derived from Phenylalanine more than 475 species of plants (Seigler, 1991). Despite the
Cyanogens Derived from Tyrosine fact that this phenomenon is widespread, the structures of
Cyanogens Derived from Valine and Isoleucine only about 60 cyanogenic compounds have been published
Cyanogens Glycosides Derived from Leucine (Nahrstedt, 1987a; Seigler, 1991). These compounds are gly-
Cyanolipids cosides of Ci-hydroxynitriles or cyanohydrins with the excep-
Cyanogenic Glucosides with a Cyclopentenoid Ring tion of a few cyanogenic lipids (see below). The glycosides
Structure are usually accompanied by a j3-glycosidase enzyme capable
Cyanogenic Compounds of Other Origins of producing the corresponding cyanohydrin (aglycone) and
Distribution a sugar (Fig. 16.1). A second type of enzyme (hydroxynitrile
j3-Glycosidases lyase) catalyzes the dissociation of the cyanohydrin to a car-
Biological Activity bonyl compound and hydrogen cyanide. Normally, the sub-
Transport of Cyanogenic Glycosides strate and enzymes are compartmentalized within the plant
Cyanogenic Compounds as Defensive Substances and cyanide release does not occur unless the plant is dam-
Cyanogenesis, Ants, and the Genus Acacia aged (Conn, 1981, 1993; Poulton, 1988; Poulton and Li,
Cyanogenesis in Millipedes and Insects 1994; Selmar, 1993).
Biological Activity of Cyanide in Plants In the family Sapindaceae (and possibly the related family
Detoxication of Cyanide Hippocastanaceae), the Ci-hydroxynitrile is esterified with a
Cyanogenesis in Crop Plants long-chain fatty acid (C 18 or C20 ) to produce cyanogenic
Nitroglycosides lipids (Fig. 16.2) (Mikolajczak, 1977).
Nitrile Glucosides Although many bacteria and fungi are cyanogenic, the
Pseudocyanogenic Glycosides cyanogenic compounds usually are labile. Glycine is the pre-
Analytical Techniques cursor of hydrogen cyanide in many instances. Labeling
References studies indicate that the C-N bond is not broken during the
biosynthesis of cyanide (Harris et al., 1987). In some cases,
cyanohydrins have been isolated, but these may result from
secondary reactions of hydrogen cyanide and other fungal
INTRODUCTION metabolites. The cyanohydrins of glyoxylic and pyruvic
acids have been reported from a snow mold fungus (Fig.
The ability to produce cyanide or cyanogenesis has long 16.3) (Bunch and Knowles, 1980; Knowles, 1988; Tapper
been recognized in plants. At least 2650 species, from more and MacDonald, 1974).
than 550 genera, and 130 families possess the ability to make Cyanogenic cyanohydrins and cyanogenic glycosides
cyanogenic glycosides (Hegnauer, 1986; Seigler, 1991). also are known to occur in animals (Duffey et aI., 1977;
Most reports of cyanogenesis are based on two simple, but Nahrstedt and Davis, 1981).
reasonably specific, color tests-the Guignard (alkaline pic- The production of cyanide in many plants is extremely

273
274 Cyanogenic Glycosides and Cyanolipids

l3-glucosidase

Hu
dhurrin (15)

H .CN o

"0><0
+HCN
hydroxynitrile lyase

OH OH

4-hydroxymandelonitrile (3)

HOH~O~CN><U" OH 0 0
H
~

j
HO ~
cyanogenic RO OR f
diglycoside ~ ~YCOSidase

8-glycosidase eN H

:;~:~><O'"
HO
~ OH
H
/
n,g,::::-----
""><u a-cyanohydrin

~
cyanogenic monoglycoside

Fig.16.1. Catabolism of cyanogenic glycosides (Seigler, 1991; modified and used with the permission of the copyright owner, Academic Press,
Orlando, FL).

variable. All individuals of some species are cyanogenic; 1992; Nahrstedt, 1981; Seigler, 1991; Seigler and Brinker,
populations of others contain plants that are cyanogenic 1993).
(ranging from very strong to weak) and plants that are not;
with other species, some populations are completely acyano-
genic, whereas others exhibit some cyanogenesis. The BIOSYNTHESIS
expression of cyanogenesis often is influenced by stress and
other environmental factors. Nonetheless, the distribution of The aglycone portion of cyanogenic compounds is derived
cyanogenic glycosides is of systematic interest as certain from amino acids. To date, most cyanogenic compounds
structural types of cyanogenic glycoside are associated with come from a relatively small number of precursors: L-tyro-
specific groups of plants (Hegnauer, 1986; Saupe, 1981). sine, L-phenylalanine, valine, isoleucine, and L-leucine. Aca-
The presence of cyanogenic compounds in food and for- Iyphin (1) (Fig. 16.14) appears to be derived from nicotinic
age plants such as cassava and sorghum creates important acid. One series of cyanogens has aglycones that contain a
health problems in many tropical countries (Davis, 1991; cyc1opentenoid ring and are derived from the nonprotein
Nahrstedt, 1993; Wilson, 1987). amino acid, (2-cyc1opentenyl)glycine.
Methods for the isolation, purification, and characteriza- Much of biosynthetic work has been done with flax
tion of cyanogenic glycosides and related compounds have (Linum usitatissimum), cherry laurel (Prunus lauroeerasus),
been reviewed (Brimer, 1988; Brinker and Seigler, 1989, and sorghum (Sorghum bieolor). Introduction of labeled
 Cyanogenic Glycosides and Cyano/ipids 275

'X'
,
o
, ..Jl
NC H 0 CH,(CH')I7CH,

NC
':c' '
I
H
0
JL
CH,(CH')I7CH,

(----0 0
OH

_ (---0
OH
--y
H CN
0
_
Ho~00.
HO~O~O)lCH,(CH')I7CH' HO OH J--;0
OH (58) (59)

Fig. 16.2. Cyanolipids.

amino acid precursors into these plants results in extremely Major intermediates include a hydroxyl amino acid, aldox-
high introduction of label (up to 25%). By double-labeling ime, nitrile, and an a-hydroxynitrile (Fig. 16.4) (Halkier and
experiments, it has been demonstrated that the C-N bond Mf2IlIer, 1990; Halkier et aI., 1988). These intermediates are
is not broken during the biosynthetic process; all intermedi- observed only at extremely low levels in the plants which
,tes in the pathway contain nitrogen. Recent work has been contain them. In the course of the biosynthetic studies, it
facilitated by the development of particulate (microsomal) was observed that the proposed intermediates were incorpo-
fractions from Sorghum bicolor, Linum usitatissimum (flax), rated at very different and inconsistent rates. These differ-
and Triglochin maritima. In several instances, removal of ences in rate were inconsistent with the order of incorpora-
the seed coat is an obligatory step; preparations made with- tion of the different intermediates. A channeled process has
out this step usually are inactive (Koch et aI., 1992). One been proposed in which the compounds are not liberated
similarity of all systems is that the product is produced with- from the enzyme surface (Conn, 1981). Channeling provides
out the accumulation of any intermediate compounds. This for the rapid and efficient flow of carbon and nitrogen atoms
phenomenon was not consistent with the kinetic parameters and at the same time perhaps protects labile intermediates
for the individual reaction steps. The enzymes of the path- from wasteful side reactions (Conn, 1981).
way are arranged in a way such that efficient flow of mole- Many of the enzyme systems of the biosynthetic process
cules from one step to the other occurs (Conn, 1983; Cutler have now been studied. A microsomal fraction from etio-
and Conn, 1982; Dewick, 1984; Mf2IlIer and Poulton, 1993). lated sorghum seedlings converted L-tyrosine (2) into p-hy-
droxymandelonitrile (3) (Halkier and Mf2IlIer, 1991; Mf2IlIer
CO,H and Poulton, 1993). The particles required ouly NADPH in
CO,H
I I addition to the substrate. N-Hydroxytyrosine (4) was shown
HO-C-CN HQ-C-CN to be an intermediate in the synthesis of the aldoxime, and an
I I N-hydroxy1ating cytochrome P-450 was identified (Dewick,
CH, H
1984; Halkier and Mf2IlIer, 1989, 1991; Halkier et aI., 1991).
The N-hydroxytyrosine (4) is converted to 2-nitro-3-(p-hy-
pyruvic acid glyoxylic acid
droxyphenyl)propionic acid (5), which is, in tum, converted
cyanohydrin cyanobydrin
to 1-aci-nitro-2-(p-hydroxyphenyl)ethane (6). The last com-
Fig. 16.3. Glyoxylic acid and pyruvic acid cyanohydrins. pound appears to be the immediate precursor to (E)-p-hy-
276 Cyanogenic Glycosides and Cyano/ipids

droxyphenylacetaldehyde oxime (7). l-aci-Nitro-2-(p-hy- ing P-450 enzymes (Koch et al., 1992). As is true in
droxyphenyl)ethane (6) exists in equilibrium with I-nitro- sorghum, nitro compounds [such as aci-1-nitro-2-methyl-
2-(p-hydroxyphenyl)ethane (8) (Hallder and Ml'lller, 1989, propane] (13) also appear to be intermediates in the synthetic
1990; Haikier et al., 1988, 1991). This nitro compound has route. 2-Cyclopentenylglycine (14) also was converted to
been isolated from culrures of cyanogenic plants, and similar cyanogenic glycosides with this enzyme system. The en-
compounds are narurally occurring. In sorghum and in cas- zyme system is located in the cotyledons and their petioles;
sava (see below), both the (E)-isomer (6) and (Z)"isomer after synthesis, linamarin and lotaustralin are transported
(9) of the oxime were accepted (Koch et aI., 1992). The rapidly to other parts of the growing seedling (Koch et al.,
apparent NADPH and oxygen requirements for the oxime- 1992).
metabolizing enzyme appear to be the same as those for Hahlbrock and Conn (1970) isolated an enzyme system
the C-hydroxylating enzyme mentioned below. Using this from flax that catalyzed the glucosylation of a series of 2-
microsomal fraction, it has been demonstrated that the nitrile hydroxynitriles. This system exhibited maximum rates with
(10) rather than the hydroxyaldoxime is involved in the syn- the acetone and methylethyiketone which would, on glyco-
thesis (Cutler and Conn, 1982; Dewick, 1984). A C-hydrox- sylation, yield linamarin (11) and lotaustralin, the cyanogens
ylating cytochrome P-450 that converted the nitrile to the of flax. This enzyme was inactive toward aromatic cyanohy-
hydroxynitrile was identified (Halkier and Ml'lller, 1991). drins. The enzyme glucosylated both the (R)- and (S)-forms
f3-Glucosyltransferases have been isolated and purified of the cyanohydrin of 2-butanone (Zilg and Conn, 1974). In
from a few cyanogenic species. They have pH optima in the most cases, however, racemic mixtures of compounds are
range 6.5-9.0 and usually lack a requirement for metal ions not encountered in plants; this suggests that, in general, only
or cofactors. Although they have an absolute specificity for one optically active form of the cyanohydrin is glucosylated
UDP-glucose, they are less specific toward cyanohydrins by the enzyme. In contrast, when the corresponding glucosy-
(Ml'lller and Poulton, 1993). lating enzyme from sorghum reacts with the cyanohydrin
A microsomal system that catalyzes the in vitro synthesis [p-hydroxy-(R,S)-mandelonitrile (3)], only dhurrin (15) is
of the aglucones of the two cyanogenic glycosides linamarin produced. When a soluble protein fraction containing the
(11) and lotaustralin has been isolated from young seedlings UDP-glucosyl transferase was added to the cyanohydrin
of cassava (Manihot esculenta, Euphorbiaceae) (Fig. 16.5). from the microsomal preparation described above, the for-
Valine (12) and isoleucine were converted in the presence mation of dhurrin occurs (Conn, 1981).
ofNADPH to linamarin and lotaustralin in a process involv- The biosynthesis of (R)-taxiphyllin (16) has been demon-

OH

--
H,N HN/

HO -oJ
-
~ CO,H
-HO
-o-FCO'H
V~
-
L-tyrosine (2) N-bydroxylyrosine (4) 2-nitro-3-(4'-hydroxyphenyl)
propionic acid (5)

HO~
yN
yH
'=I

!t
aci-t-nitro-2-p-hydroxyphenylethane

(6)
(E)-4'-bydroxypbenyl-
acetaldehyde oxlme (7)
H
(Z)-4'-hydroxyphenyl-
acetaldehyde oxime (9)
!
HO~NO, l-nitro-2-(p-bydroxypbenyl)ethaoe HO~
(8)
/" ~CN
_ (o~o H~CN p-hydroxypbenyJacetonitrile
HO~O I ~ __ HO ~~H (10)
00 00
....:;;
~N
dhurrin (15) OH p-bydroxymaodelooitrlle (3)

Fig. 16.4. Biosynthesis of cyanogenic glycosides (Halkier et aI., 1991; modified and used with penmssion of author).
 Cyanogenic GJycosides and Cyanolipids 277

~.0:H'+
Cn, CH,
--+

L-vallne (12) N·hydroxyvaline (2-cyclopentenyl)glyclne (14)

--
(E)-2-methylpropanaloxime aci-l-nitro-2-methylpropane (13) I-nitro-2-methyJpropane

HO" / C ~ glucosyl-O C=N

CH'--~-
, C~ CU;-'><:H,
linamarln (11)
2-methylpropionitrile acetone cyanohydrin

Fig. 16.5. Biosynthesis of linamarin and lotaustralin (Koch et aI., 1992; modified and used with permission of the copyright owner, Academic Press,
Orlando, PL).

strated with a microsomal preparation from etiolated Tri- glycine and one probably from nicotinic acid (Seigler, 1991).
glochin maritima. L-Tyrosine (2) was converted into p-hy- The known cyanogenic glycosides will be discussed (below)
droxyphenylacetonitrile (10) and p-hydroxymandelonitrile in groups based on their probable biosynthetic origin. The
(3) when the preparation was supplied with NADPH. When center bearing the nitrile group often is chiral. In many in-
UDP-glucose and a soluble fraction from the plant were stances, both (R)- and (S)-forms are known; occasionally,
added, taxiphyllin was formed. Incubation of the glucosyl- both epimers occur in the same plant.
transferase fraction with p-hydroxy-(R ,s)-mandelonitrile
and UDP-glucose, produced taxiphyllin (16), but not (S)- Cyanogens Derived from Phenylalanine
dhutrin (15) (Fig. 16.4) (Cutler and Conn, 1982; Dewick,
1984). It was necessary to remove the seed coats in order Several cyanogens are derived from L-phenylalanine (Fig.
to obtain an active microsomal fraction. In this case, p-hy- 16.6). Compounds derived from this precursor is found pri-
droxyphenylacetonitrile did equilibrate with exogenous ma- marily in the Rosidae and Asteridae (as defined by Cron-
terial, although other intermediates were channeled (Cutler quist). Among the compounds from this pathway, the best
and Conn, 1982; Dewick, 1984). known is amygdalin (20), which is widespread in seeds of
A similar series of compounds is converted into triglochi- members of the Rosaceae such as apples, peaches, cherries,
nin (17) (Fig. 16.10) in Trig/ochin maritima and Thalictrum and apricots. Phenylalanine-derived cyanogens include the
aquilegifolium (Ranunculaceae). 3,4-Dihydroxyphenylace- monoglucosides (R)-prunasin (19), prunasin-6-malonate
tonitrile (18) also is incorporated (Fig. 16.10) (Cutler and (21) (Nahrstedt et al., 1989), (S)-sambunigrin (22), and the
Conn, 1982; Dewick, 1984). diglycosides (R)-amygdalin (20), (R)-Iucurnin (23), (S)-epi-
Treatment of racemic mandelonitrile with UDP-glucosyl lucurnin (2-J3-primeverosyloxy-2-phenyl-2S-acetonitrile)
transferase from the fruits and leaves of black cherry, Prunus (24), (R)-vicianin (25), grayanin (26) (Shimomura et al.,
serotina, produces only (R)-prunasin (19). These prepara- 1987), prunasin 2'-glucoside (27) (Aritomi et al., 1985),
tions did not convert prunasin to the diglycoside amygdalin amydgalin 6"(4-hydroxybenzoate) (28), amygdalin 6"[4-hy-
(20) (Dewick, 1984). droxy-(E)-cinnamatel (29) (Nahrstedt et aI, 1990), oxyan-
thin [2R-J3-D-apio-D-furanosyl-(1-6)-J3-D-glucopyranosy-
loxyphenylacetonitrile (30), and the 5"-benzoate of
JllAJOR STRUCTURAL TYPES OF CYANOGENS oxyanthin (31) (Rockenbach et aI., 1992). Two complex gly-
cosides are also found in the froits of Anthemis cairica and
As mentioned above, most cyanogenic compounds are de- A. altissima. These compounds, Anthemis glycosides A and
rived from five amino acids. Other cyanogenic glycoside B, epilucumin 4"-p-(J3-D-glucopyranosyloxy)-(E)-cinna-
are derived from the nonprotein amino acid cyclopentenyl mate (32), epilucumin 4"-p-(J3-primeverosyloxy)-(E)-cinna-
 HO
'
~
4'
H~.,.~CN
I
5'
OH

"
0 0:2 3
4
~5
,,(O~O
HO~
4' : ' »I Q
: """
C

//
4~,
HO 3' OH '~ HO OH I
, 3' 8 ~6

(R)·prunasin (19)
(S)-sambunigrin (22)

HO 0
14 13 10
H ICN
f
"~~o~,
12
HO
IS
~ j 11

" 17

..-,
grayanin (26)

o 0

HO
~O
'''~' H,~ .. ICN
.... 4

4' 0 0 2 3 ~5
HO ~
HO OH ,I
3' ""-:;:::::;6
7

(a) (R)-prunasin 6'-malonate (21)

~ :~~:>\),
HOH: 3" '''OOH O~,
0 :~"\
Ic~ 4~ ""
'I ~5
~::~"0:
6"
'7~ ,

00 • I •
HO 3' OH, HOHO 7)
..--;::::; 6 OH prunasin 2'-glucoside (2
(R)-amygdalin(20) 7 ~

}-o"" "0"" "on "~~:'~'.

H ~
~ 4"
" 1
HO OH /
.-::;:::::::.

"",," , '"" "::i A }-~ II';.

5" 0 "><oCN (S)-neoamygdalin
0' ,\,

HO:O~ / ~4' ~ ~OH'" 0

(R)-amygdalin6"(4-hydroxybenzoate)(28) HO~O" H , leN

><o,.

HO 3" OH ':\ ~O. O' 3 ~

(R)-amygdalin 6"(4-hydroxy-(E)·cinnamate) (29)
HO:n~
HO 3' OH
/
.--;::::.
(b)

Fig. 16.6 (a-c). Cyanogens derived from phenylalanine.

278
 Cyanogenic Glycosides and Cyanolipids 279

."~OH,,,
HO
0

3"
'"
OH
HO
o~",0 '
HO 3'
0

OH
H CN
»j' •

•
3

"
I
""-

""
~,

7
(R)-vicianln (25)

OO~~'><o'CN
HOH~O~'H~CN 3" OH HO~~ 3' "OOH 0 r ~
3" OH" 0 ~
HO
HO OH
0 1/
I
~ (R)-Iucumln (23)

_ . do~~~:~
.--:;::;

OHOH OH.V

" O~H~'CN.
HO ~ 3"
0."

HO
0 0 3 ~
r
oxyantbin S"-benzoate (31) ..&

OH OH HO OH" ....::::::'

(e) oxyantbin (30)

Fig.16.6 (continued)

mate (33) (Fig. 16.7), contain several sugar moieties as well pounds (but have other types of cyanogens as well) (Seigler
as p-hydroxycinnarnate residues (Nahrstedt, 1987a). Pm- et aI., 1989). The diglycoside cyanogens are more restricted
nasin and sambunigrin only differ in configuration at the in distribution. Amygdalin (20) is found primarily in fruits
carbinol carbon. The families Asteraceae, Fabaceae, and Ro- of the Rosaceae, lucumin in seeds of the Sapotaceae, epilu-
saceae are characterized by the production of these com- cumin (24) and its derivatives only in the fruits of Anthemis

H('H~O
~

3",
0

OH
Hqj'O~O
6'.

~..::\.
3,.OH
_0 ~,¥
16 17

~ 11 ~~O:::\ ~0l))
_ ~.
. . .3 1 0 0
4" S"

3"
0

OHH ;o~O
q.o J' OH
H
•
eN

I ~
8 ....-:::;

Anthemis glycoside A (32)

6'. OH H 'CN
H~~o 4"'''0."o,'
~ ~
iii 17

11
~ .OHO
~o' r ~~, OH l'
3'. on 10 3" H<kO 3' OH •
.4 13
o .--::,

Anthemis glycoside B (33)

Fig. 16.7. Anthemis glycosides A and B.

280 Cyanogenic Glycosides and Cyanolipids

OH H ICN OH H~:'CN
Ho~~OHHO~ ~s
6' """ 6' / 4
o 0 ~ 0 0 OH
I
4 4'

8~~~( ~
(R}-holocalin (35) (S)-zierin (34)

4"

~ O~'IH><o./CN
"O

HOHO
~ ~ 0
OH 0 ~ OH
HO 2' fl 5
HO 3' OH I
~
zierin xyloside (36)

I
OH HO
~
6"

~o ~"O H~/CN4
14 13

HO 0 0 15 r; o
1"0 6'
HO
4'a OH 9
4"'2'"
0HO OH
~02
I' 1 ~ OH
16 17 5'" OH 3" H(lHO OH 8 '
o 3' --::::: 6

xeranthin (37)

Fig. 16.8, meta~Substituted cyanogenic glycosides.

cairica (Asteraceae) (Nahrstedt et aI., 1983), and vicianin donous plant of the genus Chloraphytum (Liliaceae). A dig-
(25) in seeds of Vicia (Fabaceae) and in Davallia (a fern) Iycoside of this group, zierinxyloside (36) [from the achenes
(prunasin also is found in ferns) (Seigler, 1991). of Xeranthemum cylindraceum (Asteraceae)] contains the
An unnamed 1,2-linked diglucoside (27) has been de- sugar primeverose, whereas an even more complex glyco-
scribed from Perilla Jrutescens var. acuta (Lamiaceae). side, xeranthin (37), has been isolated from this species (Fig,
The seeds of many Rosaceous species contain amygdalin 16.8) (Nahrstedt, 1987a).
(20) and a mixture of l3-g1ycosidases, often called emulsin.
This mixture preparation hydrolyzes both amygdalin and Cyanogens Derived from Tyrosine
prunasin (18) readily. The leaves of these plants usually con- The best known of this series is dhurrin (15), which may
tain only prunasin and the leaf enzymes hydrolyze prunasin make up to 30% of the dry weight of the leaves and coleop-
but not amygdalin (Poulton, 1983). Within seeds of Prunus tiles of etiolated sorghum seedlings (Halkier and M¢ller,
sera tina and Prunus domestica, the cyanogenic glycosides 1989; Saunders and Conn, 1977). Cyanogens derived from
amygdalin and prunasin are found in the cotyledonary paren- tyrosine [(S)-dhurrin, (R)-taxiphyllin (16), and triglochinin
chyma, whereas amydgalin and prunasin hydrolase are found (17) (probable) (Fig. 16.9) are widespread in nature, whereas
in protein bodies of the procambial cells (Poulton and Li, p-glucosyloxymandelonitrile (38) and proteacin (39) are less
1994; Swain et aI., 1992). The enzymology of cyanogenesis corrunon (Fig. 16.10). Tyrosine-derived glycosides are en-
of some Rosaceous fruits has been reviewed (Poulton, 1993). countered most commonly in monocotyledonous angio-
A eDNA clone of the flavoprotein (R)-( + )-mandeloni- sperms and in the Magnoliidae (sensu Cronquist), but are
trile lyase from Prunus sera tina has been made and a puta- found in many other plant families as well (Hegnauer, 1977;
tive Flavin Adenine Dinucleotide (FAD)-binding site was Saupe, 1981).
identified (Cheng and Poulton, 1993). Isotriglochinin (40), an isomer of triglochinin, and the
Although the biosynthetic origin has not been definitely methyl ester of triglochinin have been reported, but both
established for all meta-substituted cyanogens, (S)-zierin may be artifacts of isolation (Conn, 1981).
(34) is derived from phenylalanine (Fig. 16.8) (Nahrstedt, In addition to other cyanogenic glycosides such as p-
1992; Nahrstedt and Schwind, 1992). (R)-Holocalin (35) oc- glucosyloxymandelonitrile (38), Nandina domestica (Ber-
curs in the seeds of the legume, Halacalyx balansae. The beridaceae) contains a complex cyanohydrin, nandinin (41).
leaves of this same plant contain (R)-prunasin (19) (Nahr- This compound is the 4'-caffeoyl ester ofp-glycosyloxyman-
stedt, 1976). Holocalin is also known from a monocotyle- delonitrile (Olechno et al., 1984).
 Cyanogenic Glycosides and Cyalloiipids 281

OH
OH

0Qr
6 <' O~OHHO 8
0
o ~'S'6'
HO
0
3'
OH

8 3 -- 2' OH 2' OH
mm . m
5 CN
0
o 4 I

triglochinin (17)
isotriglochinin (40)

OH

"/.} ) < ~
'5'6'
,
6
0 ° 3' OR

3,4-dihydroxyphenylacetonitrile (18) HO OCH, M, 0" •

5 CN
4 1
o

triglochinin monomethyl ester

Fig. 16.9. Cyanogens derived by oxidative cleavage of tyrosine and 3,4-dihydroxy-phenylacetonitrile.

OH H~~CN (O~O H.. , ,eN> < o ,

.O"~ o 0
1 ~
~

OH
HOHO
~O ~
OH , I ~
OH
(S)·dhurdn (15) (R)-taxiphyllin (16)

("0
Ho~-\J'L
,,~".";;f:~;;)~~
U ~OH 0 OH
HO OH - CN 0 HO OH

p-glucosyloxymandelonitrile (38) (S)-proteacin (39)

nandinin (41)

Fig. 16.10. Cyanogens derived from tyrosine.

282 Cyanogenic Glymsidt's "lid CywlOlipids

~ O),[I.!OH:-·(-~O O~' ~'(-~O

.OH , , ,.OH

~CN
". CN I 3 OH
() 4' CN 6' 1
HOHO ,. HO ,
.,. OH ,~O~ ~O~O , ,
OH s" mf
s
linamarin (11) (S).epilotaustralin (43) (R)·lotaustralln (42)

~
" OH OH

O~, X HO~O~'
s~CN
~ ~

HO 0 , 'CN '" 0

HO '" OH'" " 0

HO
I'»A o 4 3 on 4' 0
l' 0 3
HO 3' OH HO
HO 3' OH
Ilnu,taOO (44)

~
' OH 0 ,":ollnu'taOO(45)

" "O~/"OH'
HO '1' " 5 OH
OH H OH 'CN~" 0

~
6' 0 4 ICN sarmentosin(47) 4" ~O
,. O~ HO 2' 5
5 H 3' 0 2
HOH 3' OH 0 3 OH , CN, CH,OH
isosarmentosin
sarmentosin epoxide (46)
Fig. 16.11. Cyanogens derived from valine and isoleucine.

In studies with etiolated sorghum seedlings, dhurrin (15) spread than usually thought (Nahrstedt, 1987a). (R)-Lotaus·
was found to occur primarily in epidermal tissues, whereas tralin (42) occurs in Berberidopsis beckleri, considered to
the corresponding f3-glucosidase occurred in the subtending be a primitive member of the Aacourtiaceae (Jaroszewski
mesophyll tissues (Kojima et ai" 1979). The subcellular lo- et aI., 1987b).
calization of dhurrin f3-glucosidase and hydroxynitrile lyase The corresponding gentiobiosides, Iinustatin (44) and
in the mesophyll cells and a UDP-glucose: aldehyde cyano- neolinustatin (45), have been isolated from flax seed and
hydrin f3-glucosyl transferase in epidermal plastids of certain Passiflora species (Olafsdottir et aI., 1989; Smith et
Sorghum leaf blades have been examined (Thayer and Conn, aI., 1980; Spencer et aI., 1986).
1981; Wurtele et aI., 1982). Sarmentosin epoxide (46), from Sedum cepaea (Crassula-
ceae), contains an epoxide ring and is not a glycoside of
Cyanogens Derived from Valine an a-hydroxynitrile. Upon opening of the epoxide ring, the
and Isoleucine resulting cyanohydrin liberates cyanide spontaneously
(Nabrstedt, 1987a). This compound is related to the noncy-
The cyanophoric compounds linamarin (11) (R)-Iotaus- anogenic nitrile, sarmentosin (47), from Sedum sarmen-
tralin (42), (S)-epilotaustralin (43), linustatin (44), neolinus- tosum.
tatin (45), and sarmento sin epoxide (46) (probable) are de-
rived from valine and isoleucine (Fig. 16.11). The first two Cyanogenic OIycosides Derived
compounds are widespread and probably are found in more from Leucine
plant species than any other cyanogenic glycosides. Lina- Another cluster of compounds arises from leucine. (S)-
marin and lotaustralin almost always co-occur. These gluco- Proacacipetalin (48), (R)-epiproacacipetalin (49), (S)-proa-
sides are most commonly encountered in the Asteraceae, caciberin (50), (S)-heterodendrin (51), (R)-epiheterodendrin
Euphorbiaceae, Fabaceae, and Linaceae. (52), 3-hydroxyheterodendrin (53), (S)-cardiospermin (54),
(S)-Epilotaustralin (43) was first found in Triticum mono· and the corresponding p-hydroxybenzoate, p-hydroxycinna-
coccum (Poaceae) and also occurs in Passiflora warmingii mate, and sulfate are all known (Fig, 16.12). Of these glyco-
(Olafsdottir et aI., 1989). As these (R)- and (S)-epimers are sides, proacacipetalin (48), epiproacacipetalin (49), and pro-
difficult to distinguish and most previous studies did not acaciberin (50) are known only from the genus Acacia. The
differentiate the two, (S)-epilotaustralin may be more wide- diglycoside, proacaciberin (50), which contains a vicianose
 Cyanogenic Glycosides and Cyano/ipids 283

OH
'
~
'
~
H'CN
OH H 'CN
4' 0 ~!5 4' 0 Y
"" , y ' 5
HOHO 0
H~O l' 0 2"
2' 2
3' OR
" OH 4
4

(S)-proacacipetalin (48) (R)-epiproacacipetalin (49)

OH

~
OH
~
' H'CN H 'CN

HO
o ~\ . HO 4' 0 ~'
. 4
HO 2' 0 2 HO 0,
OH 3' OR
5

(R}~epiheterodendrin (52)
(S)-heterodendrin (51)

HOHO
~
OH
o
H

0 Y O H H OHO
f
CN
5 0 !
~OH H

o y ' OSO,"
CN
5

OH OH
4 4

(S)-cardiospermin (54) (S)-cardiospermin sulfate

~ O~ ~(O~o
4'
'
'NyH OH
0
'CN
4

J H~~O~OH
HOHO l'

OH
3 OR I' 5 4
5

(a)
acacipetalin (56) sutherlandin (57)

OH
o
~
' H'CN
4' 5' 0 y ! 5
HOHO z' 0; 3 0
3' OR
4
14 13

J.-O'
(S)-cardiospermin 4-hydroxy-(E)-cinnamate (74)

H0
~
'
4
'
OH

0 o H y5 ! 0 6
'CN

\. jlOOH~OH
9

,
HO, HeN
O~!S
1211 6'

'OH 4'
4 HO l' 0
(S)-cardiospermin 4-hydroxybenzoate (55) HO 3' OH , OH

3-hydroxy-(S)-heterodendrin (53)

OR
Ls'/~
0 1" o~''oC~\
~ 0 " 5

4"
r::V
HO 3"
'OH
HOHO
4'

3'
2'
OH
4
(S)-proacaciberin (SO)

(b)

Fig. 16.12 (a & b). Cyanogens derived from leucine.

284 Cyanogenic Glycosides and Cyanolipids

unit, has been isolated from Acacia sieberiana (Brimer et Acacipetaiin (56) is probably an artifact of isolation (Et-
ai., 1981; Nartey et ai., 1981). Heterodendrin (51) and epi- tlinger et al., 1977).
heterodendrin (52) occur only in the Sapindaceae and in the Sutherlandin (57), a compound related by structure to the
Poaceae (grasses). Although cardiospermin is known only series of compounds derived from leucine, and in particular
from the Sapindaceae, the p-hydroxybenzoate (55) is known to certain cyanolipids, is weakly cyanogenic. This compound
only from Sorbaria arborea (Rosaceae: Spiraeoideae) co-occurs in Acacia sutherlandii with proacacipetalin and
(Nabrstedt, I 987a). the dimer of the two compounds (Swenson, 1986; Swenson

O Q
3 2
3 2
CN OH CN OH
4 1~'
5'
o 0 OH
4 ~'5'
5 HO 3'
o 0 OH
5 HO J'
2' OH 2' OH

tetraphyllin A (61)
deidaclin (60)

~
4'
HOHO
,OH
5'

3,OHNC
ON< l' 0
z

5
4
3
......-H

OH
~,OH
4'
HOHO
3
2'
,OHNC
0>0< l' 0
23

5
4
OH

tetraphyllin B (62) volkenin (61)

HOHo
~
4'
,OH

3'
0

OH
l' 0,

NC
>0< 23

1 4
H

080;
5

(a) tetraphyllin B sulfate (69)

HOHo
~
4'
,OH

3,OHNC
0 l' 0 N< 23

1 4
OH

H
5

epivolkenin (65) taraktophyllin (64)

~
,OH 23

HOHo
4,0

3'
2'
OH
0
0 ...
N~H
.....
- OH

HOi'IO~
.'
CH20H
~-./O, /0
~H
0
12 J
4
H

OH 3' OHNC sOH

(h) gynocardin (68) passisuberosin (70)

Fig. 16.13 (a-c). Cyanogens with a cyc1opentenoid ring structure.

 Cyanogenic Glycosides and Cyanolipids 285

!O<
1"0~' '_3 '" O~' '_3
4~~0 >0< ~~HO'"
'"
'" OH 0 .. 0 OH'" OH 00 OH
H0 3" 14 HO" , 14
"
OH 3' OH NC ,H OH 3' OH NC , H

6'·O·rhamnopyranosyJ taraktophyllin (66) 6'·O·rhamnopyranosyl epivolkenin (67)

~ >0<' H~'" >0<' ~

HO
"
CH'OHo 3 3" '''OH 'CH,OH o 3 0 ."

HORO
OH
o
Nt
l'
5
0

H
1"
0

HO
'" HOR~
6" 4'

3' OH
l' 0

NC
1
,
"
0

H
1"
2"
HO
3"

passitrifasciatin (73) passicapsin (12)

~\0II/0, 0.>0<'
3 O~
H~~ 1 4 'O~."
3' OH NC 5 H '''HO

(0) passibiOorln (71)

Fig. 16.13. (continued)

et al., 1987). As the dimeris found only in old dried material, and Spener, 1976). These compounds include deidaclin (60),
it may be an artifact. tetraphyllin A (61), tetraphyllin B (62), volkenin (epitetra-
phyllin B) (63), taraktophyllin (64) (probably same as passi-
CyanoJipids coriacin; see Seigler and Spencer, 1989), epivolkenin (65)
(possibly same as epipassicoriacin), the 6'-O-(a-L-rhamno-
Four cyanolipids occur in the seed oils of plants of the
pyranosides) of epivo1kenin and taraktophyllin (67, 66) (Jar-
Sapindaceae (Fig. 16.2) (Mikolajczak, 1977; Seigler, 1991),
oszewski etal., 1988), gynocardin (68), tetraphyllin B snlfate
although they also have been reported from the Hippocastan-
(69), passisuberosin (70), passibiflorin (71), passicapsin
caeae and Boraginaceae (Ahmad et al., 1978; Osman and
(72), and passitrifasciatin (73).
Ahmad, 1981). These lipids are derived from leucine. In
The structures of four of these compounds have been
two cyanolipids, a long-chain fatty acid is attached to an a-
established by x-ray crystallographic studies. Deidaclin has
hydroxynitri1e. Two other cyanolipids are apparent rear-
(IR)- (60), tetraphyllin A (61) has (IS)-, and tetraphyllin
rangement products and, although derived from the same
B (62) has (IS),(4S)-configuration (Nahrstedt, 1987a). The
precursors, are not cyanogenic.
structure of gynocardin (68) (from Pangium edule) was es-
Two glycosides, 3-[:l-n-glucopyranosyloxy-4-methyl-
tablished by x -ray crystal studies of the p-bromobenzenesul-
2(5H)-furanone (58) and 4-[:l-glucopyranosyloxy-3-hydrox-
fonyl derivative. By other methods, vo1kenin (epitetraphyllin
ymethylbutyronitril-2-ene (59), were isolated from the hem-
B) (63) has been shown to have a (lR),(4R)-configuration
olymph of adults of the bug, Leptocoris iso/ata (Heteropt-
(Jaroszewski and Olafsdottir, 1986; Jaroszewski et al.,
era). Th.( cyanogenic glycoside cardiospennin (54) was
1987a). The configurations of taraktophyllin (passicoria-
found in extracts of whole insects. The larvae contain one
cin?) (64) and epivolkenin (epicoriacin?) (65) also have been
of the glycosides, cardiospennin, and a mixture of cyanoli-
established (Jaroszewski et aI., 1987c).
pids. The frnit of the host plant, Allophylus cobbe (Sapinda-
These compounds occur in the families F1acourtiaceae
ceae) contains a similar mixture of cyanolipids (Braekman
(Spencer and Seigler, 1985b; Jaroszewski and Olafsdottir,
et al., 1982).
1987; Jaroszewski et al., 1988), Turneraceae (Spencer et aI.,
1985c), Passifloraceae (Olafsdottiret al., 1989), Malesherbi-
Cyanogenic mycosides with a
aceae (Spencer and Seigler, 1985a), and Achariaceae (Jensen
Cydopentenoid Ring Sbucture
and Nielsen, 1986). The two largest genera of the Passiflora-
A group of cyanogens that contain a cycIopentenoid ring ceae, Passiflora and Adenia, have different patterns of glyco-
structure (or related structure) are derived from (2-cycIopen- sides. Adenia species have mixtures of pairs of monohydrox-
tenyl)glycine, a nonprotein amino acid (Fig. 16.13) (Cramer ylated glucosides with diastereomeric aglucones [usually
 'DL
186 Cyanogenic Glycosides and Cyano/ipids

&
o
OCR

RO~O a~
" OR NC 4~,
4' 0 3 S

','
" ORRO 7 0 N
I
0
,
N
CR, CR, CR,

acalyphln (1) ricinine malIorepine

Riel"us Mallotus

CX
, N

CH,
CN

nudiftorin ricinidine
Trewio Trewia

Fig. 16.14. Acalyphin and related compounds.

tetraphyllin B (62) and volkenin (63), but in some species, Flacourtiaceae, Malesherbiaceae, Papaveraceae, Passiflora-
epivolkenin (65) and taraktophyllin (64)], and the nonhy- ceae, Poaceae, Proteaceae, Ranunculaceae, Rosaceae, Sapin-
droxylated compounds deidaclin (60) and tetraphyllin A (61) daceae, and Turneraceae. These compounds also are found
(Olafsdottir et ai" 1989). Passiflora species contain a large in bacteria, fungi, and animals.
number of compounds, including cyclopentenoid glycosides There is much variation from plant to plant with regard
as well as those derived from valine and isoleucine. Among to content of cyanogenic compounds and the f3-glycosidases
these compounds are deidaclin (60), tetraphyllin A (61), tet- necessary to hydrolyze them. In a number of taxa, only
raphyllin B (62), volkenin (63), epivolkenin (65), taraklo- 1-3% of the individuals give positive cyanide tests done by
phyllin (64), passibiflorin (71), passicapsin (72), and passi- the Feigl-Anger method. In others, virtually all individuals
trifasciatin (73) (Olafsdonir et aI., 1989). The presence of are cyanogenic (Aikman et aI., 1996).
cyclopentenoid cyanogenic glycosides has been reported In some genera, almost all species tested are cyanogenic.
from the Caricaceae but should be confirmed (Spencer and For example, few species of the genera Prunus (Rosaceae)
Seigler, 1984). or Passiflora (passifloraceae) are not cyanogenic. In other
families and genera, only a single species is cyanogenic.
Cyanogenic Compounds of Other Origins Further, there may be pronounced variation during the grow-
ing season and in the development sequences of the plant
The compound acalypbin (1) from Acalypha indica (Eu- (Hegnauer, 1986).
phorbiaceae) appears to be derived from nicotinic acid me- At present, ferns are known only to produce phenylala-
tabolism (Fig. 16.14) (Nahrstedt, 1987a). nine-derived cyanogens such as prunasin (19) and vicianin
(25). Gymnosperms produce only taxiphyllin (16) (derived
from tyrosine). The Magnoliales and their close relatives
DlSTRlBUI101'1 primarily produce cyanogens derived from tyrosine, as do
some monocotyledonous plants. There are several excep-
Cyanogenic compounds are found in all major plant groups tions to this generality, however (Nahrstedt, 1987a).
including ferns, gymnosperms, and both monocotyledonous The Euphorbiaceae consists offive subfamilies (Webster,
and dicotyledonous angiosperms. Cyanogenic compounds 1975). In the Phyllanthoideae, cyanogens are derived pri-
occur in all major subgroups of dicotyledonous angiosperms. marily from tyrosine, and, in the Crotonoideae, these com-
The distribution of cyanogenic compounds in plants has been pounds are derived from valine/isoleucine. In the Acaly-
reviewed (Saupe, 1981; Seigler, 1977, 1981a, 1981b). Fami- phoideae, acalyphin (1) appears to be closely related to the
lies in which cyanogenesis is most common are the Araceae, series of four noncyanogenic 3-cyanopyridones isolated
Asteraceae, Euphorbiaceae, Fabaceae (Seigler et aI., 1989), from this group of plants (Fig. 16.14) (Nahrstedt, 1987a).
 Cyanogenic Glycosides and Cyano/ipids 287

Although it has been thought previously that the Rosaceae Davallia trichomanoides and Vicia angustifolia hydrolyze
is relatively homogeneous with regard to cyanogenesis and (R)-vicianin (25) and (R)-amygdalin (20) at the agly-
that the major compounds present are derived from phenylal- cone-disaccharide bond, whereas the enzymes of Prunus
anine, this now appears to be true only for the subfamilies serotina and Linum usitatissimum involve stepwise removal
Maloideae and Amygdaloideae. Other members of the fam- of the sugars (Poulton, 1988).
ily contain compounds such as the p-hydroxybenzoate (55) In some instances, when an unnatural enzyme is added
and p-hydroxycinnamate (74) of (S)-cardiospermin (Fig. to a substrate and the natural enzyme then added, no reaction
16.12) and dhurrin (15). is observed. The nature of these effects is dependent on con-
The Fabaceae also are heterogeneous with regard to the centrations of enzymes and substrates as well as the order
origin of cyanogenic glycosides (Nahrstedt, 1987a; Seigler of addition. Interactions of this type for cyclopentenoid cy-
et al., 1989). The genus Acacia contains cyanogens derived anogenic glycosides and the corresponding enzyme system
from the valine/isoleucine, leucine, and phenylalanine path- have been reviewed (Spencer, 1987).
ways. Phenylalanine-derived cyanogens are found mostly in
Australian acacias of subgenus Heterophyllum and subgenus
Aculeiferum. Valine/isoleucine and leucine-derived com- BIOLOGICAL ACTIVITY
pounds are primarily found in members of subgenus Acacia.
Members of the Flacourtiaceae, Passifloraceae, Turn- Cyanogenic compounds sometimes represent a sizable pro-
eraceae, Malesherbiaceae, and Achariaceae, contain cyano- portion of the plants total nitrogen and, in some cases, this
genic compounds with cyclopentenoid rings. In many plants, nitrogen becomes available upon germination of seeds as the
these cyanogens occur as complex mixtures often consisting cyanogenic compounds are converted into other metabolites.
of epimeric, enantiomeric, and diastereomeric mixtures of For example, the cyanolipids of Ungnadia speciosa (Sapin-
compounds, a situation uncommon with most types of sec- daceae) disappear rapidly upon germination of the seeds
ondary compound. In some members of these groups, how- (Selmar et al., 1990). In other instances, however, the cyano-
ever, other types of cyanogen also occur. Valine/isoleucine genic compounds are transferred to other parts of the plant.
and cyclopentenyl glycosides co-occur in some Passiflora Linamarin (11) in lima beans (Phaseolus lunatus) appears
species (Spencer and Seigler, 1985e) and one isoleucine de- to be transferred intact to the growing seedling (Clegg et aI.,
rived cyanogen occurs in at least one member of the Flacour- 1979).
tiaceae (Jaroszewski et al., 1987b). The immature fruits of
Passiflora edulis contain prunasin (19), derived frum phe- Transport of Cyanogenic Glycosides
nylalanine (Spencer and Seigler, 1983). Leaves of Carica
Over 90% of the cyanogenic compounds of the seeds of
papaya have been reported to contain prunasin (19) and
Hevea brasiliensis (primarily linamarin) are found in endos-
cyclopentenyl glycosides (Spencer and Seigler, 1984). I have
perm tissue (Selmar et al., 1988). The major glycoside was
confirmed the presence of prunasin. This is of special inter-
linamarin (11). After storage, linustatin (44) and neolinus-
est, as the plant also contains benzyl glucosinolate and repre-
tatin (45) also were detectable (Selmar et al., 1991). During
sents one of the few plants to contain cyanogenic glycosides
germination of the seeds and development of the seedlings,
and glucosinolates (Nahrstedt, 1987a).
most of these compounds are converted into noncyanogenic
Cyanogenesis in the Asteraceae also proves to be com-
materials. No gaseous hydrogen cyanide is liberated during
plex. The pairs of epimers prunasin (19) and sambunigrin
the process. The !'I-glucosidase capable of cleaving the glu-
(22), and holocalin (35) and zierin (34) are present in most
coside [principally linamarin, although some species of
subfamilies. In the Calendulae, linamarin (11) and lotaus-
Hevea contain small amounts of lotaustralin (42)] is wide-
tralin (42) predominate. Recently, amygdalin (20), vicianin
spread in plant tissues. The greatest enzyme activity of !'I-
(25), and prunasin (19) were isolared from the below ground
cyanoalanine synthase is found in young seedling tissues.
parts of Gerbera jamesonii hybrida (Nahrstedt, 1987a).
Thus, it appears that linamarin is transferred to the young
seedling tissues and metabolized there (Lieberei et al., 1985;
Selmar, 1993). An a-hydroxynitrile lyase also has been iso-
(l-GLYCOSIDASIlS lated from Hevea brasiliensis leaves. The presence of this
enzyme greatly accelerates release of hydrogen cyanide (Sel-
In most plants, cyanogenic glycosides are accompanied by mar et al., 1989).
a corresponding !'I-glycosidase. One such enzyme system, Linamarin (11) is converted into the diglycoside linu-
emulsin, from almond seeds, has fairly broad specificity and statin (44) before transport from the endospermous tissues
has been widely studied. Most of these enzymes, however, (Selmar et al., 1988). Linustatin is not hydrolyzed by the
have more specific substrate requitements. The specificity of same !'I-glucosidase that hydrolyzes linamarin. This diglyco-
enzymes from plants containing cyclopentenoid cyanogenic side is then transported to the seedling tissues where it is
glycosides has been reviewed (Spencer, 1987). hydrolyzed by a special !'I-glycosidase. The cyanide liberated
The !'I-glycosidases from plants containing diglycosides is utilized in a manner similar to that in many other plant
also can differ in activity. For example, the enzymes from tissues (Selmar et al., 1988). Thus, in this example, glucosy-
288 Cyanogenic Glycosides and Cyano/ipids

lation is an essential step for the transport of cyanogenic ally related nitriles are responsible for the susceptibility of
glycosides from the seed to the site in the developing plant Hordeum vulgare to the pest Erysiphe gramini (Nahrstedt,
where they are metabolized. 1992; Pourmohseni et al., 1993). These glycosides are im-
Transport of linamarin (11) from the endosperm to the portant in the plant epidermis of compatible lines (those that
cotyledons of Linum usitatissimum, flax, following germina- suffer attack), whereas incompatible lines lack these com-
tion has been observed. At the stage that the embryo is small, pounds (Nahrstedt, 1992; Pourmohseni et al., 1993).
almost all of the cyanogenic glycosides present are as lina- Purified dhurrin from sorghum seedlings was shown not
marin and lotaustralin (42); after 1 week, only the diglyco- to be toxic to Locusta migratoria. The amount of cyanide
sides linustatin (44) and neolinustatin (45) are present in released was an effective plant defense and probably ac-
the large cotyledons. As apoplastic linamarase is present, counts for most of the lack of palatibility of field-grown
linustatin is a probable transport form (Selmar, 1993). sorghum to acridids in West Africa and India (Bernays,
An apoplastic enzyme that rapidly hydrolyzes prunasin 1983). The mechanism of effectiveness of cyanide in many
(19) is found in Prunus seeds; this enzyme is incapable of plants appears to be related to the release rates during chew-
breaking down amygdalin (Poulton, 1988). However, it is ing and not to the levels of cyanogenic glycosides present
not established that this apoplastic enzyme is specific for (Bernays, 1983). Other work by this group indicates that
prunasin. Many apoplastic J3-glycosidases may be related to p-hydroxybenzaldehyde may be involved in acceptance of
the formation of lignin (Selmar, 1993). In a manner similar sorghum plants by larvae (Nahrstedt, 1985, 1992).
to flax, the very young fruits contain only prunasin, but, after Several cyanogenic glycosides serve as phagostimulants.
development, the cotyledons contain amygdalin (20). Again, Amygdalin (20) serves as a phagostimulant for Malacosoma
it is probable that amygdalin is a transport form of cyano- americana, linamarin (11) and lotaustralin (42) from
genic glycosides in this plant. Phaseolus vulgaris for Epilachna varivestis (this report
should be confirmed, as cyanide is rarely found in plants of
Cyanogenic Compounds as Defensive Phaseolus vulgaris) (Nayar and Fraenkel, 1963), and dhurrin
Substances (15) from sorghum for Peregrinis maidis (Bernays, 1983).
Cyanogenic glycosides and their decomposition products However, in a study on highly cyanogenic black cherry (Pru-
almost certainly playa role as defensive substances in plants nus seratina), insect feeding was only extensive when the
(Nahrstedt, 1992). However, the exact nature of these inter- level of glycosides dropped well below the maximum
actions is complex. Hruska (1988) and Seigler (1991) (in (Schroeder, 1978). Prunasin (19) from the peach, Prunus
an improperly strong manner) have criticized many of the persica, acted as an antifeedant for the oblique banded leaf
classical papers concerning this role. Although many of roller (Nahrstedt, 1985).
these studies should be reexamined in view of newer con- The moth Yponomeuta evonymellus does not respond be-
cepts and with improved statistics, there is little question haviorally to prunasin (19) which occurs in its host plant.
that cyanogenic compounds and their decomposition prod- Other species of this genus, however, are both sensitive ane
ucts can play such a role. deterred (Stadler, 1984).
One role of cyanogenic glycosides in plants is related to In Hevea brasiliensis (Euphorbiaceae, natural rubber)
their ability to produce toxic amounts of hydrogen cyanide (and other Hevea species), mechanical injury leads to evolu-
(Conn, 1981; Nahrstedt, 1985, 1992; Poulton, 1983), which tion of hydrogen cyanide. Infection with the fungal pathogen
is extremely toxic to most organisms because it inhibits cyto- Microcyclus ulei leads to the production of a phytoalexin,
chrome oxidase and other respiratory enzymes. scopoletin (Giesemann et al., 1986) (see Chapter 8). This
Many plants that contain cyanogenic glycosides are well attack also results in a decrease of cyanogenic compounds,
known from poisonous plant literature (see, for example, and hydrogen cyanide can be detected in the leaves. In sus-
Kingsbury, 1964; Nahrstedt, 1985; Poulton, 1983). Although ceptible clones of the plant, the amount of cyanide liberated
these effects may be due to cyanide [or unrelated compounds is much greater than in resistant clones. The increased
in the plants (e.g., in Taxus species)], both cyanide and agly- amount of cyanide produced in highly cyanogenic plants
cones (aldehydes or ketones) resulting frum hydrolysis of appears to result in inhibition of the formation of scopoletin.
these compounds can be toxic to nonadapted animals, fungi, In this instance, cyanogenesis does not lead to greater, but
and other organisms (Davis, 1991). Despite this toxicity, reduced, resistance to the pathogen (Lieberei, 1986, 1988;
most animals have the ability to detoxify limited amounts Lieberei et al., 1989). In general, lines of Hevea brasiliensis
of cyanide. Intact cyanogenic glycosides do not appear to with both large amounts of cyanogenic glycosides and 13-
be highly protective against insects; these animals often are glucosidases are the most susceptible to the South American
effective in detoxification of cyanide (Bernays, 1983; leaf blight (Microcyclus ulei) (Davis, 1991; Lieberei, 1988;
Hruska, 1988). Although it has been suggested that cyano- Selmar, 1993).
genic glycosides serve as deterrents to herbivory (Jones, Intact cyanogenic glycosides appear to be relatively non-
1988; Nahrstedt, 1985), the glycosides themselves seem to toxic to mammals when injected. These compounds are ex-
be relatively nontoxic to most organisms. creted rapidly via the urine (Nahrstedt, 1987a); they are not
The presence of epiheterodendrin and a series of structur- especially toxic when taken orally unless the corresponding
 Cyanogenic Glycosides and CyanoJipids 289

/3-glycosidase is consumed at the same time (Nahrstedt, phora geniculata (Hymenoptera: Tenthredinidae), which
1987a; Poulton, 1983). feeds on Sorbus (Rosaceae), contains benzaldehyde and
Both cyanogenic and acyanogenic phenotypes of Lotus mandelonitrile (Duffield et al., 1990).
corniculatus occur. Cyanogenic forms contain linamarin The larvae of Zygaena species contain linamarin (11) and
(11) and lotaustralin (42), whereas acyanogenic plants may lotaustralin (42). These insects feed on Lotus corniculatus
lack either the cyanogenic glycosides, the /3-glucosidase, or (which contains linamarin and lotaustralin). The moths ap-
both. In general, snails preferentially graze on acyanogenic pear to synthesize the cyanogenic glycosides de novo and
forms (Jones, 1972, 1988). However, the relationship be- synthesize them even when the moths are fed on an artificial
tween cyanogenesis and herbivory by slugs on Lotus corni- diet. All stages of the insects appear to be able to synthesize
culatus and Trifolium repens appears to be complex (Dirzo these two glycosides (Nahrstedt, 1987b, 1992). Zygaena lar-
and Harper, 1982; Horrill and Richards, 1986; Kakes, 1987). vae also contain both /3-glucosidases and a-hydroxynitrile
In Lotus corniculatus, the presence of tannins (also consid- lyases needed to release cyanide from the glycosides (Miiller
ered to have antiherbivore properties) is strongly negatively and Nahrstedt, 1990).
correlated with the presence of cyanide (Ross and Jones, Nahrstedt has shown that moths of the genus Acraea and
1983). Heliconius also contain these two glycosides even when fed
Cyanolipids (Fig. 16.2) have been shown to be toxic in on species of Passiflora that contain cyclopentenoid cyano-
the diet of Callosobruchus maculatus at both the 1% and gens (Davis, 1991; Nahrstedt and Davis, 1981; Nahrstedt,
5% level; they do not appear to be particularly toxic to house- 1987a, 1987b). However, all stages of Acraea horta, which
flies. feeds on the flacourtiaceous plant Kiggelaria africana, con-
tain gynocardin (68)-the cyanogenic glycoside produced
Cyanogenesis, Ants, and the Genus Acacia by the host plant (Raubenheimer, 1989).
The larvae and imagines of Leptocoris isolata (Hemipt-
Among the American species of Acacia, a number are era) feed on fruits of Allophylus cobbe that contain cyanoli-
characteristically inhabited by ants. Among these are Acacia pids. These insects contain cardiospermin (54) as well as an
cornigera, A. hindsii, and A. chiapensis. The ants live in isomeric nitrile (Braekman et al., 1982).
swollen stipular spines and eat foliar nectaries and special
structures known as Beltian bodies that are born at the tips
Biological Activity of Cyanide in Plants
of the leaves. The ants receive most of their nutrition in this
manner, as well as provide the plant with protection against Small amounts of cyanide reversibly inhibit nitrate re-
insect, plant, and other animal infringement. Many of these ductase in plants. Furtbermore, in experiments in which mea-
Acacia species cannot survive without the ants. Most of the surements of this inhibition was used as a bioassay, it was
ants (usually of the genus Pseudomyrmex) cannot survive possible to demonstrate the presence of very small amounts
in the absence of host Acacia plants. Leaves of A. chiapensis of cyanide in a number of plants not previously known to
and A. tarnesiana proved toxic to southern army worms, have cyanide (Vennesland et aI., 1981).
Prodenia eridania, raised on Phaseolus vulgaris, whereas Because 1 mole of cyanide is produced for each mole
those of A. cornigera did not. Amygdalin (20) added to the of ethylene generated from ACC (l-amino-cyclopropane-1-
diet was also toxic. No alkaloids were present in the leaves carboxylic acid) (see Chapter 13), all plants must contain
of these tbree species, Rehr et al. (1973) concluded that non- small amounts of cyanide (Peiser et al., 1984). However, due
ant Acacia species produce cyanogenic defense compounds, to a concomitant increase in the amount of /3-cyanoalanine
whereas ant acacias do not. They reckoned A. chiapensis synthase, the ability to detoxify cyanide in the tissues is
(which produces cyanogenic glycosides) to be a case margin- much higher (Manning, 1988).
ally adapted to ant mutualisms. More recent evidence, how- Flowering in the short-day plant, Lemna paucicostata,
ever, shows that three common ant acacias, Acacia hinds;;, could be induced under continuous light by introduction of
A. globulifera, and A. chiapensis, are often strongly cyano- 2-5 iJM cyanide into culture media (Tanaka et al., 1983).
genic (Seigler and Ebinger, 1987).
Detoxication of Cyanide
Cyanogenesis in JlliIUpedes and Insects
Plants convert HCN into asparagine (75) and /3-cyano-
Compounds capable of releasing cyanide are found in alanine (76) (Fig. 16.15). When HI'CN is introduced in
a number of millipedes and insects (Duffey et aI., 1977; plants, the label is incorporated into the amide carbon of
Nahrstedt, 1985, 1988). Both millipedes and beetles of the asparagine (Davis, 1991). This suggests that the cyanogenic
Cbrysomelidae and Cicindelinae possess the ability to pro- compounds are being synthesized and catabolized, and the
duce mandelonitrile (Nahrstedt, 1988). Interestingly, one of HCN produced in the intact plant is subsequently incorpo-
these, Paropsis atomaris, contains both (R)-mandelonitrile, rated into asparagine by the action of /3-cyanoalanine syn-
a small amount of pronasin (19), and a /3-glucosidase capable thase and /3-cyanoalanine hydrolase. /3-Cyanoalanine syn-
of splitting the pronasin (Nahrstedt, 1988). thase has been shown to occur in many plants (Conn, 1981;
The larval secretion of the mountain ash sawfly, Prisot- Harborne, 1989; Nahrstedt, 1992).
290 Cyanogenic Glycosides and Cyanolipids

NH, cyanohydrin is decomposed and any free cyanide remaining

HCN H,S is eliminated (Nahrstedt, 1993).

--
~ONH, ~OOH Leaves of Manihot esculenta often contain 1-2% Iina-

- H,O ,Hz
,HNH, -

COOH
HzO ~Hz
,HNH,

COOH
marin (11) and 10taustraIin (42). Although leaves with low
cyanide-releasing ability are preferred, the acceptability of
cassava leaves for the herbivore Zonocereus variegatus de-
pends on both the physiological state of the leaves and the
insect larva, and whether other foods are available to them
cysteine 3-cyanoalanlne (76) asparagine aspartJcacld (Bernays et aI., 1977).
(75) (79) Cattle and other livestock are frequently poisoned when
allowed to eat Sorghum seedlings or material that has been
Fig. 16.15. Conversion of HCN to ~-cyanoalanine and asparagine
(modified from Conn, 1979b; used with pennission of the copyright stressed (by drought, inundation, frost, etc.) and allowed to
owner, Academic Press, Orlando, FL). resume growth (Boyd et aI., 1938; Willaman and West,
1916).

Most organisms have the ability to detoxify cyanide. One

system that appears to be common in mammals involves the
enzyme rhodanese (thiosulfate sulfur transferase) and at least NlTROOLYCOSIDES
two other sulfur-containing enzymes (Davis, 1991). This en-
zyme catalyzes the reaction of cyanide and thiosulfate to Nitro acids, alcohols, and their glycosides have been re-
produce thiocyanate and sulfite. In animal tissues, the sulfur ported from plants in a number of plant families (Seigler,
of cysteine is utilized. The thiocyanate generated is elimi- 1991). There are two main types: those derived from aro-
nated slowly and is responsible for many of the chronic ef- matic precursors and aliphatic precursors.
fects of cyanide poisoning (Conn, 1979a; Davis, 1991). The pathways to cyanogenic glycosides, nitro com-
Many other organisms contain the enzyme (3-cyanoalanine pounds, and glucosinolates in higher plants seem to be
synthase which converts hydrogen cyanide and cysteine or closely linked (Conn, 1988; Hosel et aI., 1985) (also see
serine into (3-cyanoalanine (Nahrstedt, 1985). Many fungi Chapter 17). For example, (3-phenylnitroethane (77) has
convert hydrogen cyanide and water into formamide, which been reported as the peppery and fragrant principle of edible
is relatively nontoxic (Hanis et aI., 1987). Reaction with fruits of Dennettia tripetala (Anoonaceae) (Okogun and
nitrogenase enzyme also can account for the removal of Ekong, 1969). Cell suspension cultores of Eschscholtzia cal-
small amounts of hydrogen cyanide in many plants. A num- ifornica produce low levels of HCN, ptesumably due to cy-
ber of bacteria contain a "cyanide oxygenase" system capa- anogenic glycosides. However, the source of the HCN does
ble of converting hydrogen cyanide into carbon dioxide and not appear to be triglochinin (17) or dhurrin (15) which are
ammonia (Hanis et aI., 1987). ptoduced by the intact plant. Additionally, microsomal frac-
tions from stressed cell cultores of this plant catalyze the
formation of 1-(4'-hydroxyphenyl)-2-nitroethane (8) from L-
CYAI'IOOEI'IESIS 11'1 CROP PLANTS tyrosine (2) (Fig. 16.16). A glucosyltransferase that glucosy-
lates the compound is also found in these cultores.
Many plants of economic importance are cyanogenic: ap- A number of esters of 3-nitropropionic acid (78) occur
ples, peaches, almonds, apricots, plums, cherries, macada- in the genera Astragalus, Coronilla, Indigo/era (Fabaceae),
mia nuts, forage grasses, corn, sorghum, bamboo shoots, Heteropteris, Hiptage (Malpighiaceae), Viola (Violaceae),
cassava, sugar cane, flax and lima beans. Of these, most and Corynocarpus (Corynocarpaceae) (Fig. 16.17).
human poisoning problems arise from cassava and, proba- 3-Nitroptopionic acid (78) is derived from aspartic acid
bly, the majority of animal poisoning cases from Sorghum (Fig. 16.15) in the fungus Penicillium atrovenetum, but both
species (Davis, 1991). oxygen atoms of the nitro group are derived from molecular
Cassava, Manihot esculenta, is the major starchy food for oxygen (Fig. 16.17) (Baxter and Greenwood, 1986). Further,
over 300 million people in many tropical countries of the the carbon-nitrogen bond of aspartic acid is preserved dur-
world. Plants with 20 mg HCNIl 00 g fresh weight are usu- ing the formation of 3-nitropropionic acid (Baxter et aI.,
ally considered toxic (Kingsbury, 1964). Many cultivars of 1985). In Lathyrus sativa, however, asparagine (75) gave the
cassava exceed this value. Methods of detoxication have highest rate of incorporation into the isoxazolinone moiety
been developed by the American Indians who consume the of the heterocyclic amino acid (3-(isoxazolin-5-on-2-yl)-L-
tuberous root of this plant. These involve grating or grinding alanine (Randoux et aI., 1991).
the plant, pressing the ground material to remove liquid con- Other compounds [e.g., miserotoxin (80)] involve 3-ni-
taining hydrogen cyanide and glycosides (and inadvertently tropropanol moieties. These compounds are poisonous to
mixing the (3-glucosidase and substrate), storing the product, livestock (Stermitz and Yost, 1978) and to several insects
and cooking. If the ptoduct is pressed and stored for a period and other animals (Byers et aI., 1986). 3-Nitropropionic acid
of time (often as long as overnight) and then cooked, the (78) is toxic to the hrochid Callosobruchus maculatus at
 Cyanogenic Glycosides and Cyanolipids 291

4'~NO
~l '
HO~NO'
5' 6' 5' 6'

1-(4'hydroxypbenyl)·2-nitroethane (8)
I-phenyl-2-nitroethane (77)
OH

HO~O~"
O~ IA ,
~",,,
OH
- ( J - - ! ' ,"
,'" OH
HO
HO 3"
0

OR
~
5' -
NO,
HOHO
5,,00 f' NO
_ I 2
3" OR 5'

1-(4'hydroxyphenyl)·2-nitroethane 1-(4'hydroxyphenyl)·2·nitroethane
primeveroside
glucoside

Fig. 16.16. Aromatic nitro compounds.

0.1 % when incorporated into artificial diets (Janzen et a!., tance of the root to the grass grab Costelytra zealandica
1977). However, another insect, Sparganothis sulfureana, (Harborne, 1989).
feeds on Coronilla varia, crownvetch, a plant containing 3- The eggs of many species of chrysomelid beetles contain
nitropropionate derivatives, and seems to be relatively insen- isoxazolinone glucosides. One of these compounds which
sitive to the effects of these compounds. Hiptagin (81) and contains a nitropropionate residue is highly deterrent to feed-
three other nitro derivatives of glucose occur in the root of ing of ants on the eggs (Fig. 16.18) (Pasteels et aI., 1986).
Lotus pedunculatus and appear to be responsible for resis- The defensive glands of other chrysomelid beetles possess

HO~NO' HO~NO,

3·nitropropionic acid (78) 3-nitro-l-propanol

o
o II ~NO, 0 0

~
---.[J./ '-../ O~NO, O~NO,

HOHO 0 OH HO Ao.. ( - - - ; 0 HO Ao.. ( - : OH

~O~~ ;O~
NO-,~O OH J ~NO, OH

o
coronarian cibarian 6~(3-nitropropanoyl)-j3~D-glucopyranose

o
O~NO,

NoV'rr-~
o
HO
0 OH

o OH

(a) 4,6-di(3-nitropropanoyl)-a-D-glucopyranose 4,6-di(3~nitropropanoyl)-j3-D-glucopyranose

Fig. 16.17 (a-c). Esters of 3-nitropropionic acid from plants.

292 Cyanogemc
. G/ycosides and Cyanolipids

~NO, ~O~NO'
o
~ o
0 0

O~NO'NO-'~O
' g O~NO'
HO H

HOHO 0
NO.2~O
o

karakin coronillin

corollin corynocarpin

o
O~NO,
~/'..'-../~NO, ./',.
o -....v 0
II ~
~~N00
0

./',.
o
II
NO, ~OO
r--./
H

~ ;O~ \\~
0 "'- ( - -;:- 0, /NO, 00H
NO, If
NO,~ o 0 NO 0 0
o ,

hiptagin (81) 2 3 4 6-tetra(3-nltropropanoyl)-

(b)
, , , a-D-glucopyranose

~
.' ORo
80R O "
0,

'V 'v"
' l
A ,NO, rr:o, ,. o~. A
HO~.
,. OR 0 0' 'V
•
A
'No,
• OR HORO " • OR
mfseroIoxIb{88)

Fig. 16.17. (continued)

 Cyanogenic G/ycosides and Cyanolipids 293

legumes such as the soy bean, Glycine max, the winged bean,
Psophocarpus tetragonolobus, and Erythrina species are
known to produce nitric oxide, nitrous oxide, and possibly
other nitrogen oxides (Betting, 1909; Dean and Harper,
1986; Weehuisen, 1907).
(82) (83)

l'IITRILE GLYCOSIDES

Fig. 16.18. lsoxazolinone glucosides from chrysomelid beetle eggs and Another series of noncyanogenic nitrile glycosides, with
defensive glands. structural similarities to intermediates in cyanogenic glyco-
side biosynthesis are found in members of the AquifoJiaceae,
defensive glands that contain d '-isoxazolin-5-one gluco- Boraginaceae, Crassulaceae, Menispermaceae, Fabaceae,
sides 1 (82) and 2 (83). The first of these compounds also Ranunculaceae, and Simmondsiaceae (Fig. 16.19) (Seigler,
is known from Pisurn sativum and Lathyrus odoratus (Ran- 1991).
doux et al., 1991). In the plants, d 3-isoxazolin-5-one and
UDP-glucose were precursors of the fIrst glucoside,
whereas, in the insect, labeled aspartic acid was incorporated PSEUDOCYAl'IOGEI'IIC GLYCOSIDES
into both glucosides (Randoux et al., 1991).
Both picrate and Feigl-Anger paper may give positive Glycosides based on a methyl azoxymethanol structure
tests when 3-nitropropionic acid and related compounds are occur in cycads. These glycosides are found in most mem-
present (Seigler et al., 1989). bers of this gymnospermous family, along with an enzyme
Although the origin is not known with certainty, several that effects hydrolysis of the compounds to release methyl

8HC5I:~O~:
I
H~'CN
8,
I
0 HO"
~OH
" ~OH
OH

7 • 5
H " OH
7
I : 6
H
....""'H

menisdaurin dasycarponin
H- OH H OH OH

H~'CN
8H~I'~O~:
5' OH
I O~O~OH HO~
I : I :
8 3
H :&' OH H l' OR
7 Ii """H 7 Ii ""'H

>fl
OH bauhinin H OH OH lithospennoside
(a) H OCH,

'CN~,
H~'CN
I~
'S , OH
HO
H ,
I,
5' OH
0 0 OH
o 0 OH HO
HO
I
8 3
/8 4 "OH
~ H " OH H 7 5 H
7 {; \: H CH,O •
H
OH H OCH,
H OH

(b) llex warburgii glucoside simmondsin

Fig. 16.19 (a & b). Nitrile glycosides.

294 Cyanogenic Glycosides and Cyanolipids

HO
-_\ .-0. O/N~N/CH3
HO~~
HO OH tI
o
cycasin (84)

Fig. 16.20. Pseudocyanogenic glycosides.

azoxymethanol and glucose (or other sugars). Normally, this These qualitative methods are very useful for screening
group of glycosides does not produce cyanide. Only when large numbers of samples because they are simple, fairly
treated with base do pseudocyanogenic glycosides liberate specific, and require no equipment other than vials and corks.
nitrogen, formic acid, HCN, and glucose. These qualitative methods also are useful for monitoring
Several of these plants are used as human food sources separations of cyanogenic compounds from TLC plates or
in Oceania and many are eaten by animals in many areas of chromatography paper during isolation and purification.
the South Pacific and in Australia. A form of amyotrophic Methods for the quantification of cyanide in plant tissues
lateral sclerosis (ALS or Lou Gehrig's disease) used to be involve hydrolysis of the cyanogenic glycosides and release
endemic among the Chamorros of Guam and is linked to of HCN, which is trapped in a basic solution and determined
the presence of an unusual amino acid ,,-amino-i3-propionic colorimetrically (Brinker and Seigler, 1989) by the method
acid or i3-N-methylarnino-L-alanine found in this cycad of Lambert et al. (1975) which can detect as little as 5 J.l.g
(Fowler, 1987; Spencer et al., 1987) (see Chapter 13). HCN (Lambert et al., 1975).
Methyl azoxymethanol has been shown to be carcinogenic Methods for the quantification of particular cyanogenic
(Hirono, 1972). Cycasin (84) reduces the litter size in rats glycosides have been reviewed (Brimer, 1988; Brimer and
if consumed prior to mating (Fig. 16.20). Dalgaard, 1984; Nahrstedt et al., 1981).
The hairstreak butterfly, Eumaeus atala, endemic to Flor- Emulsin, the i3-glycosidase from almonds, has been used
ida, has brilliant waming coloration. The larvae are red and for many qualitative and quantitative cyanide determina-
yellow, the adults red and black. This insect is a specialist tions. This commercially available enzyme will hydrolyze
on the cycad Zamia floridalUl, the young leaves of which several cyanogenic compounds (some very slowly), al-
contain 0.2% cycasin by dry weight, and the insect accumu- though it is inactive against others (Conn, 1993). Snail or
lates cycasin (Harborne, 1989). snail gut acetone powder has also been used to hydrolyze
many cyanogenic glycosides (Brimer et al., 1983). This com-
plex mixture of enzymes (i3-glucosidase, i3-glucuronidase,
sulfatase, and i3-D-mannosidase) is also commercially avail-
A1'IALl'11CAL TECIII'IIQUES
able. This enzyme is active against a broad spectrum of cy-
anogenic glycosides. In contrast, most other i3-glycosidases
For qualitative (presence-absence) or semiquantitative deter- have relatively specific substrate requirements (Fan and
minations of cyanide, the easiest and most reliable method Conn, 1985; Gross et al., 1982; Hosel and Conn, 1982; Hosel
is a simple test with either Feigl-Anger or sodium picrate and Nahrstedt, 1975; Hosel et al., 1987; Spencer, 1987).
paper. The reactions on which these tests are based as well
as other methods for detection of cyanogenic glycosides are
described by Hegnauer (1986).
REFERENCES
Sodium picrate paper is prepared by dipping filter paper
into an aqueous solution of 0.5% picric acid and 5% AHMAD, I., A. A. ANSARI, and S. M. OSMAN, Cyanolipids of Boragi-
Na2C03, then allowing the paper to dry. The paper may be naceac seed oils, Chern. Ind., 61-63 (1978).
stored dry but should be moistened just before use (Guig-
AlKMAN, K., D. BERGMAN, J. E. EBINGER, and D. SEIGLER, Cyano-
nard, 1906a,b; Horwitz, 1980; Kingsbury, 1964). The detec- genic variation in some Midwestern U.S. plant species, Bio-
tion limit of picrate paper with plant samples is chern. Sy,t. Bcol., 24, 637-645 (1996).
0.001-0.002% HCN by weight, which corresponds to about ARrroMl, M. T. KUMOR!, and T. KAWASAKI, Cyanogenic glycosides
0.01-0.02% prunasin (Brinker and Seigler, 1989; Hegnauer, in leaves of Perillafrutescens var. acuta, Phytochemistry, 24,
1986). 2438-2439 (1985).
Feigl-Anger paper is made by mixing equal volumes of BAXTER, R. L. and S. L. GREENWOOD, Application of the "a iso-
solutions of tetrabase [4,4'-tetramethyldiaminodiphenyl- tope shift in ISN NMR spectra to a biosynthetic problem: Experi-
methanel and copper ethylacetoacetate (Brinker and Seigler, mental evidence for the origin of the nitro group oxygen atoms
1989, 1992; Tantisewie et al., 1969). Feigl-Anger paper is of 3-nitropropionic acid, J. Chem. Soc., Chern. Commun.,
somewhat more sensitive than picrate paper (Nahrstedt, 175-176 (1986).
1980). BAXTER, R. L., E. M. ABBor, S. L. GREENWOOD, and I. J. McFAR-
 Cyanogenic GJycosides and Cyano/ipids 295

LANE, Conservation of the carbon-nitrogen bond of aspartic Biochemistry, Biochemistry of Nutrition lA, Vo!' 27 (A Neu-
acid in the biosynthesis of 3-nitropropionic acid, 1. Chern. Soc., berger and T. H. Jukes, eds.), 21-43, University Park Press,
Chern. Commun., 564-566 (1985). Baltimore, MD, 1979a.
BERNAYS, E. A., Nitrogen in defence against insects, in Nitrogen CONN, E. E., Cyanide and cyanogenic glycosides, in Herbivores
as an Ecological Factor (J. A Lee, S. McNeill, and I. H. Rorison, (G. A. Rosenthal and D. H. Janzen, eds.), 387-412, Academic
eds.), 321-344, Blackwell, Oxford, 1983. Press, New York, 1979b.
BERNAYS,E. A, R. F. CHAPMAN,E. M. LEATHER, and A R. McCAF- CONN, E. E., Cyanogenic glycosides, in Secondary Plant Products
FERY, The relationship of Zonocerus variegatus with cassava, (E. E. Conn, ed.), Vol. 7 of The Biochemistry of Plants (P. K.
Bull. Ent. Res., 67, 391-404 (1977). Stumpf and E. E. Conn, eds.), 479-500, Academic Press, New
BE'ITING, M., Mededeeling omtrent genomen proeven met York,1981.
Erythrina L., Pharm. Weekbl., 46, 1089-1092 (1909). CONN, E. E., Cyanogenic glycosides: A possible model for the
BoYD, F. T., O. S. AAMoD'l', G. BOHSTEDT, and E. ThUDO, Sudan biosynthesis of natural products, in The New Frontiers in Plant
grass management for control of cyanide poisoning, 1. Am. Soc. Biochemistry (T. Akazawa, T. Ashai, and H. Imaseki, eds.),
Agron., 30, 569-582 (1938). 11-22, Japan Scientific Societies Press, Tokyo, 1983.
BRAEKMAN, J. c., D. DALoZE, and J. M. PASTEELS, Cyanogenic CONN, E. E., Biosynthetic relationship among cyanogenic glyco-
and other glucosides in a Neo~Guinean bug Leptocoris iso/ata: sides, glucosinolates, and nitro compounds, in Biologically Ac-
Possible precursors in its host-plant, Biochem. Syst. Bcol., 10, tive Natural Products: Potential Use in Agriculture (H. G. Cu-
355-364 (1982). tler, ed.), ACS Symposium Series 380, 143-154, American
BRIMER, L., Detennination of cyanide and cyanogenic compounds Chemical Society, Washington, DC, 1988.
in biological systems, in Cyanide Compounds in Biology (D. CONN, E. E., ~-Glucosidases in plants: The matter of substrate
Evered and S. Harnett, eds.) Ciba Symposium Vol. 140, specificity, in ~-Glucosidases: Biochemistry and Molecular Bi-
177-200, John Wiley & Sons, Ltd., Chichester, 1988. ology, ACS Symposium Series 533, 15-26, American Chemical
BRIMER, L. and L. DALGAARD, Cyanogenic glycosides and cyano- Society Washington, DC, 1993.
hydrins in plant tissues. Qualitative and quantitative determina- CRAMER, U. and F. SPENER, Biosynthesis of cyclopentenyl fatty
tion by enzymatic post-column cleavage and electrochemical acids, Biochim. Biopbys. Acta, 450,261-265 (1976).
detection, after separation by high-performance liquid cbroma-
CUTLER, A J. and E. E. CONN, The synthesis, storage, and degrada-
tography, J. Cbromatogr., 303, 77-88 (1984).
tion of plant natural products: Cyanogenic glycosides as an ex-
BRIMER, L., S. BRoGOER-CHRlSTENSEN, 1. W. JAROSZEWSKI, and ample, in Cellular and Subcellular Localization in Plant Metabo-
F. NARTEY, Structural elucidation and partial synthesis of 3- lism (L. L. Creasy and G. Hrazdina, eds.), Recent Advances in
hydroxyheterodendrin, a cyanogenic glucoside from Acacia Phytochemistry Va!. 16),249-271, Plenum Press, New York,
sieberiana var. woodii,Phytochemistry,20, 2221-2223 (1981). 1982.
BRIMER, L., S. BRoGGER CHRISTENSEN, P. M~LGAARD, and F. NAR- DAVIS, R. H., Cyanogens, in Toxic Substances in Crop Plants 0.
TEY, Detennination of cyanogenic compounds by thin-layer P. F. D'Mello, C. M. Duffus, and J. H. Duffus, eds.), 202-225,
chromatography. 1. A densitometric method for quantification Royal Society of Chemistry Cambridge, 1991.
of cyanogenic glycosides, employing enzyme preparations (~­
DEAN, J. V. and J. E. HARPER, Nitric oxide and nitrous oxide pro-
glucosidase) from Helix pomatia and picrate-impregnated ion-
duction by soybean and winged bean during the in vivo nitrate
exchange sheets, J. Agric. Food Chern., 31, 789-793 (1983).
reductase assay, Plant Physiol., 82, 718-723 (1986).
BRINKER, A. M. and D. S. SEIGLER, Methods for the detection and
quantitative detennination of cyanide in plant materials, Phyto~ DEwICK, P. M., The biosynthesis of cyanogenic glycosides and
chern. Bull., 21, 24-31 (1989). glucosinolates, Nat. Prod. Rep., 1, 545-549 (1984).
BRINKER, A. M. and D. S. SEIGLER, Determination of cyanide and DlRZO, R. and J. L. HARPER, Experimental studies on slug-plant
cyanogenic glycosides from plants in Modem Methods of Plant interactions, J. Eco!., 70, 101-117, 119-138 (1982).
Analysis (H. F. Linskens and J. F. Jackson, eds.), 359-381, DUFFEY, S. S., M. S. BLUM, H. M. FALES, S. L. EVANS, R. W.
Springer-Verlag, Berlin, 1992. RoNCADoRl, D. L. TIEMANN, and Y. NAKAGAWA, Benzoyl cya-
BUNCH, A. W. and C. J. KNoWLES, Cyanide production and degra- nide and mandelonitrile benzoate in the defensive secretions of
dation during growth of the snow mould fungus, J. Gen. Micro- millipedes, J. Chern. Ecol., 3, 101-113 (1977).
bioI., 116, 9-16 (1980). DUFFIELD, R. M., A. SHAFAOATI, E. W. RIDDICK, S. BOWEN, and
BYERS, R. A, D. L. GUSTINE, B. G. MoYER, and D. L. BlERLBlN, J. W. WHEELER, Mandelonitrile in larval secretion of mountain
3-Nitropropionate in crownvetch: A natural deterrent to insects? ash sawfly, Pristiphora geniculata (Hymenoptera: Tenthredini-
in Natural Resistance of Plants to Pests (M. B. Green and P. A doe), J. Chern. Eco!., 16,1103-1110 (1990).
Hedin, eds.), ACS Symposium Series 296, 95-105, American ETI'LlNGER. M. G., J. W. JAROSZEWSKI, S. R. JENSEN, B. J. NIELSEN,
Chemical Society, Washington, DC, 1986. and F. NARTEY, Proacacipetalin and acacipetalin, Chern. Com-
ClmNo. I. and J. E. POULTON, Cloning of cDNA of Prunus serotina mun., 952-953 (1977).
(R)-( + )-mandelonitrile lyase and identification of a putative FAN, T. W. and E. E. CONN, Isolation and characterization of two
FAD-binding site, Plant Cell. Physio!., 34, 1139-1143 (1993). cyanogenic ~-glucosidases from flax seeds, Arch. Biochem. Bi-
CLEGG, D.O., E. E. CONN, and D. H. JANZEN, Developmental fate ophys., 243, 361-373 (1985).
of the cyanogenic glucoside linamarin in Costa Rican wild lima FmOL, F. and V. ANGER, Replacement of benzidine by copper ethy-
bean seeds, Nature, 278, 343-344 (1979). acetoacetate and tetra base as spot-test reagent for hydrogen
CONN, E. E., Cyanogenic glycosides, in International Review of cyanide and cyanogen, Analyst, 91, 282-284 (1966).
296 Cyanogenic G/ycosides and Cyanolipids

FOWLER, S., Plant toxin linked to "Guam dementia," New Scien- of Official Analytical Chemists, 13th ed., 433, Assoc. Official
tist, 115, 31 (13 August 1987). Analytical Chemists, Washington, DC, 1980.
GIBSBMANN, A., B. BIEHL, and R. LmBEREI, Identification of scopo- HOSEL, W. and E. E. CONN, The aglycone specificity of plant 13-
letin as a phytoalexin of the rubber tree Bevea brasiliensis, 1. glycosidases, Trends Biochem. Sci., 7, 219-221 (1982).
Phytopathol., 117, 373-376 (1986). HasEL, W. and A. NAHRSTEDT, Spezifische Glucosidasen rur das
GROSS, M., G. H. JACOBS, and J. E. POULTON, A rapid and sensitive Cyanglucosid Triglochinin-Reinigung und Charakterisierung
spectrophotometric assay for prunasin hydrolase activity em- von ~-Glucosidasen aus Alocasia macrorrhiza Schott., Hoppe-
ploying purified mandelonitrile lyase, Anal. Biochern., 119, Seyler's Z. Physiol. Chern., 356, 1265-1275 (1975).
25-30 (1982). HOSEL, W., J. BERLIN, T. N. HANZLIK, and E. E. CoNN, In-vitro
a
GUlGNARD, L., Le haricot acide cyanhydrique (Phaseolus luna- biosynthesis of 1-(4'-hydroxyphenyl)-2-nitroethane and pro-
IuS). ElUde historique, botanique et chimique. Nouveau procede duction of cyanogenic compounds in osmotically stressed cell
pour deceler l' acide cyanhydrique, Bull. Sci. Pharmacol., 13, suspension cultures of Eschscholtzia cali/ornica Cham., Planta,
401-419 (1906a). 166, 176-181 (1985).
a
GUlGNARD, L., Le haricot acide cyanhydrique, Phaseolus lunatus HOSEL, W.,1. TOBER, S. H. EcKLUND, and CONN, E. E., Characteri-
L., C. R. Hebd. Seances Acad. Sci., 142, 545-553 (1906b). zation of ~~glucosidases with high specificity for the cyano-
HAHLBROCK, K. and E. E. CONN, Purification and properties of genic glucoside dhurrin in Sorghum bicolor (L.) Moench seed-
a UDP-glucose:ketone cyanohydrin l3-glucosyltranserase from lings, Arch. Biochem. Biophys., 252, 152-162 (1987).
Unum usilalissumum, J. BioI. Chern., 245, 917-922 (1970). HRUSKA, A. J., Cyanogenic glucosides as defense compounds, J.
Chern. Ecol., 14, 2213-2217 (1988).
HALKmR, B. A. and B. L. MjilllER, Biosynthesis of the cyanogenic
glucoside dhurrio in seedlings of Sorghum bieolor (L.) Moench JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary
and partial purification of the enzyme involved, Plant Physiol., compounds to the seed-eating larvae of the bruchid beetle Callo-
90, 1552-1559 (1989). sobruchus maeulatus, Phytochemistry, 16, 223-227 (1977).
HALKIER, B. A. and B. L. M~LLER, The biosynthesis of cyanogenic JAROSZEWSKI, J. W. and E. S. OLAFSDOrrm, Natural glycosides of
glucosides in higher plants, J. Bioi. Chern., 265, 21114-21121 cyclopentenone cyanohydrins: Revised structure of so-called
(1990). epitetraphyllin B, Tetrahedron Lett., 27, 5297-5300 (1986).
HAUGER, B. A. and B. L. Mj1jLLER. Involvement of cytochrome JAROSZEWSKI, J. W. and E. S. OLAFSOOTI'IR, On the monohyroxy-
P-450 in the biosynthesis of dhurrin in Sorghum bieolor (L.) lated cyclopentenone cyanohydrin glucosides ofFlacourtiaceae,
Moench, Plant Physiol., 96, 10-17 (1991). Phytochemistry, 26, 3348-3349 (1987).
HALKmR, B. A., J. LYKKESFBLDT, and B. L. MjilLLER, 2-Nitro-3-(p- JAROSZEWSKI, J. W., E. S. OLAFso6rrm., C. CORNEIT, and K.
hydroxyphenyl)propionate and aci-l-nitro-2-(p-hydroxyphen- SCHAUMBURG, Cyanogenesis of Adenia volkensii Harms and
yl)ethane, two intennediates in the biosynthesis of the cyano- Tetrapathaea tetrandra Cheeseman (Passifloraceae) revisited:
genic glucoside dhurrin in Sorghum bieolor (L.) Moench, Proc. Tetraphyllin B and volkenin. Optical rotatory power of cyclo-
Natl. Acad. Sci. USA, 88, 487-491 (1991). pentenoid cyanohydrin glucosides, Acta Chern. Scand., B41,
410-421 (1987a).
HALKmR, B. A., C. E. OLSEN, and B. L. MjilLLER, The biosynthesis
JAROSZEWSKI, J. W., P. S. JENSEN, C. CORNETT, and J. R. BYBERG,
of cyanogenic glucosides in higher plants, J. BioI. Chern., 264,
Occurrence of lotaustralin in Berberidopsis heckleri. Relation
119487-19494 (1989).
to the chemical evolution of Flacourtiaceae, Biochem. Syst.
HAUGER, B. A., H. V. SCHELLER, and B. L. MjilLLER, Cyanogenic Ecol., 16, 23-28 (1987b).
glucosides: The biosynthetic pathway and the enzyme system
JAROSZEWSKI, J. W., J. V. ANoERSEN, and I. Bll.LESKOV, Plants as a
involved, in Cyanide Compounds in Biology (D. Evered and S.
course of chiral cyclopentenes: Taraktophyllin and epivolkenin,
Harnett, eds.), Ciba Symposium Vol. 140, 49-66, John Wiley &
new cylopentenoid cyanohydrin glucosides from Flacourtia~
Sons, Ltd., Chichester, 1988.
ceae, Tetrahedron, 43, 2349-2354 (1987c).
HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod.
JAROSZEWSKI, J. W., D. BRUUN, V. CLAUSEN, and C. CORNE'IT,
Rep., 7, 85-109 (1989).
Novel cyclopentenoid cyanohydrin rhamnoglucosides from Fla-
HARRlS, R. E., A. W. BUNCH, and C. J. KNOWLES, Microbial cyanide courtiaceae, Plant. Medica, 54, 333-337 (1988).
and nitrile metabolism, Science Progress Oxford, 71, 293-304
JENSEN, S. R and B. 1. NIELSEN, Gynocardin in Ceratiosicyos laevis
(1987).
(Achariace.e), Phytochemistry, 25, 2349-2350 (1986).
HBGNAUER, R, Cyanogenic glycosides as systematic markers in JONES, D. A., Cyanogenic glycosides and their function, in Phyto-
Tracheophyta. Plant Syst. Evol., Suppl. 1, 191-209 (1977). chemical Ecology 0. B. Harbome ed.), Academic Press, Lon-
HBGNAUER, R, Cyanogene Verbindungen, in Chemotaxonomie der don, 1972.
Pflanzen, Vol. 7, Birkhiiser Verlag, Basel, 1986. JONES, D. A., Cyanogenesis in animal-plant interactions, in Cyanide
HIRONO, I., Carcinogenicity and neurotoxicity of cycasin with spe- Compounds in Biology (D. Evered and S. Harnett, eds.), Ciba
cial reference to species differences, Fed. Proc., 31, 1493-1497 Symposium Vol. 140, 151-170, John Wiley & Sons, Ltd.,
(1972). Chichester, 1988.
HORRll.L, J. C. and A. J. RICHARDS, Differential grazing by the KAKES, P., On the polymorphism for cyanogenesis in natural popu-
mollusc Arion hortensis Fer. on cyanogenic and acyanogenic lations of Trifolium repens L. in the Netherlands. I. Distribution
seedlings of the white clover, Trifolium repens L., Heredity, 56, of the genes Ac and Li, Acta Bot. Neerl., 36, 59-69 (1987).
277-281 (1986). KrNGSBURY, J. M., Poisonous Plants of the U.S. and Canada, Pren-
HORWITZ, W. (ed.), Official Methods of Analysis of the Assoc. tice-Hall, Englewood Cliffs, NJ, 1964.
 Cyanogenic Glycosides and CyanoUpids 297

KNOWLES, C. J., Cyanide utilization and degradation by microorga- NAHRSTEDT, A .. The biology of the cyanogenic glycosides: new
nisms, in Cyanide Compounds in Biology (D. Evered and S. developments, in Nitrogen Metabolism of Plants CK. Mengel
Harnett, eds.), Ciba Symposium Vol. 140,3-31, John Wiley & and D. J. Pilbeam, eds.), Proceedings of the Phytochemistry
Sons, Ltd., Chichester, 1988. Society of Europe, 249-269, Clarendon, Oxford, 1992.
KOCH, B., V. S. NIELSEN, B. A. HALKIER, C. E. OLSEN, and B. L. NAHRSTEOT, A., Cyanogenesis and foodplants, in Phytochemistry
M~LLER, The biosynthesis of cyanogenic glucosides in seed- and Agriculture (T. A. van Beek and H. R. M. Brete1er, eds.),
lings of cassava (Manihot esculenta Crantz), Arch. Biochem. Proceedings of the Phytochemistry Society of Europe, 34,
Biophys., 292, 141-150 (1992). 107-129, Oxford University Press, Oxford, 1993.
KOJIMA, M., J. E. POULTON, S. S. THAYER, and E. E. CONN, Tissue NAHRSTEDT, A. and R. H. DAVIS, The occurrence of the cyanoglu-
distributions of dhurrin and enzymes involved in its metabolism cosides linamarin and lotaustralin, in Acraea and Heliconius
in leaves of Sorghum bicolor, Plant Physiol., 63, 1022-1028 butterflies, Compo Biochem. Physiol., 68B, 575-577 (1981).
(1979). NAHRSTEDT. A. and P. SCHWIND. Phenylalanine is the biogenetic
LAMBERT, 1. L.. J. RAMASAMY, and J. V. PAUKSTELIS, Stable re- precursor of meta-hydroxylated zierin, the aromatic cyanogenic
agents for the colorimetric detennination of cyanide by modi- glucoside of unripe akenes of Xeranthemum cylindraceum. Phy-
fied Konig reactions, Anal. Chem., 47, 916-918 (1975). tochemistry, 31, 1997-2001 (1992).
LIEBEREI, R.. Cyanogenesis of Hevea brasiliensis during infection NAHRSTEDT, A., N. ERB, and H. ZINSMEISTER, Methods of liberating
with Microcyc/us ulei, J. Phytopathol., 115,134-146 (1986). and estimating hydrocyanic acid from cyanogenic plant mate-
LIEBEREI, R., Relationship of cyanogenic capacity (HCN-c) of the rial, in Cyartide in Biology (B. Vennesland, E. E. Conn, C. J.
rubber tree Hevea brasiliensis to susceptibility to Microcyclus Knowles, J. Westley, andF. Wissing, eds.),461-471, Academic
ulei, the agent causing South American leaf blight, J. Phytopa- Press, London, 1981.
thol., 122, 54-67 (1988). NAHRSTEDT, A., P. S. JENSEN. and V. WRAY, Prunasin-6'-malonate,
LtEBEREI, R., B. BIEHL, A. GmsBMANN, and N. T. V. JUNQUElRA, a new cyanogenic glucoside from Merremia dissecta. Phyto-
Cyanogenesis inhibits active defense reactions in plants, Plant chemistry, 28, 623-624 (1989).
Physiol., 90, 33-36 (1989).
NAHRSTEDT, A" E. A. SATIAR, and S. M. H. EL-ZALABANI, Amyg-
LIEBERBI, R., D. SELMAR, and B. BmHL, Metabolization of cyano- dalin acyl derivatives, cyanogenic glycosides from the seeds of
genic glucosides in Hevea brasiliensis, Plant Syst. Evol., 150, Merremia dissecta, Phytochemistry, 29, 1179-1181 (1990).
49-63 (1985).
NAHRSTEDT, A., V. WRAY, L. GROTJAHN, L. H. FlKENSCHER, and
MANNING, K., petoxification of cyanide by plants and honnone R. iIEGNAUER, New acylated cyanogenic diglycosides from
action, in Cyanide Compounds in Biology (D. Evered and S. fruits of Anthemis cairica, Planta Medica, 49, 143-148 (1983).
Harnett, eds.), Ciba Symposium Vol. 140, 92-110, John
Wiley & Sons, Ltd., Chichester, 1988. NARTEY, F., L. BRIMER, and S. BROGGER-CmuSTBNSEN, Proacacibe-
rin, a cyanogenic glycoside from Acacia sieberiana var. wood;;,
MIKOLAJCZAK, K. L., Cyanolipids, Prog. Chem. Fats Other Lipids,
Phytochemistry, 20, 1311-1314 (1981).
15,97-130 (1977).
NAYAR, J. K. and G. FRAENKEL, The chemical basis of the host
M~LLER, B. L. and J. E. POULTON, Cyanogenic glycosides, in Meth-
selection in the Mexican bean beetle, Epilachna varivestis (Col-
ods in Plant Biochemistry, Vol. 9(p. M. DeyandJ. B. Harhome,
eoptera, Coccinellidae), Anou Ent. Soc. Am. 56, 174-178
eds.), 183-207, Academic Press, London, 1993.
(1963).
MULLER, E. and A. NAHRSTEDT, Purification and characterization
of an a-hydroxynitrile lyase from the haemolymph of the larvae OLAFSD6rrm., E. S .• J. V. ANDERSEN, and J. W. JAROSZEWSKI, Cya-
of Zygaena tri/olii, Planta Medica, 56, 611-612 (1990). nohydrin glycosides of Passifioraceae, Phytochemistry, 28,
172-132 (1989).
NAHRSTEDT, A., Prunasin in Holocalyx balansae, Phytochemistry,
15, 1983-1984 (1976). OLECHNO, J. D., J. E. POULTON, and E. E. CONN, Nandinin: An
acylated free cyanohydrin from Nandina domestica, Phyto-
NAHRSTEDT, A., Absence of cyanogenesis from Droseraceae, Phy-
chemistry, 23, 1784-1785 (1984).
tochemistry, 19, 2757-2758 (1980).
NAHRSTEDT, A .• Isolation and characterization of cyanogenic glyco- OKOGUN, J. I. and D. E. U. EKONG, The fragrant principle of the
sides, in Cyanide in Biology (B. Vennesland, E. E. Conn, C. J. fruits of Dennettia tripetaia G. Baker: A new naturally occur-
Knowles,J. Westley,andF. Wissing,eds.), 145-181, Academic ring nitro-compound, Chem. Ind., 1272 (1969).
Press, New York, 1981. OSMAN, S. M. and F. AHMAD. Forest oilseeds. in New Sources of
NAHRSTEDT, A .• Cyanogenic compounds as protecting agents for Fats and Oils (E. H. Pryde, L. H. Princen, and K. D. Mukherjee,
orgartisms, Plant Syst. Evol., 150, 35-47 (1985). eds.), 109-127, American Oil Chemical Society, Champaign,
IL,1981.
NAHRSTEDT. A., Recent developments in chemistry, distribution,
and biology of the cyanogenic glycosides, in Biologically Ac- PASTEBLS, J. M., D. DALOZE, and M. ROWELL-RAHlER, Chemical
tive Natural Products (K. Hostettmann and P. J. Lea, eds.), defence in chrysomelid eggs and neonate larvae. Physiol. Ento-
213-234, Clarendon Press, Oxford, 1987a. mol. 11, 29-37 (1986).
NAHRSTEDT, A., Flachs, Hornklee. Widderchen, und Blausaure, PEISER, G. D., T. WANG, N. E. HOFFMAN, S. F. YANG, H. Lru,
Deutsche Apotheker Zeitung, 127, 2385-2390 (1987b). and C. T. WALSH, Formation of cyanide from carbon I of 1-
NAHRSTEDT, A., Cyanogenesis and the role of cyanogenic glyco- aminocyclopropane-l-carboxylic acid during its conversion to
sides in insects, in Cyanide Compounds in Biology (D. Evered ethylene, Proc. Nat!. Acad. Sci. USA, 81, 3059-3063 (1984).
and S. Harnett, eds.), (Ciba Symposium Vol. 140), 131-150, POULTON. I. E., Cyanogenic compounds in plants and their toxic
John Wiley & Sons, Ltd., Chichester, 1988. effects, in Handbook of Natural Toxins, Vol. I (R. F. Keeler
298 Cyanogenic G/ycosides and CyanoJipids

and W. T. Th, eds.), 117-157, Marcel Dekker, Inc., New York, in Plant Biochemistry, Vol. 8 (P. M. Dey and J. B. Harbome,
1983. eds.), 51-131, Academic Press, London, 1993.
POULTON, 1. E., Localization and catabolism of cyanogenic glyco- SEIGLER, D. S. and J. E. EBINGER, Cyanogenic glycosides in ant-
sides, in Cyanide Compounds in Biology (D. Evered and S. acacias of Mexico and Central America, Southwest. Nat., 32,
Harnett, eds.), Ciba Symposium Vol. 140,67-91, John Wiley & 499-503 (1987).
Sons, Ltd., Chichester, 1988. SEIGLER, D. S. andK. C. SPENCER, Corrected structures ofpassicori-
POULTON, 1. E., Enzymology of cyanogenesis in rosaceous stone acin, epicoriacin and epitetraphyllin B and their distribution
fruits in ~-Glucosidases: Biochemistry and Molecular Biology, in the Flacourtiaceae and Passifloraceae, Phytochemistry, 28,
ACS Symposium Series 533, 170-190, American Chemical 931-932 (1989).
Society, Washington, DC, 1993. SEIGLER, D. S., B. R. MASUN, and E. E. CONN, Cyanogenesis in
POULTON, J. E. and C. P. LI, Tissue level compartmentation of the Leguminosae, in Advances in Legume Biology (C. H. Stir-
(R)-arnygdalin and amygdalin hydrolyase prevents large-scale ton and J. L. Zarucchi, eds.), Monogr. Syst. Bot. Missouri Bot.
cyanogenesis in undamaged Prunus seeds, Plant Physiol., 104, Gard., 29, 645-672 (1989).
29-35 (1994). SBLMAR. D., Apoplastic occurrence of cyanogenic ~-glucosidases
POURMOHSBNI, H., W. D. IBBNTHAL, R. MACHINBK, G. REMBERG, and consequences for the metabolism of cyanogenic glucosides,
and V. WRAY, Cyanogenic glucosides in the epidermis of in j3-Glucosidases: Biochemistry and Molecular Biology, ACS
Hordeum vulgare, Phytochemistry, 33, 295-298 (1993). Symposium Series 533, 190-203, American Chemical Society,
Washington, DC, 1993.
RANnoux, T., J. C. BRABKMAN, and D. DALOZB, De novo biosyn-
thesis of .6?-isoxazolin-5-one and 3-nitropropanoic acid deriva- SELMAR, D., R. LnmEREI, and B. BIEHL, Mobilization and utilization
tives in Chrysomela tremulae, Naturwissenschaften, 78, of cyanogenic glycosides, Plant. Physiol., 86, 711-716 (1988).
313-314 (1991). SELMAR, D., S. GROCHOLEWSKI, and D. S. SEIGLER, Cyanogenic
RAUBBNHBlMBR, D., Cyanoglycoside gynocardin fromAcraea horta lipids. Utilization during seedling development of Ungnadia
(L.) (Lepidoptera: Acraeinae), J. Chern. Ecol.,15, 2177-2189
speciosa, Plant Physiol., 93, 631-636 (1990).
(1989). SELMAR, D., R. LIEBEREI, N. JUNQUElRA, and B. BIEHL, Changes
RllHR, S. S., P. P. I'BBNY, and D. H. JANZEN, Chemical defence in in cyanogenic glucoside content in seeds and seedlings of Hevea
species, Phytochemistry, 30, 2135-2140 (1991).
Central American non-ant-acacias, J. Anim. Ecol.,42, 405-415
(1973). SBLMAR, D., R. LIEBEREI, B. BIEHL, and E. E. CONN, a-Hydroxyni-
trile lyase in Hevea brasiliensis and its significance for rapid
ROCKENBACH, J., A. NAHRSTBDT, and V. WRAY, Cyanogenic glyco-
cyanogenesis, Physio!. Plant., 75, 97-101 (1989).
sides from Psydrax and Oxyanthus species, Phytochemistry, 31,
567-570 (1992). SHIMOMURA, H., Y. SASHIDA, and T. ADACHI, Cyanogenic and
phenylpropanoid glucosides from Prunus grayana, Phytochem-
Ross, M. D. and W. T. JONES, A genetic polymorphism from tannin
istry, 26, 2365-2366 (1987).
production in Lotus corniculatus and its relationship to cyanide
SMITII, C. R., JR., D. WEISLEDER, R. W. Mn.r.ER,1. S. PALMER, and
polymorphism, Theoret. Appl. Genet., 64, 263-268 (1983).
O. E. OLSON, Linustatin and neolinustatin: Cyanogenic glyco-
SAUNDERS, J. and E. E. CONN, Subcellular localization of the cyano- sides of linseed meal that protect animals against selenium tox-
genic glucoside of Sorghum by autoradiography, Plant Physiol., icity, J. Org. Chern., 45, 507-510 (1980).
59,647-652 (1977).
SPENCER, K., Specificity of action of allelochemicals: Diversifica-
SAUPE, S. G., Cyanogenic compounds and angiosperm phylogeny, tion of glycosides, in Allelochemicals: Role in Agriculture and
in Phytochemistry and Angiosperm Phylogeny (D. A. Young Forestry (G. R. Waller, ed.), ACS Symposium Series 330,
and D. S. Seigler, eds.), 80-116, Praeger, New York, 1981. 275-288, American Chemical Society, Washington, DC, 1987.
SCHROEDER, L. A., Consumption of black cherry leaves by phytoph- SPENCER, K. C. and D. S. SEIGLER, Cyanogenesis of Passiflora
agous insects, Am. MidI. Nat., 100, 294-306 (1978). edulis, J. Agric. Food Chern., 31, 794-796 (1983).
SEIGLER, D., The naturally occurring cyanogenic glycosides, in SPENCBR, K. C. and D. S. SEIGLER, Cyanogenic glycosides of Car-
Progress in Phytochemistry, Vol. 4 (L. Reinhold, J. B. Harbome, ica papaya and its phylogenetic position with respect to the
and T. Swain, eds.), 83-120, Pergamon, Oxford, 1977. Violales and Capparales, Am. J. Bot., 71, 1444-1447 (1984).
SEIGLER, D. S., Recent developments in the chemistry and biology SPENCER, K. C. and D. S. SEIGLER, Cyanogenic glycosides of
of cyanogenic glycosides and lipids, Rev. Latinoamer. Quim., Malesherbia, Biochem. Syst. Ecol., 13, 23-24 (1985a).
12, 39-48 (198Ia). SPENCER, K. C. and D. S. SEIGLER, Cyanogenic glycosides and the
SEIGLER, D. S., Cyanogenic glycosides and lipids: Structural types systematics of the Flacourtiaceae, Biochem. Syst. Eco!., 13,
and distribution, in Cyanide in Biology (B. Vennesland, E. E. 421-431 (l985b).
Conn, C. J. Knowles, J. Westley, and F. Wissing, eds.), SPENCER, K. C. and D. S. SEIGLER, Passibiflorin, epipassibiflorin,
1133-143, Academic Press, London, 1981b. and passitrifasciatin: Cyclopentenoid cyanogenic glycosides
SEIGLER, D. S., Cyanide and cyanogenic glycosides, in Herbivores: from Passiflora, Phytochemistry, 24, 981-986 (l985d).
Their Interactions with Secondary Plant Metabolites (G. A. Ro- SPENCER, K. C., and D. S. SBIGLER, Co-occurrence of
senthal and M. R. Berenbaum, eds.), 35-77, Academic Press, valine/isoleucine-derived and cyclopentenoid cyanogens in a
San Diego, CA, 1991. Passiflora species, Biochem. Syst. Ecol., 13, 303-304 (l985e).
SEIGLER, D. S. and A. M. BRINKER, Characterization of cyanogenic SPENCER, K. C., D. S. SEIGLER, and S. W. FRALEY, Cyanogenic
glycosides. cyanolipids, nitroglycosides, organic nitro com- glycosides of the Tumeraceae, Biochem. Syst. Ecol., 13,
pounds and nitrile glucosides from plants, in Modem Methods 433-435 (1985c).
 Cyanogenic Glycosides and Cyanolipids 299

SPENCER, K., D. S. SEIGLER, and A. NAHRSTEDT, Linamarin, lotaus- short·day plant Lemna paucicostata 6746, Plant Cell, Physio1.,
tralin, Iinustatin and neolinustatin from Passiflora species, Phy- 24,705-711 (1983).
tochemistry, 25, 645-647 (1986). TAPPER, B. A. and M. A. MACDoNALD, Cyanogenic compounds
SPENCER, P. S., P. B. NUNN, 1. HUGON, A. C. LUDOLPH, S. M. Ross, in cultures of a psychrophilic basidiomycete (snow mould), Can.
D. N. Roy, and R. C. ROBERTSON, Guam amyotrophic lateral J. Microbio!., 20,319-327 (1974).
sclerosis-Parkinsonism-Dementia linked to a plant excitant THAYER, S. S. and E. E. CONN, Subcellular localization of dhurrin
neurotoxin, Science, 237,517-522 (1987). ~-glucosidase and hydroxynitrile lyase in the mesophyll cells

STADLER, E., Contact chemoreception, in Chemical Ecology ofIn- of Sorghum leaf blades, Plant Physio!., 67, 617-622 (1981).
sects (W. J. Bell and R. T. Card", eds.), 3-35, Chapman & VENNESLAND, B., E. E. CONN, C. J. KNOWLES, J. WESTLEY, and F.
Hall, New York, 1984. W,SSING (eds.), Cyanide in Biology, Academic Press, London,
1981.
STERMIlZ, F. and G. S. YOST, Analysis and characterization of nitro
compounds from Astragalus species, in Effects of Poisonous WEBSTER, G., Conspectus of a new classification of the Euphor-
Plants on Livestock (R. F. Keeler, K. R. van Kampen, and L. biaceae, Taxon, 24, 593-601 (1975).
F. James, eds.), 371-378, Academic Press, New York, 1978. WILSON, J., Cyanide in human disease, in Clinical and Experimental
Toxicology of Cyanides (B. Ballantyne and T. C. Marrs, eds.),
SWAIN, E., C. P. LI, and J. E. POULTON, Tissue and subcellular
Wright, Bristol, 1987.
localization of enzymes catabolizing (R)-amygdalin in mature
Prunus seratina seeds, Plant Physio!., 100, 291-300 (1992). WEEHUIZEN, F., Over salpeterigzuur in Erythrina L., Phann.
Weekb!.,44, 1229-1232 (1907).
SWENSON, W., Ph. D. thesis, University of Califomi a, Davis, 1986.
WILLAMAN, J. J. and R. M. WEST, Effect of climatic factors on the
SWENSON, W., J. E. DUNN, andE. E. CONN, Cyanogenesis in Acacia hydrocyanic-acid content of sorghum, J. Agric. Res., 6,
sutherlandii, Phytochemislry, 26, 1835-1836 (1987). 261-272 (1916).
TANTISEWlE, B., H. W. L. RUIJGROK, and R. HEGNAUER, Die Ver- WURTELE, E. S., S. S. THAYER, and E. E. CONN, Subcellular locali-
breitung der Blausaure bei den Connophyten. 5 Mitteilung: zation of a UDP-glucose:aldehyde cyanohydrin ~-glucosyl
Uebercyanogene Verbindungen bei den Parietales und bei eini- transferase in epidennal plastids of Sorghum leaf blades, Plant
gen wciteren Sippen, Pharm. Weekb!., 104,1341-1355 (1969). Physio!',70, 1732-1737 (1982).
TANAKA, 0., C. F. CLELAND, and Y. BEN-TAL, Effect of ferricya- ZILG, H. and E. E. CONN, Stereochemical aspects of lotaustralin
nide, ferrocyanide, and KCN on growth and flowering in the biosynthesis, 1. BioI. Chern., 240, 3112-3115 (1974).
17
Glucosinolates

Introduction sues are richer in glucosinolates than older tissues (Louda

Biosynthesis and Mole, 1991).
Distribution In many properties, these compounds resemble the cyano-
Utilization of Glucosinolate Data in Systematic Studies genic glycosides, to which they are biosynthetically related.
Biological Activity Glucosinolates are derived from many of the same amino
Properties of Glucosinolates acids as the cyanogens, although some amino acids serve as
Properties of the Hydrolysis Products a precursor for one group but not for the other. Glucosino-
Goitrigenic Derivatives in Crop Plants lates that correspond to alanine, valine, leucine, isoleucine,
Glucosinolates as AIIomones and Kairomones phenylalanine, tyrosine, and tryptophan are known (Larsen,
Glucosinolates and Oviposition by Insects 1981). In contrast to the cyanogenic glycosides, however,
Long-Distance Attraction the distribution of glucosinolates is restricted to a smaIl num-
Glucosinolates and Plant Disease Resistance ber of families, many of which are related.
Glucosinolates and AIlelopathy Glucosinolates are water-soluble, polar compounds that
References possess a common structural moiety, described correctly for
the first time in 1956 by Ettlinger and Lundeen. Glucosino-
lates occur in plants as salts, and most have been isolated
as the potassium salt because of the predominance of potas-
INTRODUCTION sium in plant cells. These compounds may also occur in
association with positively charged organic compounds and
Glucosinolates produce hydrolysis products responsible for proteins in plants. Some [e.g., sinalbin (1)] occur as salts
the pungent taste of mustard, horseradish, and other bras sica- of sinapine (2), a quaternary ammonium base composed of
ceous plants. These glycosides occur in several important choline esterified with 3,5-dimethoxy-4-hydroxycinnamic
crop plants and in several common weedy species (Ettlinger acid (Larsen, 1981). Although leaves usually contain only
and Kjaer, 1968; Louda and Mole, 1991; Tapper and Reay, about 0.1 % dry weight, as much as 10% dry weight glucosi-
1973). Approximately 100 compounds of this type have been nolates may be stored in seeds. This requires a large amount
described (Poulton and M¢IIer, 1993; Reed et al., 1988); of ionic material to counterbalance the negatively charged
some representative compounds are illustrated in Fig. 17.1. glucosinolates.
A semisystematic nomenclature for glucosinolates exists As is true for cyanogens, special i3-glucosidases, often
along with many of the earlier trivial names (Larsen, 1981). called myrosinases or thioglucosidases (BC 3.2.3.1), co-
Virtually all species of the mustard family (Brassicaceae or occur and catalyze hydrolysis of glucosinolates (Fig. 17.2)
Cruciferae) contain glucosinolates and the greatest variety (Poulton and M¢Iler, 1993). A number of isozymes of these
of glucosinolate structures is found in this family. Many enzymes occur, and some fonns of myrosinase are activated
of the species of this family are fast-growing, early-season by ascorbate. Although these enzymes show some specific-
herbaceous C3 plants (Louda and Mole, 1991). Typical ity, all known glucosinolates are substrates for these en-
amounts present are from 0.1 % to 0.6% dry weight (Bernays, zymes (Larsen, 1981; Poulton and M¢IIer, 1993). All cruci-
1983). The content of glucosinolates in seeds usually is much fer organs contain myrosinase.
higher than that of vegetative tissues. In general, young tis- On disruption of the tissues, myrosinases and glucosino-

300
 Glucosinolates 301

-
N'OSO,K +

~
CR,=CRC R,-{ OR
N'OS6'K+ OR
S~OR I S~OR
:r
OR

o-n
RO "" OR
sinigrin (15)
glucotropaeolin (6) sinalbin (1)
benzyl glucosinolate
- +
N'OSO'K OR

:r S~OR
I
""
OR

gluconasturtiin (12) progoitrin (21)

e=:rr<
glucocochlearin

~N-OSO;K+ OR N.OSO;K+ OR

/ \~OR ?"
I I
S~OR
OR
OR '" NR
benzyl isothiocyanate (32)
glucoputranjavin from glucotropaeolin
glucobrassicin (3)

OCR,

RO~o~£i(CR,J, sinapine(2)
methyl isothiocyanate
from glucocapparin

(a) OCR,

- + - +

~~~OR
UN)' OR
~~OR
UN ) OR
I I _
OCH3 neoglucobrassicin (4) S03 sulfoglucobrassicin (5)
o
t
/S~N=C=S
sulforaphene (33) sulforaphane (17)
N-OSO;K+

~S~O
RO~ ' ~N=C=S
~OR
OR
o
t
OR
2-hydroxy-3-butenylglucosinolate (16) allylisothiocyanate (26)

hirsutin (34)
o arabin (35)

t
8-methylsulfonyloctyl isothiosyanate 9-methylsulfonylnonyl isothiocyaniate
o
t cammelinin (36)
/S~N=C=S

(b) /S ~N=C = S 10-methylsulfonyldecyl isothiocyanate

Fig. 17.1 (a & b). Some representative glucosinolates.
302 Gfucos;noJates

~'NXH ~N'OS03K+
~ V V .. \S~OH
• • CO,H __ OH
9' I H ~
:::,.. NH, "y. CO,H
OH

gJuconasturtiin (12)

myrosinase or
isomerase? ~S-C=N
tbioglucosidase

an isothlocyanate a thiocyanate
+ glucose + HS04"

pH 3-6, Fe++
glucosinolate _ _ _ _ _ _ .--,~

or nitrile factor

a nitrile

Fig. 17.2. Hydrolysis of glucosinolates by myrosinase (modified from Haslam, 1974).

lates are brougbt together, resulting in the release of isothio· 1987). Indolyl glucosinolates predominate in the foliage of
cyanates through a combination of thioglucoside bond hy· many crucifers, contrary to earlier reports (fraynier and
drolysis and spontaneous rearrangement of the aglycone Truscott, 1991). These compounds occur widely in the
(Fig. 17.2). Most evidence favors a subcellular compartmen- Brassicaceae and also have been reported from the genus
talization for these compounds and enzymes (Poulton and Capparis (Capparidaceae) (Schraudolf, 1989). They are
M~ller, 1993). Isothiocyanates (mustard oils) are pungent- probably found in most other glucosinolate-containing fami-
smelling substances with a sharp taste. Under other condi- lies. Indole glucosinolates can be analyzed by HPLC of the
tions, glucosinolate hydrolysis produces thiocyanates and desulfo-derivatives followed by mass spectrometry (Goetz
nitriles (Cole, 1976; Louda and Mole, 1991). There is some and Schraudolf, 1983).
debate about the participation of enzymes in the conversion
of isothiocyanates to thiocyanates.
In order to isolate glucosinolates, the co-occurring f3·thi- BIOSl'Nl'HESIS
oglucosidases must be inactivated as quickly as possible.
Plant material is usually extracted with boiling 70% alcohol. Radiotracer studies have demonstrated that L-amino acids
Methods for paper, thin-layer (nC), and ion-exchange are the direct precursors of the known glucosinolates (poul-
chromatography have been described (Larsen, 198 I). Many ton and M~lIer, 1993). Although glucosinolates that corre-
analytical methods involve hydrolysis of the glucosinolate spond to alanine, valine, leucine, isoleucine, phenylalanine,
followed by identification of the hydrolysis products. Iso- tyrosine, and tryptophan are known (Larsen, 1981), the bio-
thiocyanates can be determined by gas chromatography or, synthesis of only a few specific compounds has been investi-
as derivatives, by nc. Methods for the isolation, purifica- gated. None of the intermediates has been properly estab-
tion, and characterization of glucosinolates by nuclear mag- lished, and the participating enzymes have neither been
netic resonance (NMR) and mass spectrometry have been isolated or characterized as components of crude prepara-
reviewed (Betz and Page, 1990; Bodnaryk, 199 I; Louda and tions (Poulton and M~lIer, 1993). It is generally assumed that
Mole, 1991; Olsson et aI., 1977; Schraudolf, 1989). these steps are catalyzed by microsomal systems analgous to
The distribution of sinapine (2) (3,5-dimethoxy-4-hy- those of cyanogenic glycoside biosynthesis, but the apparent
droxycinnamoylcholine) and aromatic choline esters in biosynthetic similarity has never been established (Poulton
many genera of the Brassicaceae have been surveyed and M~lIer, 1993).
(Bouchereau et aI., 199 I). These compounds were analyzed Isotopically labeled amino acids have been administered
by high-performance liquid chromatography (HPLC). to excised plant parts. As is troe for cyanogenic glycosides,
Indole glucosinolates are especially sensitive; many re- incorporation of administered amino acid precursors into
ports present erroneous data because these compounds are glucosinolates is often extremely high (up to 40%) (Larsen,
destroyed in isolation procedures (Slominski and Campbell, 1981). In one study, plants were fed amino acids, and biosyn-
 Glucosinolotes 303

o v
H
~COlH a1COZ- ~COlH o
HNHl - - . "'" I N -0 ~+-o-
II
II
N+
H/ 'OH
o '0-
L~pheDylalanine
+ ~
N~hydroxy~L~phenylalanine

m
(10)
(11)

~HRS_~H ~~H
"'"
I
SRH
--
N~
~O
V
~-O:- ~+-O- ..- 0
I
NOH V I
OH OH phenylacetaldoxlme (7)

+
()!
I
~ lJ..
SH UDP UDP-glucose
~S-glUCOSYI -.V..
PAPS PAP

V
~S-gIUCOSYI
"'" NOH
thiohydroxirnic acid (8)
NOH

desulfoglucosinolate (9)
V NOSO,

PAPS := 3t~phosphoadenosine~S'~phosphosulfate glucotropaeolin (6)

(benzylglucosinolate)

Fig. 17.3. Proposed mechanism for the biosynthesis of benzylglucosinolate (glucotropaeolin) (modified from Poulton and M~ner. 1993).

thetic intermediates different from those normally present of glucosinolates may take place by addition of a thiol to
in the plants examined_ Neglible amounts of "unnatural" the aci-nitro compound.
glucosinolates were formed when the amino acids were pro- Corresponding nitro compounds are known from several
vided as precursors, but high levels resulted from feeding plants [e.g., I-nitro-2-phenylethane (11)] occurs in extracts
the intermediate compounds_ This finding suggests that high of Tropaeolum majus (Tropaeolaceae). Radiotracer feeding
enzyme specificity occurs for the side-chain structure of the studies have shown that this compound is incorporated into
precursor amino acids, but not for the intermediate com- the glucosinolate (6) and that I-nitro-2-phenylethane can be
pounds (Reed et al_, 1988). L-Phenylalanine fed to nastur- derived from the aldoxime (7). Feeding of2-nitrobenzaldox-
tium plants (Tropaeolum majus) leads to the formation of ime to Brassica species leads to the accumulation of the
glucotropaeolin (6) (Fig. 17.3). Phenylacetothiohydroximate non-naturally occurring 2-nitrophenylglucosinolate (Groot-
(8) has been established as an intermediate and is subse- wassink et al., 1990).
quently glucosylated to form desulfobenzylglucosinolate (9) Several of the putative intermediates contain a C-N
by transglycosylation with UDP-glucose. Feeding doubly double bond, presenting the possibility of geometrical iso-
labeled 14C!lsN amino acids produces glucosinolates with mers. The actoal configuration of these intermediates has
essentially unaltered ratios of isotopes. Thus, the biosyn- not been examined but is assumed to be the same as observed
thesis occurs without cleavage of the carbon-nitrogen bond in the final glucosinolate (Poulton and Mf/lller, 1993). L-CyS-
(Poulton and Mf/lller, 1993). teine may serve as an efficient source of the sulfur atom in
The conversion of amino acids to thiohydroximic acids the thioglucoside moiety.
constitutes the first part of the biosynthesis of glucosinolates. S-Glucosylation of thiohydroximates catalyzed by UDP-
An early step probably involves N-hydroxylation of the G:thiohydroximate glucosyltransferases (BC 2.4.1.-) is
amino acid. This reaction is most likely catalyzed by a cyto- thought to be the penultimate stage in glucosinolate biosyn-
chrome P-450-dependent monooxygenase. An a-nitrocar- thesis (poulton and Mf/lller, 1993).
boxylic acid (10) may serve as an intermediate (Fig. 17.3) The last enzyme of the sequence, sulfation of desulfoglu-
(Poulton and Mf/lller, 1993). Such an intermediate has been cosinolates, is catalyzed by 3'-phosphoadenosine-5'-phos-
demonstrated to be an intermediate in the biosynthesis of phosulfate (PAPS) : desulfoglucosinolate sulfotranserases
cyanogenic glycosides and may represent the branch point (BC 2.8.2.-) (Fig. 17.4) (Poulton and Mf/lller, 1993; Un-
between the two pathways instead of the oxime (7) previ- derhill, 1980).
ously suggested. N-Hydroxyamino acids are easily con- Many other glucosinolates appear to arise by modifica-
verted to the corresponding amines; the reaction is stimu- tion of glucosinolates formed from protein amino acids at
lated by flavin mononucleotide (FMN) and requires the glucosinolate stage or are derived from nonprotein amino
molecular oxygen. Although oximes are easily converted to acids (Larsen, 1981).
glucosinolates, they may not be true intermediates in the An unusual feature of the biochemistry of glucosinolates
pathway. The introduction of the thioglucosidic sulfur atom is the occurrence of homologs of the aglycones which differ
304 Glucosinolates

-
OH
~CO,H

V H NH,------+
()!C0T- ~CO'H
V CO,H

vC
malonyl-eoA

T"'"v0-()
OH 0
HNH,
o:? I CO,H o:? CO,H ~
"" CO,H CO,H_

CO,
NHOH NHOH NOSO;

~CO'H_ ~H ~ ~S.glUCOSYI
lJ lJ ·lJ
gluconasturtiin (12)

-
H OH NOSO;

~s.gIUC"YI • sulfotr&nsferase

glucobarbariu

Fig. 17.4. Proposed biosynthesis of gluconasturtiin and glucobarbarin (modified from Geissman and Crout. 1969),

by one methylene group. In the case of 2-phenylethylglucos- and Tovariaceae) is conceded by most to be closely related.
inolate (gluconasturtiin) (12) in watercress (Nasturtium offi- The Papaveraceae and Fumariaceae, which some previously
cinalis and Rorippa nasturtium-aquaticum), both L-phenyl- placed with this group of families, do not have glucosino-
alanine and acetate (methyl labeled) were readily lates. Most systematists today remove these two families
incorporated into the compound with loss of their carboxyl because of a body of both chemical and morphological data.
groups (Figs. 17.2 and 17.4). Degradation studies showed The Limnanthaceae and Tropaeolaceae (Cronquist places
that the methyl group of the acetate provided the thiohydrox- these in his Geraniales) do possess glucosinolates. Most tax-
imate carbon, whereas the remaining C6 -C2 fragment came onomists do not consider these families related to the first
from phenylalanine. Furthermore, labeling studies indicated group of families, but do consider them related to each other.
that the amino nitrogen of phenylalanine was preserved in Dahlgren transferred the Limnanthacene and Tropaeolacene
the formation of glucotropaeolin (Haslam, 1974). to the same group as the Brassicaceae and then, subsequently
Homomethionine (14) is the precursor of allylglucosino- removed them (Dahlgren, 1983; Dahlgren et al., 1981).
late (sinigrin) (15). The allyl group is derived by loss of Glucosinolates are found in several other families that
the terminal sulfur function from 3-(methylsulfinyl)propyl are not particularly related to each other nor are they proba-
glucosinolate (13) (Dewick, 1984). bly closely related to the first two groups of families. Among
Feeding experiments with 3H_ and 14C-Iabeled precursors these are the Caricaceae (possibly related to the Brassica-
in leaves of horseradish (Armoracia lapathifolia) demon- ceae), the Euphorbiaceae (Putranjiva roxburghii and the
strated the stereospecific removal of the 4-pro-S-hydrogen genus Drypetes), the Gyrostemonaceae, Bretschneideraceae,
of homomethionine during the biosynthesis of sinigrin. The Bataceae, Sterculiaceae, Plantaginaceae, and Salvadoraceae
fate of the 5-hydrogens has also been established by 2H_ (Boufford et al., 1989; Louda and Mole, 1991).
NMR (Fig. 17.5) (Dewick, 1984). An overall anti process
is involved. Mutants of Arabidopsis thaliana which lack the Vtillzation of Olucosinolate Data
ability to extend the carbon chain of methionine beyond the in Systematic Studies
homomethionine stage have been reported (Davin et aI., Problems with generic delimitation and evolutionary
1988). trends in the family Brassicaceae are complex (AI-Shehbaz,
1973). Glucosinolates, however, often exhibit species-spe-
DISTRlBUDOI'l cific arrays of compounds that are of value for the study of
systematics and evolutionary studies in this group (Chew,
Glucosinolates occur primarily in II plant families. One 1988).
cluster of these families, the order Capparales (including The genus Thelypodium has been treated in several ways
the Brassicaceae, Capparidaceae, Moringaceae, Resedaceae, by taxonomists, but has been studied in a multidisciplinary
 Glucosinolates 305

NH, NOSO~
CH3S~ CO,H -+ CH,S I
~s.g1urosYI
homomethionine (14) (13) \

/"
,......,..
r
NOSO;

CHS~~
..."
O·

S·glurosyl
NOSO,'
~s.gIUCOSYI
sinigrin (15)

~
-- 8,J . . . .,rso,
H'Hd NH,

..
CHrS 5 H.
4 Co,H 'S~glucosyl
Ii H
Hb

Fig. 17.5. Biosyntbetic steps in the synthesis of sinigrin (Dewick, 1984; modified and used with pennission of the copyright owner, the Royal Society
of Chemistty, Cambridge),

manner by Al-Shehbaz (1973). The hydrolysis products of composition. Further, there seemed to be little variation
glucosinolates of the taxa of interest were examined by paper within populations of a particular species. Although the qual-
chromatography after conversion to the thioureas and oxa- itative pattern of greenhouse-grown plants was almost iden-
zolidine-2-thiones (Ettlinger and Thompson, 1962; Ettlinger tical to that of wild plants, some quantitative differences
et al., 1966). The isothiocyanates derived from hydrolysis were observed. The glucosinolates were found to be stable
were determined by gas chromatography. A variety of glu- under a number of environmental conditions and consis-
cosinolates (17 in all) were found in Thelypodium species. tently present in popUlations of each taxon. Single plants
Both seeds and vegetative material of most species were usually contained the full complement of compounds found
investigated. With the exception of a few species, the quanti- in mass collections. Distinctive qualitative and quantitative
tative as well as the qualitative patterns of glucosinolates in patterns were found for most of the taxa studied. Intraspe-
Thelypodium were species-specific. The exceptions were cific variation which correlated with morphologically recog-
five species with 2-hydroxy-3·butenyl glucosinolate (16) nized subspecies was found in Cakile edentula, C. maritima,
and T. eucosmum and T. paniculatum that were chemically and C. laneeolata. Geographic variation was not closely
indistinguishable. Almost no quantitative or qualitative dif- linked to morphological variation in Atlantic coast C. eden·
ferences in glucosinolates were found between the plants of tula ssp. edentula and in Pacific coast C. edentula where it
a given population. Although these data were useful at the appeared to be clinal (Rodman, 1974).
specific level, they did not provide significant taxonomic As indicated above, the glucosinolate complement of
information concerning the relationships of the genera in the many species appears to differ. Analysis of the hydrolysis
tribe Thelypodieae (Stanleya, Thelypodium, Thelypodiopsis, products of Draba euneifolium and D. platyearpa suggested
Caulanthus, Streptanthus, and Streptanthella) (Al-Shehbaz, that these two taxa were distinct (Hartman et al., 1975).
1973). Recent evidence indicates that insect infestations can dra-
The genus Cakile is found in both the Old World and matically alter both glucosinolate concentration and compo·
New World. Many variants of taxa of this genus were de- sition in Brassiea napus (Koritsas et al., 1989).
scribed and relationships were poorly understood until Cak·
ile was monographed by Rodman (1974). In addition to stud-
ies of the anatomy, morphology, and cytology, the BIOLOGICAL ACTIVITY
glucosinolate composition of plants of the genus was exam·
ined. Glucosinolates were isolated from seeds, hydrolyzed,
Properties of Glucosinolates
and converted to the thiocyanates and oxazolidine-2-thiones.
Analysis was similar to that above. The two morphologically Many of our common vegetables come from families
distinct seeds of fruits of Cakile species did not differ in such as the Brassicaceae that possess glucosinolates. These
J06 (;llIc(lsi"ola'('.~

plants are edible and nutritious and the glucosinolates (and Froperties of the Hydrolysis Froducts
their derivatives) seem to have little adverse physiological
The intact glucosides are probably not the active agents,
effect in humans. Some of the vegetables rich in glucosino-
but rather the isothiocyanates, thiocyanates, and nitrile deriv-
lates are mustard greens (Brossicajuncea), broccoli, brussel
atives derived from them. The initial hydrolysis products
spmuts. cabbage, cauliflower, kale, kohlrabi, collards (all
are thiohydroximate O-sulfonates, which are followed by
Brossica olerocea), horseradish (Armoracea lapathifolia), rearrangements to give isothiocyanates, and, under some
radish (Raphanus sativa), rutabaga (Brassica campestr;s), conditions, nitriles and thiocyanates are also formed (De-
turnip (Brossica rapa), a numberofiess well-known vegeta- wick, 1984). In contrast to the intact glucosinolates, most of
ble cultivars (especially in China) (Hill et aI., 1987), and the the hydrolysiS products are relatively volatile (Fig. 17.6).
oilseed crop plants crambe (Crambe abyssinica) and rape Isothiocyanates and thiocyanates are quite reactive chem-
and canola (Brassica napus). However, no mammals are ically; they react with water, alcohols, amines, and a variety
known to specialize on crucifers and excess consumption of other functional groups, either free or on macromolecules
produces numerous problems (Louda and Mole, 1991). (Larsen, 1981).
Nonetheless, many small mammals occasionally rely on cru- 4(a-L-Rhamnosyloxy)benzyl isothiocyanate (18) has
cifer seeds as a major part of their diet. been identified as an active antimicrobial agent from seeds
Indoles in vegetables of the genus Brassica inhibit carci- of Moringa oleifera and M. stenopetala (Eilert et al., 1981).
nogenesis in experimental animals. These compounds arise The seeds contain 8-10% of this compound, which inhibited
from enzymatic hydrolysis of glucobrassicin (3) (Fig. 17.1). a broad range of bacteria and fungi in cultures at concentra-
However, consumption of these compounds under other con- tions of about 40 f.Lmol/L (Eilert et aI., 1981).
ditions enhances carcinogenic activity (Beier and Nigg, Enzymic hydrolysis of3-indolylmethylglucosinolate (20)
1992). Consumption of cruciferous vegetables is linked to from fresh herbage of Brassica species leads to the formation
a decreased incidence of colo-rectal tumors in humans (Beier of 3-indoleacetonitrile (19), a compound with known auxin
and Nigg, 1992; Cassady et al., 1990). A key compound activity (Tapper and Reay, 1973).
responsible for the activity of broccoli is sulforaphane (17) In many references, it is not clear whether the hydrolysis
[( - )-I-isothiocyanoato-(4R)-(methylsulfinyl)butanel (Fig. products or the intact glucosinolates are involved. The hy-
17.1) (Zhang et al., 1992). drolysis products are volatile, whereas the glucosinolates are
Nonetheless, glucosinolates and their derivatives are bio- not. Nonetheless, both types of compound can and do have
biological activity. Some hydrolysis products of glucosino-
logically active and have pronounced effects when con-
lates that stimulate or attract herbivorous insects have been
sumed in excess or when encountered by certain other organ-
tabulated (Stiidler, 1992).
isms. Numerous instances of animal poisonings occur
because of these plants. Detoxification of the pressed or ex-
Goitrigenic Derivatives in Crop Plants
tracted seed meals of rape and crambe are important pro-
cesses because of the large quantities that are used as feed Consumption of plant materials containing glucosinolates
supplements for livestock and poultry (VanEtten and and their hydrolysis products are known to produce goiter
Tookey, 1978). in animals (Beier and Nigg, 1992). The presence of glucosi-

--
R~-OSO;K+ OH • + RNCS + So,"

-
--
R--<-OS03 K
\ HO~OH RCN+S+SOi''"
S~0---h
OH S'
RSCN+SO,"

~N:C:S

oV
HO~H
HO OH 3-jndoleacetonitrile (19)

4(a-L-rbanmosyloxy)benzyl isothiocyanate (IS)

Fig. 17.6. Hydrolysis of glucosinolates (Dewick, 1984; modified and used with pennission of the copyright owner, the Royal Society of Chemistry,
Cambridge).
 Glucosinolates 307

l-cyano-2-hydroxy·3,4-epithiobutanes

Rz Rt
t
\1 N.OSo.K+
CHrCH-C-CH,----( OH
- S~OH
OH

R. =OK, R, = K, epi-progoitrin (22) R, = H, R, = OH, progoitrin (21)

/
"'"
R, = OH, R, = H, (S)
l-cyano-l-bydroxy-3-butenes
R, = H, R, = OH, (R)

goitrins

(SR)-S-vinyloxazolidine-2-tbione (5S)-5-vinyloxazolidine-2-thione (23)

Fig. 17.7. Secondarily derived hydrolysis products (modified from VanEtten and Tookey, 1979; used with pennission of the copyright owner,
Academic Press, Orlando, FL).

nolates is especially important in two oilseed plants, nays, 1983; Louda and Mole, 1991). On the other hand,
Brassica napus (rapeseed) and Crambe abyssinica (crambe). insects that normally feed on plants containing these sub-
As the presscake from these seeds is produced in quantity stances may require their presence for continued feeding
as a by-product of oil isolation, it represents a significant (Renwick, 1988). Not all glucosinolates serve equally well
economic and waste problem. In general, rapeseed meal may in any particular case (Louda and Mole, 1991). Glucosino-
be fed at no more than 10-15% of the total diet because lates and their hydrolysis products that stimulate or attract
consumption of larger quantities of glucosinolates leads to herbivorous insects have been tabulated (Stiidler, 1992).
vesication of mouth and throat tissues (VanEtten and Butterflies of the genus Papilio normally do not feed on
Tookey, 1978, 1979). Rapeseed contains (2R)-hydroxy-3- plants that contain glucosinolates. Glucosinolates are toxic
butenylglucosinolate (progoitrin) (21) and crambe contains to the black swallowtail (Papilio polyxenes) when larvae of
the corresponding (2S)-compound (epi-progoitrin) (22) (Fig. this insect eat celery leaves that have been infiltrated with
17.7). In efforts to detoxify the meal, itis often heated. Heat potassium allylglucosinolate (sinigrin) (Fig. 17.1) (15) at a
treatment of these seed meals produces a somewhat different concentration of 0.1 % fresh weight (Chew, 1988; David and
complement of compounds than those naturally formed. In Gardiner, 1966).
this case, secondarily derived compounds are formed from Insects that normally feed on brassicaceous plants, such
glucosinolate hydrolysis products that are even more toxic as cabbage butterflies of the genus Pieris, can consume these
than the original glucosinolates. One of these compounds, glucosinolates without obvious effect. The presence of glu-
(S)-5-vinyloxazolidine-2-thione (goitrin) (23) prevents up- cosinolates is required for normal feeding by the insects.
take of iodine and leads to the formation of goiter in livestock Verschaffelt (1910) was able to induce larvae of Pieris
fed this meal. brassicae (cabbage butterfly) and P. rapae to eat plant mate-
rials they normally reject by applying potassium allylglucos-
Glucosinolates as A1lomones and inolate to the surface of the leaves. The larvae of Pieris
Kairomones brassicae will eat artificial diets when potassium allylglu-
cosinolate (sinigrin) or other glucosinolates are added (Ren-
The presence of glucosinolates in plant tissues inhibits wick, 1988). Once they have eaten this diet, the larvae
feeding by most non-coadapted insects and mammals (Ber- will starve before they will eat a diet without glucosinolates.
308 GJucosinoiates

- +
J N-OSO,K _ +
N-OSO,K
f
HO~OH ~.~~OH
CH,\ OH '-...
S
~u-------h 'I' S HO 0 OH
OH 0
OH

glucocapparin (24) glucoiberin (25)

~NH ~NH
~N) I
SASCH
'
~N} SASCH,
OCH,
(27) (28)

~NH ~N
~N) I
OASCH
' ~NH~S)lSCH'
OCH,
(29) (30)

~>-SCH'
~NHJ::O
(31)

Fig. 17.8. Some biologically active glucosinolates.

Glucocapparin (24) and glucoiberin (25) (Fig. 17.8) serve be critical for the survival of the crop under feeding pressure
as feeding stimulants for Pieris brassicae larvae and are of the flea beetle in the field.
active at about 10-6 M concentration in the diet, whereas For the diamondback moth, Plutella xylostella (syn. A.
all other glucosinolates tested were active only at 10-4 M_ maculipennis) (another crucifer-feeding insect), (2R)-hy-
When potassium allylglucosinolate (sinigrin) and linseed oil droxy-3-butenylglucosinolate (progoitrin) (21) was more ef-
were added to the diet, normal growth resulted. illterestingly, fective than other glucosinolates in inducing feeding on arti-
glucocapparin and glucoiberin, the two most effective com- ficial diets.
pounds, do not occur in cabbage or other Brassica species, The cabbage aphid, Brevicoryne brassicae, is highly spe-
but do occur in the alternate food host Tropaeolum majus. cific in its feeding behavior to cruciferous plants, although
The flea beetle Phyllotreta cruciferae has a narrow host it can be induced to feed on other plants when g1ucosinolates
range, feeding only from plants in the families Brassicaceae, are added exogenously. These aphids use the presence of
Capparidaceae, and Tropaeolaceae. ill the laboratory, the in- potassium allylglucosinolate (sinigrin) to detect an appropri-
sect fed on plants of these families and the Limnanthaceae. ate host plant. The insects land and test the plant; if potas-
All of these plants contain glucosinolates. Field experiments sium allylglucosinolate (sinigrin) is lacking, they continue
showed that allyl isothiocyanate (26) (Fig. 17.1) is a power- searching for an appropriate host plant (Harborne, 1982).
ful attractant for adults of both Phyllotreta cruciferae and
P. striolata (Feeny et aI., 1970). Glucosinolates and Oviposition by Insects
Seedlings of Sinapis alba (mustard) have high concentra- Oviposition frequently involves a complex series of stim-
tions of sinalbin (1) (20 mM in cotyledons and 10 mM in uli, many of which are chemical. Further, because many
leaves) and are resistant even to the attacks of flea beetles larvae are relatively immobile, selection of the host plant
and the bertha army worm, Mamestra corifigurata (Bodn- effectively occurs when the gravid female selects a site for
aryk, 1991). The resistance that sinalbin confers appears to oviposition (Renwick, 1988). However, many factors other
 G/ucosinolates 309

than glucosinolates are involved in host selection (Chew, The host range of the diamondback moth, Plutella xylos-
1988). For instance, nutrient availability is also related to tella, appears to be correlated with the presence of glucosino-
the selection of Brassica nigra plants for oviposition by fe- lates in the host plants, but the role of stimulants and attrac-
males of Pieris rapae (Wolfson, 1980). Orientation of this tants in oviposition seem to be important. Extracts of
insect also is based on color of the host plant. In instances Brassica oleracea (cabbage), B.juncea (mustard), and Erysi-
where stimulatory cabbage extracts were obscured from mum cheiranthoides stimulated oviposition by gravid fe-
sight by placing them in cups (with holes punched in the males on bean plants. Extracts of Erysimum cheiranthoides,
cups), there was no preference for these extracts (Renwick, Tropaeolum majus, and Capsella bursa-pastoris which de-
1988). terred oviposition by the cabbage butterfly Pieris rapae,
Some glucosinolates and glucosinolate hydrolysis prod- were not deterrent to Plutella xylostella (Renwick and
ucts that are involved in oviposition by insects have been Radke, 1990).
tabulated (Stiidler, 1992).
Glucosinolates can induce oviposition by the cabbage Long-Distance Attraction
butterfly. Gravid females of Pieris brassicae oviposit on Although there is a common view that isothiocyanates
filter paper or on Vida faba, if it is first treated with potas- are important odor components of intact brassicaceous
sium allylglucosinolate (sinigrin) (15). It has been suggested, plants, field levels are extremely low and are usually below
however, that the natural stimulant is glucobrassicin (3-indo- the level of detection of the usual analytical techniques. The
lymethyl glucosinolate) (3) (Traynier and Truscott, 1991). principal vapor component of turnip and radish is hexenyl
This compound elicited oviposition at threshold concentra- acetate (see Chapter 2). Even in the case of disrupted tissue
tions as low as 10-6 M. Sinigrin is active at 10-2 M. The of cauliflower, turnip, radish, and wallflower (Alyssum), the
markedly greater potency of glucobrassicin is consistent principal component was Z-hex-3-enyl acetate (Wallbank
with known glucosinolate profiles of crucifers which show and Wheatley, 1976). These data suggest that "green leaf
indolyl glucosinolates predominant in foliage (Traynier and volatiles" may be involved in the attraction of many species
Truscott, 1991). of insects to cruciferous plants.
In studies in which the oviposition stimulants for Pieris
rapae from nasturtium, mustard, and cabbage were fraction- QIucosinolates and Plant Disease
ated, the most attractive fractions from the three plants were Resistance
distinct; these studies indicate that the insect responds to a
different complement of compounds in different crucifers Glucosinolates via their hydrolysis products are among
(Renwick,1988). the most powerful antibiotic substances known from higher
In another study, females of Pieris rapae oviposited on plants (Louda and Mole, 1991). In cultivated Brassica spe-
plants of Cardamine cordifolia in one field, but did not ovi- cies, there is a correlation between the content of glucosino-
posit on plants of this species in an adjacent area, even lates (isothiocyanates) and disease resistance (Chew, 1988;
though both the butterfly and the plants were present (Chew, Greenhalgh and Mitchell, 1976). Wild populations of
1977). When cabbage plants were sprayed with a butanol Brassica species typically have higher levels of glucosino-
extract of Erysimum cheiranthoides, a wild crucifer normally lates and are usually more disease resistant. Selective breed-
rejected by ovipositing Pieris rapae because of the presence ing for milder flavored individuals has been a contributory
of cardenolides in the foliage, oviposition by gravid females factor to the general lack of resistance of modem varieties to
was greatly reduced (Dimock and Renwick, 1991). This in- powdery mildew and other pests (Greenhalgh and Mitchell,
sect does not oviposit on many other species of crucifers. 1976).
These results confirm that a combination of attractant and A series of antifungal sulfur-containing indole derivatives
deterrent effects are involved in oviposition behavior. Repel- (27-31), which appear to be related to glucobrassicin (3), are
lent compounds may include cardenolides, alkaloids, and produced as phytoalexins in Raphanus sativus and Brassica
campeslris in response to the fungus Leptosphaeria macu-
cucurbitacins (Renwick, 1988).
Allyl isothiocyanate (26) is the principal volatile compo- lans (Devys et aI., 1990; Barbome, 1989) (Fig. 17.8).
nent produced by damaged cabbage. This compound is at-
tractive to the cabbage root fly [Delia (Erioischia) brassi-
QIucosinolates and AUelopatby
cae) (Finch, 1978; Wallbank and Wheatley, 1976). Antennal A number of cruciferous plants inhibit the growth of com-
receptors proved sensitive to allyl isothiocyanate, whereas peting vegetation; several glucosinolates and their hydroly-
the tarsal receptors of females were sensitive only to the sis products have been isolated and demonstrated to possess
corresponding glucosinolate (Louda and Mole, 1991). The herbicidal or growth-inhibiting activity. Others inhibit seed
insect will lay eggs when provided with allyl isothiocyanate germination.
or the corresponding glycoside, allyl glucosinolate (15). Benzyl isothiocyanate (32) (Fig. 17.1) from Carica pa-
Allyl isothiocyanate is involved in attraction of the mus- paya seeds strongly inhibited germination of the seeds of
tard beetle, Phaedon cochleariae, to its host plants (Tanton, Abutilon theophrasti (velvet leaf, Malvaceae) at 4 X 10-4
1977). M (Wolf et aI., 1984).
310 Glucosinolates

Sulforaphene (4-methylsulfinyl-3-butenyl isothiocya- CoLE, R. A., Isothiocyanates, nitriles, and thiocyanates as products
nate) (33) has an EDso of approximately 2 X 10-4 M against of autolysis of glucosinolates in Cruciferae, Phytochemistry,I5,
the seedlings of velvetleat (Abutilon theophrasti, Malva- 759-762 (1976).
ceae) (Brinker and Spencer, 1993). DAlD..GRBN, R., General aspects of angiospenn evolution and ma-
A series of isothiocyanates from Rorippa indica inhibit crosyatematics, Nordic r. Bot., 3, 119-149 (1983).
lettuce hypocotyl and root growth at 0.1 mM. The corre- DAHLGREN, R., S. ROSBNDAL-IBNSBN, and B. I. NIELSEN, A revised
sponding glucosinolates hirsutin (34), arabin (35), and cam- classification of the angiospenns with comments on correlation
melinin (36) were isolated. The isothiocyanates appear to be between chemical and other characters, in Phytochemistry and
responsible for most of the aUelopathic activity of this weed Angiospenn Phylogeny (D. A. Young and D. S. Seigler, eds.),
194-204, Praeger, New York, 1981.
(Yamane et aI., 1992).
This phenomenon may involve incompatibility of brassi- DAVID, W. A. L. and B. O. C. GARDINER, Mustard oil glucosides
as feeding stimulants for Pieris brassicae larvae in a semi-syn-
caceous plants with mycorrhizal fungi (Chew, 1988). How-
thetic diet, Entomol. Exp. Appl., 9, 247-255 (1966).
ever, these authors concluded that rather than producing
compounds harmful to the mycorrhizal fungi, plants of the DAVIN, L" G. HAUGHN, D. REED, M. GIBLIN, and E. W. UNDBRHll.L,
Biochemical genetics of glucosinolates in Arabidopsis thaliana,
Brassicaceae fail to produce compounds stimulatory to the
Abstracts, Phytochem. Soc. North America, Annual Meeting,
fungi. 1988.
DBWICK, P. M., The biosynthesis of cyanogenic glycosides and
glucosinolates, Nat. Prod. Rep., I, 545-549 (1984).
REFERENCES DBVYS, M., M. BARBIER, A. KOLLMANN, T. RoUXEL, and 1. Bous-
QUET, Cyclobrassicin sulphoxide, a sulphur-containing phyto-
AL-SHBHBAZ, I. A., The biosystematics of the genus Thelypodium alexin from Brassica juneea, Phytochemistry, 29, 1087-1088
(Cruciferae), Cont. Gray Herb., 204, 3-148 (1973). (1990).
BBIER, R. C. and H. N. NIGG, Natural toxicants in foods, in Phyto- DIMOCK, M. B. and I. A. A. RBNwrCK, Oviposition by field popula-
chemical Resources for Medicine and Agriculture (H. N. Nigg tions of Pieris rapae (l.epidoptera: Pieridae) deterred by an
and D. S. Seigler, eds.), 247-376, Plenum Press, New York, extract of a wild crucifer, Environ. Ent., 20, 802-806 (1991).
1992. EILHRT, U., B. WOLTERS, and A. NAHRSTBDT, The antibiotic princi-
BBRNAYS, E. A .• Nitrogen in defence against insects, in Nitrogen ple of seeds of Moringa oleifera and Moringa stenopetala,
as an Ecological Factor (I. A. Lee, S. McNeil, and I. H. Rorison, Planta Medica, 42, 55-61 (1981).
eds.), 321-344, Blackwell, Oxford, 1983. ETI1.INGER, M. G. and K. KJABR, Sulfur Compounds in Plants, cr.
BBTZ, I. W. and S. W. PAGB, Liquid chromatographic method for 1. Mabry, R. E. AJston, and V. C. Runecldes, eds.), Vol. 1 of
the determination of intact, nonderivatized glucosinolates from Recent Advances in Phytochemistry, Vol. 1,59-144 Appleton
Brassicaceae, Planta Medica, 56, 590-591 (1990). Century Crofts, New York, 1968.
BODNARYK, R. P., Developmental profile of sinalbin (p-hydroxybe- ETI1.INGER, M. G. and A. 1. LUNDEEN, The structures of sinigrin
nzyl glucosinolate) in mustard seedlings, Sinapis alba L. and its and sinalbin; an enzymatic rearrangement, 1. Am. Chem. Soc.,
relationship to insect resistance, J. Chern. Ecol.,17, 1543-1556 78,4172-4173 (1956).
(1991). ElTLINGER; M. G. and C. P. THOMPSON, Studies of Mustard Oil
BOUCHEREAU, A., r. HAMBLIN, I. LAMOUR, M. RENARD, and F. Glucosides (II), AD No. 290747, Office of Technical Services,
LARHER, Distribution of sinapine and related compounds in U. S. Dept. Commerce, Washington, DC, 1962.
seeds of Brassica and related genera. Phytochemistry. 30, ETI1.INGER, M. G., A. KJABR, C. P. THOMPSON, and M. WAGNIERBs,
1873-1881 (1991). Veratryl isothiocyanate, a new mustard oil from Helicopia lon-
BOUFFOaD, D. E., A. KJABR, r. 0. MADSEN, and T. SKRYDSTRUP, gifolia DC. (Cruciferae), Acta Chern. Scand., 20, 1778-1782
Glucosinolates in Bretschneideraceae, Biochem. Syst. :Ecol., 17, (1966).
375-380 (1989). FEENY, P. P" K. L. PAAUWB and N. I. DBMONG, FIea beetles and
BRINKBa, A. M. and G. F. SPENCER, Herbicidal activity of sulfora- mustard oils: Host plant specificity of Phyllotreta cruciferae
phene from stock (Matthiola incana), I. Chern. Ecol., 19, and P. stridata adults (Coleoptera: Chrysomelidae), Annals En-
2279-2284 (1993). tomol. Soc. Am., 63, 832-841 (1970).
CASSADY, I. M., W. M. BAIRD, and C. CHANG, Natural products FINCH, S" Volatile plant chemicals and their effect on host plant
as a source of potential cancer chemotherapeutic and chemo- finding by the cabbage root fly (Delia brassicae), Ent. Exp.
preventative agents, 1. Nat. Prod., 53, 23-41 (1990). Appl.,24, 150-159 (1978).
CHEw, F. S.• Coevolution of pierid butterflies and their cruciferous GEISSMAN, T. and D. H. G. OOlIT, Organic Chemistry of Second-
foodplants. II. The distribution of eggs on potential foodp1ants, ary Plant Metabolism, Freeman-Cooper, San Francisco, 1969.
Evolution, 31, 568-579 (1977). GoB1"Z, 1. K. and H. SCHRANDORF, Two natural indole glucosino-
CHEW, F. S., Searching for defensive chemistry in the Cruciferae, lates from Brassicaceae, Phytochemistry, 22, 905-907 (1993).
or, do glucosinolates always control interactions of Cruciferae GREENHALGH, 1. R. and N. D. MITCHELL, The involvement of fla-
with their potential herbivores and symbionts. No! in Chemical vour volatiles in the resistance to downy mildew of wild and
Mediation of Coevolution (K. C. Spencer, ed.), 81-112, Aca- cultivated fonns of Brassica o/eracea, New Phytologist, 77,
demic Press, San Diego, CA, 1988. 391-398 (1976).
 Glucosinolates 311

GROO1WASSINK, J. W. D., J. J. BALSEVICH, and A. D. KOLENOVSKY, SLOMINSKI, B. A. and L. D. CAMPBELL, Gas chromatographic deter-
Formation of sulfatoglucosides from exogenous aldoximes in mination of indole glucosinolates-A re-examination, 1. Sci.
plant cell cultures and organs, Plant Sci., 66, 11-20 (1990). Food Agric., 40, 131-143 (1987).
HARBORNE, J. B., Introduction to Beological Biochemistry, 2nd ed., STADLER, E., Behavioural responses of insects to plant secondary
Academic Press, London, 1982. compounds, in Herbivores: Their Interactions with Secondary
HARBORNB, 1. B., Recent advances in chemical ecology, Nat. Prod. Plant Metabolites, Vol. 2 (G. A. Rosenthal and M. R. Beren-
Rep., 7, 85-109 (1989). baurn, eds.), 45-88, Academic Press, San Diego, CA, 1992.
HARTMAN, R. L., J. D. BACON, and C. F. BOHNSTBDT, Biosystemat- TANmN, M. T., Response to food plant stimuli by larvae of the
ics of Draba cuneifolia and D. p/atycarpa (Cruciferae) with mustard beetle Phaedon eoch/eariae, Ent. Exp. Appl., 22,
emphasis on volatile and flavonoid components, Brittonia, 27. 113-122 (1977).
317-327 (1975). TAPPER, B. A. and P. F. REAY, Cyanogenic glycosides and glucosi-
HASLAM, E., The Shikimate Pathway, Wiley, New York, 1974. nolates (mustard oil glucosides), in Chemistry and Biochemistry
Hn.r., C. B., P. H. Wn.r.IAMS, D. G. CARLSON, and H. L. TOOKBY, of Herbage, Vol. 2 (G. W. Butler and R. W. Bailey, eds.),
Variation in glucosinolates in oriental Brassica vegetables, J. 447-476, Academic Press, New York, 1973.
Am. Soc. Hort. Sci., 112, 309-313 (1987). TRAYNIER, R. M. M. and R. J. W. TRUSCOTT, Potent natural egg-
KORITSAS, V. M., J. A. LEWlS, and G. R. FENWlCK, Accumulation laying stimulant for cabbage butterfly, Pieris rapae J. Chern.
of indole glucosinolates in Psylliodes chrysocephala L. infested Beol.,17, 1371-1380 (1991).
or damaged tissues of the oilseed rape (Brassica napus L.), UNDERHILL, E. W., Glueosinolates, in Secondary Plant Products
Experientia, 45, 493-495 (1989). (E. A. Bell and B. V. Cbarlwood eds.), 493-511, Springer-
LARSEN, P.O., Glucosinolates, in Secondary Plant Products (E. E. Verlag, Berlin, 1980.
Conn ed.), Vol. 7 of The Biochemistry of Plants (p. K. Stumpf
VANETTBN, C. H. and H. L. TOOKBY, Glucosinolates in Cruciferous
and E. E. Conn, eds.), Academic Press, New York, 1981.
plants, in Effects of Poisonous Plants on Livestock (R. F. Keeler,
LOUDA, S. and S. MOLE, Glucosinolates: Chemistry and ecology K. R. van Kampen, and L. F. James eds.), 507-520, Academic
in Herbivores: Their Interactions with Secondary Plant Metabo- Press, New York, 1978.
lites (G. A. Rosenthal and M. R. Berenbaum, eds.) San Diego,
VANETTBN, C. H. and H. L. TOOKBY, Chemistry and biological
California 123-164 (1991).
effects of glueosinolates, in Herbivores (G. A. Rosenthal and
OLSSON, K., O. TIiEANoER, and P. AMAN, IH_NMR, "C-NMR and D. H. Janzen eds.),471-500, Academic Press, New York, 1979.
mass spectra of glucosinolates and related compounds, Carbo-
hydro Res., 58, 1-8 (1977). VERSCHAFFELT, E., The cause detennining the selection of food
in some herbivorous insects, Proc. Acad. Wet., 13, 536-542
POULIDN, J. E. and B. L. M~LLER, Glucosinolates, in Enzymes of
(1910).
Secondary Metabolism (P. J. Lea, ed.), Vol. 9 of Methods of
Plant Biochemistry (P. M. Dey and J. B. Harbome, eds.), WALLBANK, B. E. and G. A. WHEATLEY, Volatile constituents from
209-237, Academic Press, London, 1993. cauliflower and other crucifers, Phytochemistry, 15, 763-766
(1976).
REED, D. W., L. R. HOGGE, J. W. D. GROO1WASSINK, and E. W.
UNDERHn.I., Metabolic evidence for the degree of specificity of WOLF, R. B., G. F. SPENCER, and W. F. KWOLEK, fuhibition of
glucosinolate biosynthetic enzymes, Abstracts, Phytochem. velvetleaf (Abutiton theophrasti) germination and growth by
Soc. North America, Annual Meeting, 1988. benzyl isothiocyanate, a natural toxicant, Weed ScL, 32,
RENWlCK, J. A. A., Comparative mechanisms of host selection by 612-615 (1984).
insects attacking pine trees and crucifers, in Chemical Mediation WOLFSON, J. L., Oviposition response of Pieris rapae to environ-
of Coevolution (K. C. Spencer, ed.), 303-316, Academic Press, mentally induced variation in Brassica nigra, Ent. Exp. Appl.,
San Diego, CA, 1988. 27,223-232 (1980).
RENWlCK, J. A. A. and C. D. RADKE, Plant constituents mediating YAMANE, A., J. FuJIKURA, H. OGAWA, andJ. MIzuTANI, Isothiocya-
oviposition by the diamondback moth, P/utella xy/ostella (L.J nates as allelopathic compounds from Rorippa indica Hiem.
(Lepidoptera: Plutellidae), Phytophaga, 3, 37-46 (1990). (Cruciferae) roots, J. Chern. Beol., 18, 1941-1954 (1992).
RODMAN, J. E., Systematics and evolution of the genus Cakite (Cru- ZHANG, Y., P. TALALAY, C. CHO, and G. H. POSNER, A major
ciferae), Cont. Gray Herb., 205, 34-60 (1974). inducer of anticarcinogenic protective enzymes from broccoli:
SCHRAUDOLF, H., Indole glucosinolates of Capparis spinosa, Phy- Isolation and elucidation of structure, Proc. Natl. Acad. Sci.
tochemistry, 28, 259-260 (1989). USA, 89, 2399-2403 (1992).
18
Introduction to Terpenes

Introduction General procedures for the isolation, purification and

The Isoprene Rule characterization of various types of terpenes have been re-
Biosynthesis of the Cs Unit viewed (Banthorpe, 1991; GersheD20n and Croteau, 1991).
Stereochemical Features of Mevalonic Acid
Isopentenyl Pyrophosphate and Dimethylallyl
Pyrophosphate
Hemiterpenes THE ISOPREl'IE RULE
Chromenes, Benzofurans, Benzopyrans, and Precocenes
Polyterpenes Early studies indicated that terpenes were composed of five
Biosynthesis carbon units. By 1860, isoprene, a compound with the empir-
Biological Activity ical formula CsH., could be derived by pyrolysis of turpen-
Natural Rubber tine and rubber. Further, isoprene could be transformed into
Hevea brasiliensis a C IOH'6 compound that subsequently decomposed back to
Guayule the original five-carbon unit. Wallach recognized that ter-
Gutla Percha penes could be subdivided into several groups: hemiter-
Chicle penes, true terpenes, sesquiterpenes, and diterpenes. In 1887,
Dolichoprenols he proposed that monoterpenes (true terpenes) as well as
Prenylated Proteins other terpenoids were composed of isoprene units, a concept
Similar Five-Carbon Units Not Derived from Mevalonate that became known as the isoprene rule (Wallach, 1887).
References The structure of isoprene was not solved until much later
(Spurgeon and Porter, 1981).
In the 1920s, Ruzicka and co-workers systematically
studied the stroctures of many sesquiterpenes and a diter-
Il'ITKODVCTlOI'l pene, abietic acid (1959, 1973). The correct structure of cho-
lesterol was published by Wieland and Dane (1932) and by
Terpenes are the largest and most diverse group of plant others. Structures of other terpenoids, such as vitamin A,
secondary compounds; at least 15,000 terpenoids have been were solved in the 1930s. The empirical isoprene rule was
described, and thousands more are undoubtedly awaiting dis- useful for making assigrunents of structure, but the results
covery (Gershenzon and Croteau, 1991). Terpenes typically were not satisfactory in all cases. A "biogenetic isoprene
are made up of units of five carbons, Cs , C lO , CIS, ClO, C25 , rule" later was proposed by Ruzicka to explain the origin
C30 , and C40 structures (Fig. 18.1), that generally occur free of compounds formed from an aliphatic precursor made up
but may be modified, or derivatized as esters and glycosides, of isoprene units, but involving subsequent rearrangements
or attached to proteins. Terpenes are common in most plants (Banthorpe, 1991; Ruzicka, 1953, 1959, 1973).
and fungi, but they rarely accumulate in bacteria (Bell and The development and utilization of radioactively labeled
Charlwood, 1980; Poulter and Rilling, 1981). Steroids in compounds was a major step in the study of the biosynthesis
mammals are products of terpenoid metabolism. of terpenoids.

312
 Introduction to Terpenes 313

--op~ tc~~~~~-
\P: ~c:
WP I
~OPP

PPO~
A \ -+ WP \

DMAPP
.f \ a monoterpe~ne (E,E)-famesyl pyrophosphate

!
geranyl ~~;PhOSPhate I a sesquiterpene

FPP--+
¢
~
;~?P~ ~
Hl
__ ~ I ~ OPP FPP

Opp a diterpeoe
+GGPP / geranylgeraoyJ
" pyropbosphate
GGPP

~WP
*
sesterterpenes (e2S)
presqualene pyrophospbate

tetraterpenes (carotenoids) C40

Fig. 18.1. Structural types of terpenes.

610SYNfHESIS OF TIlE C5 UNIT methylglutaryl-CoA synthetase is first acetylated by acetyl-

CoA to give an acetyl-enzyme complex. This complex then
Although a number of C s precursor units for terpene had condenses with acetoacetyl-CoA to yield a covalent deriva-
been proposed over the years, studies in the 1950s indicated tive between HMO-CoA and the enzyme that is hydrolyzed
that a six-carbon compound, from brewer's yeast solubles, to produce HMO-CoA and the free enzyme (Bach et aI.,
mevalonic acid (MVA) (1), could support growth of an ace- 1990; Beale and MacMillan, 1988; Qureshi and Porter,
tate-requiring mutant strain of bacteria when acetate was 1981).
omitted from the growth medium. Subsequent feeding stud- Next, hydroxymethylglutaryl-CoA reductase (HMO-
ies indicated that mevalonic acid was incorporated into cho- CoA reductase, j3-hydroxy-j3-methylglutaryl coenzyme A
lesterol with loss of carbon dioxide. Most of the steps of reductase; E.C. 1. 1. 1.34) reduces 3-hydroxy-3-methylglu-
these basic pathways of terpenoid biosynthesis have been taryl-CoA (3) to (R)-mevalonic acid (MYA) (1) in a two-
worked out during the last 25 years, largely with yeast or step process (Fig. 18.3) (Bach et aI., 1990; Beale and Mac-
mammalian liver extracts. Millan, 1988; Harrison, 1988; Qureshi and Porter, 1981).
The first step in the formation of mevalonic acid involves This enzyme is widely considered to be the key regulatory
the formation of acetoacetyl-CoA (2) from acetyl-CoA (Fig. enzyme in the biosynthesis of cholesterol in humans and has
18.2). The enzyme responsible for this conversion, acetoace- been the target of many medically oriented studies; although
tyl-CoA thiolase (AACT, E.C.2.3.1.9) recently has been ob- plant HMOR has been less intensively studied, techniques
served in plastid preparations from the plants Parthenium for the solubilization, purification and characterization of
argentatum andPhaseolus radiatus (Bachet aI., 1990). Con- this enzyme have been developed (Bach et aI., 1990).
version of acetoacetyl-CoA to (3R)-3-hydroxy-3-methylglu- j3-Hydroxy-j3-methylglutaryl coenzyme A reductase
taryl coenzyme A (j3-hydroxy-j3-methylglutaryl coenzyme (HMO-CoA reductase) is widespread in bacteria, yeasts,
A, HMO-CoA) (3) is catalyzed by HMO-CoA synthase (3- higher plants, insects, and mammals and is present in all
hydroxy-3-methylglutaryl-CoA synthetase; j3-hydroxy-j3- tissues that synthesize isoprenoid compounds. In mammals,
methylglutaryl-CoA synthase, E.C.4.1.3.5). This enzyme this enzyme is localized in the endoplasmic reticulum; in
has been isolated from a number of plant sources, among yeasts, in the mitochondrial matrix; and, in plants, both chlo-
them orange peel and sweetpotato root tissue. 3-Hydroxy-3- roplasts and cytoplasm have HMO-CoA reductase activity
314

.-\ '-r
Introduction to Terpenes

bound
o
H
NH,
0)
H
1
alB'X'AJ -
XI~
s"

(3)
HH
"R'

pyridine nucleotide *corresponds to carbon 5

of mevalonic acid (1)

.-< ~
HAIRH Hi

~X
\1 CS/
X A "R' /N --

NH,

. X (IY
o H) H H H

I I

H)/H ) __
( A N"R'

H H H

Y
1

mevalonic acid (1)

Fig. 18.2. Mechanism of action of HMG-CoA reductase (modified from Qureshi and Porter. 1981).

(Bach et aI., 1990; Reddy and Das, 1987; Schulze·Siebert The subcellular compartmentation of terpenoid metabo-
etal., 1987; Schultz and Schulze-Siebert, 1989). The enzyme lism is a matter of controversy. Chloroplasts seem to contain
is largely membrane bound (Bach et aI., 1990). HMG-CoA the entire terpenoid biosynthetic pathway and appear to be
reductase from radish seedlings has a molecular weight of autonomous in regard to terpene biosynthesis. Carotenoid.
about 180,000, made up of units of about 45,000. A eDNA and the side chains of chlorophyll are synthesized in the
encoding an entire radish HMGR protein has been cloned chloroplasts. Sterols are synthesized in the cytoplasm (Gers-
and sequenced. This eDNA encodes a monomeric unit of henzon and Croteau, 1990).
about 63,000 molecular weight that may represent an unpro-
cessed form of the enzyme (Back et al., 1990). HMG-CoA Stereochemical Features of iIIevalonic Acid
reductase from Arabidopsis rhaliana is a protein with 592
amino acids and a molecular weight of 63,605 (Monfar et The mevalonic acid molecule (1) has three prochiral
al., 1990). This protein from potato tubers has a molecular methylene carbon atoms (and six prochiral hydrogen atoms)
weight of 110,000 and subunits of 55,000 (Beale and Mac- (Fig. 18.2). Replacement of any one of these hydrogen atoms
Millan, 1988). Factors involved in the regulation of HMG- with a tritium or deuterium atom will produce a new chiral
CoA reductase in plants have been reviewed (Gershenzon center. In strncture 1 (Fig. 18.2), HR indicates a proton that
and Croteau, 1990). will confer the (R)-configuration on the center bearing the
 Introduction to Terpenes 315

o H~
~
acetoacetyl-CoA
thiolase
j
CH,
kCH,COSCOA
CH,) (2)
-CH,COSCoA -CH,COSCoA

HMG·CoA HMG·CoA
synthetase >..
~ \.f,~OH COSCoA reductase

.O~ NADPH

3S-3-hydroxy-3-methylgIutaryl CoA (3)

H
~
OH
~,~~ NADPH
·0.,' SCoA
HO

(3R)-mevalonic acid (1)

Fig.lS.3. Biosynthesis of mevalonic acid (Qureshi et al., 1976: modified and used with pennission of the copyright owner, the American Chemical
Society, Washington, DC).

proton if replaced by an isotope of hydrogen, This proton (IPP) (4), is then isomerized to dimethylallyl pyrophosphate
is referred to as a pro·(R) proton, Replacement of the Hs' (DMAPP) (6), via the addition of a proton and abstraction
proton ipro·(S)] similarly produces the (S)·configuration at of the pro-(R)-hydrogen from C-2 (originally the 4-pro-(S)-
the new chiral center. hydrogen of MVA) (Fig, 18.4), Mechanistic studies indicate
that the oxygen from C-3 of mevalonate 5-diphophosphate
Isopentenyl Pyrophosphate and ends up in inorganic phosphate, These and other data require
Dimethylallyl Pyrophosphate a mechanism in which there is a direct enzyme-mediated
reaction between the oxygen at C-3 and ATP rather than an
The mevalonic acid molecule (1) possesses a chiral center initial transfer of phosphate from ATP to the enzyme and
(bearing hydroxyl and methyl) at C·3, Only the (3R)-enanti- then to mevalonate 5-diphosphate (Beale and MacMillan,
orner is used in isoprene synthesis, The fate of each of the 1988),
six possible (3R)-monotritiated species of MVA has been IPP and DMAPP are interconverted by isopentenyl-di-
determined in subsequent reactions of this important precur- phosphate D-isomerase (isopentenyl pyrophosphate: dimeth·
sor. These reactions are highly stereospecific (Cori, 1983; ylallyl pyrophosphate isomerase; isopentenyl pyrophosphate
Mann, 1987), ~3-~2-isomerase; EC 5.3.3.2), This enzyme has been iso-
The conversion of mevalonate (1) to isopentenyl pyro- lated from a number of plant sources, such as pumpkin.
phosphate (IPP) (4) involves two consecutive phosphoryla- orange peel, and pine seedlings, Isopenenyl-diphosphate D-
tions at position 5 by successive action of mevalonate kinase isomerase (from the fungus Claviceps) has a molecular
(EC 2,7.4.2) and a decarboxylation and dehydration of the weight of 35,000, a requirement for Mg2+, and an pH opti-
tertiary alcohol group by mevalonate 5-pyrophosphate de- mum of 6,0-85, and it is inhibited by geranyl and famesyl
carboxylase (EC 4,1.1.33) (Fig, 18.4) (Crotean & Jolinson, diphosphate,
1985; Gershenzon and Croteau, 1990), One mole of ATP Lynen and co-workers (1959) were the first to realize that
is required for each phosphorylation reaction, Mevalonate dimethylallyl pyrophosphate (6) corresponds to Ruzicka's
kinase converts mevalonic acid to (5R)-phosphomevalonate "active isoprene" and that this compound (and homologous
(5), The second phosphorylation is catalyzed by phospho- allylic pyrophosphates) is involved in repetitive condensa-
mevalonate kinase, The subsequent decarboxylation and de- tions with isopentenyl pyrophosphate (4) (Poulter and Rill-
hydration is mediated by the enzyme mevalonate diphos- ing, 1981),
phate decarboxylase (di- or pyrophosphomevalonate
decarboxylase; EC 4_1.1.3.3); this enzyme requires Mg2+ or
HElIUTEKPEl'IES
Mn 2+ and ATP for activity (Beale and MacMillan, 1988;
Harrison, 1988), All three of these enzymes are found in a Because DMAPP is such a potent electrophile, it is not sur-
number of plants, prising that the 3,3-dimethylallyl group is present in many
The product of this reaction, isopentenyl pyrophosphate secondary metabolites that derive the major part of their
316 Introduction to Telpenes

H ~H
~
mevalonate H H phosphomevalonate H H
~HOl t HOl
op kinase OPP
kinase
H H H H H H H H

mevalonic acid (1) mevalonic acid S·phosphate mevalonic acid

5·pyropbosphate (5)

~
OH~

-
f" C../ 0 0

:0 3 'O-~-O-~-OH
ATP

I
O·
I
O·
.CO,

mevalonic acid S.pyrophosphate

~ O-~-O-~-OH ~='H=R==..,,:::: k
o 0

H
I
H+
4I
H
3'
H. Hs
Y !... . . "o-LO-~-OH
O·
I I
O·
isomerase /3
H O· O·

isopentenyl pyrophosphate (4) dimethylallyl pyrophosphate (6)

DMAPP
IPP

Fig. 18.4. Origin and stereochemistry of isopentenyl pyrophosphate and dimethylallyl pyrophosphate fonnation.

skeleton from non-mevalonate-derived pathways (Fig. 18.5). Cbromenes. Benzofurans. Benzopyrans.

These reactions are catalyzed by a number of prenyltransfer- and Precocenes
ases. Products of C" N-, and O-alkylation are common, and
the dimethylallyl group is invariably located at sites that Most of the naturally occurring chromenes and benzofu-
were nucleophilic before alkylation, such as methylenes and rans (about 200 compounds) occur in the family Asteraceae
(Fig. 18.6) (Proksch and Rodriguez, 1983). These com-
phenolic oxygen atoms.
pounds are often cytotoxic and insecticidal. Both chromenes
Humulone (7), from Humulus lupulus (Cannabaceae), is
and benzofurans appear to be formed from dimethylallyl
partly responsible for the bitter flavor in beer. About 35 ppm
pyrophosphate and an acetate-derived polyketide precursor.
of hop-derived substances contibute a specific bitter taste
The terpenoid chemistry of at least 14 species of the genus
and hoppy flavor to beer. Hops also contribute for the micro-
Encelia (Asteraceae) has been examined. Most species of
biological stability and the stability of foam of beer (De
this genus are characterized by two distinct classes of terpe-
Keukeleire, 1991). However, the activity comes not from the noids: sesquiterpene lactones and benzpyransibenzofurans
native hop compounds, but from' 'iso-a<-acids" produced by (Isman et al., 1990). Analysis of the chromenes and related
boiling the wort-containing hops (De Keukeleire et ai., 1992; compounds of Encelia ventorum showed the presence of a
Bondeel et al., 1987). number of chromenes, benzopyran and benzofuran deriva-
Lysergic acid (8) occurs in ergot. The diethylamide (not tives, several of which were insecticidal. Encecalin (9) was
naturally occurring) is known as LSD. This topic is discussed moderately toxic to frrst-instar larvae of milkweed bugs and
in more detail in Chapter 35. has an LCso of 1.4 and 11.8 ....g/cm 2 (residue contact on
Hemiterpene units are also incorporated into furanocoum- glass surfaces) against the variegated cutworm (Peridromia
arins, some anthraquinones rotenoids, and other isoflavonoid saucia) and the migratory grasshopper (Melanop/us sangui-
derivatives. Often, the hemiterpene unit is further metabo- nipes, Acrididae), respectively. Encecalin also exhibits anti-
lized and may not be easily recognized. The presence of feedant properties against several species of noctuid caterpil-
intermediates sometimes serves as an indicator of mevalo- lars (Isman, 1989). Others (e.g., compound 10) were feeding
nate precursors in these pathways (see Chapter 9). deterrents to Helicoverpa zea. The benzofuran (11) from
 Introduction to Telpenes 317

o NH ~O 0 Ha02C
HN0 0 I OH ~ NCH,

o HO ~
~ I ~
"" NH

echinulin (a fungal antibiotic) humulone (7) lysergic acid (S)

HO

OCH,
OCH,
OCH, rotenone OCHJ

o o
~OH
y\,~o-\-
OCH3 CH J

lunacrine isobalfouridine

CH'OvSQ
OCHJ skimmianine dictamnine

~ HO""OO
"" -cCCl""
~ "" - r.=(X)1 I
I HO
0
""
000
I
"""00

Fig. 18.5. Hemiterpene-derived secondary metabolites.

Flourensia dentata and F. ilicifolia was toxic to milkweed been implicated (Beier and Norman, 1990; Beier and Nigg,
bugs and highly phototoxic (Harborne, 1986; Proksch et al., 1992; Kingsbury, 1964).
1983; Rodriguez, 1983). In general, benzofurans are not in- Although a ketone, tremetone (12), has been proposed to
secticidal, but antagonize or reduce the toxicity of encecalin be the compound responsible, this compound failed to pro-
to at least three species of insects (Isman, 1989). duce symptoms of the disease in test animals. Two inactive
Both eupatoriochromene and encecalin (9), from yellow fractions from Eupatorium rugosum were converted to ac-
starthistle (Centaurea solstitialis L., Asteraceae), retard seed tive agents by treatment with cytochrome P-450 enzymes
germination and reduce radicle and hypocotyl growth of (Beier and Norman, 1990). The exact structores of the active
weed and crop plant seedlings (Merrill, 1989). compounds have not been established.
Compounds with similar structure have been implicated Other representatives of this group of compounds (such as
in cases of livestock and human poisoning by Eupatorium compounds 13 and 14) from Ageratum houstonianum have
rugosum, white snakeroot, in the eastern United States antijuvenile hormone activity and cause precocious produc-
(Beier and Norman, 1990; Beier et al., 1987). This plant- tion of adult insects. Because of these properties, the com-
poisoning syndrome was once a major problem and was pounds are called precocenes. Female insects that are ex-
considered an illness, often called "milk sickness" (Beier posed to these compounds never reach sexual maturity.
and Norman, 1990; Kingsbury, 1964). The lipophilic com- External application of the compounds to second-instar lar-
pounds of white snakeroot are excreted by lactating ani- vae of the milkweed bug, Oncopeltus fasciatus, causes the
mals, usually cattle. In the southwestern United States, nymphs to molt to normal third- and fourth-instar larvae and
where white snakeroot does not grow, another species, then to precocious, sterile adults (Proksch and Rodriguez,
lsocoma wright;; (syn. Haplopappus heterophy/lus), has 1983, 1984; Rodriguez, 1983).
318 Introduction to Tnpenes

o
o
~ ~
CH3-0~O+ HO~O~
//

encecalin (9)

o o

(10)

c:Xo-<
o
o O~
~nU
CH3-0~O~
(11)

o
~
CH3-0~O--\- ~
precocene 1 (13) precocene 2 (14) tremetone (12)

Fig. 18.6. Chromenes, benzopyrans, benzofurans, and precocenes.

Precocenes induce precocious metamorphosis by causing species produce the all E-isomer (gutta percha) (Tanaka,
irreversible changes in the development of the corpus alla- 1991).
tum, a vital honnone-producing gland in insects_ Treatment Usually, either the Z- or E-isomer, but not both, is pro-
with juvenile honnone (produced by the corpus allatum) duced in any species with the major exception of chicle
temporarily reverses the effects of precocenes (Bowers, (Achras sapota, Sapotaceae). Rubber occurs in the fonn of
1988, 1991, 1992). Precocenes first are absorbed in the insect latex as minute particles which are accumulated in special-
fat body. The use of this group ofhemiterpenes for insectici- ized cells or in vessels known as laticifers.
dal purposes is limited, however, because of the specificity NMR spectroscopy is of value for the detennination of
of action. Many insects are able to detoxify and excrete pre- the structure of polyisoprenes. Gel-pennealion chromatogra-
cocenes rapidly (Bowers, 1988, 1991). When precocene 2 phy is a fast and reproducible method of detennining the
(14) was fed to the grasshopper, Me/anop/us sanguinipes, molecular-mass distribution of natural rubber (Tanaka,
it was converted to the 2-hydroxymethyl derivative and a 1991).
demethylated product. These oxidized products are less toxic
than the parental compounds and are more readily excreted Biosynthesis
(Harbome, 1989).
Rubber fonnation takes place in the latex vessels and all
intennediates of the pathway occur there. The latex of H evea
and many other plants consists of multinucleate protoplasm
POLYTERPENES in extensively anastomosed duct systems of laticifers
(Loomis and Croteau, 1980). Electron microscopy oflatici-
At least 2000 species (mostly of angiospennous plants, but fers from several plants families has shown that latex is a
including some ferns and fungi) contain polyisoprene, al- highly specialized cytoplasm containing typical structures
though the amount is small in most cases. Usually the poly- such as nuclei and mitochondria, endoplasmic reticulum, and
isoprene produced is all Z-isomer (rubber) and only a few ribosomes, as well as polyisoprene and resin particles. Latic-
 Introduction to Terpenes 319

ifers also contain lutoid particles, which have been character- Biological Activity
ized as vacuoles containing compartmentalized hydrolytic
enzymes (Loomis and Croteau, 1980). Once rubber is formed, it is not subject to remetabolism
A number of studies of the systems have examined the by the plant and it is unlikely that rubber serves as a food
production of mevalonic acid in latex and latex products reserve. Latex helps to seal wounds and its sticky properties
(Archer, 1980). HMG-CoA reductase is the rate-limiting help to protect the plant from insect attack.
enzyme in rubber biosynthesis (Bach et ai., 1990). Meva-
lonic acid (1) and isopentenyl pyrophosphate (4) are effi- Natural Rubber
ciently incorporated into latex in substantial yield into
rubber in latex of Hevea (Archer and Audley, 1973; Cor- Commercial rubber has been isolated from Castilla elas-
nish, 1993). tica (Moraceae), Manihot glaziovii (Euphorbiaceae), Lan-
The biosynthesis of rubber may be divided into three dolphia spp. (Apocynaceae), Taraxacum kok-saghyz (As-
steps: (I) initiation, which requires an allylic diphosphate teraceae), Hevea brasiliensis (Euphorbiaceae), and
molecule, (2) elongation, in which IPP units are added to Parthenium argentatum (guaynle, Asteraceae). Despite in-
a Z-I,4-polyisoprene chain, and (3) termination, in which tense interest in finding other rubber sources, virtually all
the polymer is released from the rubber transferase enzyme commercial natural rubber comes from Hevea brasiliensis
(Cornish, 1993). In plants, the elongation of Z-I,4-polyiso- or closely related species (Euphorbiaceae) (Rogers, 1981).
prene (natural rubber) requires a small E-allylic diphos- The search for alternate rubber sources is especially im-
phate initiator (less than or equal to C 20 ). Farnesyl pyro- portant as Hevea species are highly susceptible 10 certain
phosphate (FPP) is an effective initiator of polyisoprene fungal diseases (principally Microcyclus ulei). The preva-
biosynthesis (Light et ai., 1989); further, because only one lence of these diseases in Latin America prevents cultivation
molecule of FPP is needed for each molecule of rubber of Hevea in large plantations (Latin America accounted for
formed, small traces of this substance that are inadvertently less than 1% of world rubber production in 1977; Rogers,
present complicate biosynthetic studies. The E-allylic di- 1981). Although plants of Hevea are cultivated on planta-
phosphates are hydrophilic cytosolic compounds, whereas
tions in Southeast Asia and it is known that these plants are
Z-I,4-polyisoprene is hydrophobic and compartmentalized
susceptible to the fungi, the pathogenic agents have not been
in subcellular rubber particles. A soluble E-prenyl transfer-
introduced into that part of the world. If any of these patho-
ase from the latex of H evea brasiliensis serves as a farnesyl
gens were to be inadvertently introduced, major changes in
diphosphate synthase and plays no direct role in elongation
the economies of several rubber-producing and rubber-con-
of Z-I,4-polyisoprene (Cornish, 1993). Because the hydro-
phobic rubber molecule is produced inside a rubber particle suming nations wonld occur. Partly because of the demand
but is formed from hydrophilic precursors from the cyto- for radial tires, natural rubber still accounts for about one-
plasm, the polymerization reaction must take place at the third of the world's rubber usage (Rogers, 1981).
particle surface.
The Z-I,4-prenyl transferase is associated with rubber Hevea brasiliensis
particles in Hevea brasiliensis and other rubber-producing
plants. This enzyme is membrane bound (Cornish, 1993). In Hevea brasiliensis, rubber is largely formed and stored
This enzyme requires Mg2+ or Mn2+, but will not synthesize in the bark, in rings of latex vessels interspersed with the
rubber unless a second component of the system, an allylic sieve tubes of the secondary phloem of the trunk, branches,
diphosphate, is present (Fig. 18.7). or roots. Laticifers also are present in the leaves, flowers,

,
~opp ~opp
+ 2 '::::"" __
R~ I ~
Opp

opp
n

to a molecular weight of about 500,000 to 1,000,000

Fig. 18.7. Proposed origin of Z-polyprene compounds.

320 Introduction to TeJpenes

and fruit. In the cortex, the latex vessels run at a slight angle Guayule
to the longitudinal axis of the trunk or branches and originate
from longitudinally contiguous rows of cells, from which As observed above, there has been much interest in find-
the cross-walls are resorbed. Anastomoses between adjacent ing a nontropical rubber source. So far, one of the most
vessels in each ring allow the latex to drain from a large area promising candidates is guayule, Partheniurn argentaturn
of the cortex on tapping, but there are few or no connections (Asteraceae). This plant, which grows in the northern part
between the rings. Hevea is particularly useful as a source of Mexico and the southwestern United States, has long been
of rubber, as the latex can be drained from a large area of known to produce latex. Although there is some minor pro-
bark when the vessels are cut. The rubber regenerates rapidly duction in Mexico, guayule rubber is not currently produced
and rubber trees can be tapped on alternate days, yielding a in the United States.
constant or increasing amount of rubber for a period of 30 Guayule rubber is found in individual thin-walled cells
or more years (Benedict, 1983). throughout the plant with the exception of the leaves (Rubis
H evea latex is a suspension in which rubber particles are et aI., 1981).
stabilized by an adsorbed film of phospholipids and proteins;
triglycerides together with l3-sitosterol and related com-
pounds are the principal constituents of the neutral lipids of Gutta Percba
the rubber phase.
Hevea synthesizes both rubber and low-molecular-weight Gutta percha or gutta is all E-polyisoprene. The biosyn-
E-compounds such as triterpenes from the same precursors thesis of gutta does not seem to have been investigated. The
in the latex (Light and Dennis, 1989). molecular weight of this polymeric material is usually lower

tiglicacid (15) angelic acid (16)

heliosupine senecionine
(heliotridine + angelic (retronecine +
and echimidinic acids) senecic acid)
Boraginaceae Asteraceae

ri"'""K
o

Xll
NC H 0 (CH2)18CH, NC H 0 (CHil18CH,

NZ ro~ (CHil18CH,

HI~')l.
0 (CH2)18CH,

Fig. 18.8. TigHe and angelic acids and other five-carbon nonterpenoid compounds from plants.
 Introduction to Terpenes 321

than that of natural rubber. The all E-double-bond system SIMILAR. FIVE·CARBON Vl'IITS NOT DERIVED
results in a product with much different physical properties FROM MEVALONATE
than those of natural rubber. Gutta tends to be thennoplastic
or hard at ordinary temperatures. Plants that produce gutta Some commonly encountered five-carbon units are not de-
are known from the Celastraceae, Eucomiaceae, Euphor- rived from mevalonic acid. These moieties are found in such
biaceae, Garryaceae, Poaceae, and Sapotaceae. Two species diverse groups as pyrrolizidine alkaloids and cyanolipids.
dominate as commercial sources: Palaquium gutta and Mi- Tiglic (15) and angelic (16) acids and their derivatives are
musops balala (both Sapotaceae). The latex does not flow widespread (Buckles et al., 1955). These compounds are
as does that of Hevea and the isolation procedures are de- derived in plants from leucine (Fig. 18.8).
structive to the trees. Gutta percha has largely been replaced
by synthetics (Loomis and Croteau, 1980).
REFERENCES

ARCHER, B. L., Polyisoprene, in Secondary Plant Products (E. A.

Chicle, a mixture of 2- and E-isomers of polyisoprene is Bell and B. V. Charlwood, eds.), Vol. 8 of Encyclopedia of
isolated from Manilkara (Achras) sapota (Sapotaceae). The Plant Physiology, 309-327, Springer-Verlag, Berlin, 1980.
biosynthesis of this material does not appear to have been ARCHER, B. L. and B. O. AUDl.EY, Rubber, gutta percba and chicle,
examined. There is little doubt that there are two stereospe- in Phytochemistry, Vol. 2 (L. P. Miller, ed.), 310-343, Van
cific enzyme systems metabolizing IFP in chicle latex, one Nostrand Reinhold, Princeton, NJ, 1973.
producing rubber (2) and the other yielding gutta (Charl- BACH, T. J., T. WEBER, and A. MOTEL, Some properties of enzymes
wood and Banthorpe, 1978). Chicle is one of the few known involved in the biosynthesis and metabolism of 3-hydroxy-3-
examples where a mixture of the two structures occurs (Ar- methylglutaryl-CoA in plants, in Biochemistry of the Mevalonic
cher and Audley, 1973). Chicle, which is commonly used Acid Pathway to Terpenoids (0. H. N. Towers and H. A. Staf-
ford, eds.), Recent Advances in Phytochemistry, Vol. 24), 1-82,
in chewing gum, has been partially replaced by synthetic
Plenum Press, New York, 1990.
substitutes (Loomis and Croteau, 1980).
BANlHORPE. D. Y., Classification of terpenoids and general proce-
dures for their characterization, in Terpenoids (B. V. Charlwood
and D. V. Banthmpe, eds.), Vol. 7 of Methods in Plant Biochem-
istry (J. B. Hatborne and P. M. Dey, eds.), 1-41, Academic
DOLICHOPRENOLS Press, London, 1991.
BEALE, M. H. and I. MAcMn.LAN, The biosynthesis of Cs-c'o
Dolichoprenols (C80 to C IOS ) are derived from 2 elongation terpenoid compounds, Nat. Prod. Rep., 5, 247-264 (1988).
of E,E-famesyl pyrophosphate and are universal components BEIER, R C. and H. N. NWG, Natural toxicants in foods, in Phyto-
of eukaryotic cells. These compounds function in glycosyl chemical Resources for Medicine and Agriculture (H. N. Nigg
and D. S. Seigler, eds.), 247-367, 1992.
transfer reactions (Waechter and Lennarz, 1976).
BEIER, R C. and I. O. NORMAN, The toxic factor in white snakeroot:
Identity, analysis, and prevention, in Proceedings of the Sympo-
sium on Public Health Significance of Natural Toxicants in
Animal Feeds fW. R Keller, V. R. Beasley, and J. F. Robens,
PREl'IYLATED FROTEIl'IS eds.), Vet. Human Toxicol., 32 (Supp!.), 81-88 (1990).
BBlBR, R C., J. O. NORMAN, T. RIRVIN, and D. A. WITZEL, Micro-
Recent evidence indicates that various prenyl groups can be somal activation of constituents of white snakeroot (Eupatorium
attached to proteins. Both famesyl and geranylgeranyl rugo.um Houtt.) to form toxic products, Am. I. Vet. Res., 48,
583-585 (1987).
groups have been isolated from proteins (Epstein et al.,
1990). BELL, E. A. and B. V. CHARLWOOD (eds.), Secondary Plant Prod-
ucts, Vol. 8 of Encyclopedia of Plant Physiology, Vol. 8,
Cellular ras genes encode a family of proteins tenned
Springer-Vedag, Berlin, 1980.
p21ras. These, in turn, can acquire the ability to transform
BENEDICf. C. R., Biosynthesis of rubber, in Biosynthesis of Iso-
cells through structural mutations that alter the ability of the
prenoid Compounds, Vol. 2 (J. W. Porter and S. L. Spurgeon,
protein to bind or hydrolyze guanidine triphosphate (GTP).
eds.), 355-369, Wiley, New York, 1983.
Altered ras genes are the most frequently identified onco-
BONDEEL, M. A., D. DE KEUKELEIRE, and M. VBRZELB, The photo-
genes in human tumors (Casey et al., 1989). Processing of lysis of trans-isohumulone to dehydrohumulinic acid, a key
these ras proteins involves the addition of a famesyl moiety, route to the development of sunstruck flavour in beer, J. Chern.
near a carboxy-tenninal cysteine, followed by proteolytic Soc. Trans. I, 2715-2717 (1987).
processing. Famesylation may be important for membrane BOWERS, W. S., Cytotoxic defensive strategies of plants: Chemistry
association and transfonning ability of ras proteins (Casey and mode of action, in Endocrinological Frontiers in Physiologi-
et aI., 1989). cal Insect Ecology (F. Sehnal, A. Zobza, and D. L. Denlinger,
322 lnrrodut'tion to Terpenes

cds.), 19-26, Wroclaw Technical University Press, Wroclaw, and H. A. Stafford, eds.), Recent Advances in Phytochemistry,
1988. Vol. 24), 249-264, Plenum Press, New York, 1990.
BOWERS, W. S., Insect hormones and antihormones in plants, Ter- KINGSBURY, J. M., Poisonous Plants of the U.S. and Canada, Pren-
penoids, in Herbivores: Their Interactions with Secondary Me- tice-Hall, Englewood Cliffs, NJ, 1964.
tabolites, Vol. 1 (G. A. Rosenthal and M. R. Berenbaum, eds.), LIGHT, D. R. and M. S. DENNIS, Purification of a prenyltransferase
431-456, Academic Press, San Diego, CA, 1991. that elongates cis-polyisoprene rubber from the latex of Hevea
BOWERS, W. S., Insecticidal compounds from plants, in Phytochem- brasiliensis, 1. BioI. Chem., 264, 18589-18597 (1989).
ical Resources for Medicine and Agriculture (H. N. Nigg and LIGHT, D. R., R. A. LAZARUS, and M. S. DENNIS, Rubber elongation
D. S. Seigler, eds.), 227-235, Plenum Press, New York, 1992. by farnesyJ pyrophosphate synthases involves a novel switch in
BUCKLES, R. E., G. V. MOCK, and L. LOCATELL, JR., Tiglic and enzyme specificity, 1. BioI. Chem., 264, 18598-18607 (1989).
angelic acids, Chem. Rev., 55, 659-677 (1955). LOOMIS, W. D. and R. CROTEAU, Biochemistry of terpenoids, in
CASEY, P. 1., P. A. SOLSKI, C. 1. DER, and J. E. Buss, p21ras is Lipids, Structure and Function (P. K. Stumpf, ed.), Vol. 4 of
modified by a farnesyl isoprenoid, Proc. NatI. Acad. Sci. USA, The Biochemistry of Plants (P. K. Stumpf and E. E. Conn, eds.),
86, 8323-8327 (1989). 363-418, Academic Press, New York, 1980.
CHARLWOOD, B. V. and D. V. BANTHORPE, The biosynthesis of LYNEN, F., B. W. AGRANOFF, H. EGGERER, U. HENNING, and E. M.
monoterpenes, in Prog. Phytochem" Vol. 5 (L Reinhold, J. M6sLEIN, Dimethyl-allyl-pyrophosphat und Geranyl-pyrophos-
B. Harbome, and T. Swain, eds.), 65-125, Pergamon, Oxford, phat, biologicshe Vorstufen des Squalens, Angew. Chern., 71,
1978. 657-663 (1959).
CORI, 0., Enzymic aspects of the biosynthesis of monoterpenes in MANN, J., Secondary Metabolism, Oxford University Press, Ox-
plants, Phytochemistry, 22,331-341 (1983). ford, 1978; 2nd edition, 1987.
CORNISH, K., The separate roles of plant cis and trans prenyl trans- MERRILL, G. B., Eupatoriochromene and encecalin, plant growth
ferases in cis-l,4-polyisoprene biosynthesis, Eur. 1. Biochem., regulators from yellow starthistle (Centaurea solstitialis), 1.
218,267-271 (1993). Chem. Eco!., 15, 2073-2087 (1989).
CROTEAU, Rand M. A. JOHNSON, Biosynthesis of terpenoid wood MONFAR, M., C. CAELLES, L. BALCELLS, A. FERRER, F. G. HEGARDT,
extractives, in Biosynthesis and Biodegradation of Wood Com- and A. BORONAT, Molecular cloning and characterization of
pounds (T. Higuchi, ed.), 379-439, Academic Press, Orlando, plant 3-hydroxY-3-methyIglutaryl coenzyme A reductase, in
FL,1985. Biochemistry of the Mevalonic Acid Pathway to Terpenoids
DE KEUKELEIRE, Photochemistry of beer, The Spectrum, 4, 1-7 (G. H. N. Towers and H. A. Stafford, eds.), Recent Advances
(1991). in Phytochemistry Vol. 24, 83-97, Plenum Press, New York,
DE KEUKELEIRE, D" 1. VINDEVOGEL, R Szucs, and P. SANDRA, The 1990.
history and analytical chemistry of beer bitter acids, Trends POULTER, C. D. and H. C. RILLING, Prenyl transferases and iso-
Anal. Chem., 11, 275-280 (1992). merase, in Biosynthesis ofIsoprenoid Compounds, VoL 1 (1. W.
EpSTEIN, W. W., D. C. LEVER, and H. C. RILLING, Prenylated pro- Porter and S. L. Spurgeon, eds.), 161-224, Wiley-Interscience,
teins: Synthesis of geranylgeranylcysteine and identification of New York, 1981.
this thioether amino acid as a component of proteins in CHO PROKSCH, P. and E. RODRIGUEZ, Chromenes and benzofurans of
cells, Proc. Natl. Acad. Sci. USA, 87, 7352-7354 (1990). the Asteraceae, their chemistry and biological significance, Phy-
GERSHENZON, J. and R. CROTEAU, Regulation of monoterpene bio- tochemistry, 22, 2335-2348 (1983).
synthesis, in Biochemistry of the Mevalonic Acid Pathway to PROKSCH, P. and R. RODRiGUEZ, Distribution of chromenes and
Terpenoids (G. H. N. Towers and H. A. Stafford, eds.), Recent benzofurans in Encelia cali/ornica, Biochem. Syst. Bcol., 12,
Advances in Phytochemistry Vol. 24, 99-160, Plenum Press, 179-181 (1984).
New York, 1990. PROKSCH, P., M. PROKSCH, G. H. N. TOWERS, and E. RODRIGUEZ,
GERSHENZON, J. and R. CROTEAU, Terpenoids, in Herbivores: Their Phototoxic and insecticidal activities of chromenes and benzo-
Interactions with Secondary Metabolites, Vol. 1 (G. A. Rosen- furans from Enceiia, J. Nat. Prod., 46, 331-334 (1983).
thal and M. R. Berenbaum, eds.), 165-219, Academic Press, QURESHI, N. and 1. W. PORTER, Conversion of acetyl-Coenzyme
San Diego, CA, 1991. A to isopentenyl pyrophosphate, in Biosynthesis of Isoprenoid
HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod. Compounds, Vol. 1 (1. W. Porter and S. L. Spurgeon, eds.),
Rep., 4, 323-344 (1986). 47-94, Wiley-Interscience, New York, 1981.
HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod. REDDY, R. and DAS, Chloroplast autonomy for the biosynthesis of
Rep., 7, 85-109 (1989). isopentenyl diphosphate in guayule (Parthenium argentatum,
HARRISON, D. M., The biosynthesis of triterpenoids, steroids, and Gray), New Phytologist, 106, 457-464 (1987).
carotenoids, Nat. Prod. Rep., 5, 387-415 (1988). RODRiGUEZ, E., Cytotoxic and insecticidal chemicals of desert
ISMAN, M. B., Toxicity and fate of acetylchromenes in pest insects, plants, in Plant Resistance to Insects (P. A. Hedin, ed,), ACS
in Insecticides of Plant Origin (1. T. Amason, B. J. R. Philogene, Symposium Series 208, 291-302, American Chemical Society,
and P. Morand, eds.), ACS Symposium Series 387, 44-58, Washington, DC, 1983.
American Chemical Society, Washington, DC, 1989. ROGERS, T. H., Natural rubber, in Resource Materials (T. A. Mc-
ISMAN, M. B., P. PROKSCH, and C. CLARK, Terpenoid anti-herbivore Clure and E. S. Lipinsky, eds.), Vol. 2 of Handbook of Biosolar
chemistry of Encelia species (Asteraceae), in Biochemistry of Resources (0. R. Zaborsky, ed.), 387-396, CRC Press, Boca
the Mevalonic Acid Pathway to Terpenoids (0. H. N. Towers Raton, FL, 1981.
 Introduction to Telpenes 323

RUBIS, D. D., L. CASSENS, and M. PRICE, Guayule, in Handbook A Biacs, K. Gruiz, T. Kremmer, eds.), 313-319, Akademiai
of Biosolar Resources Vol. 2 (0. R. laborsky, ed.), 231-244, Kiado, Budapest, 1989.
CRC Press, Boca Raton, FL, 1981. SPURGEON, S. L. and J. W. PORTER, Introduction, in Biosynthesis of

RUZICKA, L.. The isoprene rule and the biogenesis of terpenic com- Isoprenoid Compounds, Vol. I (J. W. Porter and S. L. Spurgeon,
pounds, Experientia, 9, 357-367 (1953). eds.), 1-46, Wiley-Interscience, New York, 1981.
RUZICKA, L., Faraday Lecture (History of the isoprene rule), Proc. TANAKA, Y., Rubber and related polyprenols, in Terpenoids (B. V.
Chern. Soc. (Lond.), 341-360 (1959). Charlwood and D. V. Banthorpe, eds.), Vol. 7 of Methods in
Plant Biochemistry 0. B. Harhome and P. M. Dey, eds.),
RUZICKA, L. t In the borderland between bioorganic chemistry and 519-536, Academic Press, London, 1991.
biochemistry, Annu. Rev. Biochem., 42, 1-20 (1973).
WAECHTER, C. J. and W. J. LENNARZ, The role of polypreno I-linked
SCHULZE-SIEBERT, D., AlfEINTzE, and G. SCHULTZ, Substrate flow sugars in glycoprotein synthesis, .Annu. Rev. Biochem., 45,
from photosynthetic carbon metabolism to chloroplast isopren- 95-112 (1976).
oid syuthesis in spinach: Evidence for a plastidic phosphoglyc- WALLACH, 0., Zur KentniB der Terpene uDd dec atherischen Oele,
erate mutase, Z. Naturforsch., 42 C, 570-580 (1987). Liebig's Ann. Chern., 239, I-54 (1887).
SCHULTZ, O. and SCHULZE-SmBERT, Chloroplast isoprenoid synthe- WIELAND, H. and E. DANE, Untersuchungen iiber die Konstitution
sis in spinach and barley: Direct intermediate supply by photo- der Gallensauren, Hoppe-Seyler's Z. Physiol. Chern., 210,
synthetic carbon fIxation, in Biological Role of Plant Lipids (p. 268-281 (1932).
19
Monoterpenes

Introduction (Charlwood and Charlwood, 1991a). Monoterpenes are also

Biosynthesis present in fungi and some bacteria. Many monoterpenes also
Formation of Geranyl Pyrophosphate occur in animals, particularly in insects (Nahrstedt, 1982).
Interconversion of GPP and NPP Monoterpenes are often partially responsible for the essence
Studies with Labled Precursors of Monoterpenes or odor of piants and are the major odoriferous compounds
Acyclic Monoterpenes of many flowers and fruits. The heartwood of many trees,
Cyclic Monoterpenes especially conifers, is rich in terpenoid materials. In general,
Secondary Transfonnations sapwood of trees contains few terpenoids, but can saturate
Catabolism of Monoterpenes with resin in response to wounding (Croteau and Johnson,
Compounds Derived by Alkylation with Geranyl-OPP 1985).
Monoterpene-Derived Compounds of Marijuana The volatile fractions of many plants are isolated by steam
Hydroquinone Derivatives in Cordia Species distillation, distillation, or by solvent extraction (Banthorpe,
Chemosystematic Use of Monoterpene Chemical Data 1991). The resulting volatile compounds that make up the
Chemosystematic Study of Citrus Species essence or aroma of plants are called essential oils. Although
Chemosystematic Study of Gymnospenns essential oils are comprised of many types of compounds,
Chemosystematic Studies in Mints monoterpenes are major among them. These oils may also
Biological Activity contain metabolically modified fatty acids, aldehydes, hy-
Monoterpenes as Allomones, Kairomones, and drocarbons, esters, phenylpropanoid compounds, acetylenic
Synomones compounds, volatile alcohols, volatile alkaloids, phenylpro-
Allomones panoids, and other shikimic acid derivatives.
Allomones Produced by Arthropods, Especially Insects About 1000 monoterpenes, belonging to about 38 differ-
Kairomones ent skeletal types, are known (Fig. 19.1) (Charlwood and
Aggregation Pheromones
Charlwood, 1991a; Gershenzon and Croteau, 1991; Har-
Trail-Marking Pheromones
borne, 1982). These compounds usually occur free, but
Alarm Pheromones
sometimes occur as glycosides, especially in the iridoid se-
Synomones
ries. Essential oils are usually associated with specialized
Allelopathy
storage structures in plants. These lipophilic materials are
Uses of Monoterpenes by Hnmans
Medicinal Uses of Monoterpenes frequently found in secretory canals, cavities, glandular tri-
Flavors chomes, or in special oil cells (Croteau, 1981; Croteau and
Insect repellents Johnson, 1984; Gershenzon and Croteau, 1991; Johnson and
Irregular Monoterpenes Croteau, 1987; Rodriguez et aI., 1984). Monoterpenes are
References usually synthesized in the epithelial cells lining the ducts or
cavities in which they are eventually sequestered (Croteau
and Johnson, 1985). The production of monoterpenes often
INTRODUCTION
occurs in nonphotosynthetic tissues and is highly dependent
Monoterpenes are widespread in plants. These C IO com- on imported carbohydrates for both carbon and energy (Cro-
pounds occur in algae, bryophytes, ferns, gymnospenns, and teau and Johnson, 1985).
both monocotyledonous and dicotyledonous angiospenns Essential oil components appear isolated from the rest of

324
 MonOlelpenes 325

~ ~ 2 22
~ ~"~1'mcri
p-cymene (81) sabinene (22) tbujeue (23) camphene (IS)
«-pheUandrene (79)

~l!e~200~
'~: ~~ 4'-~:~~
£" "1
Hnalool (6) thymol l,8-dneole (10) borny. acetate (77) plperitone

$" crt (tt

2 25:
(a) earvooe (48) thujone (ll) camphor {33} fencbone verbenone (65)

Eft { 00

~~OO2" 2 $00
a-fenebene E. or B-oclmene (47) terplnoleoe (12) «-terpinene dtnmellol (7)

5
geraniol (4) nerol (5) I,B-cineole (10) ascaridol (87) borneol (32)

oo-0~-?J ~~':(
~I°IPP
~ ~ CO,H

(b) p-thujaplicin (49) p-thujapUcinoi (46) tbujic aeld (SO) fenchyl acetate Iinaly. pyrophosphate (8)

Fig. 19.1 (a & b). Representative monoterpenes.

326 Monoterpenes

the plant; there is evidence that despite reports of tnrnover Regnlation of monoterpene biosynthesis is controlled by
in cuttings, there is a lack of rapid tnrnover of monoterpenes many factors. Morphological differentiation of the plant is
in intact plants (Mihaliak et ai., 1991). of paramount importance. Monoterpene formation occurs
Most essential oils are complex mixtnres involving from only in cells of specialized secretory structures during the
a few to several hundred compounds. The techniques for relatively brief periods when these cells are metabolically
collecting floral fragrances and other similar mixtnres of active (Gershenzon and Croteau, 1990). Feeding stndies with
essential oils have been reviewed (Charlwood and Charl- isotopically labeled precursors have demonstrated that the
wood, 1991a; Gershenzon and Croteau, 1990, 1991; Hills site of monoterpene formation is physiologically isolated
and Schutzman, 1990). Extraction with supercriticalliquids from the mainstream of plant metabolism, suggesting that
and headspace trapping are useful to avoid decomposition monoterpene biosynthesis is subject to control by the avail-
of essential oil components. ability of assimilate. Cellular compartmentation and regula-
General chromatographic techniques for monoterpenes tion at the enzyme level are likely involved in overall regula-
have been reviewed (Banthorpe, 1991; Charlwood and tion of monoterpene biosynthesis but have been inadequately
Charlwood, 1991a; Coscia, 1984; Gershenzon and Croteau, stndied. HMG-CoA reductase appears to catalyze a key regu-
1991). As most monoterpenes are volatile and relatively non- latory step in the biosynthesis of all plant terpenoids, includ-
reactive, capillary gas chromatography (GC) and mass spec- ing monoterpenes (Gershenzon and Croteau, 1990).
trometry are most widely used techniques for investigation Biosynthetic reactions leading to the formation of monot-
of essential oils (Banthorpe, 1991; Charlwood and Charl- erpenes largely involve formation of geranyl pyrophosphate
wood, 1991a; Hills and Schutzman, 1990; Shibamoto, 1981). (GPP), cyclization of GPP to one of a number of skeletal
However, because monoterpenes usually contain 10 carbon types, and secondary transformations of the initial cyclic
compounds, many have identical parent ions and similar product (Gershenzon and Croteau, 1990).
fragmentation patterns. Nuclear magnetic resonance (NMR)
techniques also are quite useful in determining structnres of Formation of OeranyI Pyrophosphate
these compounds (Atta-ur-Rahman and Ahmad, 1992;
Monoterpenes are the product of the union of two acti-
Charlwood and Charlwood, 199Ia).
vated Cs units, the biological equivalents of isoprene (Fig.
19.2). The reaction is catalyzed by prenyltransferases (BC
BIOSYNTHESIS 2.5.1.1) that effect condensation of sopentenyl pyro-
phosphate (IPP) with an allylic pyrophosphate, such as
In contrast to some other plant metabolites, most species of dimethylallyl pyrophosphate (DMAPP), giving an ally-
monoterpene-producing plants do not produce and accumu- lic pyrophosphate with five additional carbon atoms.
late monoterpenes in tissue cultnre (Charlwood and Charl- Prenyltransferases catalyze the first committed step to the
wood, 1991b). This lack of monoterpene synthesis may be isoprenoid pathway (Gershenzon and Croteau, 1990). Mo-
associated with special requirements for compartmentaliza- noterpene biosynthesis requires a specific prenyl transferase,
tion. However, a few such cultnres do produce monoter- geranyl pyrophosphate synthetase, that occurs in cells near
penes. Callus cultnres of Perilla Jrutescens produce up to the specialized storage sites of monoterpenes. Such a geranyl
2 mg of volatile oil per gram fresh weight of tissue. The pyrophosphate synthase, that synthesizes geranyl pyrophos-
composition of the oil is similar to that of the parent plant. phate (3) as the sole (>95%) product from dimethylallyl
In other plants, such as Jasminum officinale and Rosa da- pyrophosphate (1) and isopentenyl pyrophosphate (2), has
mascena, the enzymes of biosynthesis have been demon- been isolated from the leaves of sage (Salvia officinalis,
strated to be present, although there is no accumulation of Lamiaceae) (Croteau, 1987; Croteau and Puckett, \989). In
monoterpenes (Beale and MacMillan, 1988; Ellis, 1988). contrast to farnesyl pyrophosphate synthetase, geranyl pyro-
Cell-free extracts of callus or suspension cultnres of several phosphate synthetase is restricted in distribution within the
species that did not accumulate monoterpenes were shown plant (Croteau, 1981, 1987; Croteau and Johnson, 1985; Cro-
to convert mevalonate and isopentenyl pyrophosphate into teau and Purkett, 1989).
geranyl, neryl, and farnesyl pyrophosphates at rates much DMAPP (1) is an excellent aikylating agent. Although it
greater than those of the parental plant (Beale and MacMil- would appear to suffer nucleophilic attack at C-1 with loss
lan, 1988). of pyrophosphate, a good leaving group, ionization provides
Cells thought to be involved in monoterpene synthesis the C-1 cation generally regarded as the active electrophile.
typically possess an abundance of nonpigmented plastids Inversion of the center of DMAPP-bearing pyrophosphate
known as leucoplasts that lack an organized system of thyla- occurs; Mg2+ appears to be involved in the condensation. In
koids and have few other internal membranes. Leucoblasts cell-free extracts of Tanacetum vulgare (Asteraceae), the
frequently have been observed to contain droplets of lipid- pro-(4R)-hydrogen of MVA [the pro-(2S)-hydrogen of IPPl
like material. Monoterpene-synthesizing secretory cells also and both tritium atoms from [5-3 H21MVA (C-1 of IPP) are
contain a well-developed network of smooth endoplasmic retained in geraniol (4) (Banthorpe et al., 1978a).
reticulum that may be involved in monoterpene biosynthesis The stereochemistry is well established, and many ques-
(Gershenzon and Croteau, 1990). tions concerning the overall mechanism of the condensation
 MonOferpenes 327

-0
.
JlXA 3
.
PH, 0
II
O-P-O-P-OH
0
11 __

4 I I
0- O·
mevalonic acid

y
mevalonic acid 5-pyrophosphate

~
# 2
o
II
0
II -HR oo
II
o-i-o-i-
H 3 .,' O-P-O-P-OH -----"- 3~ 1\
4~. I I ~
Isomerase 2 OH
H HR HS 0- 0-
H 0- 0-

isopentenyJ pyrophosphate (2)

dimethylallyl pyrophosphate (1)
IPP
(a) DMAPP

(1)

OPP

(b) E-geranyl pyrophosphate (3)

Fig. 19.2 (a & b). Biosynthesis of geranyl pyrophosphate.

have now been resolved (Connforth et a!., 1966). Pamesyl- The fonnation of a 2,3-E- or 2,3-Z-double bond is a con-
pyrophosphate synthetase (BC 2.5.1.1) is the key enzyme in sequence of the syn- or anti-conformation of the double
the biosynthetic pathways for several classes of terpenes. bonds of the precursors (Cori, 1983). Most, if not all, mono-
This enzyme catalyzes 1'-4 condensation between IPP and terpenes appear to arise from GPP (geranyl pyrophosphate)
DMAPP, or geranyl pyrophosphate, polymerizations that (with a 2,3-E-double bond) and most of the numerous ter-
constitute the major building steps of terpenoid biosynthesis penes in nature are based on the E-configuration mentioned
(Pig. 19.3) (Poulter and Rilling, 1978; Poulter et al., 1978, above. Loss of the pro-S-proton to fonn compounds of Z-
1981). The condensation reaction may be subdivided into confonnation (mostly polyisoprenoids) occurs in some
three phases: ionization, condensation, and elimination. Ioni- plants (e.g., Hevea brasiliensis) (Cori, 1983) (see Chapter
zation of the pyrophosphate group of DMAPP to generate 18).
the allylic cation is the first step of condensation of IPP.
Studies by Poulter and co-workers indicate that cleavage Interconversion of OPP and l'IPP
of the carbon-oxygen bond of geranyl pyrophosphate is a
discrete step, yielding a geranyl cation-pyrophosphate ion Neryl pyrophosphate (NPP) (with a 2-Z-double bond)
pair and that this reactive species subsequently alkylates the also has been suggested as a product of the reaction of IPP
double bond in IPP (presumably the same is true for and DMAPP. Tracer studies show that the pro-(4S)-hydro-
DMAPP). These workers concluded that ionization and con- gen of mevalonic acid [the pro-(2R)-hydrogen of IPP] is lost
densation are not concerted (Poulter et a!., 1981). Studies in both the fonnation of geraniol (4) and nerol (5). This
with [1-180]geranyl pyrophosphate indicate that exchange evidence is in accord either with the prior fonnation of gem-
of oxygen does not occur during the reaction and that a nyl-OPP and subsequent conversion to neryl-OPP (see
highly structured ion pair exists (Mash et al., 1981). below), or with the presence of two transferases which cata-
328 Monoterpenes

~.~~. --
:8·
-...~H
/ ' .. + "--H :8·

OPI'"

~.
l OP~H
'~
-----+- H -.. H

~ H H

H H H8'·
OPI'"
OPI'"

Fig, 19.3. Proposed cationic mechanism for monoterpene biosynthesis.

Iyze the simultaneous fonnation of geranyl-OPP and neryl- whole plants (Loomis and Croteau, 1980). In cell-free sys-
OPP. There is some evidence in support of each hypothesis tems, somewhat greater incorporation of label is observed,
(Banthorpe and Modawi, 1978b). although the levels are still less than those observed for many
Nerol (5) and related compounds with a 2-Z double bond other secondary metabolites. Increased rates of incorporation
occur in many plants. Although GPP is a precursor to many of biosynthetic precursors for monoterpenes in Tanacetum
other groups of terpenoid compounds and is highly reactive vulgare (Asteraceae) in vivo occurred during the plant's dor-
in biological systems, despite earlier reports, neither neryl mant phase. Rates 75 times greater than those of the growing
and linalyl pyrophosphates serve as intennediates in monot- phase were observed (Banthorpe et al., 1977b). This may
erpene biosynthesis (Croteau, 1984). Present evidence sug- partially explain the low incorporations in some stodies.
gests that geranyl pyrophosphate (GPP) is the preferred pre- In any case, studies with specifically labeled precursors
cursor of most compounds of this series (Croteau, 1984). indicate that MYA is an obligate precursor of monoterpenes
Enzymes in cell-free systems from carrot (Daucus carota, (Pig. 19.4). Incorporation of CO2 and glucose is much better
Apiaceae) and peppermint (Mentha piperita, Lamiaceae) (Charlwood and Banthorpe, 1978), but the large number of
catalyze the reversible fonnation of nerol and neryl phos- products obtained complicates analysis. It seems probable
phate from geraniol and geranyl phosphate, respectively. that MYA (often introduced as the 8-lactone) is not trans-
Geranyl phosphate appears to have been isomerized directly ported efficiently to the site at which monoterpene biosyn-
to the neryl derivative without the intervention of an alde- thesis occurs.
hyde intennediate. The presence of a flavin, thiol, and light The high rate of incorporation of MYA into geraniol (4)
was required for the reaction (Croteau, 1984). and nerol (5) and their ~-D-glucosides in flower petals of
Rosa dilecta (up to 22%) represents an exception to the usu-
Studies with Labeled Precursors ally low rates of incorporation which are observed (Charl-
of Monoterpenes wood and Banthorpe, 1978).
Often as little as O.DJ-OJ % incorporation is observed in CuriOUSly, when complete degradation of labeled monot-
attempts to introduce labeled MYA into monoterpenes in erpenes has been carried out, in many studies, label from
 Monoterpenes 329

MVA or from l4C02 resides almost entirely in the portion mine the basic character of the monoterpene end products
of the molecule derived from isopentenyl pyrophosphate (Gershenzon and Croteau, 1990). Monoterpene cyclases are
(IPP) (2) but is not significantly present in the portion of key regulatory enzymes in the synthesis of monoterpenes.
the molecule derived from dimethylallyl pyrophosphate Intermediates are seldom isolated and, usually, do not appear
(DMAPP) (1). This appears to be a general phenomenon. It to be released during synthesis. Multiple cyclases, each pro-
is likely that a pool of DMAPP exists and labeled IPP reacts ducing a different skeletal arrangement from the same acy-
with this pool of inactive DMAPP before significant isomer- clic precursor, often occur in higher plants; single cyclases
ization can occur. The amount of labeled DMAPP produced which synthesize a limited variety of skeletal types also are
is probably small with regard to the pool size, and the amount known (Croteau, 1987).
oflabel introduced would be slight. The source of this endog- A hypothetical biogenetic scheme was proposed by Ruz-
enous DMAPP is not known, although it is probably due to icka and co-workers (1953) that explains the origin of many
the fact that the equilibrium of the isomerase favors DMAPP structural types of monoterpene. The final structures are the
over IPP, and the next step in the pathway, the prenyltransf- result of an initial cyclization followed by chemical modifi-
erase, is a relatively slow step, allowing buildup of DMAPP. cation, skeletal rearrangements, and functional group intro-
Such a pool may be observed only when incorporation times duction (Fig. 19.5). A carbocation or ion pair is formed from
are brief, or when incorporation rates of exogenous precur- the allylic substrate. The positive charge developed adds to
sors are low, as is frequently the case with monoterpenes. a double bond and a new carbocation is developed. The final
product is formed through regiospecific and stereospecific
Acyclic Monoterpenes elimination of a proton. Other intramolecular additions, hy-
dride shifts, and skeletal rearrangements must occur before
Acyclic, monocyclic, and bicyclic monoterpenes are
the final proton elimination in the biosynthesis of bicyclic
found in many plants, fungi, and certain insects, but rarely
monoterpenes.
in other animals. Acyclic compounds are derived from both
The formation of the cyclohexanoid ring from an acyclic
geranyl-OPP and from neryl-OPP, whereas cyclic species
precursor requires a syn- or Z-conformation either in the
are almost always derived from geranyl-OPP.
ground state or in the activated complex. On a basis of this
argument, formerly it was thought that NPP, the Z-isomer of
Cyclic Monoterpenes
GPP, was the most likely precursor of cyclic monoterpenes.
The cyclization of geranyl-OPP (3) to produce monocy- However, it is now known that GPP is the preferred precur-
clic and bicyclic monoterpenes proceeds via enzyme-bound sor. Recent studies with cell-free systems indicate that GPP
intermediates that are probably similar to those postulated as can be converted into cyclic products without detectable con-
intermediates in Wagner-Meerwein rearrangements. These version into any other acyclic intermediates (Croteau, 1984,
reactions (catalyzed by strong acid) result in the formation 1987; Johnson and Croteau, 1987).
of many types of monoterpenes when compounds similar to Geranyl pyrophosphate is cyclized without preliminary
the precursors of cyclic monoterpenes are used as reactants. conversion to NPP or linalyl pyrophosphate (LPP) (8). An
These enzymes, called monoterpene cyclases, largely deter- isomerase system is not involved (Croteau, 1984). This im

-0 ~lOH •
,
OH
.~opp
-+PPO~I
I
.- +
.2"'~
I
+'
:
-+

. .
[2_ 14Clmevalonic acid ~

6 -~D~oD
~ ~ .~.
(+)-sabinene (+)-Z-sabinol (+ )-3-thujone

Fig. 19.4. Labeling of monoterpenes.

330 Monoterpenes

plies a multistep cyclization process whereby the cyclase the demonstration that geranyl pyrophosphate (3) was pre-
itself converts GPP into a bound intermediate stereochemi- ferred to neryl pyrophosphate (9) as substrate (Croteau,
cally suitable for cyclization. The mechanism of cyclization 1984) (Fig. 19.6). Cyclization of(IR)- and (lS)-[l-3H]gera-
appears to involve initial ionization of the pyrophosphate nyl pyrophosphate and neryl pyrophosphate by partially pu-
moiety, in which a divalent ion appears to assist, followed rified d- or (+ )-bornyl pyrophosphate synthase (a cyclase
by stereospecific syn-isomerization to a linalyl intermediate from Salvia ojJieinalis) was investigated, as the C-l of gera-
with rotation about the C-2-C-3 single bond and subsequent nyl pyrophosphate should exhibit net retention of configura-
cyclization of the cisoid rotamer in the anti-endo conforma- tion, whereas neryl pyrophosphate should exhibit net inver-
tion. The chiral a-terpinyl cation so produced may then sion of configuration. The bornyl pyrophosphate obtained
undergo electrophilic addition to the remaining double bond from each precursor was converted to camphor and the tri-
as well as hydride shifts and rearrangements to provide the tium label was located by selective exchange of the exo- and
various other cationic cyclic products (Croteau, 1988; John- endo-a-hydrogens by established techniques. Tritium was
son and Croteau, 1987). in the predicted position in all cases (Fig. 19.6). With 1- or
Partial purification of the d-bomyl pyrophosphate cyclase ( - )-bornyl pyrophosphate cyclase (from Tanaeetum vul-
from sage leaves (Salvia ojJicinalis, Lamiaceae) and removal gare), exactly the opposite results were obtained. These re-
of containing phosphatases and pyrophosphatases allowed sults establish the rotation of the C-2-C-3 bond and net

~'
r<!- ~
~_~w ~ pyrophosphate
(-)-3R-LPP ~ (3) + / (+)-3S-LPP

+opp~ _ opp+

~~~/~
~~
>-0- h
T-wr:ne~
t
~
PPO- ~ ~
lJ ~
(+)-a-plnene (14) (-)-a-pinene

t+.t
+ -y
PPo-

~-:CfY
myrcene (17)

~~1r,-~-. I j

~;£~ ~ ~
(+)-sablnene (22)
OPP
(+)-bornyl
pyrophosphate
(-)-endo-rencbol (-)-bornyl pyrophosphate (-)-camphene (15)

Fig. 19.5. Wagner-Meerwein rearrangements of geranyl-OPP (modified from Croteau, 1987; used with permission of the copyright owner, the
American Chemical Society, Washington. DC).
 Monoterpenes 331

Fig. 19.6. A model for the cyclization of GPP and NPP to d- or (+)- and /- or (- )-bomyl pyrophosphate based on studies with lR- and IS-labeled
precursors (Croteau, 1984; modified and used with pemrission of the copyright owner, Marcel Dekker, Inc., New York).

inversion of configuration at C-l of geranyl pyrophosphate, ates (Charlwood and Banthorpe, 1978; Croteau and Karp,
and the lack of rotation and net retention of configuration 1977). A number of cyclases including 1,8-cineole, 'Y-terpi-
in neryl pyrophosphate. The detailed mechanism by which nene, endo-fenchol, bornyl pyrophosphate, and d- and I-pi-
geranyl pyrophosphate is cyclized and the relationship to the nene cyclases have been isolated, purified, and partially
cyclization of neryl pyrophosphate (9) and linalyl pyrophos- characterized from this system. Most of these produce a sin-
phate (8) is an important question related to monoterpene gle product from the acyclic precursor.
biosynthesis (Beale and MacMillan, 1988; Croteau, 1984; In a subsequent examination of the cyclases that formed
Croteau and Johnson, 1985). (+ )- and ( - )-pinene, cyclase I, 96,000 MW, converted ger-
Cyclase preparations have been obtained which catalyze anyl pyrophosphate to (+ )-a-pinene (d-a-pinene) (14), but
the cyclization of geranyl pyrophosphate to essentially all produced smaller amounts of ( + )- or d-camphene (15) and
major structural classes of monoterpenes. These cyclases are ( + )- or d-limonene (11) as side products (Johnson and Cro-
membrane bound in the synthesizing cells, but are readily teau, 1987). Cyclase II, 55,000 MW, transformed geranyl
solubilized (Charlwood and Charlwood, 1991a; Croteau, pyrophosphate into (- )-(3-pinene (l-(3-pinene) (16) along
1987). with smaller amounts of I-a- or ( - )-a-pinene, (-)- or 1-
A cell-free system from Salvia officinalis (Lamiaceae) camphene, (-)- or I-limonene, and myrcene (17) as copro-
that synthesizes 1,8-cineole (10), Iimonene (11), terpinolene ducts (Fig. 19.7) (Croteau, 1984; Johnson and Croteau,
(12), and a-terpineol (13) from GPP (NPP also serves as a 1987). Extensive purification of each enzyme and differen-
precursor) has been established. These cyclic monoterpenes tial inactivation studies ensured that each set of stereochemi-
were formed by independent routes from the precursor and cally related products was synthesized by a single, distinct
not as free intermediates of a common reaction sequence enzyme.
(Loomis and Croteau, 1980). Fractionation of the soluble Involvement of tight ion pairs in monoterpene cycliza-
preparation allowed separation of 1,8-cineole, a-terpineol, tions is indicated by the absence ofPa-P(3 interchange (i.e.,
and limonene cyclase activities. Thus, a series of competing, the two ends of the pyrophosphate group are not inter-
but distinct, cyclases exist in relatively crude preparations changed during the reaction) and lack of I·O-isotope ex-
with the ability to synthesize cyclic monoterpenes in this change of the pyrophosphate moiety. These factors establish
plant. A single protein seemed to be involved in the forma- a remarkably tight restriction on the motion of the transiently
tion of 1,8-cineole (10) from an acyclic precursor and there generated pyrophosphate anion with regard to its cationic
was no evidence for any free intermediates between GPP terpenyl reaction partner (Johnson and Croteau, 1987).
and 1,8-cineole. It is possible that some sort of channeled A mixture of (3R)-[8,9- 14Cjlinalyl diphosphate and
process occurs that precludes entry of exogenous precursors (IE,3RS)-[I-'Hdlinalyl diphosphate was incubated with the
to the enzyme of the cell-free system or release of intermedi- two cyclase preparations. The borneol from the Tanacetum
332 Monoterpenes

Ce
(+~-pinene (16)
ee--p
(+)-a-pinene (14)
~ (-l-a-pinene

pinene n CYcla~ finene I cyclase

t~r~~~2
~~/.~. ~ ,~, ~ i~
_0" / I' ll(
Ce (+p-pinene (16)
~ ~=cfJ (-)-camphene
(I-camphene)

Fig. 19.7. Formation of (+ )~a.·pinene and (- )-~-pinene (modified from Croteau and Johnson, 1985; used with pennission of the copyright owner,
Academic Press, Orlando. FL).

vulgare system had a 3H: I'C ratio of 31 : 1, indicating an At least 20 cyclases have been isolated, and as many
almost exclusive use of the (3S)-enantiomer, whereas the as 50 may exist. Most catalyze the fonnation of olefinic
borneol of the Salvia ojJicinalis system had a ratio of 4: I, compounds, but a few involve oxygenated products (Chari-
indicating a preference for the (3R)-enantiomer. (3R)-Lina- wood and Charlwood, 1991a).
Iyl pyrophosphate (18) preferentially gave rise to the (+ )-
olefms (generated by cyclase I), and (3S)-linalyl pyrophos- Secondary Transformations
phate (19) preferentially gave rise to the ( - )-olefins (gener- Although a relatively small number of cyclases must de-
ated by cyclase II) (Fig. 19.8) (Beale and MacMillan, 1988; termine the basic structural character of the monoterpenes
Johnson and Croteau, 1987). produced, the great variety of derivatives of each structural
Preparations from Mentha piperita (peppennint, La- type encountered suggests that numerous enzymes must be
miaceae) convert the acyclic precursors geranyl and neryl involved in the secondary transfonnations of the parent
pyrophosphate into limonene (11). The bulk of the cyclase cyclic compounds (Croteau, 1984). As a group, the enzymes
activity is in the soluble-enzyme fraction (Croteau, 1984). catalyzing secondary transfonnations are not well studied.
In stodies with a geranyl pyrophosphate: (- )-endo-fen- The fonnation of oxygenated derivatives usually involves
chol cyclase from fennel (Foeniculum vulgare, Apiaceae), oxygenation of the parental hydrocarbons. The essential oil
(3R)-linalyl pyrophosphate (18) had a Km lower than that of Artemisia absinthum contains d-sabinyl acetate (42%)
obtained with geranyl pyrophosphate (3) and a relative ve- (20), d-3-thujone (32%) (21), d-sabinene (12%) (22), and
locity nearly three times higher. These results suggest that l-a-thujene (3%) (23). Sabinene, fonned by cyclization of
the isomerization step is rate limiting (Johnson and Croteau, geranyl pyrophosphate with a soluble enzyme, is the key
1987). cyclic precursor of 3-thujone and other C-3-oxygenated
 0 yt
Monotelpenes 333

0pp 10pp
tansy -----.

I -- I opp
geranyl OPP (3) (+)-(3S)-linalyl OPP (19)
(-)-bornyIOPP

(-)-(3R)-linalyl OPP (18)

~ (+)-bornyIOPP
OPP

Fig. 19.8. Biosynthesis of bomanes (Croteau, 1987; modified and used with pennission of the copyright owner, Copyright 1987, the American
Chemical Society, Washington, DC).

compounds of this structural group (Croteau, 1984)_ Under

aerobic conditions, geraniol is incorporated into thujane de-
rivatives_ Sabinene is fIrst converted to cis-sabinol (24) by
a membranous, NADPHl02 , P-450-dependent mixed-func-
tion oxidase (Croteau and Johnson, 1985; Gershenzon and
Croteau, 1990)_ Under conditions where oxygen is limiting,

ot) --
most of the labeled geraniol precursor was incorporated into d~sabinene (22) d-cis-sablnol (24) d~cis..sabIDyl acetate (20)
sabinene_ d-[1O-3H1Sabinene was incorporated into both d-
sabinyl acetate (20) and d-3-thujone (21) (Croteau, 1984)_
These data support a pathway based on the intermediacy of
+-
d-sabinene (Fig_ 19_9), but not a-thujene (23), a-terpineol
(13), and terpinen-4-ol (25), as had been previously sug-
gested (Croteau, 1984)_
~
d-3-thuJone (21) d-sablnone 1-3-isothujone (51)
Evidence for conjugate reduction as a key step in monot-
erpene biosynthesis has been obtained from studies of the
oxygenated monoterpenes of Mentha piperita (Fig_ 19_10)
(Croteau, 1984)_ The pathway from isopiperitenone to the
menthol esters was deduced largely by time-course studies
and direct feeding experiments_ Other evidence supports the
intermediacy of I-Iimonene (11) as the fIrst cyclization prod-
uct of GPP in this plant. This is followed by the allylic
a-thuJene (23) (-)-trans-isopiperitenol (34) (-)-trans-carveol (35)
oxidation of the olefin and subsequent isomerization and
reduction of the double bonds of isopiperitenone to the men-
thones (such as 26)_ Furthermore, stereospecifIc dehydro-
genases responsible for the synthesis of I-menthol (27) and
d-neomenthol (28) have been isolated (Croteau, 1984)_
( + )-Pulegone (29) arises from ( - )-isopiperetinone (30)
via (+ )-cis-isopulegone (31) in leaf disks and enzyme
preparations of Mentha piperita_ The enzyme responsible (-)-perillyl aleobol (36)
for the reduction of the enone double bond had a molecular
Fig. 19.9. Proposed pathway for the biosynthesis of C-3-oxygenated
weight of 60,000 (Croteau and Venkatachalam, 1986)_ thujane monoterpenes from sabinene (modified from Croteau, 1984;
In sage, geranyl pyrophosphate (3) is converted to ( + )- modified and used with pennission of the copyright owner, Marcel
bomyl pyrophosphate, which is subsequently hydrolyzed by Dekker, Inc., New York).
334 Monoterpenes

geranyl OPP (3) I·limonene (11) isopiperitenol I·isopiperitenone (30) d·cis·isopulegone (31)

d·isomenthol d.isomenthone d·pulegone (29) I-menthone (26)

60H
A
d-neoisomenthol d-neomenthol (28) I-menthol (27)

Fig. 19.10. Origin of oxygenated monoterpenes in Mentha species (Croteau, 1984; modified and used with permission of the copyright owner, Marcel
Dekker, Inc .• New York).

Avo ~o\
ttJ -lV O
-
tG -glUCOSYl

o -..
O·glucosyl
transported to roots

camphor (33) 1,2-campholide

transported

to rhizome

6=
I-menthone (26) d-neomenthol (28) d-neomenthyl glucoside

modified J3-oxidation

/ .+te

A
. 0 acyl lipids

PhytosterOI:;n rh;zome

d-neomenthol (28) I-menthone (26) /-3,4-menthone lactone (37)

Fig. 19.11. Catabolism of monoterpenes (Croteau, 1988; modified and used with permission of the copyright owner, Elsevier Science, Amsterdam).
 Monoterpenes 335

cytochrome PA50-dependent mixed-function oxygenases

o~ (Karp et al., 1990). The three limonene hydroxylases exhib-

ited a requirement for O2 and a reduced pyridine nucleotide.

~
Although there have been many previous suggestions that
the enzymes involved in the secondary transformations of
lloJlAoJ,o monoterpenes are relatively nonspecific, most of the work
cited above suggests that these enzymes are quite specific
(Croteau, 1984; Karp et al., 1990).
bergamottln (38)

FIg. 19.12. Bergamotin. Catabolism of Monoterpenes

The content of monoterpenes in peppermint after flower-
a distinct pyrophosphatase to ( + )-borneol (32), followed by ing is 35-50% less than in younger plants. This decrease
the NAD-dependent dehydrogenation to ( + )-camphor (33) begins when flower buds appear. The ratio of ( - )-menthone
(Croteau and Johnson, 1985; Gershenzon and Croteau, (26) to menthol (27) decreases. UDP-glucose reacts with
1990). ( + )-neomenthol (28) in the presence of ( + )-neomenthyl
The enzymes responsible for the hydroxylation of monot- glucosyl transferase (a soluble enzyme with 46,000 MW) to
erpenes such as ( - )-limonene (11) from peppermint (Men- form ( + )-neomenthyl glycoside, which is transferred to the
tha piperita), spearmint (Mentha spicata), and perilla (Per- roots. There the glycoside is hydrolyzed to neomenthol, oxi-
illa Jrutescens) have been isolated and characterized (Karp dized to menthone, and subsequently converted to 1-3,4-
et al., 1987, 1990). Microsomal preparations from the epider- menthone lactone (37) (Fig. 19.11) (Croteau, 1988; Gershen-
mal oil glands of these plants catalyze the NADPH and Oz- zon et al., 1989).
dependent allylic hydroxylation of (- )-(4S)-limonene (the
major olefinic constituent of each of the three species) at C- Compounds Derived by Alkylation with
3, C-6, and C-7, respectively, to produce ( - )-trans-isopiper- (ieranyl·OPF
itenol (34), ( - )-trans-carveol (35), and ( - )-perillyl alcohol
(36) (Fig. 19.9) (Karp et al., 1990). These transformations Geranyl pyrophosphate is a potent alkylating agent that
are the key steps in the biosynthesis of oxygenated monoter- reacts with compounds like DMAPP to produce compouods
penes in the respective species. The enzymes appear to be similar to hemiterpenes such as bergamotin (38) (Fig. 19.12)

geranyl OPP (3) olivetol

t55
I
" ---
HOHI
HO" C,Hll
-+

CsHu

a1.tetrahydrocannibinol
(40)

Al(fi).tetrabydrocannibinol cannabidiol R=H (42) cannibinol R=H (43)

(41) cannabidiolic acid R=COzH (44) cannabinolic acid R=COzH (45)

Fig. 19.13. Proposed biosynthesis of 1l 1-tetrahydroc8IUlabinol.

336 Monoterpenes

and with isopentenyl pyrophosphate to produce FPP and then nyl pyrophosphate and olivetol are major precursors (Kajima
sesquiterpenes (Geissman and Crout, 1969). and Piraux, 1982; Crombie, 1986) (Fig. 19.13).
Geranylhydroquinone (39) (Fig. 19.14), a compound re-
sponsible for photodermatitis from Plulcelia crenulata, ap- Hydroquinone Derivatives in Cordia
pears to be derived in a similar manner (Rodriguez, 1983). Species
A series of hydroquinone terpenoids in Cordia alliodora
lIIonoterpene-Derived Compounds (Boraginaceae) are derived from a geranylphenol precursor
oflllarijuana (Fig. 19.14). The presence of a number of compounds de-
Cannabis sativa (Cannabaceae) has been used as a drug rived from this pathway in this species suggests that a simple
plant for millenia. The plant also is important as a fiber and reaction of the phenolic nucleus is followed by cyclization,
oilseed crop in some countries and is widely cultivated for rearrangement, and subsequent oxidation (Manners and
that purpose as well as for drug purposes. It is estimated Jurd, 1977).
that 200-300 million people use marihuana, hashish, bhang,
daga, and so forth. The major psychoactive component is
(- )-A-1-tetrahydrocannabinol (A-1-THC) (40) (Kajima ClmlIIOSYSTElIIADC USB OF 1II01'l0TERPBNB
and Piraux, 1982) (Fig. 19.13). ClmmCAL DATA
Varying amounts of Al(6)_THC (41) (also psychoactive),
cannabidiol (42, R = H), cannibinol (43, R = H), cannabidi- The analysis of volatile monoterpenes has been of considera-
olic acid (44 R = C02H), and cannabinolic acid (45, R = ble value for the resolution of systematic problems at the
C02H) co-occur in most plants of the species. generic, specific, and infraspecific level (Adams, 1977; Heg-
Labeling studies have not been overly successful because nauer, 1978; Irving and Adams, 1973; von Rudloff, 1975),
of poor incorporation of precursors, but it appears that gera- but is oflimited value for the resolution of phyletic problems

cymopol
~ OH
geranylbydroquinone (39)
PhaeeUa crenulata

,ry0-l~.A
HO~- - ,

cordiaebromenes

CH,OH

cordiachromes allioqulnols cardiols

Fig. 19.14. Geranylhydroquinone and the proposed biosynthesis of related compounds from Cordia alliodora (Manners and Jurd, 1977; modified and
used with pennission of the copyright owner, the Royal Chemical Society, Cambridge).
 MonOlerpenes 337

at the family level and above. In several, if not most cases, (17). Linalool (6) was variable during fruit development.
a few genes appear to be involved in the control of the struc- Less limonene occurred in hybrids.
tnre and amount of monoterpenes produced. The comple- Hybrids of diploid and tetraploid Citrus species exhibited
ment of monoterpenes for many plants is often unique, but essential oil composition that varied widely from those of
the individual compounds are widely distributed and the the parental stock. In many cases, the tetraploid and diploid
pathways leading to them are found in many unrelated taxa parents contributed almost equally in terms of essential oil
of plants. Further, plant-tu-plant, populational, and specific composition, despite the fact that two-thirds of the chromo-
variation often is sufficient to pose problems in utilization somal material was from the tetraploid parent. The trends
of monoterpenes as characters for systematic stndies and observed in leaf oil composition often did not coincide with
must be evaluated in any study of this type. Under some those of the rind oil (Scora et al., 1970).
circumstances, stressful environments cause enhanced allo-
cation of fixed carbon into essential oils (Ross and Som- Chemosystematic Study of Gymnosperms
brero, 1991).
The use of essential oils for chemosystematic stndy of
Although the value of specific compounds and pathways
North American conifers has been reviewed and many of the
is of limited phylogenetic value, it is clear that certain groups
problems of sample collection, analysis, and interpretation
of plants accumulate monoterpenes and possess the morpho-
of data discussed (von Rudloff, 1975). In addition, specific
logical features for storing them. Monoterpenes often are
treatments of Abies, Chamaecyparis, Juniperus, Picea,
found in gymnosperms (Coniferopsida), the Annoniflorae
Pinus, Pseudotsuga, Thuja, and Tsuga are summarized.
(order Annonales), Apiaceae, Araliaceae, Asteraceae, Cyp-
Sampling of these coniferous trees was found to be most
eraceae, Euphorbiaceae, Geraniaceae, Juglandaceae, La-
reproducible during the dormant period from fall through
miaceae, Myrtaceae, Orchidaceae, Pittosporaceae, Rosa-
winter.
ceae, Rutales (Anacardiaceae, Burseraceae, Meliaceae,
Among members of the genus Picea (spruces), examples
Rutaceae, and Simaroubaceae), Verbenaceae, and Zingiber-
are found in which there is so little variation that no signifi-
aceae. These volatile compounds are encountered, but occur
cant measure of introgression can be made, some in which
sporadically or in small amounts, in many other families the degree of variation is sufficient to study both intraspecific
(Gibbs, 1974; Hegnauer, 1962-1986). and interspecific problems and others in which tree-tu-tree
The accumulation of large amounts of monoterpenes in variation is so large that systematic studies are impractical
the Annonales, but not in the Berberidales, Nelumbonales, with this type of character. Picea mariana and P. rubens
and Nympheales, supports segregation of that order. An (black and red spruce, respectively) exhibit little intrapopula-
overall unity of the superorder is demonstrated by the pres- tional or interpopulational variation in essential oil composi-
ence of benzylisoquinoline alkaloids throughout (Seigler, tion. These species vary only slightly throughout their
1977). ranges, and the complement of monoterpenes in the two
Dahlgren (1977) placed the Pittosporaceae in his Arali- species is quite similar. This confirms the close relationship
iflorae, in agreement with chemical data previously reviewed that has been proposed for these species. Picea breweriana
by Hegnauer (1969). Additionally, Dahlgren segregated the is similar to black and red spruces in monoterpene composi-
Araliaceae and Apiaceae from the Comiflorae and placed tion and less so to white, Sitka, blue, and Engelmann spruces.
them in his Araliiflorae along with the Pittosporaceae. Both The northwest complex of white (Picea glauca), Sitka (P.
the Araliaceae and Apiaceae lack the iridoid monoterpenes sitchensis), and Engelmann spruces (P. engelmannii) has
commonly found in the Corniflorae. several regions where the species overlap and hybridization
Hegnauer (1969, 1978) has pointed out that the Hippuri- is reported to occur in all of these. This is especially true
daceae differs from the Haloragidaceae and Gunneraceae in for white and Engelmann spruce. Although the essential oil
chemistry and morphology and most Closely resembles the composition for Picea glauca is relatively constant over
Bignoniaceae. The Verbenaceae and Lamiaceae are qnite most of the range of the species, in areas of proximity to P.
similar chemically; both accumulate many types of mono-, engelmannii much more variation is observed, suggesting
sesqui-, and diterpenes and both contain iridoid monoter- that introgression occurs. The essential oil composition of
penes. These two families are chemically distinct from the blue spruce (P. pungens) resembles that of Picea glauca.
Boraginaceae and Hydrophyllaceae, but they are placed in The ranges of the two species do not overlap. Chemical
the same superorder by Thorne (1976). evidence suggests, however, that blue spruce and Engelmann
spruce do hybridize in some areas (von Rudloff, 1975).
Cbemosystematic Study of Citrus Species Over 50 species and varieties of the genus Pinus occur
in North America. These have previously been studied by
Monoterpenes from a series of Citrus species and certain Mirov and were reviewed by von Rudloff in addition to his
closely related genera have been studied [mostly by Scora own studies on Canadian members of the genus. An espe-
and coworkers (1970)]. The essential oils of rinds of 4 Citrus cially large difference was noted in the essential oils of Pinus
parents, 2 Pondrus parents, and 17 hybrids were examined ponderosa and P. jeffreyi. The essential oil of the latter con-
and found to contain principally limonene (11) and myrcene sists of n-heptane. Hybrids of the two species can easily be
338 Monoterpenes

recognized by chemical analysis. Differences in essential and "pure C. sargentii" were selected. In 25 out of 35 trees,
oil composition between P. banksiana Gack pine) and P. the terpenoid composition identified the tree as either C.
contorta var. latifolia (lodgepole pine) were large enough sargentii or C. macnabiana. Of these trees, 12 were identi-
to permit studies of introgression in populations in Alberta. fied as C. sargentii and 13 as C. macnabiana or a ratio of
This phenomenon was also observed in the composition of about 1 : 1 of pure genotypes. Ten trees could be identified as
the leaf oils (von Rudloff, 1975). intermediate and hybrids. The complexity of this population
The species Pseudotsuga menziesii (Douglas fir) consists suggests that extensive hybridization had been occurring for
of a coastal and an inland taxon. These meet in British Co- some period of time (Lawrence et aI., 1975).
lumbia and many intermediate plants exist. The two taxa The composition of essential oils of Juniperus scopu-
also have distinct essential oil composition from leaf oil and lorum (Cupressaceae) was shown to differ between young
cortical oleoresins. Samples from the Cascades of southern leaves and mature leaves. In general, hydrocarbon terpenoids
British Columbia and northern Washington are intermediate were higher in concentration in new leaves, and oxygenated
between the coastal and the inland taxa (von Rudloff, 1975). compounds were higher in old leaves. This pattern has also
Many systematic aspects of the genus Abies (firs) are been observed in a number of other gymnosperms. The
difficult to understand. It is probable that hybridization and amount of variation in young leaves was less than in older
introgression have played a role in this. The essential oil leaves with regard to essential oil content.
composition of eastern and western balsam fir (Abies balsa- Oil samples from Juniperus scopulorum trees taken at
mea) in Canada was found to differ. In addition, where A. different times of the day differed significantly in composi-
balsamea and A. lasiocarpa (alpine fir) meet or overlap, the tion. Oxygenated terpenes and sesquiterpenes tended to in-
essential oil of balsam fir assumes many of the features of crease during the day, whereas sabinene (22) decreased until
alpine fir oil. The wide distribution of these characters in late evening and increased during early morning. Diurnal
populations of balsam fir in Saskatchewan and Manitoba variation was much less in winter than in summer (Adams,
suggests that this is an ancient phenomenon. Many other 1979; Adams and Hagerman, 1976, 1977).
troe firs show variation in areas where overlap of species is Analysis of populations of Juniperus ashei throughout
thought to occur (von Rudloff, 1975). Texas for areas of population differentiation and possible
An extensive survey of the essential oils from populations hybridization with J. virginiana and J. pinchotii by both
of Juniperus virginiana from Texas to Ontario indicates that morphological and essential oil features failed to detect evi-
elinal variation occurs. Further, the proposed introgression dence of hybridization (Adams, 1975; Adams and Tumer,
between J. virginiana and J. ashei which had been thought 1970).
to occur widely in Texas in areas of overlap between the
two species does not appear to exist, based on analysis of
the essential oils. This was also confirmed by analysis of Chemosystematic studies in lIfints
micromorphological characters (von Rudloff, 1975).
Irving and Adams (1973) found that essential oil compo-
In species of the Pinaceae, three general trends concern-
nents of Hedeoma drummondii and H. reverchonii var. re-
ing the inheritance of essential oils in relation to hybridiza-
verchonii (Lamiaceae) were additive or complementary in
tion are observed. In F, hybrids, essential oils often are com-
F, hybrids of the two species. In F2 hybrids, plants were
positionally intermediate between those of the parents.
intermediate in chemistry, some of which resembled each
Although they are commonly not exactly intermediate, F,
parent, and a few with novel chemistry were observed (irv-
hybrids rarely possess oils with composition outside the
ing and Adams, 1973).
range of the two parental species. In other instances, a weak
linkage between morphological characters of the plant and
essential oil composition has been observed in some popula-
tions of conifers. In general, however, there is no necessary
BIOLOGICAL ACTIVITY
correlation of essential oil composition and the various mor-
phological features of plants, and in backcrosses, plants re-
sembling one parent morphologically resemble the other in Most monoterpenes are cytotoxic to plant and animal tissues,
essential oil chemistry. The variation observed has been use- causing a drastic reduction in the number of intact mitochon-
ful in determination of the presence of hybridization and dria and Golgi bodies, impairing respiration, and photosyn-
backcrossing in populations of mixed parentage (von Rud- thesis, and decreasing cell membrane permeability (Charl-
loff, 1975). wood and Charlwood, 199Ia). At the same time, they are
In studies of Cupressus sargentii and C. macnabiana in volatile and many serve as chemical messages for insects
California, Lawrence and co-workers (1975) noted that the and other animals. Furthermore most monoterpenes serve as
essential oil composition of the two parental types was rela- signals of relatively short duration, making them especially
tively constant and differed in a number of features. In a useful for synomones and alarm pheromones. Several types
population where the ranges of the two species overlapped, of semiochemical functions of monoterpenes are described
trees which could be identified as typical C. "macnabiana" below.
 Monoterpenes 339

lIIonoterpenes as A11omones. Kairomones. tions of defensive substances in many species of plants. For
and Synomones example, bark beetle feeding and the presence of their fungal
symbionts causes increased accumulation of terpene-con-
Essential oils are comprised of complex mixtures of ter-
taining resin in a locallzed area surrounding the site of attack
penes, phenylpropanoid-derived compounds, and a number
(Gershenzon and Croteau, 1991).
of esters, alcohols, aldehydes, ketones, acids, and hydrocar-
Ironically, terpenes serve both as major deterrents to feed-
bons derived from fatty acid metabolism. The constituent
ing and as attractants for animals (Hedin et al., 1974). Vola-
compounds usually are of low to medium molecular weight
tile compounds are often involved in long-distance interac-
and not highly oxygenated (Fig. 19.15). Many of these vola-
tions and are especially important as attractants and
tile compounds are involved in interactions with animals and
repellents (Kogan, 1975).
are major components of the "taste" and "odor" of plants.
Many monoterpenes are biologically active (Fig. 19.15)
A11omones
(Herout, 1970; Seigler, 1983) and representatives of this
group of compounds are toxic to a large number of organ- Monoterpenes serve as a solvent that permits other resin
isms. For example, thujaplicinol (46) (Fig. 19.1) is a useful constituents (particularly diterpenes) to flow into wounds,
antibacterial adenyltransferase inhibitor (Croteau and John- an important defense of plants against insect, fungal, and
son, 1985). Among the biologically active terpenoids, mo- bacterial attack. Changes in the composition of secondarily
noterpenes and sesquiterpenes predominate. formed resins have been correlated with an increase in anti-
Biotic stresses can have a significant effect on plant terpe- fungal and insect-repellent properties. Biological effects also
noid levels. Herbivory often induces increased concentra- are highly dependent on the enantiomeric composition of

limonene (11) nerol (60)' geraniaJ (61)· citronellal (62)

(e
a-pinene (14)
\ (57)
0\ 0.0 (58) (59)

methyl chaviool (56) carvone (48) canooe oxide (80) E.limonene oxide (78)

2
a phenylpropanoid

a;-phellandrene (79) borny. acetate (77) n-phellandrene (63) 2-terpinolene (64)

•citral (mixture of geranial and neral)

Fig. 19.15. Some biologically active monoterpenes.

340 Monoterpenes

terpenoid mixtures (Croteau and Johnson, 1985). For exam-

ple, (+ )-limonene (11) (common in Citrus oils) strongly
inhibits germination and sperm tube growth of Diplodea
pinea, a pathogen of pines. ( - )-Limonene, which occurs in
many Pinus species, was relatively nontoxic.
Volatile monoterpenes are common components of the
"taste" and "odor" of plants. A significant proportion of
herbivorous insects are "specific" (monophagous or oli- ochtodene (54) cbondrocole A (55) dtondrocole C
gophagous), feeding on plants of a few genera or a single
Fig. 19.16. Monoterpenes from algae.
family (Bernays and Graham, 1988; Jermy, 1984). This
choice often is determined by the ptesence of plant second-
ary compounds, among which monoterpenes are important. respectively, when applied as l00-mglml soil drenches
Most insects do not eat most plants; in part, this is because (Chitwood, 1992).
of the presence of allomones or compounds that benefit the Few mammals appear to be able to adapt tu large quan-
emitting organism. In general, a successful herbivore is tities of monoterpenes in the diet, but have an aversion to
adapted to the secondary compounds of a particular plant. terpenoid odors and plants rich in terpenoid constituents
\3-0cimene (47) is a repellent to the leaf culter ant Alta (Harborne, 1991).
cephalotes in both field and laboratory experiments (Har- The leaves of the California bay laurel (Umbellularia
borne, 1987). Experiments with the aphid Cavariella aego- cali/ornica, Lauraceae), sometimes used as a spice, contain
pod;;, which feeds on umbellifer species in summer, indicate 0.5-4% of an irritating oil of which 40-60% is umbellulone
that the aphid can be captured in traps baited with carvone (53). The vapors of this compound produce severe headache,
(48), but are repelled by linalool (6) (Chapman et al., 1981; skin irritation, and, in some cases, unconsciousness. This
Harborne, 1987). Carvone occurs in the essential oils of sev- plant is one of several avoided by deer. Essential oils inhibit
eral plants of the Apiaceae. microbial activity in ruminants and disrupt digestive pro-
A number of monoterpenes and methyl esters of fatty cesses (MacGregor et al., 1974; Buttery et al., 1974).
acids were evaluated for their repellent and attractant proper- Several species of algae produce halogenated secondary
ties toward Ips, Dendroctonus, and Hylurgops species. Al- metabolites. For instance, the red alga Ochtodes secundira-
though activity was observed in the laboratory, none of the mea, which is highly resistant to herbivory, produces ochto-
compounds tested were active in field tests (Seigler, 1983). dene (54) and chondrocole A (55) (Fig. 19.16). Octodene
The wood of western red cedar (Thuja plicata) is re- was highly repellent toward marine fishes, but only the unre-
nowned for its ability to withstand insect attack in tim- solved monoterpene mixture from the plant was deterrent to
berstands and as lumber. This may be attributed to the mo- herbivorous amphipods (panl et al., 1987).
noterpenes that it contains. \3-Thujaplicin (49) is toxic to Cymopol, a terpenoid bromohydroquinone frum the
larvae of the old house borer (Hylotrupes bajulus), a com- green alga Cymopolia barbata, significantly reduced feeding
mon pest in Europe. Both oc- and \3-thujaplicin showed low by reef fishes in feeding trials, but significantly stimulated
activity against the termite Reticulothermes Jlavipes. Inter- feeding by an herbivorous sea urchin species of the genus
estingly, methyl thujate (52) (thujic acid, 50) was much more Diadema (Fig. 19.16) (Hay et al., 1987).
active. This compound is active against a number of other
insects. (+ )-3-Thujone (21) and (- )-3-isothujone (51), AlIomones Froduced by Arthropods.
which make up most of the leaf essential oil of this plant Especially Insects
(80-90%), are feeding deterrents to the white pine weevil Monoterpenes are encountered commonly in a number
(Pissodes strobil (Alfaro et al., 1981). of insects (Nahrstedt, 1982). Many of these compounds are
Bay leaves (Laurus nobilis, Lauraceae) and cineole (10), identical to those found in plants and, in any given case,
geraniol (4), and piperidine (which occur in bay leaves) pos- it may be difficult to decide whether the compounds are
sess repellent properties tuward cockroaches (Verma and sequestered from plants in the diet, synthesized de novo by
Meloan, 1981). the insect, or produced by symbiotic gut microorganisms.
Female Caribbean fruit flies, Anastrepha suspensa, lay The lower terpenoids are relatively nonspecific toxicants
their eggs readily in ripe grapefruit, but do not oviposit in produced in the defensive secretions of many insects (Fig.
immature grapefruit or in ripe or immature oranges because 19.17) (Herout, 1970; Harborne, 1982). These compounds
of the presence of linalool (6). This compound is toxic to are volatile and this may be important in the mechanism of
the eggs and larvae of the insect. Limonene (11), found in action. The vapors often have an irritating action.
sour oranges (Citrus aurantium), is toxic tu adult bean wee- Larvae of the sawfly, Neodiprion setifer (Hymenoptera)
vils (Callosobruchus phasecoli), but highly attractive to discharge an oily effluent when disturbed. This material is
male Mediterranean fruit flies (Jacobson, 1982). identical in composition to the resin of the host plant Pinus
Two monoterpenes, citra! (sic) and geraniol (4) inhibited sylvestris and the larvae appear to sequester the resin from
reproduction of the root-rot nematode by 52% and 86%, the plant. Some of the components are oc- and \3-pinene (14
 Monoterpenes 341

~ Ce
myrcene (17) a-pinene (14)
-
in vivo

~"
verbenone (65)

p~ ~
resin compound resin compound beetle pheromone

(e
0
0

~-plnene (16) car-3-ene (71) exo-brevicomin (72) frontalin (73)

beetle pheromone

~oo ~~ ~ H
~~--
I ~ OH

(+)-eis-verbenol (74) Ipsdienol (75) (-)-Ipsenol (76)

Fig. 19.17. Biologically active compounds from arthropods.

and 16) and a number of diterpenes. The diterpenes appear ceae locate the plants by the presence of methyl chavicol
to act as a fixative for the more volatile components (Eisner (56) andlorcarvone (48) (Fig. 19.17). If filter paper is soaked
et aI., 1974). with these compounds, the insects will eat it (Herout, 1970).
A number of monoterpenes are used as defensive com- A number of factors control the feeding preferences of
pounds by the walking stick insect Anisomorpha bupresti- most animals. In the case of the silkworm, Bombyx mori,
odes and the ant Acanthomyops daviger. The compounds which feeds almost exclusively on the mulberries Morus
are apparently synthesized within the insect. Other allomonal alba and M. nigra, a range of factors is involved. Most of
compounds (57-59) produced by ants are probably terpene the olfactory components are monoterpenes such as citra! [a
derived. ex-Pinene (14) is used by some tennites for defense. mixture of neral (60) and geranial (61)] linalool (6), and
linalylacetate. Bombyx larvae are sensitive to the monoter-
Kairomones pene mixtures when they get within 3 cm of a leaf.
In other instances, compounds called kairomones are re- Upon infestation ofliroa bean leaves (Phaseolus lunatus)
leased by plants (or other organisms) and benefit the receiv- with two-spotted spider mites (Tetranychus urticae), induc-
ing organism. A number of these compounds serve as both tion of (E)-j3-ocimene (47) and a homomonoterpene, E-4,8-
repellent and attractant substances; most kairomones proba- diroethyl-l,3,7-nonatriene, occurs. A homosesquiterpene,
bly originated as allomones. In any instance, it is possible (3E, 7E)-4,8,12-trimethyl-l,3,7,l1-tridecatetraene also has
for a compound to be an allomone with regard to many been observed (Dicke et al., 1990). These compounds attract
insects and a kairomone to others. More than 4000 com- another mite, Phytoseiulus persimilis, which is a predator of
pounds have been evaluated for their ability to attract several the two-spotted spider mite, a phenomenon known as indi-
kinds of insects (Wright, 1966). Plant volatiles are only one rect defense (Dicke et al., 1990; Takabayashi et al., 1991).
factor in the attraction of animals to plants. Visual cues may When uninfested leaves are placed on moist cotton that had
also be extremely important (Stfuller, 1983). previously come into contact with leaves that had been in-
Essential oils are involved in the feeding response of sev- fested (but had the mites removed), production of these com-
eral Papilio species to members of the Apiaceae (Feeny et pounds again occurred, providing evidence that induction
al., 1983). Many species of butterflies that feed on the Apia- of these kairomones occurs (Takabayashi et al., 1991). The
342 Monoterpenes

homomonoterpene appears to arise by cleavage of a sesqui- the constitutive resin. Healthy trees respond by a hypersensi-
terpene (Gabler et a!., 1991). tive response; that is, the cells around the area of attack die
and deprive the pathogen of nutrients and compartmentalize
Aggregation Pheromones the attack. In addition, adjacent wood parenchyma differen-
tiates and secretes a secondary resin into the wound site.
Many of the monoterpenes found in essential oils of Bark beetles respond by trying to clear the wound site or
plants also occur as aggregation pheromonal substances in abandoning attack. Fungal growth is also inhibited by the
insects. Examples of some compounds found both in plants secondary resinosis. Elicitors from the fungi may modify
and insects are the monoterpenes citronellal (62), citronellol the monoterpene content of the resin (Johnson and Croteau,
(7), geraniol (4), myrcene (17), citral [a mixture of geranial 1987).
and neral (60, 61)], l3-phellandrene (63), limonene (11), 2- The terpenoid fraction of many conifers consists of
terpinolene (64), a-pinene (14), l3-pinene (16), 1,8-cineole 20-50% volatile monoterpenes and sesquiterpenes and
(10), and verbenone (65). 50-80% nonvolatile diterpene acids. In general, there is little
The male cotton boll weevil (Anthonomus grandis) is variation in the diterpene fraction among trees of a particular
attracted to the cotton plant by a number of monoterpenes species, although the monoterpene fraction often varies
and sesquiterpenes, among them are a-pinene (14),limonene greatly. Alterations in the monoterpene content often deter-
(11), and caryophyllene (66) (Fig. 19.18). The boll weevil mine whether attack by bark beetles will occur and be suc-
then produces a series of metabolites that attract both male cessful. The monoterpenes may be attractants, repellents, or
and female individuals. Although these terpenes can come toxins of not only the insects, but also the pathogenic fungi
from the plant, the beetles can also synthesize the com- involved in these systems. The conversion of geranyl pyro-
pounds de novo. At least four (67-70) have been isolated phosphate to the parent compounds of the various cyclic
and characterized (Vanderwel and Oehlschlager, 1987). types is catalyzed by monoterpene cyclases (see the subsec-
In some cases, the production of monoterpene aggrega- tion biosynthesis, above).
tion pheromones by insects is quite complex. The production The effect of enantiomeric monoterpenes on bark beetle
of pheromones by bark beetles (mostly of the genera Den- activity is very significant. In some cases, as little as 2-5%
droctonus, Ips, and Scolytus) has been reviewed (Gershen- of an enantiomeric product will completely inhibit the effect
zan and Croteau, 1991; Vanderwel and Oehlschlager, 1987). of the active form and greatly upset the normal pheromonal
Death of the host tree usually follows successful colonization system of the insect. Fungi are also sensitive to these effects.
(Johnson and Croteau, 1987). Tree mortality is usually nec- For example, (+ )-a-pinene (14) is three times more inhibi-
essary if the beetles are to survive because excavation of tory than (- )-a-pinene against the blue stain fungus, Cerat-
egg galleries and subsequent larval development can only ocystis spp. The differential expression of the cyclases re-
take place after host defensive reactions have stopped. Pi- sponsible for coproduction of enantiomeric monoterpenes
oneer beetles may encounter constitutive (preformed) resin may determine the highly selective resistance response coni-
on initial attack. This often leads to emission of pheromonal fers exhibit toward bark beetles and their fungal symbionts
substances. Mass attack usually leads to rapid depletion of (Johnson and Croteau, 1987).

limonene (11) a-pinene (14) caryophyllene (66)

(68) (69) (70)

(67)

Fig. 19.18. Proposed biosynthesis of attractive compounds from cotton for the male boll weevil and bon weevil metabolites (Vanderwel and
Oehlschlager, 1987; modified and used with pennission of the copyright owner, Academic Press. Orlando, Fl.).
 Monoterpenes 343

In the case of the Western pine beetle (Dendroctonus tion of ( + )-ipsdienol by the males of Ips paraconfusus in-
brevicomis) and the ponderosa pine (Pinus ponderosa), uti- hibits colonization by two other species of bark beetles, Ips
lization of terpenes involves several stages. The oleoresin pini and Dendroctonus brevicomis (Harborne, 1986).
of pine bark is rich in volatile terpenes and traces of the In the related species, Dendroctonus frontalis, the com-
vapor exude from the tree, especially if it has been damaged pound frontalin (73) is primarily attractive to male bees and
(Wood, 1982). The tree contains (3-myrcene (17), (3-pinene the female does not release a female attractant. The first
(16), and 3-carene (71) (Fig. 19.17). The female beetles are males to arrive ingest resin and synthesize and release a male
attracted to the tree and begin to feed. Once the females inhibitor, verbenone (65). It seems quite likely that the male
are established, they begin to attract males for reproductive beetles have the ability to convert a-pinene into verbenone.
purposes. The females employ a mixture of three compo- Parasitoids and predators of many of these bark beetles
nents: myrcene (17), which is sequestered from the oleoresin are attracted to the monoterpenes and derived substances.
and used directly, and two bicyclic compounds, exo-brevi- In this way, these insects locate their insect hosts (Wood,
comin (72) and frontalin (73), which appear to be synthe- 1982).
sized de novo by the insect. Brevicomin and frontaIin proba- Bornyl acetate (77) and verbenyl acetate mimic the activ-
bly are products of fatty acid metabolism. The males, ity of the American cockroach sex pheromone (Bowers,
attracted by the pheromone, arrive at the site, copulate with 1985).
the females, and a new generation of beetles emerge. The Honey bee swarm attractants include geraniol (4) and
female beetles appear to control the sex ratio by varying the (E)-citral (neral ?), monoterpenes that are found in the Naso-
proportions of the sex pheromone mixture exuded at any nov glands of living honey bees (Pickett, 1991).
given period of time. If the mixture contains all three compo-
nents, a I: 1 mixture of males and females are attracted, Trail.Marking Pheromones
whereas if frontaIin is omitted, the ratio changes to 2: 1 in Trail markers are used almost exclusively by social in-
favor of the males. When the area of infestation reaches its sects, such as bees, ants, and termites. An insect that has
optimum size, there is only food for the existing beetles. located a new source of food returns to the nest, marking his
The female then stops pheromone prodUction and begins to trail by releasing these substances. At the nest, recruitment
form a male repellent, verbenone (65). This compound repels is initiated by perception of these trail pheromones. Some
males and keeps them from reaching the infestation site. representative trail pheromones are presented in Fig. 17c.
Verbenone is formed from one of the major bark terpenes,
a-pinene (14). The addition of myrcene (17) to the fron- Alarm Pheromones
talin-brevicomin mixture doubles the effectiveness of the
attractant. Furthermore, myrcene is essential and cannot be Many insects produce volatile compounds that are deliv-
substituted by the closely related compound limonene (11). ered from the mandibular or anal glands or from the sting
A mixture of frontalin and 3-carene (71) is highly attractive apparatus. Production of these compounds is often related
to females, but not to males (Fig. 19.19) (Harborne, 1982; to the production of defensive compounds. Alarm is commu-
Wood, 1982). Production of verbenone (65) inhibits coloni- nicated to other members of the society by diffusion of the
zation by the bark beetles Ips paraconfusus, and the produc- compound into the air (Harborne, 1982).
tion of ( + )-ipsdienol (75) inhibits colonization by Ips pini Wasps of the genus Vespa spray venom containing an
(Harhorne, 1986). alarm substance, whereas honey bees leave traces of isoamyl
Many of the monoterpenes from host trees are toxic to acetate at the sting site which induced other bees to sting at
the beetles that feed on the trees. The ability to oxidize these the same location. In some insects, the same compound may
compounds may have originated as a detuxification mecha- serve for both alarm and defense. This is true of formic
nism and, secondarily, been selected for reproductive pur- acid produced by the genus Formica. Most of these alarm
poses (Vanderwel and Oehlschlager, 1987). Investigation of compounds have a relatively simple structure. Essential oil
the stereochemistty of these oxidation and hydration re.ac- components including citronellol (7), citral (a mixture of
tions indicates that most are highly specific. neral and geranial) (60, 61), a-pinene (14), terpinolene (12),
A bacterium has been isolated from the gut of Ips confu- and Iimonene (11) have been implicated in various examples
sus that can oxidize a-pinene (14) to verbenol (74), whereas studied (Fig. 19.19). Ants use neral (61) and geranial (60)
a symbiotic fungus of Dendroctonus has been shown to fur- as alarm signals.
ther oxidize verbenol to verbenone (65) (Fig. 19.19). The
male of the insect Ips paraconfusus, which feeds on Pinus Synomones
ponderosa, produces a frass which contains terpenoid-de- Synomones are compounds that are involved in interac-
rived compounds ipsdienol, ipsenol (76), and cis-verbenol tions that are of net benefit to both the producer and the
(74) (Wood, 1982). This frass attracts both male and female receiver. The role of monoterpenes in pollination has been
insects to the tree. It is quite possible that microorganisms reviewed (Vogel, 1983).
playa major role in the survival of the beetle on a success- The pollination of orchids (Orchidaceae) involves many
fully colonized tree (Harborne, 1982; Wood, 1982). Produc- species-specific interactions and exhibits many interesting
344 Monoterpenes

and benzyl acetate is used (Dodson et aI., 1969). Although

most bees are attracted to many of the pure compounds,
fewer are attracted to mixtures in almost all cases.
The complement of volatile substances produced by most
species of orchids is highly specific and this specificity ap-
pears to preclude hybridization in many cases (Hills et aI.,
1972). Although some species of orchids are pollinated by
geraniol (4) geranial (60) '" geranic acid several species of euglossine bees, most are pollinated by a

A Am. A_
single species of bee (Ackennan, 1983).
There is controversy about the exact role of the odorifer-
ous compounds collected by the male bees, but it is likely

~~OH ~ ~
that they are involved in attraction of other euglossine bees
or converted into sexual (aggregation) pheromones. It has
been suggested that males use the compounds and various
aspects of behavior to attract other males. This makes a small
nerol(5) neral(61) '" nerolieacid swarm of bees, and the females are attracted. This is similar
'" a mixture of geranial and neral is called citral to lek behavior in birds (Harborne, 1982, 1989).
Although the orchids are visited only by euglossine bees,
Fig. 19.19. AIann pheromones.
the bees are known tu visit flowers of plants of other families
with similar chemistry (Williams and Dodson, 1972). Al-
though the bees have undoubtedly been important in deter-
types of evolutionary problems. The color, shape, and odor
mining the course of evolution in the orchids, facturs in
of the flower are all important. Orchids are pollinated var-
addition to those associated with the orchids have been im-
iously by butterflies, moths, flies, mosquitos, birds, and bats,
portant in the evolution of the bees (Ackennan, 1983).
but most commonly, by bees.
In orchids pollinated by five species of Eulaema and three
Several species of orchids are pollinated by eUglossine
species of Euglossa in Panama, carvone oxide (80) is the
bees (Dodson et aI., 1969; Hills, 1989; Williams and Whit-
dominant volatile compound in II of 12 plants pollinated.
ten, 1983). In one example, pollination appears to involve
Although 11 of these plants are orchids, Dalechampia
a relatively simple monoterpene. The essential oil comple-
spathulata (Euphorbiaceae) also produces carvone oxide and
ment of Stanhopea pulla, which consisted mostly of E-Iimo-
is pollinated by the same bees (Harborne, 1987, 1989).
nene oxide (78) (Fig. 19.17), 1,8-cineole (10), and an uniden-
One of the important monoterpenes in bark beetle-coni-
tified monoterpene, varies both quantitatively and
fer interactions, ipsdienol (see above), is also a major compo-
qualitatively depending on the time of day. Maximum fra-
nent of several tropical orchid species and is attractive to a
grance production occurs early in the morning. The euglos-
number bees of the genus Euglossa (Whitten et al., 1988).
sine bee involved responds to E-Iimonene oxide (78) in mix-
Many Ophrys species (Orchidaceae) are pollinated by
tures of E- and Z-limonene oxide (Fig. 19.17) (Hills, 1989).
bees of the genera Andrena and Colletes. Pseudocopnlation
In other instances, the parameters that detennine pollina-
and the presence of highly specific flower scents are in-
tion are complex. Many of these orchids attract only the
volved in this process. Linalool (6), citronellol (7), citronel-
male bee. When a flower is placed in an opaque container,
lal (62), geraniol (4), and geranial (61) occur in the scents
the bees still find it, indicating that the bees use odor and
of Ophrys species (Harborne, 1989). In Ophrys lutea var.
not vision to locate the flowers. Even when the flower was
lutea, famesol (a sesquiterpene), geraniol (4), and geranial
removed, the bees continued to be attracted to the container.
are primarily responsible for release of odor-guided mating
Most of the orchid species produce a combination of 7-10
behavior at the O. lutea labellum and are general attractants
compounds. Among these are d-carvone (48), l,8-cineole
for many species of Andrena (Harborne, 1987, 1989). For
(10), eugenol, bornyl acetate (77), anethole, geraniol (4),
discussion of other Ophrys-bee interactions, see Chapter 21.
methyl cinnamate, methyl salicylate, myrcene (17), vanillin,
and indole. In pure fonn, 1,8-cineole, methyl salicylate, eu-
genol, methyl cinnamate, bornyl acetate, a-phellandrene
(79), myrcene (17), piperonal, indole, vanillin, and benzyl ALLELOPA'DIY
acetate serve as attractants. Neither a- or j3-pinene nor oci-
mene (47) are attractive to the bees tested. Combinations of As previously discussed (see Chapter 8), many plant second-
these compounds (in proportions similar to those found in ary compounds are involved in plant-plant chemical warfare
various orchid species) lead to greater specificity of attrac- called allelopathy. Terpenoids often are involved in interac-
tion. For example, when a-pinene (14) is added to a mixture tions of this nature (Fig. 19.20). Many monoterpenes inhibit
of l,8-cineole (10) and benzyl acetate in the proportions gennination of seeds and growth of seedlings. Exposure of
found in the orchid Stanhopea tricorn is, fewer bees of two plants to the monoterpene vapors produces developmental
species are attracted than when a mixture of only l,8-cineole and physiological changes of several types (Fischer, 1986,
 Monoterpenes 345

£0 S: 2
1,8·cio..l. (10) camphor (33) a-pinene (14) p·pio.o. (16) camphene (15)

1
0
H£CH3COO£
0 u
o
0 u
0
:::...1
:::... :::...
Ihuj.oe (21) (+ ).puleg.o. (29) epievodone (85) calimintbone (86) p-cymene (S1)

piquer.1 A (82) piquer.1 B (83) p-menthane--3,S-cis-dlol (84)

Fig. 19.20. Terpenes involved in a1lelopathy.

1990; Vaughn and Spencer, 1993a, 1993b). Among the most duced by the effects of monoterpenes liberated from the
active monoterpenes are a-pinene (14), (3-pinene (16), cam- plants. These workers carefully eliminated other factors such
phene (15), limonene (11), a-phellandrene (79), p-cymene as shade, lack of water, nutrient conditions, the slope of
(81), 1,8-cineole (10), borneol (32), pulegone (29), and cam- the ground, root competition, herbivory, and water·soluble
phor (33). In a comparison of many structural types of mo- compounds. However, other factors may playa significant
noterpenes, the greatest phytotoxic effects were observed role in the creation of these bare zones. Small rodents that
with cyclic a,(3-unsaturated ketones (Fischer, 1990). Several consumed seeds and shoots also are implicated (Bartholo-
volatile monoterpenes were effective in inhibiting sprouting mew, 1970).
of potato tubers; 1,4·cineole, 1,8-cineole (10), and fenchone The allelopathic activity present is associated with vola-
were among the most effective (Vaughan and Spencer, 1991, tile, lipophilic compounds. Monoterpenes occur in high con-
1993b). centrations in the leaves and a vapor cloud hangs around the
Pioneering plants of disturbed habitats often produce ac- plants. A similar complement of monoterpenes occurs in the
tive monoterpenes. ( + )-PuJegone (29) was four times, and soil surrounding the plants. These terpenes remain in the dry
( + )- and ( - )-camphor two times more toxic to seeds and soil until rain encourages the growth of microorganisms that
seedlings of Raphanus sativus (Brassicaceae) than HCN degrade them. Fires, which periodically sweep through the
(Asplund, 1968). The Mexican plant Piqueria trinervia (As- chaparral, decompose the compounds. These monoterpenes
teraceae) produced monoterpene diols, piquerol A (82) and can be transported into plant cells through the waxy coatings
B (83), that strongly inhibited root and stem growth of com- of seeds or roots. Componnds from the plants have a signifi-
peting plants at 50-200 ppm. cant effect on the germination of seeds of annuals that grow
One of the best known studies of allelopathy was done in the adjacent grassland areas (Muller and Chou, 1972).
in the California chaparral, in an area where two of the most The most effective toxins of Salvia leucophylla are a-
important shrubs are Salvia leucophylla and Artemisia cali· and (3-pinene (14 and 16) and camphene (15) (Fig. 19.20).
fornica. Bare zones of soil from 1 to 2 m in width surround Those of Artemisia californica are 1,8-cineole (10) and cam-
each shrub. Beyond the bare zones are areas of stunted phor (33).
growth where a few herbs show limited development. Muller In the allelopathic effects of Eucalyptus camaldulensis,
and Chou (1972) have postulated that these zones are pro- cineole (10) and a·pinene (14) proved to be the most active
346 Monoterpenes

compounds (del Moral and Muller, 1970). The major allelo- preparation, effects vasodilation of coronary vessels and the
pathic monoterpene from Eucalyptus citriodora proved to femoral vascular bed (Hikino, 1985).
be p-menthane-3,8-cis-diol (84). The ( + )-cis-enantiomer in-
hibited germination and hypocotyl growth of several test Flavors
species but showed no effect on E. citriodora (Cutler, 1992;
Man has long used plant materials as spices and herbs.
Fischer, 1990).
More recently, essential oils have been substituted for many
In a sand hill area of Florida, monoterpenes from Con-
radina canescens and Calamintha ashei (both Lamiaceae) purposes. The chemistry of most of these materials is com-
plex, but in a few fruit or plant flavors, a single compound
are important in inhibition of competing vegetation. Satu-
is responsible for most of the characteristic properties. Those
rated aqueous solutions of monoterpenes from Conradina
canescens inhibited germination and radical growth of two of apple, peach, coconut, and pear are largely single-compo-
native grasses (Weidenhameret al., 1993; Williamson et al., nent flavors. Others, such as banana, are comprised of rela-
1989). The triterpene ursolic acid appears to increase toxicity tively simple mixtures. Many others, such as apricot, are
of the monoterpenes, probably by enhancing the solubility more complex. The flavors of black currants, strawberries,
coffee, and chocolate have not been successfully reproduced;
of the monoterpenes in water. However, monoterpenes are
sufficiently soluble in water to exhibit biological activity the last two have yielded over 700 components on analysis.
(Weidenhamer et al., 1993). A mixture of the monoterpenes Trace components are often important and may contribute
more significantly to the flavor than major ones. In lemon,
epievodone (85) and caliminthone (86) and the sesquiterpene
for example, limonene (11) makes up about 70% of the oil,
caryophyllene oxide (from Calamintha ashei) totally inhib-
but it is the content of citra!, a mixture of geranial (61) and
ited germination of little bluestem (Schizachyrium scopa-
rium), but had only minor effects on lettuce (Fischer, 1990; neral (60) (less than 5% of the oil), that produces the lemon
flavor. An even more effective example is that of cucumber
Fischer et al., 1988). This fmding is of importance because
lettuce seeds have often been used as an assay system for smell produced by nona-2,6-dienal. This substance has an
phytotoxic compounds. odor threshold of 0.0001 ppm (Harborne, 1982).
Thujone (21), a major component of wormwood, Artemi-
sia absinthum, was once a major flavoring of the liquor ab-
sinthe (Arnold, 1989; Harborne, 1991). At doses of 30 mg/kg
USES OF JIIOI'lOTERPEl'IES BY IIUJIIAl'IS of body weight, thujone produces convulsions associated
with lesions of the cerebral cortex. Thujone is also a major
Many monoterpenes have antimicrobial activity. This prop- constituent of cedar leaf oil (Thuja occidentalis) and an im-
erty is linked to many human uses of monoterpenes, includ- portant component of sage (Salvia officinalis) (Alfaro,
ing medicinal and food uses (Beier and Nigg, 1992). The 1981).
odor-taste properties of many are considered desirable and
are used for flavorings and perfumery ingredients. 1,8-Cine- Insect Repellents
ole (10) is commonly used as a flavoring and an antioxidant
Oil of citronella (the essential oil of Andropogon nardus,
in foodstuffs (Charlwood and Charlwood, 199Ia).
Poaceae) has long been used as a mosquito repellent. This
oil is mostly composed of geraniol with lesser amounts of
JIIedicinai Uses of JIIonoterpenes citronellol (7), citronellal (62), and borneol (32). Other es-
Monoterpenes and other volatile terpenes have a number sential oils also have been used to repel insects (Levin, 1973;
of widespread medicinal uses (Beier and Nigg, 1992; Dnke, Seigler, 1983).
1992 and references cited therein). Compounds such as cam-
phor and menthol are used as "counterirritants" or "cool·
ing," analgesic, and antiitching agents and as components IRREGULAR JIIOI'lOTERPEl'IES
of liniments.
Many monoterpenes have been used for anthelminthics. A number of monoterpenes do not fit the "isoprene rnle"
Arnong these are sabinol, cadinol (a sesquiterpene), pinene, and are called irregular monoterpenes. There are three major
ascaridole (87), p-cymene (81), citronellol (7), caryophyl- structoral types of irregular monoterpenes: the chrysan-
lene (a sesquiterpene), limonene (11), phellandrene, cadi- themyl (89), artemisyl (90), and santolinyl (91) skeletons
nene (a sesquiterpene), camphor (33), pyrethrin, carvone (Charlwood and Banthorpe, 1978; Charlwood and Charl-
(48), linalool (6),linalyl alcohol, tagelone, thymol (88), and wood, 1991a). These compounds are found primarily in the
1,8-cineole (eucalyptol) (10). Asteraceae (they are especially well known from the genus
Paeoniae radix, prepared from the roots of Paeonia lac- Artemisia), although a few are found in the Apiaceae and
tif/ora, is an antispasmodic, analgesic, mitigative, and astrin- Lamiaceae (Poulter, 1990). As two skeletal types often occur
gent drug. A series of monoterpene glycosides appear to be in the same plant, it is reasonable to suspect that a common
responsible for much of the activity. For example, paeon- biosynthetic pathway exists and it is possible that the three
iflorin, a monoterpene glycoside isolated from this medicinal types arise from a common catiouic species (Fig. 19.21).
 MonOlerpenes 347

bond opens between

-_-
bond opens between
santolinyl b" ' /

~M lavandulyl

bond opens between

<a)
~--~ artemisyl rothrockyl

~~hr~(89)J~
artemisyl skeleton (90)
santollnyl skeleton (91) lavandulyl skeleton

it1: --
rothrockyl skeleton
rolbrockene\

PPO~ PPO
1
+
1
1
HO

~Hb.J I
~ """ " -
H,.110
h

/ lavandulyl skeleton

_H~_H~
HOCH, H HO,C H
chrysantbemyl skeleton chrysanthem.yl alcohol (92) chrysantbemic acid (93)

OH

~ H
'</
/
~'X:"""""'fOH
....,...

santoUna alcohol yomogi alcohol

4+ ~
~';x' ..<;>-, /_:'X:" ~ /~'X:" ~ /
'V'" "0"+'," to.;'1 r "I '</

artemisia alcohol
.'</

artemisia ketone
(95) (94)
<b) artemisyl skeletons

Fig. 19.21 (a & b). Proposed biosynthesis of irregular monoterpenes (modified from Epstein and Poulter, 1973; used with permission of the copyright
owner, Elsevier Science Ltd., The Boulevard Langford Lane, Kidlington OXS 1GB, UK).
348 Monoterpenes

chrysanthemlc acid (93)

Fig. 19.22. Pyrethrins and related compounds.

It is of interest that in some nonenzymatic stodies, a small properties. These compounds also are highly toxic to fish.
percentage of head:to-head condensation products are ob- Pyrethrins interfere with the sodium channel, upset sodium
served. Chrysanthemyl alcohol (92) is an analog of presqua- and potassium balance, and cause rapid, recurrent excite-
lene alcohol and prephytoene (see Chapters 23 and 26), but ment of nerve endings followed by paralysis, Pyrethrins are
the fissions postolated to lead to the irregular monoterpenes rapidly decomposed by polysubstrate monooxygenase
have no counterparts for the higher classes (Charlwood and (PSMO) [mixed-function oxidase (MFO)] enzymes. Syner-
Charlwood, 1991a). gists are often used to slow detoxication and make the insec-
Label from (3R,4R)[4- 3 H]mevalonic' acid but not from ticidal preparations more effective. Pyrethrins account for
(3R,4S)[4-3 H]mevalonic acid was incorporated into chrysan- about one-third of world's insecticide use (Pickett, 1988).
themic acid (93). Chrysanthemyl alcohol is an intermediate A number of synthetic insecticides have been modeled
in the biosynthesis of chrysanthemic acid (Pattenden et al., on the structores of natorally occurring pyrethrins. Several
1975). of these are less biodegradable than pyrethrins and avoid
Artemisia ketone (94), derived from IPP and DMAPP in the problem of too rapid decomposition. In addition, these
Santolina chamaecyparissus, is asymmetrically labeled; that synthetic derivatives have better knockdown and toxicity
is, the label from IPP is incorporated at much greater levels properties.
than that from DMAPP (Allen et al., 1977). A dehydrogen-
ase system rather than a mixed-function oxidase (MFO) sys-
tem based on cytochrome P-450 appears to be involved in REFERENCES
formation of artemisia ketone.
In cell-free extracts of Artemisia annua and Santolina ACKERMAN, J. D., Specificity and mutual dependency of the or-
chamaecyparissus, chrysanthemyl alcohol and its pyrophos- chid-euglossine bee interaction, BioI. J. Linn. Soc., 20,
phate are incorporated into artemisia ketone and alcohol. 301-314 (1983).
Artemisia alcohol (95) is converted into artemisia ketone ADAMS, R. P., Numerical-chemosystematic studies of infraspecific

(94) and trans-chrysanthemic acid in the preparation from variation in Juniperus pinchotii, Biochem. Syst. Bcol., 3, 71-74
(1975).
S. chamaecyparissus (Banthorpe et al., 1977a), IPP and
ADAMS, R. P., Chemosystematics-Analysis of populational dif-
DMAPP are incorporated into irregular monoterpenes
ferentiation and variability of ancestral and modem Juniperus
whereas geranyl and neryl-OPP are not. An enzymatic sulf-
ashei, Ann. Mo. Bot. Gard., 64, 184-209 (1977).
hydryl group is involved.
ADAMS, R. P., Diurnal variation in the terpenoids of Juniperus
Pyrethrins (such as 96 and 97, Fig. 19.21 and 19.22) are
scopulorum (Cupressaceae)-Summer Versus Winter, Am. J.
of special interest because of their insecticidal properties Bot., 66, 986-988 (1979).
and ecomonic importance (Mabry and Gill, 1979). These
AnAMS, R. P. and A. HAGERMAN, A comparison of the volatile
compounds are isolated from the genus Chrysanthemum oils of mature versus young leaves of Juniperus scopulorum.
(sometimes segregated as species of Pyrethrum, or trans- Chemosystematic significance, Biochem. Syst. Ecol., 4, 75-79
ferred to the genus Tanacetum). Several species are culti- (1976).
vated and the flowers harvested and extracted with lipid sol- ADAMS, R. P. and A. HAGERMAN, Diurnal variation in the volatile
vents (such as petroleum ether) to yield a crude pyrethrin terpenoids of Juniperus scopulorum (Cupressaceae), Am. 1.
fraction. Bot., 64, 278-285 (1977).
In general, these compounds have relatively low mamma- ADAMS, R. P. and B. L. 'TURNER, Chemosystematic and numerical
lian toxicity but are highly toxic to insects. Pyrethrins are studies of natural populations of Juniperus ashe; Buch., Taxon,
especially useful because they have good "knockdown" 19,728-751 (1970).
 Monotelpenes 349

ALFARO, R. I., H. D. PIERCE, JR., J. H. BORDEN, and A. C. of Modern Methods in Plant Biochemistry (J. B. Harborne and
OEHLSCHLAGER, Insect feeding and oviposition deterrents from P. M. Dey, eds.), 43-98, Academic Press, London, 1991a.
western red cedar foliage, J. Chern. Ecol., 7, 39-48 (1981). CHARLWOOD, B. V. and K. A. CHARLWOOD, Terpenoid production
ALLEN, K. G., D. V. BANTHORPE, B. V. CHARLWOOD, and C. M. in plant cell cultures, in Ecological Chemistry and Biochemistry
VOLLER, Biosynthesis of artemisia ketone in higher plants. Phy- of Plant Terpenoids (J. B. Harborne and F. A. Tomas-Barberan,
tochemistry, 16, 79-83 (1977). eds.), Proceedings of Phytochemistry Society of Europe Vol.
ARNOLD, W. N., Absinthe, Sci. Am., 260, 112-117 (June 1989). 31,95-132, Oxford University Press, Oxford, 1991b.
ASPLUND, R. D., Monoterpenes: Relationship between structure and CHI1WOOD, D. J., Nematicidal compounds from plants, in Phyto-
inhibition of gennination, Phytochemistry, 7, 1995-1997 chemical Resources for Medicine and Agriculture (H. N. Nigg
(1968). and D. S. Seigler, eds.), 185-204, Plenum Press, New York,
ArrA-UR-RAHMAN and V. U. AHMAD, Monoterpenes and Sesquiter- 1992.
penes, Vol. I, I3C_NMR of Natural Products, Plenum Press, CORNFORTH, J. W., R. H. CORNFORTH, C. DONNINGER and G. Pop-
New York, 1992. JAK, Studies on the biosynthesis of cholesterol XIX. Steric
BANTHORPE, D. V., Classification of terpenoids and general proce- course of hydrogen elimination and of C-C bond formations
dures for their characterisation, in Terpenoids (B. V. Charlwood in squalene biosynthesis, Proc. Roy. Soc. (B), 163, 492-514
andD. V. Banthorpe, eds.), Vol. 7 of Modern Methods in Plant (1966).
Biochemistry 7 (1. B. Harborne and P. M. Dey, eds.), 1-41, CORI, 0., Enzymic aspects of the biosynthesis of monoterpenes in
Academic Press, London, 1991. plants, Phytochemistry, 20, 331-341 (1983).
BANTHORPE, D. V., O. EKUNDAYO, and M. G. ROWAN, Evidence COSCIA, C. J., 1'ERPBNOIDS, in Handbook of Chromatography, Vol.
for geraniol as an obligatory precursor of isothujone, Phyto- 1 (G. Zweig and J. Shenna, eds.), CRC Press, Boca Raton, FL,
chemistry,17, 1111-1114 (1978a). 1984.
BANTHORPB, D. V., B. M. MODAWI, I. PooTS, and M. G. ROWAN, CROMBffi, L., Natural products of cannabis and khat, Pure Appl.
Redox interconversions of geraniol and nerol in higher plants, Chern., 58, 693-700 (1986).
Phytochemistry,17, 1115-1118 (1978b). CROTEAU, R, Biosynthesis of monoterpenes. in Biosynthesis of
BANTHORPE, D. V., S. DOONAN, and J. A. GUTOWSKI, Biosynthesis Isoprenoid Compounds, Vol. 1 (J. W. Porter and S. L. Spurgeon,
of irregular monoterpenes in extracts from higher plants, Phyto- eds.), 225-282, Wiley, New York, 1981.
chemistry, 16, 85-92 (1977a). CROTEAU, R. Biosynthesis and catabolism of monoterpenes, in
BANTHORPB, D. V., J. MANN, and I. POOTS, 1,2 Hydrogen-shifts in Isopentenoids in Plants (W. D. Nes, G. Fuller, and L. Tsai, eds.),
the biosynthesis of the thujane skeleton, Phytochemistry, 16, 31-64, Marcel Dekker, Inc., New York, 1984.
547-550 (1977b). CROTEAU, R, Biosynthesis and catabolism of monoterpenoids,
BARTHOLOMEW,B., Bare zone between California shrub and grass- Chern. Rev., 87, 929-954 (1987).
land communities: The role of animals, Science, 170, CROTEAU, R, Catabolism of monoterpenes in essential oil plants,
1210-1212 (1970). in Flavors and Fragrances: A World Perspective (B. M. Law-
BEALE, M. H. and J. MAcMn.r.AN, The biosynthesis of C,-C20 rence, B. D. Mookherjee, and B. J. Willis, eds.), 65-84, Else-
terpenoid compounds, Nat. Prod. Rep., 5, 247-264 (1988). vier, Amsterdaro, 1988.
BEIER, R. C. and H. N. NIGG, Natural toxicants in foods, in Phyto- CROTEAU, R and M. A. JOHNSON, Biosynthesis of terpenoids in
chemical Resources for Medicine and Agriculture (H. N. Nigg glandular trichomes, in Biology and Chemistry of Plant Tri-
and D. S. Seigler, eds.), 247-367, Plenum Press, New York, chomes (E. Rodriguez, P. L. Healey and I. Mehta, eds.),
1992. 133-185, Plenum Press, New York, 1984.
BERNAYS, E. and M. GRAHAM, On the evolution of host specificity CROTEAU, R. and M. A. JOHNSON, Biosynthesis of terpenoid wood
in phytophagous arthropods, Ecology, 69, .886-892 (1988). extractives, in Biosynthesis and Biodegradation of Wood Com-
BOWERS, W. S., Phytochemical resources for plant protection, in ponents cr.
Higuchi, ed.), 379-439, Academic Press, Orlando,
Recent Advances in the Chemistry of Insect Control (N. F. FL,1985.
Janes, ed.), 272-292, Royal Society of Chemistry, London, CROTEAU, R. and F. KARP, Biosynthesis of monoterpenes: Partial
1985. purification and characterization of I.S-cineole syntetase from
BUTfBRY, R. G., D. R. BLACK, D. G. GUADAGNI, L. C. LrNG, G. Salvia ojJicinalis, ATch. Biochem. Biophys., 179, 257-265
CONNOLLY, and R. 'fERANIsm, California bay oil. I. Constitu- (1977).
ents, odor properties, 1. Agric. Food Chern., 22, 773-777 CROTEAU, R. and P. 1'uRKB:rr, Geranyl pyrophosphate synthase:
(1974). characterization of the enzyme and evidence that this chain-
CHAPMAN, R. F., E. A. BERNAYS, and S. J. SIMPSON, Attraction and length specific prenyltransferase is associated with monoterpene
repulstion of the aphid, Cavariella aegopodii by plant odors, 1. biosynthesis in sage (Sa/via officinalis), Arch. Biochem. Bio-
Chern. Ecol., 7, 881-888 (1981). phys., 271, 524-535 (1989).
CHARLWooD, B. V. and D. V. BANTHORPE, The biosynthesis of CROTEAU, R. and K. V. VBNKATACHALAM, Metabolism of monoter-
monoterpenes, in Progress in Phytochemistry, Vol. 5 (L. Rein- penes: Demonstration that (+ )-cis-isopulegone, not piperiten-
hold, J. B. Harborne, and T. Swain, eds.), 65-125, Pergaroon one is the key intennediate in the conversion of ( - )-isopiperi-
Press, Oxford, 1978. tenone to the (+ )-pulegone in peppermint (Mentha piperita),
CHARLWOOD, B. V. and K. A. CHARLWOOD, Monoterpenes, in Ter- Arch. Biochem. Biophys., 249, 306-315 (1986).
penoids (B. V. Charlwood and D. V. Banthorpe, eds.), Vol. 7 Cun.BR, H. G., Herbicidal compounds from higher plants, in Phyto-
350 Monoterpenes

chemical Resources for Medicine and Agriculture (H. N. Nigg Interactions with Secondary Plant Metabolites, Vol. I (G. A.
and D. S. Seigler, eds.), 205-226, Plenum Press, New York, Rosenthal and M. R. Berenbaum, eds.), 165-219, Academic
1992. Press, San Diego, CA, 1991.
DAHLGREN, R., A commentary on a diagrammatic presentation of GERSHBNZON, J., M. MAFFEI and R. CROTEAU, Biochemical and
the angiospenns in relation to the distribution of character states, histochemical localization of monoterpene biosynthesis in the
Plant Syst. Evol., Suppl. 1,253-283 (1977). glandular trithomes of spearmint (Mentha spicata), Plant Phys-
DEL MORAL, R. and C. H. MULLER, The allelopathic effects of iol., 89, 1351-1357 (1989).
Eucalyptus camaldulensis, Am. Midi. Nat., 83, 254-282 (1970). GmBs, R. D., Chemotaxonomy of Flowering Plants, McGiII-
DICKE, M., T. A. VAN BBEK, M. A. POSTHUMUS, N. BEN DOM, H. Queen's University Press, Montreal, 1974).
VAN BOKHoVEN, and A. DE GROOT, Isolation and identification HARBORNE, J. B., Introduction to Ecological Biochemistry, 2nd ed.,
of volatile kairomone that affects acarine predator-prey interac- Academic Press, New York, 1982.
tions, J. Chern. Ecol., 16, 381-396 (1990). HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod.
DODSON, C. H., R. L. DRESSLER, H. G. Hn.Ls, R. M. ADAMS, and Rep., 4, 323-344 (1986).
N. H. WILLIAMS, Biologically active compounds in orchid fra- HARBORNE, J. B., Chemical signals in the ecosystem, Ann. Bot.•
grances, Science, 164, 1243-1249 (1969). 60, (Suppl. 4), 39-57 (1987).
DUKE, 1. A., Natural medicines are natural pesticides. in Phyto- HARBoRNE,1. B., Recent advances in chemical ecology, Nat. Prod.
chemical Resources for Medicine and Agriculture (H. N. Nigg Rep., 7, 85-109 (1989).
and D. S. Seigler, eds.), 237-245, Plenum Press, New York, HARBORNE, J. B., Recent advances in the ecological chemistry of
1992. plant terpenoids, in Ecological Chemistry and Biochemistry of
EISNER, T., J. S. JOHNESSBE, J. CARVELL, L. B. HENDRY, and J. Plant Terpenoids (J. B. Harbome and F. A. Tomas-Barberlin,
MEINWALD, Defensive use by an insect of plant resin, Science, eds.), Proceedings of the Phytochemistry Society of Europe Vol.
184,996-999 (1974). 31),399-426, Oxford University Press, Oxford, 1991.
ELLts, B. E., Natural products from plant tissue culture, Nat. Prod. HAy, M. E., W. FBNICAL, and K. GUSTAFSON, Chemical defense
Rep., 5, 581-612 (1988). against diverse coral-reef herbivores, Ecology, 68, 1581-1591
EPSTEIN, W. W. and C. C. POULTER, A survey of some irregular (1987).
monoterpenes and their biogenetic analogies to presqualene al- HEDIN, P. A., F. G. MAxwm.r., and J. N. JENKINS, Insect plant
cohol, Phytochemistry,l2, 737-747 (1973). attractants, feeding stimulants, repellents, deterrents, and other
FEENY, P., L. ROSENBERRY, and M. CARTER, Chemical aspects of related factors affecting insect behavior, "in Proc. Summer Inst.
oviposition behavior in butterflies, in Herbivorous Insects (S. Bioi. Control of Plant Insects and Diseases (F. G. Maxwell and
Ahmad, ed.), 27-76, Academic Press, New York, 1983. F. A. Harris, eds.), 494-557, University of Mississippi Press,
Jackson, 1974.
FISCHER, N. H., The function of mono and sesquiterpenes as plant
gennination and growth regulators, in The Science of Allelopa- HEGNAUER, R., Chemotaxonomie der Pflanzen, Vols. 1-10, Birk-
thy (A. Putnam and C. Tang, eds.), 203-218, Wiley, New York, hauser Verlag, Basel, 1962-1992.
1986. HEoNAUER, R., Chemical evidence for the classification of some
FISCHER, N. H., Plant terpenoids as allelopathic agents, in Ecologi- plant taxa, in Perspectives in Phytochemistry (J. B. Harborne
cal Chemistry and Biochemistry of Plant Terpenoids (J. B. Har- and T. Swain, eds.), 121-138, Academic Press, London, 1969.
borne and F. A. Tomas-Barber..., eds.), Phytochemistry Society HEGNAUER, R., The Importance of Essential Oils in Plant Classifi-
of Europe Vol. 31, 377-398, Oxford University Press, Oxford, cation, Dragoco Rept., No. 10, 1978, pp. 203-230.
1990. HEROUT, V., Some relations between plants, insects, and their iso-
FISCHBR, N. H., N. TANRlSEVBR, and G. B. Wn.LIAMSON, Allelopa- prenoids, in Progress in Phytochemistry, Vol. 2 (L. Reinhold
thy in the Florida scrub community as a model for natural herbi- and Y. Liwschitz, eds.), 143-202, Interscience, London, 1970.
cide actions, in Biologically Active Natural Products: Potential HIKINo, H., Recent research on oriental medicinal plants, in Eco-
Use in Agriculture (H. G. Cutler, ed.), ACS Symposium Series nomic and Medicinal Plant Research, Vol. I (H. Wagner, H.
380, 232-249, American Chemical Society, Washington, DC, Hikino, and N. R. Farnsworth, eds.), 53-85, Academic Press,
1988. London, 1985.
GABLER, A., W. BOLAND, U.l'Rmss, and H. SIMON, Stereochemical Hn.Ls, H. G., Fragrance cycling in Stanhopea pulla (Orchidaceae,
studies on homoterpene biosynthesis in higher plants; mechanis- Stanhopeinae) and identification of trans-limonene oxide as a
tic, phylogenetic, and ecological aspects, Helv. Chim. Acta., major fragrance component, LindIeyana, 4, 61-67 (1989).
74, 1773-1789 (1991). Hn.Ls, H. G. and B. SCHUTZMAN, Considerations for sampling floral
GEISSMAN, T. A. and D. H. G. CROUT, The Organic Chemistry of fragrances, Phytochem. Bull., 22, 2-9 (1990).
Secondary Plant Metabolism, Freeman Cooper, San Francisco, Hn.LS, H. G., N. H. Wn.LlAMS, and C. H. DODSON, Floral fragrances
1969. and isolating mechanisms in the genus Catasetum (Or-
GERSHENZON, J. and R. CROTEAU, Regulation of monoterpene bio- chidaceae), Biotropica, 4, 61-76 (1972).
synthesis in higher plants, in Biochemistry of the Mevalonic IRVING, R. S. and R. P. ADAMS, Genetic and biosynthetic relation~
Acid Pathway to Terpenoids (G. H. N. Towers and H. A. Staf- ships ofmonoterpenes, in Terpenoids: Structure, Biogenesis and
ford,eds.), Recent Advances Phytochemistry, Vol. 24), 99-160, Distribution (V. C. Runeckles and T. J. Mabry, eds.), Recent
Plenum Press, New York, 1990. Advances in Phytochemistry Vol. 6, 187-214, Academic Press,
GERSHENZON, J. and R. CROTEAU, Terpenoids, in Herbivores: Their New York, 1973.
 MonOferpenes 351

JACOBSON, M., Plants, insects, and man-Their interrelationships, PAUL, V. J., M. E. HAY, J. E. DUFFY, W. FENICAL, and K. Gus-
Econ. Bot., 36,346-354 (1982). TAVSON, Chemical defense in the seaweed Ochtodes secundira-
JERMY, T., Evolution of insect/host plant relationships, Am. Nat., mea (Montagne) Howe (Rhodophyta): Effects of its monoter-
124,609-630 (1984). penoid components upon diverse coral-reef herbivores, J. Exp.
Mar. BioI. Ecol., 114, 249-260 (1987).
JOHNSON, M. A. and R. CROTEAU, Biochemistry of conifer resis-
tance to bark beetles and their fungal symbionts, in Ecology PICKETT, J. A., Chemical pest control-The new philosophy,
and Metabolism of Plant Lipids (G. Fuller and W. D. Nes, eds.), Chern. Br., 137-142 (1988).
ACS Symposium Series 325, 76-92, American Chemical Soci- PICKEIT, J. A., Lower terpenoids as natural insect control agents,
ety, Washington, DC, 1987. in Ecological Chemistry and Biochemistry of Plant Terpenoids
KAJIMA, M. and M. PIRAUX, The biogenesis of crumabinoid in Can- (J. B. Harborne and F. A. Tomas-Barbenin, eds.), Proceedings
nabis sativa; Phytochemistry, 21, 67-69 (1982). of The Phytochemistry Society of Europe Vol. 31, 297-3 I3,
Oxford University Press, Oxford, 1991.
KARP, F., J. L. HARRIS, and R. CROTEAU, Metabolism of monoter-
penes: demonstration of the hydroxylation of ( + )-sabiene to POULTER, C. D., Biosynthesis of non-head-to-tail terpenes. Forma-
( + )-cis-sabinol by an enzyme preparation from sage (Salvia tion of 1'-1 and 1'-3 linkages, Ace. Chern. Res., 23, 70-77
officinalis) leaves, Arch. Biochem. Biophys., 256, 179-193 (1990).
(1987). POULTER, c. D. and H. C. RILLING, The prenyl transfer reaction.
KARP, F., C. A. MIHALIAK, J. L. HARRIS, and R. CROTEAU, Monoter- Enzymatic and mechanistic studies of the l'A coupling reaction
pene biosynthesis: Specificity of the hydroxylations of ( - )- in the terpene biosynthetic pathway, Acc. Chern. Res., 11,
limonene by enzyme preparations from peppennint (Mentha 307-313 (1978).
piperita), spearmint (Mentha spicata), and perilla (Perilla POULTER, C. D., I. C. ARGYLE, and E. A. MASH, Famesyl pyrophos-
Jrutescens) leaves, Arch. Biochem. Biophys., 276, 219-226 phate synthetase, J. BioI. Chern., 253, 7227-7233 (1978).
(1990). POUL1'ER, c. D., P. L. WIGGINS, and A. T. LE, Famesylpyrophos-
KOGAN, M., Plant resistance in pest management, in Introduction phate synthetase. A stepwise mechanism for the 1'-4 condensa-
to Insect Pest Management (R. L. Metcalf and W. H. Luckman, tion reaction, I. Am. Chern. Soc., 103, 3926-3927 (1981).
eds.), 103-146, Wiley, New York, 1975. RODlUGUEZ, E., Cytotoxic and insecticidal chemicals of desert
LAWRENCE, L., R. BARTSCHOT, E. ZAVARIN, andJ. R. GRIFFIN, Natu- plants, in Plant Resistance to Insects (P. A. Hedin, ed.), ACS
ral hybridization of Cupressus sargentii and C. macnabiana and Symposium Series 208, 291-302, American Chemical Society,
the composition of the derived essential oils, Biochem. Syst. Washington, DC, 1983.
Ecol., 2, 113-119 (1975).
RODRiGUEZ, E., P. L. HEALEY and I. MEHTA (eds.), Biology and
LEVIN, D., The role oftrichomes in plant defense, Quart. Rev. Biol., Chemistry of Plant Trichomes, Plenum Press, New York, 1984.
48,3-15 (1973). Ross, J. D. and C. SOMBRERO, Environmental control of essential
LOOMIS, W. D. and R. CROTEAU, Biochemistry of terpenoids, in oil production in Mediterranean plants, in Ecological Chemistry
Lipids, Structure and Function (P. K. Stumpf, ed.), Vol. 4 of and Biochemistry of Plant Terpenoids (1. B. Harborne and F.
The Biochemistry of Plants (P. K. Stumpf and E. E. Conn, eds.), A. Tornas-Barbenin, eds.), Proceedings of the PhytOChemistry
363-418, Academic Press, New York, 1980. Society of Europe Vol. 31, 83-94, Oxford University Press,
MABRY, T. J. and J. E. GILL, Sesquiterpene lactones and other Oxford, 1991.
terpenoids, in Herbivores (G. A. Rosenthal and D. H. Janzen, RUZICKA, L., A. ESCHENMOSER, and H. HEUSSER, The isoprene rule
eds.), 502-537, Academic Press, New York, 1979. and the biogenesis of terpenic compounds, Experientia, 9,
MACGREGOR, J. T., L. L. LAYTON, and R. G. BUTIERY, California 357-396 (1953).
bay oil. II. Biological effects of constituents, 1. Agric. Food SCORA, R. W., J. W. CAMERON, and J. A. BERG, Rind and leaf oils
Chern., 22, 777-780 (1974). of tetraploid interspecific Citrus hybrids and their diploid and
MANNERS, G. D. and L. JURD, The hydroquinone terpenoids of tetraploid parents, Taxon, 19, 752-761 (1970).
Cordia alliodoro, J. Chern. Soc., Perkin I, 405-410 (1977). SEIGLER, D. S., Plant systematics and alkaloids, in The Alkaloids,
MASH, E. A., G. M. GURRIA, and C. D. POULTER, Farnesylpyrophos- Vol. 16 (R. H. F. Manske, ed.), 1-82, Academic Press, New
phate synthetase. Evidence for a rigid geranyl cation-pyrophos- York, 1977.
phate anion pair, J. Am. Chern. Soc., 103, 3927-3929 (1981). SEIGLER, D. S., Role of lipids in plant resistance to insects, in Plant
MrHALlAK, c. A., J. GERSHENZON, and R. CROTEAU, Lack of rapid Resistance to Insects (P. A. Hedin, ed.), ACS Symposium Series
monoterpene turnover in rooted plants: Implications for theories 208, 303-328, American Chemical Society, Washington, DC,
of plant chemical defense, Oecologia, 87, 372-376 (1991). 1983.
MULLER. c. H. and C. CHOU, Phytotoxins: An ecologiCal phase of SHIBAMOTO, T., Analysis of essential oils in Application of Glass
phytochemistry, in Phytochemical Ecology (I. B. Harborne, Capillary GC, (W. G. Jennings, ed.) (Chromatography Science
ed.), 201-216, Academic Press, London, 1972. Series 15),455-509, Marcel Dekker, Inc., New York, 1981.
NAHRSTEDT, A., Strukturelle Beziehungen zwischen planzlichen STADLER, E., Attractants, arrestants, feeding, and oviposition stimu-
und tierischen Sekundiirstoffe, Planta Medica, 44, 2-14 (1982). lants in insect-plant relationships: Applications for pest control,
PATTENDEN, G., C. R. POPPLESTONE, and R. STORER, Investigation in Natural Products for Innovative Pest Management (D. L.
of the role of chrysanthemyl, lavandulyl, and artimisyl alcohols Whitehead and W. S. Bowers, eds.), 243-258, Pergamon Press,
in the biosynthesis of chrysamthernic acid. J. Chern. Soc., Chern. Oxford, 1983.
Commun., 290-291 (1975). T AKABA YASHI, J., M. DICKE, and M. A. POSTHUMUS, Induction of
352 Monotelpenes

indirect defence against spider-mites in uninfested lima bean studies of North American conifers, Biochem. Syst. Ecol., 2,
leaves, Phytochemistry, 30, 1459-1462 (1991). 131-167 (1975).
THORNE, R. P., A phylogenetic classification of the Angiospennae, WEIDENEAMER, J. D., F. A. MACIAS, N. H. FISCHER, aod G. B.
Evol. BioI., 9, 35-106 (1976). WILLIAMSON, lust how insoluble are monoterpenes? I. Chern.
VANDBRWEL, D. and A. C. OBHLSCHLAGER, Biosynthesis of phero- Ecol.,19, 1799-1807 (1993).
mones and endocrine regulation of pheromone production in WHIITEN, W. M., H. G. Hn.Ls, aod N. H. WILLIAMS, Occurrence of
Coleoptera, in Pheromone Biochemistry (G. D. Prestwich and ipsdienol in floral fragraoces, Phytochemistry, 27, 2759-2760
G. J. Blomquist, eds.), 175-215, Academic Press, Orlaodo, FL, (1988).
1987. WILLIAMS, N. H. and C. H. DoDSON, Selective attraction of male
VAUGHN, S. F. aod G. F. SPENCER, Volatile monoterpenes inhibit euglossine bees to orchid floral fragrances and its importance
potato sprouting, Am. Potato J., 68, 821-831 (1991). in long distaoce pollen flow, Evolution, 26, 84-95 (1972).
VAUGHN, S. P. and O. P. SPENCER, Volatile monoterpenes as poten- WILLIAMS, N. H. aod W. M. WHIITEN, Orchid floral fragrances
tial parent structures for new herbicides, Weed Sci., 41, and male euglossine bees: Methods and advances in the last
114-119 (1993a). sesquidecade, BioI. Bull., 164, 355-395 (1983).
VAUGHN, S. F. and O. F. SPENCER, Naturally-occurring aromatic WILLIAMSON, G. B., N. H. FISCHER, D. R. RICHARDSON, and A.
compounds inhibit potato tuber sprouting, Am. Potato J., 70, DE LA PEN-A, Chemical inhibition of fire-prone grasses by fire-
527-533 (1993b). sensitive shrub, Conradina canescens, I. Chern. Ecol., 15,
VERMA, M. and C. E. MELOAN, A natural cockroach repellent in 1567-1577 (1989).
bay leaves, Am. Lab., 13, 64-69 (1981). WOOD, D. L., The role of pheromones. kairomones, and allomones
VOGEL, S., Ecophysiology of zoophilic pollination, in Physiological in the host selection and colonization behavior of bark beetles,
Plaot Ecology III (0. L. Laoge, P. S. Nobel, C. B. Osmond, Aonu. Rev. Entomol., 27, 411-446 (1982).
and H. Ziegler, eds.), 559-624, Springer-Verlag, Berlin, 1983. WRIGHr, R. H., Primary odors aod insect attraction, Cao. Entomol.,
VON RUDLOFF, E., Volatile leaf oil analysis in chemosystematic 98, 1083-1093 (1966).
20
Iridoid Monoterpenes

Introduction but many fail to produce the same complement of com-

Biosynthesis pounds found in the parental plants (Ellis, 1988). However,
Fonnation of Deoxyloganic Acid from Geraniol various cultures from Gardenia jasminoides (Rubiaceae)
Fonnation of 8-epi-Deoxyloganin and Related Iridoids and Valeriana and related genera (Valerianaceae) produce
Iridoids Derived from 10-Hydroxycitronellol a similar complement of iridoid monoterpenes to the parent
Conversion of Loganin to Secologanin plants. Cultures of Fedia cornucopiae and Valerianella 10-
Secologanin as a Precursor for Indole Alkaloids custa (both Valerianaceae) produce greater amounts of iri-
Distribution of Iridoid Monoterpenes doids than do the roots of the parental plants. Transfonned
Biological Activity hairy root cultures of Valerianella discoidea produce essen-
Medicinal Uses of Iridoid Compounds tially the same valpotriates as the original plant (Charlwood
References and Charlwood, 1991). Indole alkaloids are produced in a
number of plant cell cultures, indicating that iridoid monot-
erpene biosynthesis must be occurring (Ellis, 1988).
II'ffKODUCTIOI'I Iridoid monoterpenes can be purified by chromatography
on a variety of substrates; general chromatographic tech-
At least 500 compounds (-300 iridoid glucosides, 100 sec- niques have been reviewed (Bowers, 1991; Coscia, 1984;
oiridoid glucosides, and 100 nonglycosidic iridoids) (Bow-
Inouye, 1991; Junior, 1990). Thin-layer chromatography
ers, 1991; Croteau, 1981; Inouye, 1991; Inouye and Uesato,
(TI..C) (Bowers, 1991; Inouye, 1991; Junior, 1990; van der
1986), derived from a specialized monoterpenoid pathway
Sluis, 1985) and high-perfonnance liquid chromatography
and usually characterized by a 5-membered carbocyclic ring
(HPLC) (Foerster et aI., 1984; Inouye, 1991; Junior, 1990)
fused to a 6-membered heterocyclic ring containing an acetal
have proven especially useful for the separation and purifica-
linkage, are known (Fig, 20.1). Although the tenn iridoid is
tion of iridoid compounds. Other physical properties and
based on iridomynnecin (1) and iridodial (2), two monoter-
spectral data, such as the optical rotation, melting point, ul-
penes that occur in the defensive secretions of ants of the
traviolet, 'H- and l3C-spectral measurements are valuable
genus lridomyrmex, most compounds of "iridoid" structure
for detennination of structure (Boros and Stennitz, 1990;
are found in plants. These compounds are frequently in-
volved in plant-insect interactions and are sequestered, El-Naggar and Beal, 1980). A number of known compounds
sometimes after modification, and apparently synthesized have been studied by 13C_NMR (nuclear magnetic reso-
independently by some insects (Rimpler, 1991). In contrast nance) spectroscopy; the characteristic shifts observed are
to other monoterpenes, iridoid monoterpenes in plants are useful in detennination of substitution patterns and stereo-
usually not associated with specialized oil glands or other chemistry (Boros and Stennitz, 1990; Chaudhury et aI.,
such structures, and occur in all plant parts, typically as 1980; Damtogt et aI., 1981). Mass-spectral measurements
water-soluble glycosides, sometimes comprising several also are valuable for characterization of iridoid monoter-
percent of the dry weight of the plant (Bowers, 1991; Ster- penes (Junior, 1990). The plant sources of many iridoid mo-
mitz, 1988). In some instances, up to 22-25% of plant mate- noterpenes also are indicated in these references.
rials may consist of iridoid compounds. Monoterpene and indole alkaloids are frequently derived
Some suspension cultures produce iridoid monoterpenes, from iridoid monoterpene precursors (Inouye, 1991).

353
 q:
354 Iridoid Monoterpenes

CHO

H CHO ~

iridodial (2) aetinidine (45) iridomyrmecin (I)

1R
OH lIC0 2CH3

oo~
OH

oct) ~
~
~ 6 4~ 3
HO 7 H O-glucosyl
~ 0 • 1 0 -
O-glucosyl 10 H O-glu_yl CH,O,C :::,... 0
HO H O-glucosyl

unedoslde aucubio (19) loganio (6)

w<t
the major alkal0?Jld from
Valeriana o.fjicinalis

H j
~ I~
~~ ~
CHO

g)'CH'
o H CHO
o I
nepetalaetone (29) dolicbodial (27) Il....ytanthine dolicbolaetone (28) :::,...
(a) OH

Fig. 28.1 (a & b). Some naturally occuning iridoid compounds.

BIOSYNTHESIS at positions 3 and 11 is involved and the product is epi-deoxylo-

ganic acid (20), in a manner paralleling the first route ofbiosyn-
The biosynthesis of iridoid monoterpenes is complex and thesis. In other examples, the pathway is identical up to epi-
much work was required to establish the precursors and path- iridotrial (8), where glucosylation takes place, followed by oxi-
dation of C-1I to the carboxyl stage (Jensen, 1991).
ways leading to iridoid monoterpenes. Label from mevalonic
acid, in contrast to other monoterpenes, is incorporated into Route 3 begins with lO-hydroxycitronellol (9), but also involves
the intermediacy of iridodial (2) (Fig. 20.4) (Jensen, 1991).
the entire molecule. Incorporation stodies in this group of
compounds often are complicated by the presence of side
branches to the main pathways that allow incorporation of Thus, contrary to earlier expectations, various routes lead-
nonobligatory precursors (Inouye, 1991). In most situations, ing to the iridane skeleton in the biosynthesis of iridoids and
dialdehydes may exist in equilibrium with the corresponding secoiridoids are not identical (Inouye and Uesato, 1986).
dihydropyran forms. These dihydropyran forms are impor-
tant for the formation of many of the known glycosides. Formation of Deoxyloganic: Acid
Although glycosides at other positions also are known, the li'om Geraniol
glycosides of dihydropyran forms usually are !3-D-glucopyr-
Iridodial (2) has been established to be a major intermedi-
anosides (Jensen, 1991).
ate in the biosynthesis of indole alkaloids in Rauvolfia ser-
There seem to be at least three routes to iridoid com-
pounds (Bowers, 1991; Jensen, 1991):
pentina (Inouye and Uesato, 1986). Subsequently, the con-
version of lO-oxogeranial (10) to iridodial (2) was
demonstrated. Geraniol (11), lO-hydroxygeraniol (3), and
Route 1 derives from lO-hydroxygeraniol (3), via iridodial (2)
and iridotrial (4), and culminates in deoxyloganic acid (5) (Fig. iridodial (2) (and, in some instances, nerol and lO-hydrox-
20.2). Scrambling of label between positions 3 and II occurs. ynerol) are efficient precursors of deoxyloganic acid (5) and
Deoxyloganic acid is converted to loganin (6) and, fmally, to loganin (6) (Fig. 20.2) (Balsevich, 1985; Balsevich and
secoiridoids (Jensen, 1991). Kurz, 1983; Inouye, 1991; Jensen, 1991). In the formation
Route 2 also involves lO-hydroxygeraniol (3) as a major precur- of loganin in Catharanthus roseus, the first intermediate
sor, but proceeds via the intermediacy of epi-iridodial (7) and seems to be 10-hydroxygeraniol (3); 9-hydroxygeraniol is
epi-iridotrial (8) (Fig. 20.3). In some plants, scrambling oflabel not incorporated well. ColO hydroxylation of geraniol (and
 too
Iridoid Monoterpenes 3SS

= cop, o 0

W COlCH.

0;R~0~.:." ;D.:.
?' "'"
R, H 0
Rz O-glucosyl
o
foliamenthin ~ secoJoganin (18) sweroside R=CHzOH, R=OH, monotropein
R=OH, R=CHzOH, gardenoside (26)
I H O-g1ucosy)
HO~HO : O-CO
iH"",
HO~O~CH'
~ ~ 0

~b o CH,sCOO H O-g1ucosy)

O-g1u..sy) IH O-g1ucosy)
oleuropln hydrangenoside A paederoside (48)

O·g1ucoay)

ipecoside
O-g1ucosy)

(b) vincoside

Fig. 20.1. (continued)

nerol) is accomplished by a cytochrome P-450-mediated In certain other iridoid monoterpenes, positions 3 and 1 I
system. do not become equivalent. In Deutzia crenata (Saxifraga·
Although the exact mechanism of ring closure is not ceae), lO-hydroxygeraniol (3), iridodial (2), and its glucoside
known, that proposed by Escher et aI. (1970) and Battersby also are converted to deutzioside (13) (Fig. 20.2). When
et aI. (1970) is reasonable in view of other studies (Fig. iridodial glucoside was used as the precursor, no scrambling
20.5). In formation of the cyclopentane ring ofioganin, posi- of label was observed (Jensen, 1991). In old plants of Ver-
tions C-9 and C-lO of geraniol (C-3 and C-11 of loganin) bena officinalis, introduction of 2- 14C-Iabeled mevalonic
become equivalent (Beale and MacMillan, 1988). acid resulted in the formation of iridoid compounds and the
The conversion of 10-oxogeraniai (10) (and lO-oxoneral) production of verbenalin (comin) (14), in which no scram-
to iridodial (2) has been demonstrated; a novel monoterpene bling was observed (Fig. 20.6). Presumably, the pathway
cyclase from Rauvolfia serpentina that catalyzes formation involved iridodial glucoside (12) and iridotrial glucoside
of iridodial has been partially purified and characterized (15) (Fig. 20.2) (Jensen, 1991).
(Uesato et aI., 1987). NADPH accelerates the cyclization. Both [1-3H]IO-hydroxygeraniol (3) and [1O-3H]iridodiai
The molecular weight of the cyclase and its subunit are (2) are incorporated into vindoline (17) in Catharanthus ro-
118,000 and 28,700, respectively (Uesato et aI., 1987). seus and secologanin (18) in Lonicera morrowii (Caprifolia-
Scrambling of radioactive label between positions 3 and ceae), whereas (R,S)-[1O-3H]hydroxycitroneIIol (9) was not.
11 in deoxyloganic acid (5) indicates that iridotrial (4) [not Thus, the cyclization pathway involving conversion of 10-
iridodial glucoside (12)] is an intermediate in the formation oxogeranial (10) (and lO-oxoneral) is common, not only in
of deoxyloganic acid (5) (Fig. 20.3) (Jensen, 1991). plants producing iridoid glycosides but also in plants produc-
356 lridoid Monole/penes

~
~HO
---+ ~+~
HO eHO
eHO

0
lO HH

H
"'"

0
O-glucosyl
geraniol (11) 10-hydroxycitronellol (9) deutzioside (13)

~~ ~
~
HO
OH
----+
~!
~
#
eHO
eHO
----+
<;=CeHO

eHO
--..
yO"'" <;Q"'"
H
O--+-
H
eHO

O-glucosyl O-glucosyl

10-hydroxygeraniol (3)10-oxogeranial (10) iridodial (2) iridodial glucoside (12) iridotrial glucoside (15)

~ 1 11
l
,
OH eHO eHO
CHO _ ?-glucosyI

~ ~ eHo
~
HO
OH----..
9' eHO
-----.
~eHo ~
eHO
10 HO
~'_<=pf
~
H
• #
C0 2 H
0

iridotrial (4) deoxyloganic acid (5)

Fig. 20.2. Biosynthesis of deoxyloganic acid (Jensen, 1991; modified and used with pennission of the copyright owner, Oxford University Press,
Oxford).

~~q:~ro-q5
HO ~ ! HO ! HO
10-hydroxygeraniol (3) 8-ep;-iridodial (7) 8-epi-iridotrial (8)

OH

~
~

\ 0 -

HO H O-glucosyl H
O-glucosyl
geniposidic acid (37) deoxygeniposidic acid aucubin (19) epi-deoxyloganic acid (20)

WOH
OH >=pH
CO,eH,
Plantago major
Scrophularia racemOSQ

, ~ ~ Ho_~H'
o ;
i H
o
.
° :>r+-r~
~ HO! H
O-glucosyJ ! O-gJucosyl O-glucosyl
antirrhinoside (23) ipolamiide (21) lamiide (22)
Anti"hinum majus Hebenstretia dentala Hebenstretia dentola

Fig. 20.3. Biosynthesis of epi-deoxyloganic acid (Jensen, 1991; modified and used with pennission of the copyright owner, Oxford University Press,
Oxford).
 Iridoid Monoterpenes 357

~0¢if-~

HO
OH?
~

HO
OH

---+- H C~
CHO

H CHO
CHO

l()..hydroxygeraniol (3) l()"hydroxycltrolellol (9) irldodlal (2) dollehodlal (27)

WC;_~O~~H
0:0
H
H
0
OH
H
0
0

~~ ir°"-~~'
~0~H3
teuerein (30) +
10 0
nepetalactone (29)

y¢~
photocltralA dolleholaetone (28)

Fig. 10.4. Biosynthesis of nonglycosidic iridoid monoterpenes derived from lO-hydroxycitronellol (Inouye and Uesato, 1986; Jensen, 1991; modified
and used with pennission of the copyright owner, Springer-Verlag, Vienna).

OH_~:O_~CHO__
~ ~
I7 geraniol (11) H+
9' OH

10
10-oxogeraniai (10) iridodlaI(2)

OHC
~~HO---
H

""" OH
HO

O-glucosyl
HO
-yp H
CO'CH3

"""
0=
HO'" 7

O-glucosyl
H
w~
,
l! ~.gl:COSYI
'.-6'3
4
COCH
11 2 3

IridotrlaI(4) )oganin (6)

Fig. 20.5. Proposed cyclization of iridoid monoterpenes (modified from Escher et aI., 1970; used with pennission of the copyright owner, the Royal
Society of Chemistry, London).
 t;:,. t
358 lridoid Monoterpenes

•
,t~
•
IOH
-+
OH_
CHO
CHO
10 --+
I'
3
9' CHO
, 1

q:'
OH
, 10 CHO
geraniol (11) 10-hydroxygeraniol (3) 10-oxogeranial (10)

10 10 • O-glucosyl 10 • O-glucosyl

~-
CHO Ho.".. q;l,
';,I,: 0
v;;y, s..H
/tCH: ; -
H 7 U J
H CHO - -
•
10 CHO 11 C02H U C02CHJ
iridodial(2) iridotrial(4) deoxyloganic acid (5) loganin (6)

¢6' O-g1ucosyl

secologanin (18) vindoline (17) verbenaJln (14)

(cornin)

Fig. 20.6. Biosynthesis of secologanin and the indole alkaloid vindoline (modified from Inouye and Uesato. 1986; used with pennission of the
copyright owner, Springer~Verlag, Vienna).

ing secoiridoid glucosides and indole alkaloids (Fig. 20.6) deoxy10ganin was not a precursor (Fig. 20.7) (Inouye and
(Inouye, 1991; Inouye and Uesato, 1986; Uesato et al., Uesato, 1986; Jensen, 1991).
1986).
lridoids Derived from lO.Hydroxydtronellol
Formation of 8.epJ.Deoxyloganin and
Related lridoids The biosynthesis of some nonglycosidic iridoids such as
dolichodial (27), dolicholactone (28), and nepetaIactone (29)
A second series of iridoid monoterpenes is derived from involve pathways somewhat different from those of glyco-
the epimeric series of precursors (Figs. 20.3 and 20.7). Intro- sidic iridoids and secoiridoids (Inouye and Uesato, 1986;
duction of [9- 13C]-IO-hydroxygeraniol (3) in Plantago Jensen, 1991). (S)-( - )-Citronellol and (S)-( - )-IO-hydrox-
major gives incorporation into aucubin (19) with scrambling ycitronellol (9), but not lO-hydroxygeraniol (3) nor other
of label. Furthermore, 8-epi-iridodial (7) and 8-epi-iridotrial 2,3-unsaturated analogs, can serve as intermediates for doli-
(8) and the corresponding glucosides were all incorporated chodial (27), dolicholactone (28), and teucrein (30) in Teu-
with scrambling of the label at C-1I (Fig. 20.7) (Jensen, crium rnarum (Lamiaceae), and for nepetaIactone (29) and
1991). Because both glycosides are incorporated with scram- dihydronepetaIactone (31) in Nepeta cataria (Lamiaceae)
bling of label, hydrolysis must occur prior to incorporation. (Fig. 20.4) (Inouye and Uesato, 1986).
In addition, 8-epi-deoxyloganic acid (20) is a precursor for
ipolamiide (21) and lamiide (22) in Rebenstretia dentata Conversion of Loganin to Secologanin
(Globulariaceae), for aucubin (19) in Scrophularia racemosa
(Scrophulariaceae), and for antirrhinoside (23) in Antirrhi- Loganin (6) is a key intermediate in the biosynthetic path-
num rnajus (Scrophulariaceae) (Fig. 20.3) (Inouye and way leading to other iridoid monoterpenes as well as com-
Uesato, 1986). plex indole alkaloids. Many of these additional compounds
Gardenia jasminoides (Rubiaceae) cell cultures incorpo- are derived by conversion of this bicyclic iridoid monoter-
rated epi-iridotrial (8) and 7,8-dehydroiridotrial glucosides. pene into secologanin (18). The mechanism by which ring
8-epi-Iridodial (7) and boschnaloside (24) are readily incor- cleavage occurs to yield secologanin is not well understood,
porated into tarennoside (25) and gardenoside (26), but epi- but apparently the cleavage happens after oxidation of the
 /ridoid Monoterpenes 359

..
~
,;::--

HO
OH
-- «-=~ro --
I ! HO

!
lO·hydroxygeraniol (3) epi-iridodial (7)
'CHO CHO

\P --~ q)
CHO

__ 5Q
~
~.
0

O·glucosyl
HO H 01llucooyl
HO~I~COOYI s O-glucosyl

~
A
tarrenoslde (25) boscbnaloslde (24)

H
,

HO~6
HO'" O·glucooyl

gardenoside (26)

Fig. 20.7. Iridoid monoterpenes derived from 8-epi-deoxyloganin (Jensen, 1991; modified and used with pennission of the copyright owner, Oxford
University Press. Oxford),

methyl group and fonnation of a phosphate ester (Fig. 20.8) SecologllDin as a Frecursor for Indole
(Inouye, 1991). Alkaloids
Some evidence supports the idea that distinct biosynthetic
Secologanin undergoes condensation with tryptamine in
pathways lead to iridoid g1ycosides and to secoiridoids.
some groups of plants to yield strictosidine (isovincoside)
Equal labeling of positions C-3 and C-lI is observed in
(36), the immediate precursor of indole alkaloids.
secoiridoids and in indole alkaloids.
Secologanin (18) is, in torn, the parent of a group of
compounds called secoiridoids of which gentiopicroside
(32) and sweroside (33) are representative members. These DIS1RIBUI'IOI'l OF IRIDom JIIOI'lOTERPEl'IES
compounds arise in the sequence sweroside, swertiamarin
(34), and gentiopicroside (32) (Fig. 20.8) (Inouye and Although not all steps of the biosynthesis of iridoid monoter-
Vesato, 1986). When [4R-4- 3HJ-, [4S-4- 3HJ-, or [2S-2-3H] penes have been examined in detail, the probable pathways
mevalonic acid (MYA) together with [2_14C]MYA were ad- leading to this group of compounds appear to be lengthy
ministered to Swertia caroliniensis (Gentianaceae) and the and complicated. Thus, their distribution should be limited
products loganic acid (35) and gentiopicroside (32) were and it seems improbable that iridoid monoterpenes would
studied by degradation, it was found that both C-4 pro-R have arisen often in the course of evolution. The distribution
hydrogens of two molecules of MYA were retained in 10- of iridoid mouoterpenes among higher plants is restricted
ganic acid, whereas only one remained in gentiopicroside to about 50 families, most of which belong to Wettstein's
(Fig. 20.8); neither of the two C-4 pro-S hydrogens was Tubiflorae (Gershenzon and Mabry, 1983). The families that
present in either compound. Both of the C-2 pro-R hydro- possess these compounds belong primarily to two major
gens of MYA were found at C-3 and C-7 in loganic acid groups as viewed by most phylogenists. Dahlgren observed
(35); one pro-S hydrogen was at C-3, whereas the other was that the presence of iridoid compounds is strongly correlated
essentially eliminated from the hydroxylated C-7 position. with the presence of certain significant morphological fea-
Gentiopicroside (32) from [2S-3H2 , 2_14C]MYA incorpora- tures (Dahlgren et al., 1981). The placement of a number of
tion had the same 'H! 14C ratio as loganic acid (Inouye and families in these groups and removal of others has been
Vesato, 1986). Randomization of label from [2- 14CJMVA discussed by Dahlgren and reviewed concisely by Gershen-
between C-3 and C-lI usually was observed in iridoid and zon and Mabry (1983). A number of families such as the
secoiridoid g1ycosides and in indole alkaloids (Inouye and Escalloniaceae, Grossulariaceae, Hydrangeaceae, and Saxi-
Vesato, 1986). fragaceae were thought to be closely related to the Cornaceae
360 lridoid Monoterpenes

HO
.A.. _
J 1 ~OlH
HO,'
W •

10
8

H
ll
C:CH3

1
",'

0..........

O·glucosyl
H_ 0
V
5Q~01CH3
H
"":::

0 ----.

O.glucosyl
CI(r0':?

H3COOC~
H

0
O·glucosyl
----...

mevalonic acid :J

Q&
loganin (6) secologanin (18)
10

~O_gIUCOSYI OH H

1f.·-
o !~ 0 O.glUCOSYI,

~~~
o o
o r
HO::r:v

oo-g5
gentioplcroslde (32) swertiamarln (34) sweroside (33) secologanic acid

O-glucosyl
O-glucosyl

loganic acid (35)

Fig. 20.8. Proposed formation of secologanin and related compounds (modified from Inouye and Uesato, 1986; modified and used with pennission of
the copyright owner, Springer-Verlag, Vienna).

(Bate-Smith et al., 1975), whereas there does not appear to mation, the presence of secoiridoids and of compounds de-
be a close relationship of the Comaceae to the Apiaceae carboxylated at C-11 is mutually exclusive. Most of the iri-
(Umbelliferae) and the Araliaceae with which they are doid compounds found in the Comanae (orders Comales,
placed by many phylogenists. The Apiaceae and Araliaceae Eucommiales, and Dipsacales), Loasanae (Loasales), and
completely lack iridoid compounds. The presence of alka- Gentianananae (Gentialales, Goodeniales, and Oleales) are
loids in the Icacinaceae and the Nyssaceae based on an iri- synthesized by the biosynthetic route involving formation
doid monoterpene structure suggests that these plants also of secoiridoids (Jensen, 1991). Those of the Lamianae (La-
may be related to the Comaceae. miales and Hippuridales) are synthesized by a second route
Dahlgren felt that because of the combination of charac- (see the subsection biosynthesis, above) involving decarbox-
ters above, these groups should be combined with the other ylation at C-l1. Data related to the Ericanae and other iri-
iridoid producing families (Gershenzon and Mabry, 1983). doid-containing taxa are not as easily interpretable. In a few
Many of the iridoid compounds of the Comaceae and related instances, compounds such as geniposidic acid (37) are inter-
families are shared with the Rubiaceae, Dipsacaceae, and mediates in both routes of biosynthesis. More detailed inves-
other families, suggesting a link between the two groups. tigation of these biosynthetic pathways will be needed to
Dahlgren and co-workers (1981) transferred the family Fou- establish relationships more clearly (Jensen, 1991).
quieraceae to the Comiflorae because it was the only family Plants in the Comanae, Gentiananae, and Loasanae, in
in the Solaniflorae that contained iridoid monoterpenes (Jen- general, contain both normal iridoid monoterpenes and sec-
sen and Nielsen, 1982). oiridoids. Plants in the Lamianae contain not only many
Most of the plants concemed were in the superorders normal iridoid monoterpenes but also those in which C-l1
Ericanae, Comanae, Loasanae, Gentiananae, and Lamiana- is decarboxylated. They lack secoiridoids. Plants of the Eri-
nae, but a few other sources have been reported (Jensen, canae do not exhibit a clear pattem. Iridoid compounds with
1991). In a reevaluation of the application of chemical evi- 8-J3-stereochemistry were assumed to derive from pathways
dence to the systematics of this group of plants, Jensen ob- associated with secoiridoid biosynthesis, whereas those with
served that data based on the biosynthetic pathways provide 8-a-stereochemistry were associated with the route involv-
a sounder basis for these placements. Based on present infor- ing C-l1 decarboxylation (Jensen, 1991). Some exceptions
 Iridoid Monoterpenes 361

exist: Decarboxylated iridoids also are found in the Aucuba- The iridoid glycosides geniposide (38) and geniposidic
ceae, Eucommiaceae, and Garryaceae (Cornanae), as well acid (37) and their aglycones have have been shown to in-
as in the Ericanae. Secoiridoid structures are known from hibit growth of wheat embryos (Cameron et al., 1984).
the Cornanae, Gentiananae, and Loasanae, but also from the The iridoid glycoside plumieride (39) [from Plumeria ob-
Sarraceniaceae and Stylidiaceae (Ericanae) (Jensen, 1991). tusifolia (Apocynaceae)] has been demonstrated to have
The Cornanae include the ComaIes (Adoxaceae, Alangia- plant growth-inhibiting activity. Although gibberellin-stim-
ceae, Aralidiaceae, Aucubaceae, Cornaceae, Curtisiaceae, ulated growth was inhibited, there was no effect on indole-
Davidiaceae, Escalloniaceae, Garryaceae, Griselinaceae, acetic acid (lAA)-stimulated growth (Adam et al., 1979).
Hydrangiaceae, Icacinaceae, Mastixiaceae, Menyanthaceae, lridoid monotetpenes are involved in a number of
Montiniaceae, Montiniaceae, Nyssaceae, Sambucaceae, plant-animal interactions (Bowers, 1988, 1991; Rimpler,
Stylidiaceae, Symp10caceae, Torricelliaceae, and Vibuma- 1991). Many insects sequester iridoid monotetpenes in a
ceae), Dipsacales (Calyceraceae, Caprifoliaceae, Dipsaca- variety of tissues, including all stages of the insect and the
ceae, Moringaceae, Triplostegiaceae, and Valerianaceae), silk produced. Uptake of iridoids may be selective or nonse-
Loasales (Loasaceae), Goodeniales (Goodeniaceae), and lective; they may be modified before or after uptake. These
Oleales (Oleaceae) (Jensen, 1991). compounds also are excreted in the frass in many instances
The order Gentianales (Apocynaceae, Gentianaceae, Lo- (Rimpler, 1991).
ganiaceae, Rubiaceae, and Theligonaceae) contains normal Many iridoids can act as antifeedants. To mammals, iri-
iridoid monotetpenes and secoiridoids, as well as iridoid- doids often have an intensely bitter taste. Growth rates of
derived alkaloids in families other than the Asclepiadaceae. generalist feeders such as Spodoptera eridiana are reduced
Multiple types of alkaloids are found in the Apocynaceae with iridoid-containing diets (Rimpler, 1991). In contrast,
(indole and steroidal types), the Loganiaceae, and Rubiaceae Lymantria dispar, the gypsy moth, tolerates the "taste" of
(emetine, quinine, and indole types). iridoid glycosides and excretes catalposide unchanged in the
The iridoid families that constitute the Lamianae, include frass (Harborne, 1989). Nonetheless, this insect grows and
the Lamiales [which includes the Acanthaceae, Bigno- survives less well on diets containing iridoids than on iri-
niaceae, Buddlejaceae (possibly misplaced here), CaIIitrica- doid-free control diets (Rimpler, 1991). The vapors of many
ceae, G1obulariaceae, Lamiaceae, Lentibulariaceae, Marty- volatile iridoid monotetpenes repel insects such as ants and
niaceae, Myoporaceae, Pedaliaceae, Plantaginaceae, beetles.
Retziaceae, Scrophulariaceae (including Orobanchaceae), Moths and butterflies that sequester iridoid compounds
Selaginaceae, Stilbaceae, and Verbenaceae), and the Hippur- are unpalatable to birds that might be potential predators.
idales (Hippuridaceae) (Jensen, 1991; Jensen et al., 1988). Adults of Euphydryas phaeton that fed on Chelone glabra
The relatively large family Gesneriaceae apparently lacks (Scrophulariaceae) during larval development are emetic to
iridoids. lridoids are found in the Thunbergiaceae, but not birds that eat a piece of the butterfly (Rimpler, 1991).
in the Mendonciaceae (Jensen et al., 1988), both sometimes The compounds iridomyrmecin (1) and iridodial (2) serve
merged with the Acanthaceae. In general, members of the as defensive compounds in ants of the genus Iridomyrmex.
Larniaceae that accumulate iridoid monoterpenes do not ac- Nepetalactone (29), from Nepeta cataria, Lamiaceae, is in-
cumulate normal monotetpenes and vice versa (Waterman secticidal. Eisner (1964) demonstrated that this compound
and Gray, 1987). is repellent to 17 out of 24 insect species studied. NepetaIac-
Among the iridoid-containing families of the Ericanae tone also has been reported to have hallucinogenic properties
are the Ericales (Actinidiaceae, Epacridaceae, Ericaceae, in humans. NepetaIactone and a similar compound, boschni-
Monotropaceae, and Pyrolaceae), the Stylidiales (Stylidia- alactone (40) from Boschniakia rossica (Orobanchaceae),
ceae), Sarraceniales (Sarraceniaceae), and Fouquieriales have attractive activity toward cats (Herout, 1970) (Fig.
(Fouquieriaceae) (Jensen, 1991). 20.9). These compounds occur free, in contrast to most iri-
lridoid monotetpenes also are found in several other doid compounds.
groups of plants not considered closely related to the above Xylomollin (41), a secoiridoid isolated from the bitter
by most systematists or phylogenists [for example, the Daph- unripe fruit of Xylocarpus moluccensis (or more likely X.
niphyllaceae, Eucommiaceae, Hamamelidaceae, Malpighia- granatum) (Meliaceae), is an effective deterrent to feeding
ceae (Sainty et al., 1981), Meliaceae (however, see Jensen, by the African army worm, Spodoptera exempta at the 100-
1991), and Sarraceniaceae. ppm level in leaf disks and strongly inhibits respiration in
rat liver mitochondria (Kubo and Nakanishi, 1977; Mabry
and Gill, 1979; Nakanishi, 1977; Nakane et al., 1978). How-
BIOLOGICAL ACTIVITY ever, there is some doubt about the source of this compound;
reinvestigation failed to reveal the reported compounds (Jen-
Many iridoid monoterpenes possess biological activity, al- sen, 1991).
though the role of these compounds in biological interactions The interaction of the catalpa sphinx moth and Catalpa
has not been stodied as much as many or other groups of is very complex and involves a mixture of at least 15 iridoids,
secondary metabolites. including cataIpol (42) and catalposide (43) in the feeding
362 lridoid Monoterpenes

attraction system (Bowers and Puttick, 1986; Harborne, and neonepetalactone, actinidine (45) and actinidiolide, and
1982; Nayar and Fraenkel, 1963). dihydroactinidiolidel. Cats (Felidae) responded to the pres-
Catalpa speciosa, Bignoniaceae, a tree native to the ence of these compounds (Inouye, 1991). In addition, male
south-central United States, is pollinated by bumblebees, adults of lacewings (Chrysopa septempunctata, Chrysopi-
carpenter bees, and some moths. In contrast to many other dae) also are attracted strongly to the leaves and fruits of
flowers, those of catalpa attract few nectar thieves. This is this plant. Furthermore extracts of the plant proved to be
surprising, as the flowers contain a large amount of sugar attractive. When these extracts were chromatographed
compared to other bee-pollinated flowers, and catalpa trees (TLC), the active zone on thin-layer plates could be deter-
produce a large floral display. In Michigan (outside of the mined by observing the pattern of bites of the adult males
native range of the plant), ants that visited the extrafloral on the plates. Adult males responded to 10-6 ,.M of neoma-
nectaries and a small butterfly (Poanes hobomok) that fed tatabiol (46) and isoneomatatabiol (47), 10-3 ,.M of mata-
on nectar of catalpa flowers consumed less (compared to tabiol and dehydroiridodiol, and 1 ,.M of iridodiol, 5-hy-
sugar solution) and developed behavioral abnormalities. The droxymatatabiether, 7 -hydroxydihydromatatadiether, and
iridoid glycosides catalpol (42) and catalposide (43) were allomatatabiol.
present in the nectars of both flowers and extrafloral nectar- Jridomynnecin (1) and isoiridomyrmecin (44) are used
ies of Catalpa speciosa (Stephenson, 1981, 1982). by ants such as Iridomyrmex humilis, I. nitidis, and Doli-
Plants of Actinidia polygama (Actinidiaceae) have an ex- choderus scabridus as defensive compounds. These lactones
citatory effect on cats and other Felidae. This plant is also also were found in Aphis polygama. Thus, the structures of
used in folk medicine in Japan. Several iridoid monoterpenes the lacewing attractants correspond to the reduced forms of
have been isolated [iridomynnecin (1), isoiridomynnecin the defensive compounds of ants (Sakan et al., 1970).
(44), dihydronepetalactone (31), isodihydronepetalactone Although many aphids (the major insect pests of northern

yO q:r··W-Ml
~ o I OH

nepetaiaeione (29) bosclml.lactone (40) xylomollin (41) neomatatabiol (46)

:~~ catalpol (42)

HO...... O-glucosyl
isoiridomyrmecin (44)

M~ -'~ ;+<
~ LJ:?. ".. rn~~
!

isoneomatatabiol (47) iridomyrmecin (1) dihydronepetalactone (31) pnederoside (48)

~'<:::~
i O_gl':"y~

~ ~
COZH
H COzCH,
'<:::
~ 0 ~ '<::: ~ 0
~ 0
H '\'\ 0
HO H O-glucosyl
O-gluc:osyl H
CH,COO HO O-glucosyl HO H O-glucosyl

(a) asperuloside geniposide (38) genoposidic acid (37) 6-0-glucosylaucubin (49)

Fig. 20.9 (a & b). Biologically active iridoid monoteIpenes.

 W
lridoid Monoterpenes 363

~ ~
o
HO
COZCHJ

~
0
q) H
CO,H
~

o
CHJCOO
roOHH
~

0
H H O.glucosyl
H H
O.glucosyl O·gluoosyl O·glucosyl

cy
iridodial glucoside (12) loganln(6) deoxyloganic acid (5) lamioside

HO

(4aS,7S,7aR)·nepetaJactol (2) (4aS,7S,7aR).nepetalactone (29)

COZCHJ

a?
CO'CH~H l O H
~ OH ~
o
o H. 0
o 0 H
o o H ~ O.glucosyl

(b) plumlerlde (39) allamandln (50) scabroside

Fig. 20.9. (continued)

European agriculture) reproduce asexually during the sum- found in the larval secretions of chrysomelid beetles of the
mer, at certain times of the year, the male aphid locates the tribe Chrysomelini (Fig. 20.10) (Rowell-Rabier and Pasteels,
female by means of sex attractant pheromones. The phero- 1986). These iridoids appear to be synthesized by the insects
mones for the vetch aphid Megoura viciae are nepetalactone and do not come from the host plants. The host plants include
(29) and iridodial (nepetalactol) (2). Neither of these com- Alnus, Rumex, Ranunculus, Brassica, Nasturtium, Salix, and
pounds is known to occur in the host plant Vicia laba (Daw- Juglans (Pasteels et al., 1982). Eggs and newly hatched lar-
son et al., 1987, 1990; Pickett, 1991). Iridodial (nepetalactol) vae are not protected by these compounds (Rowell-Rabier
alone is the pheromone of the greenbug, Schizaphis grami- and Pasteels, 1986).
num. Again, this iridoid does not occur in the host plant Iridoid glycosides appear to be feeding stimulants for
(Dawson et al., 1988). These compounds are closely related checkerspot butterflies (Euphydryas) and butterflies of the
to known lacewing attractants and may act as kairomones genus Anartia. These butterflies are restricted to iridoid gly-
in enabling lacewings to locate populations of aphids when coside-containing plants (Bowers, 1988, 1991). Larvae of
aphids are scarce (Pickett, 1988). Other aphids that produce E. cynthia sequester iridoid compounds from the host plant
iridoid sex pheromones are the black bean aphid, Aphis Plantago lanceolata (Harborne, 1989). Two of these com-
labae, the pea aphid, Arcthosiphon pisum, and the peach- pounds, aucubin (19) and catalpol (42) occur in the host
potato aphid, Myzus persicae (Dawson et al., 1990). plant, whereas the third, 6-0-glucosylaucubin (49), probably
The damson-hop aphid Phorodon humuli, a pest resistant is formed by conjugation within the insect (Harborne, 1989).
to all registered insecticides in England, must produce sexual Larvae of E. chalcedona were attracted to and ate much
forms to leave hops (Humulus lupulus) for its primary host more of diets containing known iridoid monoterpenes or
(Prunus spp.) when the hops are harvested. A single active plants that contained them than diets that lacked them. These
compound similar to nepetalactol, but presently unidentified, compounds seem to playa major role in host-plant specific-
serves as a sexual pheromone (pickett, 1991). ity in these insects (Bowers, 1983, 1988). Individuals of
The aphid Acyrsiphon nipponicus secretes paederoside Euphydryas anicia sequester iridoids from Besseya alpina,
(48) from the host plant, Paederia scandens (Rubiaceae). B. plantaginea, and Castilleja integra (Gardner and Ster-
This secretion apparently protects the aphid from the lady- mitz, 1988; Stermitz et al., 1986b). The ratio of iridoids in
bird beetle Harmonia axyridis (Harbome, 1989; Nishida and the insects often differs from that of the host plants. Catalpol
Fukami, 1989). esters are hydrolyzed by the insect that sequesters the result-
A number of methylcyc1opentanoid monoterpenes are ant catalpol and excretes the acid. A similar situation prevails
364 lridoid MonoteJpenes

~
<x~ Ct~o
H
CHO

"'H
~~CHO Hr
HCHO
H

chrysomelidial epichrysomelidial plagiodial

Cy Co cy
0

H 0 H i 0
i

plagiolactone epiplagiolactone gastrolactone

Fig. 20.10. Methylcyc}opentanoid monoterpenes from larval secretions of Chrysomelini.

in the interactions of the buckeye butterfly, Junonia coenia Gentian (the dried rhizome and roots of Gentiana lutea,
(Nymphalidae) and plants of the families Acanthaceae, Gentianaeeae) has long been used as a bitter touic in treat-
Plantaginaceae, Scrophulariaceae, and Verbenaceae (all iri- ment of anorexia and dyspepsia. This preparation contains
doid-containing families) (Bowers, 1984). about 2% gentiopicrin. These products are also used to pre-
Many species of Castilleja (Scrophulariaceae) contain iri- pare alcoholic beverages (Tyler et al., 1981).
doid monoterpenes (Arslanian et al., 1985) as well as pyrrol- An iridoid glycoside, allarnandin (SO), with antileukemic
izidine, quinolizidine, and monoterpene iridoid alkaloids properties has been isolated from AI/amanda cathartica
(Stermitz et al., 1986a). Both pyrrolizidine and quinolizidine (Apocynaceae) (Kupchan et aI., 1974).
alkaloids appear to be taken from the hosts of these hemipar- A number of the pharmacological activities of iridoids
asitic plants (see Chapter 30). The larvae of Platyptilia pica are due to the aglycone, because the activity appears only
(Pterophoridae) excrete but do not sequester iridoids. The after treatment with [:I-glucosidase, or only when the pure
adult moths contained rhexifoline, an iridoid monoterpene aglycone is tested (Stermitz, 1988).
alkaloid (Stermitz et al., 1986a).
The iridoid glycoside catalpol (42) is sequestered by lar-
vae of Meris alticola feeding on Penstemon virgatus, and
Neoterpes graefiaria on P. barbatus. The larvae have similar REFERENCES
larval patterns, but the adults, which contain little of the
iridoids, are cryptic (Stermitz et al., 1988). Antirrhinoside ADAM, G., N. H. KHOJ, C. BERGNER, and N. T. LmN, Plant growth
(23) is sequestered by the larvae of the highly aposematic inhibiting properties of plumieride from Plumeria obtusifolia,
larvae of the moth Meris paradoxa and two moth species Phytochemistry, 18, 1399-1400 (1979).
of Lepipolys that feed on Maurandya antirrhinijlora (Scro- ARsLANIAN, R. L., G. H. HAruus, and F. R. STERMITZ, Some iridoid
phulariaceae) (Boros et al., 1991). These larvae accumulate glucosides, including the new 6-deoxycatalpol, from Indian
from 3% to 11 % dry weight of the glycoside. Adults do not paintbrush species related to Castilleja miniata, J. Nat. Prod.,
contain antirrhinoside and are cryptic (Boros et aI., 1991). 48,957-961 (1985).
BALSEVICH, J., Biotransfonnation of IO-hydroxygeraniol and re-
lated compounds by a cell suspension culture of Catharanthus
JllEDiCIl'IAL USES OF IRIDOID COl'll"OVNDS roseus: The formation of reduced products, Planta Medica, 47.
128-132 (1985).
Valerian consists of the dried rhizome and roots of Valeriana BALSEVICH, J. and W. G. W. KURZ, The role of 9- andlor 10-
officinalis (Valerianaceae). Valeriana wallichii and V. edulis oxygenated derivatives of geraniol, geranial, nerol. and neral in
also serve as commercial sources of the drug. Species from the biosynthesis of loganin and ajmalacine, Planta Medica, 49,
related genera also are known to contain the active princi- 79-84 (1983).
ples. Valerian has been used as a calmative in nervousness BATE-SMITH, E. C., I. K. FERGUSON, K. HUTSON, S. R. JENSEN, B.
and hysteria for at least 1000 years. A series of water-soluble J. NIELSEN, and T. SWAIN. Phytochemical interrelationships in
glycosides, collectively called valepotriates, are responsible the Comaceae, Biochem. Sys!. Beol., 3, 79-89 (1975).
for the mild tranquilizing activity. These preparations are BATIERSBY, A. R., S. H. BROWN, and T. G. PAYNE, Biosynthesis
widely available in Europe (Charlwood and Charlwood, of loganin and the indole alkaloids from hydroxygeraniol-hy-
1991; Tyler et al., 1981). droxynerol, J. Chern. Soc., Chern. Cornrnun., 827-828 (1970).
 Jridoid Monoterpenes 365

BEALE, M. H. and J. MAcMILLAN, The biosynthesis of C,-C20 DAWSON, G. W., D. C. GRIFFITHS, L. A. MERRITT, A. MUDD, J.
terpenoid compounds, Nat. Prod. Rep., 5,247-264 (1988). A. PICKEIT, L. J. WADHAMS, and C. M. WOODCOCK, Aphid
BOROS, C. A. and F. R. STERMl'IZ, Iridoids. An updated review. semiochemicals-A review, and recent advances on the sex
Part I., J. Nat. Prod., 53, 1055-1147 (1990). pheromone, J. Chern. Ecol., 16, 3019-3030 (1990).
BOROS, C. A., F. R. STERMTIZ, and N. McFARLAND, Processing of EISNER, T., Catnip: Its raison d'etre, Science, 146, 1318-1320
iridoid glycoside antininoside from Maurandya antirrhiniflora (1964).
(Scrophulariaceae) by Meris paradoxa (Geometridae) and Lepi- ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod.
polys species (Noctuidae), J. Chern. Ecol., 17, 1123-1133 Rep, 5, 581-612 (1988).
(1991). EL-NAGGAR, L. J. and J. J. BEAL, Iridoids. A review, J. Nat. Prod.,
BOWERS, M. D., The role of iridoid glycosides in host-plant speci- 43,649-797 (1980).
ficity of checkerspot butterflies, J. Chern. Ecol., 9, 475-493 ESCHER, S., P. LoEW, and D. ARIGONI, The role of hydroxygeraniol
(1983). and hydroxynerol in the biosynthesis ofloganin and indole alka-
BOWERS, M. D., Iridoid glycosides and host-plant specificity in loids, J. Chern. Soc., Chern. Commun., 823-825 (1970).
larvae of the buckeye butterfly,Junonia coenia (Nymphalidae), FOERSTER, W., H. BECKER, and E. RODlUGUEZ, HPLC analysis of
J. Chern. Ecol.,10, 1567-1577 (1984). valepotriates in the North American genera Plectritis and Vale-
BOWERS, M. D., Chemistry and coevolution: Iridoid glycosides, riano, Planta Medica, 50, 7-9 (1984).
plants, and herbivorous insects, in Chemical Mediation of Coe- GARDNER, D. R. and F. R. STERMTIZ, Host plant utilization and
volution (K. C. Spencer, ed.), 133-165, Academic Press, San iridoid glycoside sequestration by Euphydryas anicia (Lepidop-
Diego, CA, 1988. tera: NymphaIidae), J. Chern. Ecol., 14, 2147-2168 (1988).
BOWERS, M. D., Iridoid g1ycosides, in Herbivores: Their Interac- GERSHENZON, J. and T. J. MABRY, Secondary metabolites and the
tions with Secondary Plant Metabolites, Vol. I (G. A. Rosenthal higher classification of angiosperms, Nordic 1. Bot., 3, 5-34
and M. R. Berenbaum, eds.), 297-325, Academic Press, San (1983).
Diego, CA, 1991.
HAREoRNE, J. B., Introduction to Biochemical Ecology, 2nd ed.,
BOWERS, M. D. and G. M. PurnCK, Fate of ingested iridoid glyco- Academic Press, New York, 1982.
sides in lepidopteran herbivores, J. Chern. Ecol., 12, 169-178
~lIARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod.
(1986).
Rep., 7, 85-109 (1989).
CAMERON, D. W., G. I. FEUTRILL, P. 1'ERLMurrBR, and J. M. SASSE,
Iridoids of Garrya e/liptica as plant growth inhibitors, Phyto- HEROUT, V., Some relations between plants, insects and their iso-
chemistry, 23, 533-535 (1984). prenoids, in Progress in Phytochemistry, Vol. 2 (L. Reinhold
and Y. Liwschitz, eds.), 143-202, Interscience, London, 1970.
CEARLWOOD, B. V. and K. A. CHARLWooD, Terpenoid production
in plant cell cultures, in Ecological Chemistry and Biochemistry INOUYE, H., Iridoids, in Terpenoids (B. V. Charlwood and D. V.
of Plant Terpenoids (J. B. Harborne and F. A. Tomas-Barbenin, Banthorpe, eds.), Vol. 7 of Modern Methods of Plant Biochem-
eds.), Phytochemical Society of Europe Vol. 31, 95-132, Clar- istry (J. B. Harborne and P. M. Dey, eds.), 99-143, Academic
endon, Oxford, 1991. Press, London, 1991.
CEAUDHURY, R. K., F. U. AFIFI-YAZAR, and O. STICHER, .,c NMR INOUYE, H. and S. UESATO, Biosynthesis of iridoids and secoiri-

spectroscopy of naturally occuning iridoid glucosides and their doids, Fortschr. Chern. Org. Naturst., 50, 169-236 (1986).
acylated derivatives, Tetrshedron, 36, 2317-2326 (1980). JENSEN, H. F. W., S. R. JENSEN, and B. J. NIELSEN, Chemotaxonomy
COSCIA, C. J., ThRI'ENOIDS, in Vol. 1 of Handbook of Chromotogra- of the Acanthaceae. Iridoids and quaternary amines, Phyto-
phy (G. Zweig and 1. Sherma, eds.), CRC Press, Boca Raton, chemistry, 27, 2581-2589 (1988).
Fl., 1984. JENSEN, S. R., Plant iridoids, their biosynthesis and distribution in

CROTEAU, R. t Biosynthesis of monoterpenes, in Biosynthesis of angiospenns, in Ecological Chemistry and Biochemistry of

Isoprenoid Compounds, Vol. 1 (J. W. Porter and S. L. Spurgeon, Plant Terpenoids (1. B. Harborne and F. A. Tomas-Barbenin,
eds.), 225-282, Wiley, New York, 1981. eds.), (Phytochemistry Society of Europe, Vol. 31), 133-158,
Clarendon, Oxford, 1991.
DAHLGREN, R. M. T., S. R. JENSEN, and B. J. NIELSEN, A revised
classification of the angiospenns with comments on correlation JENSEN, S. R. and B. 1. NIELSEN, Iridoid glucosides in Fouquieria-
between chemical and other characters, in Phytochemistry and ceae, Phytochemistry, 21,1623-1629 (1982).
Plant Phylogeny (D. A. Young and D. S. Seigler, eds.), JUNIOR, P., Recent developments in the isolation and structure elu-
194-204, Praeger, New York, 1981. cidation of naturaIly occurring iridoid compounds, Planta Med-
DAMTOGT, S., S. R. JENSEN, and B. 1. NmLSEN, .,c and 'H-NMR ica, 56, 1-13 (1990).
spectroscopy as a tool in the configurational analysis of iric,loid KUBO, J. and K. NAKANISHI, Insect antifeedants and repellents from
glucosides, Phytochemistry, 20, 2717-2732 (1981). African plants, in Host Plant Resistance to Pests (p. A. Hedin,
DAWSON, G. W., D. C. GRIFFITHS, N. F. JANES, A. MUDD, J. A. ed.), ACS Symposium Series 62,165-178, American Chemical
PICKEIT, L. J. WADHAMS, and C. M. WOODCOCK, Identification Society, Washington, DC, 1977.
of an aphid sex pheromone, Nature, 325, 614-616 (1987). KUPCHAN, S. M., A. L. DESSERUNE, B. T. BLAYLOCK, and R. F.
DAWSON, G. W., D. C. GRIFFITHS, L. A. MERRITT, A. MUDD, J. BRYAN, Isolation and structure elucidation of allamandin, an

A. PICKEIT, L. 1. WADHAMS, and C. M. WOODCOCK, The sex anti-leukemic iridoid lactone from Allamanda cathartica, J.
pheromone of the greenbug, Schizaphis graminum, Ent. Exp. Org. Chern., 39, 2477-2482 (1974).
Appl.,48, 91-93 (1988). MABRY, T. J. and J. E. GllL, Sesquiterpene lactones and other
366 Iridoid Monoterpenes

terpenoids, in Herbivores (G. A. Rosenthal and D. H. Janzen, verstein and M. Nakajima, eds.), 237-247, Academic Press,
eds.), 502-537, Academic Press, New York, 1979. New York, 1970.
NAKANE, M., C. R. HUTCHINSON, D. VANENGEN, and J. CLARDY, STEPHENSON, A. G., Toxic nectar deters nectar thieves of Catalpa
Revision of the structure of xylomollin, J. Am. Chern. Soc., speciosa, Am. Midl. Nat., 105, 381-383 (1981).
100, 7079-7080 (1978). STEPHENSON, A G., lridoid glycosides in the nectar of Catalpa
NAKANISID, K., Insect growth regulators from plants, in Natural speciosa are unpalatable to nectar thieves, J. Chem. Ecol., 8,
Products and the Protection of Plants (G. B. Marini-Bettolo, 1025-1034 (1982).
ed.), Elsevier, Amsterdam, 1977. STERMITZ, F. R., lridoid glycosides and aglycone. as chiral syn-
thons, bioactive compounds, and lepidopteran defenses, in Bio-
NAYAR, J. K. and G. i'RAENKEL, The chemical basis of the host
logically Active Natural Products: Potential Use in Agriculture
selection in the catalpa sphinx Ceratomia cata/pae (Lepidop-
(H. G. Cutler, ed.), ACS Symposium Series 380, 397-402,
tera, Sphingidae), Ann. Ent. Soc. Am., 56,119-122 (1963).
American Chemical Society, Washington, DC, 1988.
NISHIDA, R. and H. FuKAMI, Host plant iridoid-based chemical de- STERMITZ, F. R., G. H. HARRIS, and W. JING, lridoids and alkaloids
fense of an aphid, Acyrthosiphon nipponicus, against ladybird from Castilleja host plants for Platyptilia pica. Rhexifoline con-
beetles, J. Chern. Ecol., 15, 1837-1845 (1989). tent of P. pica, Biochem. Syst. Ecol., 14, 499-506 (1986a).
PASTEBLS, J. M., J. C. BRAEKMAN, D. DALOZE, and R. OrrINGER, STERMITZ, F. R., D. R. GARDNER, F. J. ODENDAAL, and P. R. EHR-
Chemical defence in chrysomelid larvae and adults, Tetrahed- LICH, Euphydryas anicia (Lepidoptera: Nymphalidae) utiliza-
ron, 38, 1891-1897 (1982). tion of iridoid monoterpenes from Castilleja and Besseya (Scro-
PICKE1T, J. A, Integrating use of beneficial organisms with chemi- phuJariaceae) host plants, J. Chern. Ecol., 12, 1459-1468
cal crop protection, Philos. Trans. Roy. Soc. Lond. B, 318, (1986b).
203-211 (1988). SWRMITZ, F. R., D. R. GARDNER, and N. McFARLAND, lridoid gly-

PICKEIT,1. A., Lower terpenoids as natural insect control agents, coside sequestration by two aposematic Penstemon~feeding

in Ecological Chemistry and Biochemistry of Plant Terpenoids geometrid larvae, J. Chern. Ecol., 14, 431-437 (1988).
(J. B. Harborne and F. A. Tomas-Barberan, eds.), Phytochemis- TYLER, V. E., 1.. R. BRADY, and J. E. ROBBERS, Pharmacognosy,
try Society of Europe Vol. 31, 297-313, Clarendon, Oxford, 8th ed., Lea and Febiger, Philadelphia, 1981.
1991. UBSATO, S., H. IKBoA, T. FUJITA, H. iNOUYE, and M. H. ZENK,
Elucidation of iridodial formation mechanism-Partial purifi-
RlMPLER, H., Sequestration of iridoids by insects, in Ecological
Chemistry and Biochemistry of Plant Terpenoids 0. B. Har-
cation and characterization of the novel monoterpene cyclase
borne and F. A. Tomas-Barberan, eds.), Phytochemistry Society
from Rauvolfia serpentina cell suspension cultures, Tetrahedron
Lett., 28, 4431-4434 (1987).
of Europe Vol. 31,314-330, Clarendon, Oxford, 1991.
UBSATO, S., S. KANOMI, A IDA, H. iNOUYE, and M. H. ZENK,
ROWELL-RAHIBR, M. and J. M. PASTEELS, Economics of chemical Mechanism for iridane skeleton fonnation in the biosynthesis
defense in Chrysomelinae, J. Chern. Ecol., 12, 1189-1203
of secologanin and indole alkaloids in Lonicera tatarica, Cath~
(1986).
aranthus roseus, and suspension cultures of Rauwolfia ser-
SAINTY, D., F. BAILLEUL, P. DELAVEAU, andH. JAQUEMIN, Malpigh- pentina, Phytochemistry, 25, 839-842 (1986).
a
iac""s: Nouvelle fantille iridoides. Etude du Stigmatophyllon VAN DER SLUIS, W. G., Chemotaxonomical investigations of the
sagittaturn, J. Nat. Prod., 44, 576-578 (1981). genera Blackstonia and Centaurium (Gentianaceae), Plant Syst.
SAKAN, T., S. !sOB, and S. B. HYEON, The chemistry of attractants Evol., 149, 253-286 (1985).
for Chrysopidae from Actinidia polygama Miq., in Control of WATERMAN, P. G. and A I. GRAY, Chemical systematics, Nat.
Insect Behavior by Natural Products (D. L. Wood, R. M. Sil- Prod. Rep., 4,175-203 (1987).
21
Sesquiterpenes

Introduction Il'fI'KODvcnON
Biosynthesis
Biosynthesis of (E, E)-Farnesyl Pyrophosphate Sesquiterpenes are the most numerous of all terpenoid com-
Interconversion of 2-E-6-E- and 2-Z-6-E-Farnesyl pounds. The approximately 5000 known compounds of this
Pyrophosphate type mostly can be grouped into about 30 major skeletal
Acyclic Sesquiterpenes types, but at least 200 less common skeletal types are known
Cyciization of Acyclic Precursors (Fig. 21.1). The distribution of sesquiterpenes in plants is
The Origin of Major Groups of Cyclic Sesquiterpenes essentially the same as monoterpenes. Sesquiterpene hydro-
Biological Activity carbons are common essential oil components. Although
AIlelopathy only a few fungi accumulate monoterpenes, many accumu-
Medicinal Properties late sesquiterpenes. Abscisic acid, a plant growth-regulating
Fungal Pheromones or Chemotactic Agents compound, is synthesized as a sesquiterpene in fungi. De-
<'Iant-Fungal Interactions spite the fact that this compound often is considered a sesqui-
Phytoalexins terpene, in higher plants abscisic acid is derived from the
Phytotoxins
breakdown of xanthophylls and should be considered as a
Plant-Animal Interactions
tetraterpene derivative (see Chapter 26).
Plant allomones
In the sesquiterpene series, a-, mono-, bi-, trio, and tet-
Kairomones
racyclic compounds are known (Fraga, 1991). Of these, bi-
Synomones
cyclic and tricyclic compounds predominate. Most sesquiter-
Insect semiochemicals
penes occur free, although glycosides of this series are
Allomones or Defensive Compounds in Insects
known (Wong et al., 1991).
Insect pheromones
Insect Juvenile Hormones and Plant Juvenile Hormone Basically, the same phylogenetic conclusions that can be
Mimics derived from studies of the structural types and distribution
Essential Oil Components of monoterpenes are true for sesquiterpenes. Despite their
Sweeteners diversity, these compounds have proved to be of liruited
Picrotoxins value in establishing the phylogeny of higher plants.
Sesquiterpene Lactones Sesquiterpenes are catabolized by poorly understood
Biogenesis pathways (Loomis and Croteau, 1980).
Chemosystematic Studies Involving Sesquiterpene Techuiques for the isolation and purification of sesquiter-
Lactones penes include extraction, followed by various types of chro-
Biological Activity matography (Baerheim Svendsen and Scheffer, 1985; Cos-
Plant Growth Regulator Activity cia, 1984; Fraga, 1991). Mass spectrometry and nuclear
Allelopathic Effects magnetic resonance (NMR) spectroscopy are the most com-
Plant AIlomones monly used methods for characterization of sesquiterpenes.
Chemotactic Agents from Parasitic Plants Accumulation of sesquiterpenes is associated with spe-
Medicinal Uses cialized secretory structures, and even though both monoter-
References penes and sesquiterpenes may be synthesized within oil

367
368 Sesqu;terpenes

~OH
~ 9J1
'"
0
I h i h' "
; 6 ;0S 0
",OH yjy""
A HO

emmotinA S-elemene
7-hydroxyealamenene furanoeudesma-l,3-diene

drimenol cedrol guaiol ex-copaene (58)

longirolene (22) P-santalol p-caryophyllene (11) germncrene D (67)

1 •

~~o
:,..

12 o 0 HO ~

P~
costunolide (119) germaerene A y-lirlodenolide , 0
epox/de (20)
H

HO~"" ,,,p ~
goYazeOSOvlide(l20)
...
"-._",, , :,..

....
o _""...,,"" :,.. ""

gynomitr-8(12)·en.9-ex-ol (1) gymnomitr-8(12)-enone (2) patchonlol (14)

humulene (10)

Fig. 21.1. Representative sesquiterpenes.

glands, accessibility of the sites of synthesis to the precursor terpene cyclase activities, although several studies show that
molecules differ (Banthorpe and Charlwood, 1980; Croteau key biosynthetic enzymes are operationally soluble (Gers-
and Johnson, 1984; Loomis and Croteau, 1980). In Pinus henzon and Croteau, 1990).
pinaster needles, synthesis of sesquiterpenes occurred for Production of sesquiterpenes in cell cultures has been
the length of the needle, whereas monoterpenes were ouly of value for study of biosynthesis (Ellis, 1988); in several
synthesized at the base of young needles (Croteau and John- instances, the amount of products was enhanced by challeng-
son, 1985). Rates of incorporation of exogenous precursors ing the culture with fungal pathogens. In general, however,
often are low, but the use of specifically labeled precursors the levels of sesquiterpenes produced in culture is low
and degradation studies indicates that sesquiterpenes are la- (Charlwood and Charlwood, 1991). Bryophytes are unique
beled by mevalonic acid (MYA), isopentenyl pyrophosphate in that terpenoids are stored in intracellular oil bodies. In
(IPP), dimethylaliyl pyrophosphate (DMAPP), and geranyl Calypogeia granulata, both gametophytic cells and suspen-
pyrophosphate (GPP) (Loomis and Croteau, 1980). sion cultures contain dark blue bodies in which the trinorses-
Most evidence supports synthesis of sesquiterpenes in quiterpene, 1,4-dimethylazulene, accumulates (Charlwood
the cytosol/microsomal compartment of plant cells (Threlfall and Charlwood, 1991). The main components of the oil from
and Whitehead, 1991). The endoplasmic reticulum appears cultures of Reboulia hemisphaerica are gymnomitr-8(12)-
to be the site ofFPP synthase (prenyltransferase) and sesqui- en-9a-ol (1) and gymnomitr-8(l2)-en-9-one (2). Differen-
 Sesquiterpenes 369

tiated cultures of liverworts have proven to be useful for the ecule of isopentenyl pyrophosphate (IPP) and dimethylallyl
accumulation of sufficient material for study (Charlwood pyrophosphate (DMAPP) to produce an enzyme-bound gera-
and Charlwood, 1991). nyl pyrophosphate. That intermediate, in tum, reacts with
A number of alkaloids that possess sesquiterpene skele- another unit of isopentenyl pyrophosphate (Fig. 21.2) (Cane,
tons are discussed under Sesquiterpene Alkaloids in Chapter 1985, 1990; Croteau, 1987; Croteau and Johnson, 1985).
36. Geranyl pyrophosphate normally does not accumulate dur-
ing this process. The reaction occurs with net inversion of
configuration (Cane, 1990).
BIOSYNI'HESIS A prenyl transferase, FPP-synthetase (BC 2.5.1.1), from
extracts of germinating castor bean seedlings, has been
Sesquiterpenes are derived from a IS-carbon intermediate, highly purified; both forms obtained synthesized all E-farne-
farnesyl pyrophosphate (3), which is assembled by head-to- syl pyrophosphate (porter and Spurgeon, 1981). FPP synthe-
tail fusion of isoprene units (Cane, 1990). Biogenetic paths tases also have been demonstrated from Gossypium hirsutum
to most of the important structural types of sesquiterpenes and from Pisum sativum (Croteau and Johnson, 1985). E,E-
have been proposed, but few have been studied in detail. Farnesyl pyrophosphate synthetase is proposed to employ
The diversity of sesquiterpenes is much greater than that a stepwise ionization-condensation mechanism involving a
of monoterpenes, but also exceeds that of diterpenes and rigid pyrophosphate-carbonium ion pair as an intermediate
triterpenes. This situation presumably reflects the much (Poulter et al., 1981).
greater conformational flexibility of the 10- or II-membered The stereochemistry is well established, and many ques-
rings as compared with that of 6-membered rings in the last tions concerning the overall mechanism of the condensation
two classes. have now been resolved. Farnesyl pyrophosphate synthetase
(EC 2.5.1.1) is the key enzyme in the biosynthetic pathways
for several classes of terpenes. This enzyme catalyzes 1'-4
Biosynthesis of (E.E)-Farnesyl
condensation between IPP and DMAPP, or geranyl pyro-
Pyrophosphate
phosphate, polymerizations that constitute the major build-
Each of the known skeletal types is derived from (E,E)- ing steps of terpenoid biosynthesis (Fig. 21.2) (Poulter and
farnesyl pyrophosphate (FPP) (3), formed by union of a mol- Rilling, 1978; Poulter et al., 1978, 1981). The condensation

OH
~ OH_ H~

,
Ho,e HO,Cr>4.X'7 ~ B /-J
OH'" ) ( "'-./ 'OPP -+ OPP
,~ ,~
mevalonic acid IPP

~
APP

A)(!'s
:::,... + J
~OPP
inversion
--+ "" lis B,,~
OPP
OPP
DMAPP B"HR ""
IPP

I"... I".. I"..

~OPP
Hs Us

GPP
R I .y

OPP

(E,E)-farnesyJ pyrophosphate (3)

Fig. 21.2. Formation of (E,E)-farnesyJ pyrophosphate.

370 SesquUerpenes

reaction may be subdivided into three phases: ionization, ceae) tissue culture leads to loss of the pro-S hydrogen atom
condensation, and elimination. Ionization of the pyrophos- from C-l of Z,E-famesol (6) (Fig. 21.3).
phate group of DMAPP and activation of the allylic moiety
is the first step of both hydrolysis and condensation of IPP. Acyclic Sesquiterpenes
Studies by Poulter and co-workers indicate that cleavage
of the carbon-oxygen bond of geranyl pyrophosphate is a Most acyclic sesquiterpenes are derived from famesol
discrete step, yielding a geranyl cation-pyrophosphate ion and nerolidol (7) or from the corresponding pyrophosphate
pair and that this reactive species subsequently alkylates the esters. Although the number of acyclic sesquiterpenes is rela-
double bond in IPP. These workers concluded that ionization tively small, Z- and E-famesol (5 and 6), nerolidol (7), and
and condensation are not concerted (Poulter et al., 1981). the corresponding olefins are relatively common in essential
Studies with [1- 18 01geranyl pyrophosphate indicate that ex- oils. The famesenes and a few other compounds, such as 13-
change of oxygen does not occnr dnriog the reaction, and sinesal (8), are derived in a straightforward manner, but most
that a highly structured ion pair exists (Mash et al., 1981). acyclic sesquiterpenes are more complex in origin (Fig.
The biosynthesis of sesquiterpenes in plants appears to 21.4).
be isolated from that of either monoterpenes or diterpenes. A
prenyl transferase isolated from pumpkin (Cucurbita pepo, Cyclization of Acyclic Precursors
Cucnrbitaceae) converts Cs units into FPP, but not into gera-
Cyclases transform 2-E-6-E-famesyl pyrophosphate into
nylgeranyl pyrophosphate (the precnrsor to diterpenes). An-
cyclic sesquiterpenes via ionization and electrophilic attack
other enzyme from the same source forms C20 terpenoids
of the resultant allylic cation on either the central or distal
from Cs units, but does not accumulate lower homologs (also
double bonds (Cane, 1990). The stereochemistry of bond
see Chapters 19 and 22). However, because famesyl pyro-
scission, of ring closure, and of the migration of hydrogens
phosphate synthetase is a branch-point metabolite for the
and alkyl groups involves overall anti stereochemistry. Such
synthesis of sesquiterpenes, triterpenes, and sterols, this en-
stereochemistry probably is observed because the cyclase
zyme is ubiquitous in plants (Croteau and Johnson, 1985).
acts as a template for the acyclic precnrsor, allowing it to
Asynunetric labeling of sesquiterpenes by labeled meva-
adopt the conformation from which the series of stereospe-
lonic acid or other precnrsors, presnrnably caused by an en-
cific transformations proceed with greatest facility (Cane,
dogenous pool of DMAPP, has been reported for a few ses-
1990; Loomis and Croteau, 1980). The central theme of the
qniterpenes. Label primarily is introduced into the portions
biosynthetic hypotheses for sesquiterpene biosynthesis is an
of the molecules derived from IPP (Charlwood and Banth-
intramolecular attack by electrons of either the central or
orpe, 1978).
distal double bond of the famesyl molecule on the carbon
The formation of 2-E-6-E-famesyl pyrophosphate (3)
bearing the pyrophosphate (Cane, 1985; Croteau, 1987; Cro-
from (4S)- and (4R)-[4-3Htl-MVA in systems from Pinus
teau and Johnson, 1985). Orbital aligument also is an impor-
and from Citrus species, is accompanied by loss of the pro-
tant consideration in sesquiterpene cyclizations that are initi-
4S hydrogen and retention of the pro-4R hydrogen (the pro-
ated by double-bond protonation rather than pyrophosphate
4S proton of mevalonic acid becomes the pro-2R proton of
ionization. Enzymatically controlled initial intramolecular
isopentenyl pyrophosphate). The possibility of interconver-
attack governed by either electronic or steric effects pro-
sion of 2-E-6-E-famesyl (2-trans-6-trans-famesyl) (3) and
duces a monocyclic system. Subsequent 1,2- and 1,3-hydride
2-Z-6-E-famesyl (2-cis-6-trans-famesyl) (4) pyrophosphate
shifts, oxidations, cyclizations, and reductions lead to the
has been ruled out in these systems.
observed diversity of structures. Predictable cationic inter-
mediates are useful for explaining the lines of compounds
Interconversion of 2-.8-6-.8- and 2-Z,6-.8-
that arise from various points in the biosynthesis (Fig. 21.5)
FarnesylPyropbospbate
(Cane, 1990; Loomis and Croteau, 1980; RUcker, 1973).
Both 2-E-6-E-famesyl (2-trans-6-trans-famesyl) (3) and The first step in the enzymatic formation of sesquiter-
2-Z-6-E-famesyl (2-cis-6-trans-famesyl) (4) pyrophosphate penes is the ionization of 2-E-6-E-famesyl pyrophosphate
have been suggested as intermediates for various types of (3) to the corresponding transoid allylic cation-pyrophos-
cyclic systems (Fig. 21.3), but, at present, there are no unam- phate anion pair. This ion pair can undergo several transfor-
biguous examples of conversion of 2-E- to 2-Z-famesyl py- mations. Nucleophilic attack on the back face of the allylic
rophosphate. The direct conversion of the 2-E compound to cation by the neighboring 10,11 double bond will result in
cyclic intermediates without intermediacy of the 2-Z com- net anti displacement of the pyrosphate moiety from C-l
pound has been suggested. This may possibly occnr because and formation of a new C-C bond (Cane, 1990). On the
of the conformational flexibility of the lO-carbon ring other hand, collapse of the ion pair will either regenerate
(Banthorpe and Charlwood, 1980; Loomis and Croteau, the starting material or the transoid conformer of nerolidol
1980). The status of a 2-Z-famesyl pyrophosphate synthetase pyrophosphate (9) by a net suprafacial allylic rearrangement.
is uncertain (Croteau and Johnson, 1985). The tertiary allylic pyrophosphate ester can undergo simple
The isomerization of E,E-famesol to Z,E-famesol by an rotation about the newly generated 2,3 single bond followed
enzyme system from Andrographis paniculata (Acantha- by reionization to the corresponding cisoid allylic cat-
 Sesquiterpenes 371

Q 'R.
Andrographis paniculata

ceJl free e)[tract

R
(5)

Andrographisptmicu/sla

ceJlfreeexh'ad

I OH (5)
H,

(3)
Opp

(4)

Opp

Fig. 21.3. Isomerization of 2-E-6-E-famesol and 2-Z-6-E-famesol (modified from Mackie and Overton, 1977; used with pennission of the copyright
owner, European Journal of Biochemistry Zurich).

ion-pyrophosphate anion pair. The nature of the products The cyclases isolated to date are operationally soluble
eventually formed are a function of the stereochemistry and proteins in the molecular weight range of 40,000-100,000.
conformation of the intermediates (Cane, 1990). Cyclases Several are monomers, whereas at least two are homodimers.
may serve as the rate-controlling enzymes in sesquiterpene The enzymes are moderately lipophilic and require no cofac-
biosynthesis. tor other than a divalent metal ion, Mg2+ usually being pre-
In order for cyclization of the intermediates to occur, the . ferred. The role of these enzymes appears to be to bind the
1T-orbitals of the relevant double bonds must be properly substrate in a conformation appropriate for cyclization and
aligned to achieve the required geometry for interaction. to catalyze the ionization of the allylic pyrophosphate ester
This condition is met only if the individual double bonds of (Cane, 1990).
the allylic substrate are mutually perpendicular to a common Two of the most common sesquiterpenes, humulene (10)
plane. Further, C-l to C-4 and the attached methyl group, and J3-caryophyllene (11), were synthesized by a soluble
C-IS, are constrained to a common plane, as are C-S to C- enzyme preparation from leaves of sage (Sa/via ojficinalis,
8 andC-14, and C9 to C-13. To cyclize, C-l must be brought Lamiaceae). Partial purification of the extracts allowed reso-
to within bonding distance of either the central or distal dou- lution of the two cyclizing activities. Both cyclases had mo-
ble bond with displacement of pyrophosphate from C-l of lecular weights of S8,000 and depended on the presence of
famesyl pyrophosphate occurring with net anti stereochem- a divalent cation; Mg2+ was preferred (Cane, 1990).
istry (Cane, 1990). For those cases which require the inter- The origin of eremophilane sesquiterpenes by migration
mediacy of an nerolidol pyrosphate-like intermediate, the of a methyl group from C-IO to C-S in an eudesmane inter-
allylic displacement of the pyrophosphate moiety will take mediate has been postulated to reconcile the apparent devia-
place in an anti sense from a cisoid confonnation (Cane, tion of eremophilanes from the isoprene rule (Cane, 1990).
1990). ( - )-Aristolochene (12), an eremophilane-type sesquiter-
372 Sesquiterpenes

R I -;?

farnesyl pyrophosphate (3)

opp

nerolidol (7) p-sinesal (8)

davanone ipomeamarone (38)

o
freelingyne

Fig. 21.4. Representative acyclic sesquiterpenes.

pene has been isolated from several plants; among these on the 4,5 double bond to form the bicyclic eudesmane cat-
are Aristolochia indica (Aristolochiaceae) and Bixa orellana ion, which can rearrange by sequential 1,2-hydride and
(Bixaceae), as well as in the defensive secretions of soldiers methyl migrations to yield ( - )-aristolochene (12, Fig. 21.6).
of the termite Synfermes na. According to the proposed The reaction is catalyzed by a single enzyme, with no evi-
mechanism, cyclization of farnesyl pyrophosphate by elec- dence for the release of free intermediates (Cane, 1990).
trophilic attack at C-1O of the distal double bond, followed In tobacco and in potato cell cultures, the levels of cy-
by loss of a proton from one of the two adjacent methyl c1ases are increased by treatment with fungal elicitors; at the
groups will generate the known sesquiterpene germacrene same time, the level of squalene synthase activity declines
A (13). The latter intermediate can undergo further cycliza- (Gershenzon and Croteau, 1990; Loomis and Croteau, 1980).
tion by protonation at C-I and attack of the resultant cation The 5-epi-aristolochene cyclase is involved in the formation

12

13
15

opp

/
~-\rb-8 neroJidyl pyrophosphate (9)

:if
farnesyl pyrophosphate (3)

~-~/
Fig. 21.5. Isomerization of E,E~famesyl pyrophosphate to the corresponding alllylic cation-pyrophosphate ion pair, and subsequent modifications
(modified from Cane, 1990; used with permission of the copyright owner, Copyright 1990, American Chemical Society, Washington, DC),
 Sesquiterpenes 373

__
H+

" --+-

-H

11
farnesyl pyrophosphate (3) germacrene A (13)
H

~ --+-

~
13

eudesmane cation (-)-arisotolochene (12)

Fig. 21.6. Biosynthesis of ( - )-aristolochene (modified from Cane, 1990; used with pennission of the copyright owner, Copyright 1990, American
Chemical Society. Washington, DC).

of products such as capsidio!. TItis enzyme consisted of two exlracts catalyze the conversion of farnesyl pyrophosphate
proteins with molecular weights of 61,500 and 63,500, re- (3) to patchoulol (14) along with a mixtore of cyclic olefins
spectively (Cane, 1990). such as a-, (3-, and 'Y-patchoulene (15, 16, 17) as well as a-
A cyclase from Pogostemon cablin (syn. P. patchouli hulnesene (18) and a-guaiene (19). However, sesquiterpene
and P. heyneanus) leaves is capahle of converting farnesyl olefins do not participate as free intermediates in the trans-
pyrophosphate to (- )-patchoulol (14, Fig. 21.7). Cell-free formation of farnesyl pyrophosphate to patchoulol (Cane,

Fig. 21.7. Biosynthesis of patchoulol (Cane, 1990; modified and used with pennission of the copyright owner, Copyright 1990. American Chemical
Society, Washington, DC).
374 Sesquiterpenes

1990; Croteau et al., 1987). Furthermore, studies with and 4 (Fig. 21.8). The reactive cyclodecadiene system
[12,13- 14C;6-3Hlfarnesyl pyrophosphate as a precursor iodi- has been hypothesized to be transferred readily ioto other
cate that deprotonation-protonation steps to form bound ole- mono-, bi-, and tricyclic sesquiterpenes and, thus, the germa-
finic iotermediates io the biosynthesis of patchoulol do not cranes often are suggested as key iotermediates in the synthe-
occur, providiog support for a hydride shift mechanism (Cro- sis of other sesquiterpenes. However, io many iostances, it is
teau et al., 1987). Patchoulol synIhase had a molecular probable that no olefinic iotermediates occur, and the actual
weight of 80,000 and consisted of two identical subunits of iotermediates may be only transient cations. Other major
40,000. types of sesqniterpenes are the selioanes (eudesmanes), guai-
In this iostance, the sesquiterpene olefin cyclase activity anes, pseudoguaianes, elemanes, and eremophilanes (Fig.
could not be separated from the patchoulol synthase activity 21.8) (Riicker, 1973). Guaianes, eremophilanes, and pseu-
and showed similar pH and metal ion requirements, suggest- doguaines are especially common io the Asteraceae, the last
ing that the olefinic products are derived by diversion of the type occurring only as lactones and in that family (Herz,
various carbocation iotermediates (Cane, 1990). 1977).
The biosynthesis of many relatively complex sesquiter-
Tbe Origin of lIIigor Groups of Cyclic penes, many of them fungal metabolites, has been reviewed
Sesquiterpenes (Cane, 1981). Much biosynthetic study has been carried out
Germacranes often are portrayed as beiog derived from a with fungi because of the greater ease of labeliog and culture
2-E-6-E-iotermediate. These sesquiterpenes possess a 4,10- of the organism.
dimethyl-7 -isopropyl cyclodecane system and often have In the past, it has been considered Ihat a 2-Z-6-E-precur-
double bonds io positions 1 (io germacrane A epoxide, 20) sor gives rise to monocyclic sesquiterpenes with one 6-

a-humulene

~ CHOH
':'OHIII ~~
0 H
NIIH I 0
HOI
h -
carotol sirenin (30) 2 tricbothecin OCOCH=cHCH3
(a)

Fig. 21.8 (a & b). Sesquiterpenes derived from a 2-E-6-E-famesyl pyrophosphate precursor.
 Sesquiterpenes 375

4)~7 5)A~ ~i- YQy""'i

I Yr,C(HOiH I'
!
~ opp \o..0Pp!
~ a maaliane
•
an aristolane

P~~->B-
~ ~.~
\,;{/)f?y~
I·~ i

rH~
I
• +
a se:inane or eudesmane 0 al
a pseudogu ane

~H
~!
0yPy~~
~O 0
, H , ! 9' ! a germacranolide

ro =
-::- + - peremophilane
?an ~COCH3

rth.
1i H

(b)
~~-~
(+).loDgifolene (22) a germacrane

Fig. 21.8. (continued)

anelemane
CHO

membered ring (bisabolenes) and systems with ai,7-di- The monocycIic humulanes (11 -membered ring) and
methyl4-isopropyl skeleton (Fig. 21.8). Among these ses- bicyclic caryophylIanes (cyclobutane-cyclononane) may
quiterpenes are the cadinenes, cubebanes, copabomanes, and arise from a common precursor. In Salvia officinalis,
ylangenes (Rucker, 1973). However, most (if not all) sesqui- humulene (10) and j3-caryophylIene (11) appear to have
terpenes are now thought to arise via the intennediacy of 2- a common intennediate (a humulyl cation), although
E-6-E-farnesyl pyrophosphate (Cane, 1990). The cyclase for they may be synthesized by separate enzymes, humu-
bisabolene derivatives of Andrographis paniculata has been lene cyclase and caryophyllene cyclase (Croteau and
examined (Cane, 1990). Although the origin of the cadinenes Gundy, 1984; Dehal and Croteau, 1988). Both enzymes
[e.g., 'Y-cadinene (21») usually has been explained by a 2- have a molecular weight of 57,000, and prefer Mg2+
Z-6-E-precursor, there is considerable evidence for a biogen- as the divalent cation. Both humulene and caryophylIene
esis involving the all E-precursor (Fig. 21.9) (Arigoni, 1975; are widely distributed in nature (Riicker, 1973). The
Loontis and Croteau, 1980). same precursor that leads to caryophylIene also leads to
The sesquiterpene alkaloid dendrobine (87, Fig. 21.20) longifolene (22), longibomeol, and 10ngicycIene, all of
also appears fonnally to be derived from a 2-Z-6-E-precur- which are found in pine resins (Riicker, 1973). 'Y-Bisabo-
sor, but is labeled by the E,E-isomer and not by the 2-Z-6- lene (23) occurs commonly in plants, usually accompanied
E-compound (Loontis and Croteau, 1980) (see sesquiterpene by compounds of similar structure and apparent biosyn-
alkaloids Chapter 36). thetic origin.
376 Sesquiterpenes

--,Y~l{.
GJ -1'1(
r;I
H

--
r-cadinene (21)

Fig. 21.9. Proposed mechanism for the conversion of E,E-famesyl pyrophosphate to 'Y-cadinene.

BIOLOGICAL ACTIVITY genus (a coenocytic, filamentous aquatic fungus) which re-

produces both sexually and asexually, the gamete-bearing
A number of sesquiterpenes have pronounced biological ac- generation produces small, orange male gametangia and
tivity both in the plants themselves and toward other organ- larger, colorless female gametangia (gametes are haploid sex
isms. Illudane sesquiterpenes such as ptaquiloside (24) from cells that unite to produce a zygote). The gametes released
bracken and several other ferns are responsible for the car- are motile, uniflagellate cells that lack cell walls and are
cinogenic activity of these plants, including some used as surrounded by a double membrane. Male gametes are highly
food products (Fig. 21.10) (Saito et al., 1990). motile and are strongly attracted to the female gametes.
Under the microscope, the male gametes can be observed
Allelopatby to cluster in the vicinity of unopened female gametangia
(the stroctures that bear the gametes). Male gametes also
Several sesquiterpenes [as well as sesquiterpene lactones approach and cluster against a dialyzing membrane that sep-
(see the subsection later in chapter)] inhibit the growth of arates them from a supernatant in which female gametangia
plants and the germination of seeds (Fischer, 1991a; Fischer have discharged female gametes. The active compound was
et al., 1989a). The sesquiterpene aldehydes (+ )-vitrenal christened sirenin (Mascarenhas, 1978; O'Day and Horgen,
(25), ( - )-Iepidozenal (26), and isobicyclogermacrenal from 1981). Parisin, a compound of similar but undefined stroc-
the liverwort Lepidozia vitrea and plagiochilin (27) from tore, which serves as an attractant for the female gametangia,
Plagiochila ovalifolia inhibit growth of other plants. Plagio- is produced by the sperm cells (pommerville et al., 1990).
chilin inhibits the growth of rice seedlings at 50 ppm (Fi- In order to obtain large quantities of material for study,
scher, 1991a). A mixtore of ~-bisabolene (23), a-guaiene hybrids of A. arbuscula and A. javanicus that produce about
(19), a-bulnesene (18), ~-patchoulin (16), and bergamotene 95% either male or female gametangia were produced. Hy-
(28) from Ambrosia artemisifolia strongly inhibited germi- brids that gave rise to mostly female gametangia produced
nation of onion, oat, rye grass, and Amaranthus palmeri sirenin. Of several analogs tested, I-sirenin (30) (the natu-
seeds (Fig. 21.10) (Fischer, 1991a). rally occurring form) was the most active.

Medicinal Properties
Gossypol (29, Fig. 21.14) has been evaluated as a male PLAl"IT-ftJNGAL Il'I'fERACDOl'IS
contraceptive. This compound also has direct effects on
sperm and may have potential as a vaginal spermicide. Gos- Many sesquiterpenes are produced and accumulated by both
sypol is active orally, but sometinles has serious side effects. plants and fungi. A variety of different types of biological
In vivo tests reveal that only ( - )-gossypol is active (Anon., activity is expressed. In plant pathogen-plant interactions,
1984; Bingel and Fong, 1988). sesquiterpenes play both defensive and offensive roles.

Fungal Pberomones or Cbemotactic Agents Pbytoalexins

Sirenin (30, Fig. 21.8) is a sexual pheromone from AlIo- Several sesquiterpenes are phytoalexins (Brooks and
myces macrogynus (Jaenicke, 1991). In this phycomycete Watson, 1985; Gross, 1977; Kuc, 1992). These compounds
 Sesqu;terpenes 377

HO~
Xb
OH 0

HO OH

ptaquiloside (24) (+)-vitrenal (25) (-)-Iepidozenal (26)

~
:
,
CH,COO H0

I OCOCH,
~",,/
I
CH,COO

plaglocbllin (27) a-goalene (19) «-bulnesene (18)

B-PBkboulin (16) bergamotene (28) longicyclene

longlborneol

Fig. 21.10. Biologically active sesquiterpenes.

are especially common in plants of the Solanaceae and Con- ceous plants (Harbome, 1986a; Kuc, 1992; Nahrstedt, 1979).
volvulaceae. The biosynthetic pathway leading to rishitin involves a vet-
Capsidiol (31), an eudesmane sesquiterpenoid, is fonned ispirane intennediate, although other similar compounds
from E,E-famesyl pyrophosphate by the pepper Capsicum such as aubergenol (35) and aubergenone (36) are nonnal
annuum when inoculated with the spores of Moniliafruticola eudesmanes (Beale and MacMillan, 1988).
(Threlfall and Whitehead, 1991). The mode of incorporation A sesquiterpene from Nicotiana debneyi, debnyol (37),
of acetate suggests a biosynthetic scheme in which the angu- is highly fungitoxic to spores of the fungal pathogen Peron-
lar group has undergone a 1,2 shift, rather than involving ospora tabacina (Kuc, 1992).
spiran intennediates, as are involved in the biosynthesis of Ipomeamarone (38) is a fungicide or phytoalexin pro-
other solanaceous phytotoxins (Brooks and Watson, 1985; duced by the sweet potato (Ipomoea botatas, Convolvula-
Harbome, 1982; Stoessl, 1982). ceae) in response to the black-rot fungus, Ceratoeystis fim-
A number of sesquiterpene phytoalexins accumulate in briatum (Fig. 21.11) (Kuc, 1992). Some of the enzymes
potato (Solanum tuberosum) in response to attack by Monol- involved in the fonnation of this compound have been iso-
iniafruticola, Phytophtlwra infestans, and Alternaria solani. lated and studied (Threlfall and Whitehead, 1991). Ipo-
Among these are rishitin (32), lubimin (33), phytotuberin meamarone is not produced by healthy sweet potatoes
(34), and several other sesquiterpenes (Brooks and Watson, (Brooks and Watson, 1985; Coxon, 1982; Threlfall and
1985; Kuc, 1992). Cell walls of P. ilifestans contain an elici- Whitehead, 1991), but is thought to be elicited by weevil-
tor of phytoalexin accumulation in potato (Kuc, 1982). Simi- derived polygalacturonase activity (Croteau and Johnson,
lar phytoalexins are synthesized by a number of other solana- 1985). Although sweet potatoes are an importaut food for
378 Sesquiterpenes

pbytotuberin (34) rishitin (32) phytotuberol bentigossypol (39)

lubintin (33) solavetivone anhydro-jl-rotunol aubergenone (36)

01 .~ .. ?Y, . ~" ~
~~~C~H
o.
debneyol (37) capsidiol (31) ipomeamarone (38) .ubergenol (35)

Fig. 21.11. Sesquiterpenoid phytoalexins.

humans and livestock, approximately 35% of the annual crop Other phytotoxins contain the 12,13-epoxytrichothecene
is lost by spoilage. Phytoalexins, such as ipomeamarone, nucleus (such as 45). Most compounds of this group are
often are produced; these compounds are hepatotoxic. Re- potent skin irritunts and are acutely toxic to mammals at low
lated compounds produce lung edema. The actual levels of concentrations. Trichothecenes are powerful mycotoxins
toxins are not predictable by visual examination, and normal and often have been implicated in fatal human and animal
cooking does not reduce the levels of bioactive compounds diseases (Fenwick and Morgan, 1991; Stoessl, 1981). Ali-
(Beier and Nigg, 1992). mentary toxic aleukia, caused by trichothecenes in cereal
A number of sesquiterpenoid phytoalexins related to gos- grains, killed tens of thousands of people in the former Soviet
sypol (29, Fig. 21.14) have been isolated from members of Union, especially in the aftermath of World War II (Fenwick
the genus Gossypium (Malvaceae) (Coxon, 1982; Gershen- and Morgan, 1991). T-2 toxin (46) from moldy brewers'
zon and Croteau, 1991). These compounds inhibit germina- grain has produced death of livestock as well as humans
tion of conidia of the fungus responsible for verticillium wilt when consumed. At least 40 beer drinkers in Quebec, the
in cotton at levels well below that of fungicidal activity United States, and Belgium were killed when moldy grain
(Mace et al., 1985). The principal phytoalexin produced is was used to prepare beer (Beier and Nigg, 1992). Other trich-
hemigossypol (39) (Brooks and Watson, 1985). othecenes have been reported to possess antitumor activity
(Cordell, 1978). Similar compounds have been isolated from
Pbytotoxins Fusarium, Myrithecium, and Trichothecium species (Ballio,
1981;· Herout, 1973).
Prehelminthosporal (40), a phytotoxin produced by the The cyclases from trichodiene systems have been exam-
fungus Helminthosporium sativum, causes necrosis in cereal ined (Cane, 1990).
grains (Fig. 21.12) (Ballio, 1981; Stoessl, 1981). Another Trichothecenes and related sesquiterpenes have been iso-
compound, helminthosporal (41), probably an artifact, lacks lated from the Brazilian plants Baccharis megapotamica and
significant biological activity. The related compound, hel- Baccharis cordifolia (Asteraceae) (Fig. 21.12). The com-
minthosporol, has gibberellin-like properties. The phytotox- pounds are chemically identical to fungal metabolites pro-
ins of Helminthosporium sacchari are three isomeric sesqui- duced by the soil fungi (Jarvis et al., 1981, 1985, 1988,
terpene glycosides (42-44) (Harborne, 1986b; Macko et a!., 1991). Although it had been suggested that these sesquiter-
1983). The phytotoxins of several Phoma species also are penes are taken up by the plant and chemically modified,
sesquiterpenes (Harborne, 1986b). more recent evidence indicates that they are synthesized by
 Sesquiterpenes 379

the plant (Jarvis et al., 1988). Removal of the seed coats of PLAl'IT-Al'IIl'IAL INTERACTIONS
these species precluded germination of the seeds; the addi-
tion of trichothecene roridins or baccharinoids (10- 6 fJ.g/ml) Plant aIlomones
resulted in germination (Kuti et al., 1990). these com-
pounds had pronounced anticancer activity. Only two Many plants produce sesquiterpenes that are inhibitory
species of Baccharis (out of 100 tested) contained these com- to growth or repellent to at least some groups of insects (Fig.
pounds. 21.13) (Hedin et al., 1974; Kelsey et al., 1984; Mabry and
Trichothecenes are potent mycotoxins. In the period fol- Gill, 1979). Several examples from the literatnre are dis-
lowing the Vietnam War, one such compound achieved no- cussed below.
toriety as a chemical warfare agent called "yellow rain" Several sesquiterpene antifeedants, such as the drimane
(Jarvis et al., 1981, 1985). However, traces oftrichothecenes dialdehydes, warburganal (47) and polygodial (48), have
are common in natnre, making identification of the source been isolated from the East African trees Warburgia stuhl-
difficult. mann;; and W. ugandensis (Canellaceae) (Gershenzon and
Mass spectrometry has been useful for the characteriza- Croteau, 1991). Polygodial (48) also appears to be wide-
tion of compounds of this group (Plattner et al., 1989). spread in the genus Polygonum (Polygonaceae). Warburga-
A number of other sesquiterpenoid phytotoxins and my- nal is a less general antifeedant than azadirachtin (see Chap-
cotoxins have been reviewed (Herout, 1970, 1973; Stoessl, ter 25) and is active against the army worm (Spodoptera
1981). littoralis), but it does not deter locust feeding. Warburganal

HO
~. . ~H'&
0
CHO
~HO
R
helminthosporal (41)
prehe1minthosporal (40) an artifact of workup

rl~o/OH
OH

-::?"Y--f
o
(CH'hCHCH,lLo:u

CH,COO OCOCH,
a trichothecene (45) T-2 toxin (46)

H H

~0"
o OH

) 0

O~
Ol'
,. 011' 10'

OH
3'
S'
4'
H
8'
17,
.'
o
H
baceharinoid 3 roridin E roridinA

RO (y~ RO'M RO M

YJnOR ~OR ~OR

(42) (43) (44)

Helminthosporium sacehari pbytotoxins

R = gal(f) (1 ---->5) gal (f)
Fig. 21.12. Some sesquiterpenoid phytotoxins.
380 Sesquiterpenes

~'
OCOCH3

shiromodial diacetate (SO) lasadiol angelale (51) caryophyllene oxide (52)

warburgaoal (47) polygodlal (48) muzigadial (49)

Fig. 21.13. Sesquiterpene antifeedants.

is highly cytotoxic (0.01 fLglml against a KB cell culture and other herbivores and is responsible for several toxicity
line) and antifeedant (0.1 ppm/cm2 against the African anny effects when cottonseed meal and other cotton products are
worm). Warburgana1 (47) and muzigadial (49) are toxic to utilized by humans and livestock (Gershenzon and Croteau,
snails that are vectors for schistosomiasis at 5-10 ppm (2 1991). Gossypol is a major component of the resistance fac-
h) (Famsworth et aI., 1987; Fraga, 1991; Henderson et aI., tors of cotton; low gossypol lines are more susceptible to
1987). These compounds have a hot taste to humans and insect attack. An array of CIS' C2S , and C30 compounds are
the plants are used as a spice (Harbome, 1982; Kubo and found; all are structuraJly related to gossypol. The Cos com-
Nakanishi, 1977; Mabry and Gill, 1979). A sesquiterpene pounds are toxic to Helicoverpa species. These C2S com-
dialdehyde blocks the sugar receptor of S. exempta for 10-20 pounds appear to arise by the addition of ClO units to gossy-
min after two contacts of 2 min (Stadler, 1984). pol or related compounds (Fig. 21.14) (Mabry and Gill,
Polygodial (48) also occurs in the mantle of a dorid nudi- 1979). Caryophyllene oxide (52), another sesquiterpene
branch Dendrodoris limbata and has antifeedant properties present in the pigment glands, also is toxic to the tobacco
against a number of marine and freshwater fish. This sesqui- budworm. However, this compound synergizes the effects
terpene defensive compound is synthesized by the nudi- of gossypol against this species. The boll weevil, a specialist
branch, in contrast to the protective compounds of most nu- on cotton, is insensitive to gossypol at the concentrations
dibranchs that are taken from the sponges on which they in any common cotton cultivars (Gershenzon and Croteau,
feed (Harbome, 1986b). 1991).
Other antifeedants such as shiromodiol diacetate (50) Myoporum deserti (Myoporaceae), a widespread Austra-
have been isolated from Parabenzoin trilobum (syn. Lind- lian shrub, has been responsible for serious livestock losses
era, Lauraceae). These compounds are active against feeding in that region. These plants cause liver and kidney lesions.
by polyphagous and oligophagous insects at the 0.05-0.1 % Mydesmone (53) is one of the active compounds (Fig. 21.15)
level in artificial diets (Mabry and Gill, 1979; Munakata, (Mabry and Gill, 1979).
1977). Famesol (5) has been demonstrated to be a feeding deter-
Leaf cutter ants, abundant from Texas to Argentina, are rent for gypsy moth larvae (Lymantria dispar ) (Doskotch et
polyphagous herbivores, but will not attack several plants. aI., 1980; Seigler, 1983).
The ant, Alta cephalotes, for example, does not feed on Lasi- A number of sesquiterpenes (as well as a variety of other
anthaea fruticosa (Asteraceae). The active repellent sub- terpenes) have been isolated from marine algae. These differ
stance is lasidiol angelate (51) (Wiemer and Ales, 1981). in large part from those of terrestrial plants (De Rosa, 1991;
Three of four other inhibitory compounds to ant feeding, Hay and Fenical, 1988). A number of sesquiterpenes from
nerolidol (7), caryophyllene epoxide (52), kolavenol (a diter- the green alga Cau/erpa ashmeadii appear to strongly inhibit
pene), and caryophyllene (not active), either produced dra- herbivory by grazing fish and some invertebrates in tropical
matic effects on the ants or the fungus cultivated by the ants waters (Fig. 21.16) (De Rosa, 1991; Paul and Fenical, 1987;
(Howard et aI., 1988). Paul et aI., 1987). Both sesquiterpenes such as flexilin (54)
Cotton plants bear glands that contain gossypol (29), a and diterpenes such as trifarin occur in Caulerpa species.
dimeric cadinene-type sesquiterpene. This compound is as- These two metabolites contain the 1,4-diacetoxy-1,3-diene
sociated with resistance to bollworm larvae, rabbits, rodents, (his-enol acetate) moiety, which is now known to be wide-
 Sesquiterpenes 381

spread among compounds derived from green algae (Paul ~-bergamoten-12-oic acid (61), from leaf exudates of Lyco-
and Fenical, 1987). Algae of the family Udoteaceae have persicon hirsutum are the principal oviposition stimulants
been shown to contain similar sesquiterpenoid compounds for Helicoverpa zea in the leaves of this wild tomato species
such as rhipocephaiin (55) and rhipocephanal (56) (Paul and (Coates et al., 1988; Douglass et al., 1993).
Fenical, 1987). Several of the sesquiterpenes and diterpenes
of these algae contain acetylenic linkages. Many of these Synomones
terpenoid compounds are active in a variety of bioassays
and are quite likely responsible for the antiherbivore proper- Many essential oils contain sesquiterpenes that are in-
ties of many green algae toward fish and sea urchins (Paul volved in pollination and seed and froit dissemination. One
and Fenical, 1987). Terpenoid compounds appear to limit interesting example is the pollination of the orchid genus
herbivory by large mobile herbivores, but not against the Ophrys by wild solitary bees of the genus Andrena. The
small and generally sedentary polychaetes and amphipods shape and color of the orchid flower closely resemble that of
that may indirectly be protected by the compounds (Hay et the female bee. The male descends on the plant performing
aI., 1988). "pseudocopulation" and pollinating the flower in the pro-
A number of halogenated sesquiterpenes [e.g., dehy- cess. It is thought that the visual lure of the orchid flower
drochloroprepacifenol (57)] and related compounds (usually shape is closely associated with an olfactory attraction and
with chamigrane skeletons) have been isolated from red that the orchid scent mimics the sexual odors of the female
algae of the genus Laurencia (De Rosa, 1991). bee, ensuring pollination by the male (Borg-Karlson, 1990).
(E,E)-Farnesol (5) and several monoterpenes are major at-
Kairomones tractants, especially in one group of Ophrys species (section
Fusci-Iuteae) and to certain bees of the genus Andraena.
Many sesquiterpenoid plant-derived kairomones have The major component of the flower extracts of O. insectifera
been reported (Hedin et al., 1974; Kelsey et al., 1984). In was cyclosativene (62). "y-Cadinene (21) is the major odorif-
addition to monoterpenes, many sesquiterpenes that are not erous compound of orchids of Ophrys section Fusci-Iuteae).
highly oxygenated also are found in essential oils. Several T -Muurolol (63) and cubenol (64) were present in section
are known to possess kairomonal activity. (+ )-a-Copaene Fusci-Iuteae, as well as in O. tenthredinifera and O. bombyli-
(58), from orange froits, is an attractant for the Mediterra- flora. (E,E)-Farnesol (5) and the related esters farnesyl ace-
nean froit fly, Ceratitis capitata (Teranishi et al., 1987). a- tate and farnesyl octanoate were found ouly in O. lutea, O.
Farnesene (59), from the skin of several varieties of apple sph. litigiosa, and, in traces, in O. insectivera (Borg-Karlson,
and pear froits, attracts newly hatched larvae of the codling 1990). Borg-Karlson (1990) has suggested division of the
moth, Laspeyresia pomonella, a serious pest of pome froits genus into six groups based on the composition of floral
(Jacobson, 1982; Sutherland et al., 1977). volatiles.
( + )-(E)-a-Santalen-12-oic acid (60) and ( + )-(E)-endo- Several pollinator species do show a typical sexual behav-

CHO OH HO CHO
HO OH

HO

r
gossypol (29)

HO~CHOOH
CHO 0
HO~CHOOH
9"
~ I I
"'" HO~I
l Hi 9" I I
HO HO ""
HO ""
o o
o
I
hellocideHl beliocideH4
heliocideHl

Fig. 21.14. Gossypol and related terpenoids from cotton.

382 Sesquiterpenes

mydesmone (53) farnesol (5)

~I::'" CH,O-hexanoyl
~ (a monoterpene)

farnesyl hexanoate geranyl hexanoate

female Arulrena scents

~~~'¥8
(E,E)-a-farnesene (59)

r(--
(+)-(E)-a-santalen-12-oie add (60) (+)-(E)-endo-8-bergamoten-12-oie add (61)

H OH OH!

~~. ~ ~
cyclosativene (62) T·muurolol (63) cubenol (64)

Fig. 21.15. Sesquiterpene allomones, kairomones. and synomones.

ior when exposed to the odors of flowers or female insects This compound is also known from the tree Torreya nuci-
(Borg-Karlson, 1990). Compounds such as (E,E)-famesol lera, a gymnosperm.
(5) and primary aliphatic alcohols have been demonstrated Blister beetles (family Meloidae) contain considerable
to be the most important compounds in the pollinating sys- amounts of cantharidin (66) (sometimes known as Spanish
tems of Ophrys sections Fusci-luteae and Araneiferae. fly), a powerful irritant once thought to possess aphrodisiac
Ophrys flowers do not appear to compete with virgin females activity. This compound is synthesized by male beetles and,
or males. Rather, they produce a set of "second class attracti- during'mating, is transferred to the female. Mating stimnlates
vity compounds" that successfully attract only a fraction of de novo synthesis of the compound by the male beetle. The
the male population. Many of the behavior-releasing com- lethal dose in man is about 0.5 mg/kg of body weight and
pounds also are present in other flowers and insects. The cantharidin has been responsible for human fatalities (Cutler,
possibility of chemical mimesis has been suggested (Har- 1992). The beetle releases the compound by reflex bending
borne, 1982; Kullenberg and Bergstrom, 1975, 1976). from the knee joints and cantharidin appears to act as a feed-
ing deterrent to predators of the insect. The insects contain
Il'ISECT SEl'UOCHEl'UCALS 0.2-2.3% of the compound. Famesol serves as a precursor
for cantharidin, whereas geraniol does not (Fig. 21.17)
A1lomones or Defensive Compounds (Beale and MacMillan, 1988; Cane, 1981; McCormick and
in Insects Carrel, 1987; Peter et al., 1977a,b. Meloid beetles contain
Dendrolasin (65), an acyclic sesquiterpene, is known as sufficient cantharidin to be toxic to livestock (Capinera et
a defensive compounds from ants of the genus Dendrosius. al.,1985).
 Sesquiterpenes 383

Insect Pheromones trichomes that liberate (E)-J3-famesene (in addition to a

sticky exudate produced by other types oftrichomes). How-
Most sexual pheromones of insects appear to be derived ever, it is not known whether this substance is produced by
from either fatty acid or mevalonic acid metabolism. In a the plant as a defense against aphids (Harborne, 1986b).
number of instances, modified monoterpenes or sesquiter- (Z,E)- and (E,E)-a-famesene (59) and related homoses-
penes play roles as aggregation pheromones (Prestwich and quiterpenes have been reported to serve as trail pheromones
Blomquist, 1987). for the imported fire ant of the southern United States
Germacrene D (67) provides an active mimic of the (Vander Meer, 1983).
American cockroach pheromone, periplanone B (68) (Bow-
ers, 1985).
Insect Juvenile Hormones and Plant
The hair pencil (an extrusable brush-like structure that
Juvenile Hormone Jllimics
serves for pheromone dissemination during courtship) secre-
tion (69) of the Monarch butterfly (Danaus plexippus) con- The juveuile hormones of many insects are sesquiter-
sists of sesquiterpenes (Herout, 1970) (Fig. 21.17). The sex- penes (Bowers, 1991, 1992; Fraga, 1991; Law, 1983; Menn
ual pheromones (69 and 70) of this insect also appear to be and Beroza, 1972).
terpenoid in origin. The first knowledge that plants produce mimics of insect
The alarm pheromones of aphids often are sesquiterpenes juvenile hormones was acquired largely by accident. When
such as (E)-J3-famesene (71) and germacrene A (72) (Bow- the Czechoslovak entomologist, Karel Shima, came to Har-
ers, 1985). Upon release of these compounds, aphids drop vard University, he tried to cultivate his research organism,
from the plant on which they have been feeding. The com- the European bug, Pyrrhocoris apteris. Strangely, the insect
pounds are very labile and break down in only a few minutes. refused to grow normally and obstinately remained in the
This is important to prevent the aphids from being in a con- fifth larval stage. After some time, it was discovered that
stant state of alarm (Bowers, 1985). Although this sesquiter- the filter paper on which the bugs were grown had been
pene elicits response in many species of aphids, the turnip replaced with paper towels (made in the United States).
aphid Lipaphis (Hyadaphis) erysimi responds only weakly. When the towels were again replaced with filter paper, the
Addition of plant-derived isothiocyanates to J3-famesene has insects began to grow normally. Further study revealed that
a synergistic effect and greatly enhances the effect of 13- U.S. newspapers, but not Japanese or European ones, inhib-
famesene (71) (Dawson et ai., 1987; Harborne, 1989). ited growth of the insect. American paper is largely made

c;c:y
The wild potato species, Solanum berthaultii, possesses from Abies balsamea, the balsam fir, and a compound, ju-

~~CHO
OAC

~ OA~ I
OAc I ~
Ac OAc
OAc
caulerpenyne

CY "'.~ ~rno
~ ~~ I\.~
~~ CHO

CHO OAc onchidal

~
CI
'" '" I
tlexilin (54) OAc

rhipocephalin (55)
OAc dehydrochloroprepaclfenol (57)

Fig. 21.16. Sesquiterpenes from marine plants.

384 Sesquiterpenes

HO~C02H

E,E·I0·hydroxy·3,7~dimethyl~2,6·decadienoic acid

HOC~C02H H02C~CH20H
2

(69) (70)
monarch butterny pheromones

E·p·farnesene (71) germacrene A (72)

germacrene D (67) periplanone B (68)

~o
dendrolasin (65)

crt)o
o

~opp_~:_
- 0
o
portions of the farnesyl opp cantharidin (66)
molecule that are incorporated
into cantharidin

Fig. 21.17. Insect pheromones and allomones.

vabione (75), was isolated from the paper. This compound have been isolated from plants. Juvenile hormone III (76),
appears to be active only against one family of insects. the a juvenile honnone found in insects, also occurs in leaves
Pyrrhocoridae (Slama and Williams, 1965). of the sedge Cyperus iria accompanied by methyl (2E, 6E)-
Another example of a sesquiterpene-derived insect juve- famesoate. Nymphs of the grasshopper, Melanoplus sangui-
nile hormone is that of the giant silkworm moth, Hyalophora nipes, that fed on Cyperus iria grew at a similar rate to
cecropia (Fig. 21.18). Several juvenile hormone mimics controls, but showed pronounced morphogenetic effects
have been found in plants (Bowers, 1985, 1991, 1992; Fraga, when they molted to adults (Harborne, 1989).
1991; Menn and Beroza, 1972). Compounds with anti-juvenile-hormone activity have
When juvenile hormone is present in excess, no metamor- been found in Ageratum houstonianum, Asteraceae (see
phosis occurs and adult insects are not formed. Under natural Chapter 18). In this case, the insects molt precociously and
conditions, synthesis of the hormone ceases at the end of the produce adults one or two stages too early. The females
juvenile stages of development and metamorphosis occurs. produced usually are sterile.
Although feeding studies indicate that (73) incorporates
three molecules of MVA, (74) incorporates only two and ESSBNTIAL OIL COMPOl'lBl'ITS
probably (77) only one. Compound (74) incorporates 1 mole
of propionate and 8 moles of acetate. These cumpounds are Among the most prized of essential oils are those of sandal-
synthesized by the insect. The hormone is synthesized in wood, which contains the santalenes (78 and 79), and pa-
the corpora allata glands, which regulate its release into the tchouli, which contains patchouli alcohol (80) (Fig. 21.19).
blood. Other components such as j3-ionone (81) (actually a tetrater-
Other types of compounds with juvenile hormone activity pene or carotenoid degradation product) and ct-( - )-bisabo-
 E"
Sesquiterpenes 385

•~ Opp

o O~ ---..

homomevalonate

(77) mevalonate

~C02C"~C02C",
(73) (74)

~C02C"3

farnesol (5) juvabione (75)

~C02C",

juvenile hormone I juveniJe hormone II

juvenile hormone III (76)

Fig. 21.18. Insect juvenile honnones and juvenile honnone mimics (modified from Jennings et al., 1975; modified and used with pennission of the
copyright owner, the Royal Society of Chemistry, Cambridge),

101 (82) are encountered in flower odors and are important biaceae (Hyenanche), and Orchidaceae (Dendrobium)
in pollination as well as for their commercial importance as (Cane, 1981; Coscia, 1969). One of these compounds, melli-
perfumery ingredients. toxin (85), is a contaminant of honey in New Zealand, where
the source plant Coriaria arbarea is common. The plant
SWEETEl'IEKS contains the sesquiterpene tutin (86). A leaf hopper (Scaly-
papa australis) that feeds on the plant is believed to hydrox-
The sesquiterpene hernandulcin (83) from Lippia dulcis ylate tutin and excrete the metabolite in its honeydew. Bees
(Verbenaceae), a plant used as a sweetener by the Aztecs, collect this material and use it for making honey. Mellitoxin
is more than 1000 times sweeter than sucrose (Compadre et also is known to occur in the euphorbiaceous plant Hyen-
ai., 1985, 1987). anche glabasa (Coscia, 1969).
A series of alkaloids belonging to this group of sesquiter-
PICKOTOXINS penoid compounds occurs in the genus Denbrabium of the
Orchidaceae (Cane, 1981; Coscia, 1969). Denbrobine (87)
A complex type of sesquiterpenes known as picrotoxins are is one of the most common of these alkaloids.
highly toxic and contain a lactone ring. Pictrotoxins, such This series of compounds is derived from copaborneol
as picrotoxinin (84), are found in the Menispermaceae (Men- (88) (Newman, 1972). Copaborneol may arise via the bioge-
ispermum, Cocculus, and Anamirta), Coriariaceae, Euphor- netic scheme proposed by Coates (1976) (Fig. 21.20).
386 Sesqu;terpenes

(78) (79) (80)

a·H·bisabolol (82) p~ionone (81) hernandulcin (83)

Fig. 21.19. Sesquiterpenes and other essential oil components.

SESQUITERPEl'IE LACTOl'IES macranolide lactones are, by far, the most common type
(Fischer, 1990, 1991b). The majority of sesquiterpene lac·
Most of the approximately 3500 known sesquiterpene lac· tones from higher plants contain a·methylene''Y·lactone
tones occur in members of the Asteraceae. This family, in groups in which H·7 is, without exception, a·oriented. How·
tum, may be the largest of all plant families, with at least ever, several sesquiterpene lactones from liverworts possess
1443 genera and 32,160 species (Hendrych, 1985). These enantiomeric compounds (Fischer, 1990).
compounds occur only infrequently in lower plants and in The methods for isolation, purification, and characteriza·
a few other higher plant families (Fischer, 1989a, 1989b, tion of sesquiterpene lactones have been reviewed (Fischer,
1990, 1991a, 1991b; Gershenzon and Croteau, 1991; Pic, 1991b; Yoshioka et al., 1973).
man, 1986a; Seaman, 1982; Yoshioka et al., 1973). Sesqui·
terpenes may constitute up to 5% of the dry weight of the
plant. Because of their taste properties, sesquiterpene lac· BIOOEl'IESlS
tones are sometimes referred to as bitter principles (as are
many other types of bitter·tasting compounds). Sesquiter· Biogenetic schemes for most of the major groups of sesqui·
pene lactones may occur throughout a plant, but they are terpene lactones have been outlined (Fischer, 1990; Seaman
most commonly associated with leaves and flower parts and 1982). Based on stmctural information, many biomodifica·
glandular trichomes (Gershenzon and Croteau, 1991; Mabry tions in the advanced biosynthetic stages of sesquiterpene

fu'
and Gill, 1979; Picman, 1986a; Yoshioka et al., 1973). Ger· lactones involve oxidative reactions. The isolation of a large

+ •

. fi
'R
A

copaborneol (88) picrotoxinin (84) tutin (86) mellitoxin (85) dendrobine (87)

Fig. 21.20. Biogenesis of picrotoxins (modified from Coates. 1976; used with pennission of the copyright owner, Springer-Verlag, Vienna).
 Sesqu;telpenes 387

ct\- ct\- ct:)f-

cbr-c6y- ~.·. " .". . CHO
6oxygeuatioJl
~ CO,H

rYL:;~oo~
~~~~
py~ 17
H ---+.......
17
H
Co,H
(89) +
germacrene A (13) germacranolides 0 0

Fig. 21.21. Proposed biogenesis of lactone ring of sesquitezpene lactones (modified from Geissman and Crout, 1969).

number of sesquiterpene lactones containing epoxide groups species), Bombacaceae, Burseraceae, Canellaceae, lllicia-
and the natural occurrence of hydroperoxide-bearing sesqui- ceae, Lamiaceae, Lauraceae, Maguoliaceae (5 species),
terpenes suggests the involvement of epoxides in sesquiter- Menispermaceae, Polygonaceae, and Winteraceae (Fischer,
pene cyclizations and singlet oxygen-type reactions in allylic 1991b; Herz, 1977; Picman, 1986a). Some reports for ses-
oxidations (Fischer, 1990). Initial cyclization of E,E-farnesyl quiterpene lactones in the Scrophulariaceae are based on
pyrophosphate to germacrene A (13) yields a compound in misinterpretation of Veronica (Scrophulariaceae) for Ver-
which all nonolefinic protons (except. at C-8) are activated nonia (Asteraceae). A number of other families contain lac-
for allylic oxidations. Conversion of the isopropenyl double tones described previously under picrotoxins.
bond to an epoxide, followed by opening of the epoxide and The distribution of sesquiterpene lactones often is re-
oxidation to an acid group (or by an alternative oxidative ferred to as an indication of affmity between the Asteraceae
pathway) would give an acid (89) that, after introduction of and the Apiaceae (Hegnauer, 1977). These two families
oxygen into the 6- or 8-position, can serve as a precursor for share polyacetylenes, coumarins, and flavonoids, as well as
germacranolides (Fig. 21.21). Many biosynthetic questions, other types of compounds. The stereochemical features of
especially those related to the sequence of events, remain the sesquiterpene lactones found in these families provide
open (Fischer, 1990). additional support for the relationship (Holub et al., 1987).
Although many details of the biosynthesis of sesquiter- The occurrence of sesquiterpene lactones in the tribes of
pene lactones are not established, guaianolides, eudesmanol- the Asteraceae is of taxonomic interest. These compounds
ides, and pseudoguaianolides in the Asteraceae appear to are common and complex in the Heliantheae (including
be derived from germacranolide precursors (Fischer, 1978, some Helenieae); of intermediate frequency in the Anthemi-
1990; Fischer et al., 1979). Biomimetic reactions leading to deae, Cichorieae, Cynareae, Senecioneae, Inuleae, and Ver-
the formation of these and other types of sesquiterpene lac- nonieae; infrequent and biogenetically less advanced in the
tones have been proposed (Fischer, 1990). Eupatorieae; and not known in the Mutisieae, Astereae, Ta-
geteae, and Arctoteae-Calenduleae (Dahlgren et al., 1981;
CtreIIIOSYSTEJllATIC STUDIES INVOLVIl'IO Waterman and Gray, 1987).
SESQUflERPEl'IE LACTOl'IES Structural studies of sesquiterpene lactones have proven
useful for understanding systematics and evolution in the
Sesquiterpene lactones are known to occur in the following genus Ambrosia (Asteraceae). Populations of Ambrosia psi-
families of angiosperms: Acanthaceae, Amaranthaceae, Api- lostachya from the coastal islands off Texas produce only
aceae (12 species), Aristolochiaceae, Asteraceae (about 450 the dilactone compounds psilostachyin (90), psilostachyin B
388 Sesquiterpenes

o~o~o~
p.nostacltyin (90) p.Uostachyin B (91) 0 psUostachyin C (92)

HO~ ~'H ~~H

HO 0 0
o 0 0
0 0 0
ambroslol (%) coronopilin (93) parthenin (94)

~~ ~oo nG-.
~ o
~O~~O 0
confertiftorin (95) desacetylconfertiflorin (99) tomentosin (133)

Fig. 21.22. Sesquiterpene lactones of systematic significance.

(91) and psilostachyin C (92). Collections of the same spe- Ambrosia chamissonis is similar to most other species of
cies from the adjacent mainland contained only monolac- ragweeds in that it synthesizes a number of sesquiterpene
tones, primarily ambrosiol (96), coronopilin (93), and par- lactones-in this case, a series of structurally related genna-
thenin (94) (Fig. 21.22). ht one area of south Texas with cranolides. This species also is extremely variable in tenns
ecology quite similar to the islands, dilactone populations of morphology as well. Ambrosia chamissonis occurs on the
also were encountered. It has also been observed that a re- Pacific coast of both North America and South America
lated species, Ambrosia cumanensis, from Mexico contains (Chile) where it was apparently introduced about 100 years
the same three dilactones. There appears to be a close rela- ago. Five morphological groups have been recognized.
tionship between the coastal populations of A. psilostachya North of San Francisco, there is little correlation of chemis-
and A. cumanensis of Mexico. It is quite probable that the try and morphology, whereas south of that point, there is a
Mexican plants served as ancestors for the island races which greater relationship. It has been suggested that the southern
were established 3000-5000 years ago (Mabry, 1970). Stud- populations have been more recently introduced from the
ies of the volatile essential oils confirmed this relationship. north and a founder effect still exists. Furthennore the chem-
Populations of Ambrosia confertiflora, a ragweed distrib- istry of the populations from Chile is consistent in sesquiter-
uted widely through Texas and Mexico, reveal complex ses- pene lactone chemistry and leaf morphology. This illustrates
quiterpene lactone chemistry. Most populations from south- how a weedy species can rapidly colouize a new region; all
central Texas are characterized by a chromosome number n of the Chilean plants may have come from a single introduc-
= 66 and the pseudoguaianolides confertiflorin (95) and tion from California. Both the chemistry and morphology
desacetylconfertiflorin (99), whereas plants from northeast- are quite sintilar to those populations near San Francisco,
ern Mexico yield germacranolides and a few eudesmanol- suggesting that the source of the genetic line introduced into
ides. All of those studied have chromosome number n = Chile may have come from that area (Mabry, 1973).
54. A dilactone race (with both n = 54 and n = 66) occurs
further west in Texas and in Mexico along the western slopes BIOLOGICAL AC11VITY
of the Sierra Madre into the area where Ambrosia cuma-
nensis occurs. A fourth race characterized by the presence A number of sesquiterpene lactones are known to have var-
of gennacranolides occurs in northwestern Mexico (Mabry, ious types of biological activity (Bohlmann, 1986; Fischer,
1972). 1991 b; Mabry and Gill, 1979; Picman, 1986a; Rodriguez et
 Sesquiterpenes 389

al., 1976; Stevens, 1984). Part of the cytotoxic, allergenic, antagonist for auxin. Heliangin (102) from Helianthus tub-
and antibiotic properties of this group of compounds are erosus also is known to be an antagonist in the Avena coleop-
attributable to the fact that they can form Michael addition tile test. Vemolepin (103) from Vernonia hymenolepis has
products by the addition to active-site sulfhydryl groups in been reported to be the strongest inbibitor of auxin. It has
key enzymes, as well as desoxyribonuclease and ribo- been suggested that vemolepin affects an essential step in
nuclease (Hall et al., 1977). This property is dependent on cell wall synthesis or extensibility and is not merely a toxic
the presence of the a,l3-unsaturated "V-lactone moiety. Be- principle (picman, 1986a).
cause of their taste properties, sesquiterpene lactones are
sometimes referred to as bitter principles. Cnicin (97) from Allelopatbic Effects
Cnicus benedictus and absinthin (98) from Artemisia ab-
sinthe are the most important bitter principles of the bitter Many plants with sesquiterpene lactones have been sus-
liqueurs made from these plants. Lactucin (100) is the bitter pected to possess allelopathic properties. An American plant,
principle of the dried juice of wild lettuce or Lactucarium Parthenium hysterophorus, was introduced into Australia
(Fischer, 199Ib). A number of Asteraceae and some liver- and India and has subsequently been reported to affect nu-
worts of the genus Frullania are known to produce allergic merous crop plants (Picman, 1986a). The compounds par-
contact dermatitis in humans (Fig. 21.23) (Evans and thenin (94) and coronopolin (93) probably are responsible
Schmidt, 1980; Picman, 1986a). for this activity. Interestingly, the pollen of this plant, which
also contains these compounds, appears to be responsible
Plant Growth Regulator Activity for the inbibition of pollen germination and pollen tube
Xanthinin (101), a sesquiterpene lactone from many Xan- growth on stigmas of other species (Picman, 1986a).
thium (cocklebur, Asteraceae) species, is known to be an The compound arbusculin-A (104), from Artemisia spe-

~ ~
AC

d>-~
O_dgIYI 0

HO.&I 0

/fJ~o o
o
o
hecagenin xanlbinln (101) beJcnaJin (Il2)

~
'!
o
~I!!
00 o
i

HO
-...'0

0
0
HO
~
0
H'
0

tenoon (Ill) ambrosin arbusculin-A (104)

#' ,
01'11 0H H0tf>-~ ~
lrn1-{ -Yul~o
~ o
vernolepin (103) bymenovin (Il3) parthenin (94)

0-S-
~
o ~
o ~
o OAc
o
gJaucolide-A (Il4) ainntoJactone (lOS)
(al
Fig. 21.23 (8 & b). Some sesquiterpene lactones with biological activity.
390 Sesquiterpenes

~~ ~
O!,
- .
00

o 0 HO ~
eremanthine (U8) costunolide (119) - , go~ze"oHde (120)

~~
~ :0-~o
~~ o

:~:<~( pM
santamarin (123) reynosin (124) plenolin (131)

e1epbantin (132) vernolide (127) yingzbaos. A (129)

(b)

Fig. 21.23. (continued)

cies, is responsible for some of the allelopathic effects of Sesquiterpene lactones from Parthenium species were
that plant (Fig. 2 I .22) (Mabry and Gill, 1979). Alantolactone strongly inhibitory to growth of larvae of Helicoverpa zea
(105), found in many members of the Asteraceae, inbibits and Spodoptera exigua (Isman and Rodriguez, 1983). Tenu-
seed germination and growth of the common weedy species lin (111), the major sesquiterpene lactone of Helenium
Amarathus retroflexus (Amaranthaceae) and Chenopodium amarum, imparts a bitter taste to milk and is toxic to live-
strictum var. glaucophyllum (Chenopodiaceae) (picman, stock. Helenalin (112) from Helenium microcephalum is
1986b). Other examples of allelopathic effects have been toxic to several mammalian species in oral doses as low as 85
summarized by Picman (1986a). jJ.glkg. Hymenoxys odorata, from which a toxin, hymenovin
The sesquiterpene lactones burrodin (106), confertiflorin (113), is isolated, is economically siguificant, as it is very
(95), desacetylconfertiflorin (99), dihydroparthenolide toxic to sheep and goats. A similar type of poisoning has
(107), parthenin (94), and 7oc-hydroxy-3-desoxyzaluzanin C been observed with South African species of Geigera.
(108) inbibited or promoted the germination of seeds of 16 Three species of Vernonia that grow in the southeastern
dicot and 9 monocot species, depending on concentration United States differ in their sesquiterpene lactone content.
and species at concentrations as low as 1 r.W1 (Fischer et Vernonia gigantea and V. glauca contain glaucolide-A
aI., 1989a). A sesquiterpene lactone (109) from Iva axil/aris (114), whereas the third, Vernonia flaccidifolia, produces
seeds inbibited germination of Abutilon theophrasti seeds at almost none of these compounds. The insect-feeding-deter-
10- 3 M. Although axivalin (110) did not inhibit seed germi- rent properties of these compounds were studied with six
nation, this compound profoundly affected growth of seed- insects, all of which occurred in the same area as the plants.
ling radicles (Spencer et aI., 1984). Three of these insects, the yellow-striped armyworm
(Spodoptera ornithogalli), cabbage looper (Tricoplusius nil,
Plant Allomones and the yellow wooly bear (Diacrisia virginica), feed in
nature on Vernonia species that contain glaucolide-A. A
Many sesquiterpene lactones have antifeedant properties fourth species, the saddle-back caterpillar (Sibine stimulea),
against animals. Numerous compounds have been evaluated feeds on V. flaccidifolia. The polyphagous southern army
against insects, especially economically important pests. worm (Spodoptera eridania) and fall armyworm (S. [rugi-
These reports have been summarized and many of the spe- perda) occur in the same area, but normally are not seen on
cific effects reviewed (Fischer, 1991b; Picman, 1986a). Vernonia species. In feeding tests, the fall, southern, and
 Sesquiterpenes 391

yellow-striped annywonns and the saddle-back caterpillar (Yoshida et aI., 1976). The molluscicidal activity of sesqui-
all avoided Vernonia species with glaucolide-A. The wooly terpene lactones has been investigated (Marchant et aI.,
bear and cabbage looper seemed to prefer the plants that 1984).
contain glaucolide-A (114). ht feeding trials with young in- Several members of the Asteraceae such as Vanillos-
sect larvae, those insects that occur on Vernonia species in mopsis, Eremanlhus, and Moquinea are widely used in Bra-
the field tended to be the least affected in trials (Mabry and zil for fencing. The wood is highly resistant to insects and
Gill, 1979). ht feeding trials, rabbits (Sylvifagusfloridana) fungi. The active compounds [eremanthine (118), costunol-
preferred V. flaccidifolia over species that had glaucolide- ide (119), and goyazensolide (120)] are sesquiterpenes (Gil-
A. Similar results were obtained with whitetail deer (Odo- bert, 1977).
coileus virginianus).
Despite the usual antifeedant or toxic effects observed Chemotactic Agents from Parasitic Plants
with other insects, larvae of the sunflower moth Homoesoma Several parasitic weeds of the genera Slriga (Scrophulari-
eleclel/um, Lepidoptera: Pyralidae), a specialist on species aceae) and Orobanche (Orobanchaceae) attack grasses,leg-
of sunflower (Helianthus), are able to feed on flower parts urnes, and other plants (see Chapter 2). Seeds that are in the
that contain appreciable amounts of sesquiterpene lactones soil remain viable for long periods of time, and they only
(Gershenzon and Croteau, 1991). germinate when a stimulant from the host plant is present.
Many sesquiterpene lac tones are toxic to fish (picman, A sesquiterpene, strigol (121), recently has been isolated
1986a). Buddlejin A, B, and C (115, 116, and 117) possess a from the roots of colton plants that effects gennination of
caryophyllane skeleton and are piscicidal (Fig. 21.24). These Striga seeds in concentrations down to the 10 - 13 M level.
compounds are found in Buddleja davidii (Buddlejaceae) A structurally similar compound from synthetic sources

0% % ~o
buddlejlo A (115) buddl_Jio B (116) buddl_jlo C (117)

o ii i
0
0
0
sanloolo (130) alnnlolo<loo_ (105)

Q=5{0 CQ=: 0~o r~

HO H
-y-o
'f0\.
yo
.
.trigol (121) (122) isoalantolactone (126)

~
o

m~
![J~o
,
h
•. ""O~
0

HO 0 0

(aJ
o
Fig. 21.14 (a & b). Additional biologically active sesquiterpene lactones.
 %.~"'O
392 Sesquiterpenes

,;Y
OH II ./
Yl
0 OH
~:7
~ ~ OH
./'
~ HO 0 O~
HO 0 0 0

enieln (97) laetucin (100) dihydropartbenollde (107)

~ ~
"!l ' O-tlgloyl
:0-0

, H """-\
o o
o o
o
abslntbin (98) artemisinin (quinghaosu) (128) heliangin (102)

(b) 7a-bydroxy-3-desoxyzaluzanin C (108) burrodln (106) axivalin (110)

Fig. 21.24. (continued)

(122) is almost as active (10-9 M). This compound also metronidazole, an antiamoebic drug (Borris and Schaeffer,
promoted gennination of Alectra (Scrophulariaceae), a para- 1992).
site of cowpeas, and several Orobanche (Orobanchaceae) Helenalin and a series of related compounds are responsi-
species. These compounds now are being tried to control ble for the cardiotonic properties of Arnica flowers. This
parasitic plants in cultivated crops (Johnson, 1978; Pep- drug has long been known for its heart stimulant and analep-
perman and Blanchard, 1985)_ tic qualities (Wagner, 1988). Helenalin (112), tenulin (111),
Sesquiterpene lactones such as the eudesmanolides san- and eupahyssopin had antiarthritic activity (50-70% inhibi-
tamarin (123) and reynosin (124) strongly enbance germ- tion of edema) at concentrations of 2.5 mglkg (Wagner and
ination of the seeds of Striga asiatica. Confertiflorin (95), Proksch, 1985). The crude drug Atractylodis rhizoma, from
desacetylconfertiflorin (99), parthenin (94), and dihydropar- Atractylodis macrocephala (Asteraceae), and several other
thenolide (107) from Ambrosia species also have high activ- related medicinal plants is used clinically as a diuretic and
ity. Sesquiterpene lactones may serve as alternatives to stri- for analgesic purposes. The plant also has anti-inflammatory
gol; these compounds are nontoxic to sorghum at the properties. Active compounds include (+ )-eudesma-4(14)-
concentrations employed (10- 9 M) (Fischer et a1., 1989b; 7(1l)-dien-8-one and atractylenolide I (Hikino, 1985).
Fischer, 199Ia). An effective antimalarial constituent, artemesinin (128),
was isolated from a Chinese medicinal herb, Artemisia
Medicinal Uses annua (Asteraceae). A promising antimalarial, but only
weakly toxic sesqniterpene lactone, was isolated from this
A number of sesquiterpene lactones show antibacterial, plant (Fischer, 1991b; Klayman, 1985, 1989; Lewis, 1992;
antifungal, and antiprotozoan activity (Fischer, 1991b; Pic- Picman, 1986a). A similar compound, yingzhaosu A (129),
man, I 986a). Alantolactone (105) and isoalantolactone (126) from Artabotrys uncinatus (Annonaceae) also has efficacy
have been used as urinary antiseptics. Several compounds as an antimalarial agent (Borris and Schaeffer, 1992).
of this group have activity against Entamoeba histolytica Santonin (130), a sesqniterpene lactone from Artemisia
and related organisms. Parthenin (94) from Parthenium hyst- maritima (Asteraceae) has been used as an ascaricide, and
eropherus and vernolide (127) from Vernonia colorata in- is especially active against roundworms (Ascaris spp.).
hibit Entamoeba histolytica at concentrations comparable to A number of sesquiterpene lactones have been demon-
 Sesquiterpenes 393

strated to possess activity against the trematode Schistosoma. Recent Advances in the Chemistry of Insect Control (N. F.
Infection by the cerceriae of this organism are responsible Janes, ed.), Special Publication No. 53, 272-292, Royal Society
for a disease called schistosomiasis. Certain snails serve as of Chemistry, London, 1985.
an intermediate host for the trematode. Several of the active BOWERS, W. S., Insect hormones and antihonnones in plants, terpe-
sesquiterpene lactones kill the vector snails and their eggs noids, in Herbivores: Their Interactions with Secondary Plant
Metabolites, Vol. 1 (G. A. Rosenthal and M. R. Berenbaum,
but are not toxic to fish (picman, 1986a).
eds.), 431-456, Academic Press, San Diego, CA, 1991.
Many sesquiterpenes have pronounced antitumor proper-
BOWERS, W. S., Insecticidal compounds from plants, in Phytochem-
ties (Bohlmann, 1986; Cordell, 1978; Picman, 1986a;
ical Resources for Medicine and Agriculture (H. N. Nigg and
Rodriguez et aI., 1976). In general, the conjugated exocycJic
D. S. Seigler, eds.), 227-235, Plenum Press, New York, 1992.
double bond in the lactone group is necessary for activity,
BROOKS, C. 1. W. and D. G. WATSON, Phytoalexins, Nat. Prod.
although at least one antitumor compound lacks this feature.
Rep., 2, 427-459 (1985).
The ability to inhibit DNA synthesis directly appears to be
CANE, D. E., Biosynthesis of sesqlliterpenes, in Biosynthesis of
a common feature among sesquiterpene lactones with cyto-
Isoprenoid Compounds, Vol. I (J. W. Porter and S. L. Spurgeon,
toxic and antitumor activity. Most of these compounds are eds.), 283-374, Wiley, New York, 1981.
from plants of the Asteraceae. Some examples are a -santo- CANE, D. E., Isoprenoid biosynthesis. Stereochemistry of the cycli-
nin (130), vemolepin (103), pJenolin (131), and elephantin zation of al1ylic pyrophosphates, Ace. Chern. Res., 18, 220-226
(132) (Mann, 1987). A summary of active sesquiterpene 1ac- (1985).
tones and references is given by Bohlmann (1986) and Pic- CANE, D. E., Enzymatic formation of sesquiterpenes, Chern. Rev.,
man (1986a). 90, 1089-1103 (1990).
CAPINERA, J. L., D. R. GARDNER, and F. R. STERMrrz, Cantharidin
levels in blister beetles (Coleoptera: Meloidae) associated with
REFERENCES alfalfa in Colorado, J. Econ. Entornol., 78, 1052-1055 (1985).
CHARLWOOD, B. V. and D. V. BANTHORPE, The biosynthesis of
ANON., Contraceptives, Chern. Br., 970 (1984). monotetpenes in Progress in Phytochemistry Vol. 5 (L. Rein-
hold, J. B. Harborne, and T. Swain, eds.), 65-125, Pergamon,
ARIGONI, Stereochemical aspects of sesquiterpene biosynthesis,
Oxford, 1978.
Pure Appl. Chern., 41, 219-245 (1975).
CHARLWOOD, B. V. and K. A. CHARLWOOD, Terpenoid production
BAERHEIM SVENDSEN, A. and 1. J. C. SCHEFFER (eds.), Essential
in plant cell cultures, in Ecological Chemistry and Biochemistry
Oils and Aromatic Plants, Nijhoff/Junk, Dordrecht, 1985.
of Plant Terpenoids (J. B. Harborne and F. A. Tomas-Barberan,
BALLIO, A., Structure-activity relationships, in Toxins in Plant Dis- eds.), Phytochemistry, Society of Europe Vol. 31, 95-132, Ox-
ease (R. D. Durbin, ed.), 395-441, Academic Press, New York,
ford University Press, Oxford, 1991.
1981.
COATES, R. M., Biogenetic-type rearrangements of terpenes,
BANTHORPE, D. V. and B. V. CHARLWOOD, The terpenoids in Sec- Fortschr. Chern. Org. Natursl., 33, 73-220 (1976).
ondary Plant Products, (E. A. Bell and B. V. Charlwood, eds.),
COATES, R. M., J. F. DENlSSEN, 1. A. JUVIK, and B. A. BABKA,
Encyclopedia of Plant Physiology Vol. 8), 185-220, Springer-
Identification of a-santalenoic and endo-f3-bergamotenoic acids
Verlag, Berlin 1980.
as moth oviposition stimulants from wild tomato leaves, l Org.
BEALE, M. H. and 1. MACMn.LAN, The biosynthesis of C.s-C20 Chern., 53, 2186-2192 (1988).
terpenoid compounds, Nat. Prod. Rep. 5, 247-264 (1988).
COMPADRE, C. M., J. M. PEZZUTO, A. D. KINGHORN, and S. K.
BEIER, R C. and H. N. NIGG, Natural toxicants in foods, in Phyto- KAMATH, Hemandulcin: An intensely sweet compound discov-
chemical Resources for Medicine and Agriculture (H. N. Nigg ered by review of ancient literature, Science, 227, 417-419
and D. S. Seigler, eds.), 247-367, Plenum Press, New York, (1985).
1992. COMPADRE, C. M., R. A. HUSSAIN, R. L. L. DE COMPADRE, J. M.
BINGEL, A. S. and H. H. S. FONG, Potential fertility-regulating PEzzUTO, and A. D. KINGHORN, The intensely sweet sesqlliter-
agents from plants, in Economic and Medicine Plant Research, pene hemandulcin: Isolation, synthesis, characterization, and
Vol. 2 (H. Wagner, H. Kikino, and N. R. Farnsworth, eds.), preliminary safety evaluation, 1. Agric. Food Chern., 35,
73-118, Academic Press, London, 1988. 273-279 (1987).
BOHLMANN, F., Structure and biological activity of compounds CORDELL, G. A., Anticancer agents from plants, in Progress in
from the Compositae, in Advances in Medicinal Phytochemistry Phytochemistry Vol. 5 (L. Reinhold, J. B. Harborne, and T.
(D. Barton and W. D. Ollis, eds.), 37-46, Libbey, London, Swain, eds.), 273-316, Pergamon, London, 1978.
1986. COSCIA, C. J., Picrotoxin, in Cyclopentanoid Terpene Derivatives
BORG-KARLSON, A., Chemical and ethological studies of pollina- (W.!. Taylor and A. R. Battersby, eds.), 147-201, Marcel Dek-
tion in the genus Ophrys (Orchidaceae), Phytochemistry, 29, ker, Inc., New York, 1969.
1359-1387 (1990). COSCIA, C. 1., Terpenoids, Vol. 1 of Handbook of Chromatography
BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from (G. Zweig and 1. Sherma, eds.), CRC Press, Boca Raton, FL,
plants, in Phytochemical Resources for Medicine and Agricul- 1984.
ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum COXON, T., Phytoalexins from other families, in Phytoalexins (l
Press, New York, 1992. A. Bailey and 1. W. Mansfield, eds.), 106-132, Wiley, New
BOWERS, W. S., Phytochemical resources for plant protection, in York, 1982.
394 Sesquiterpenes

CROTEAU, R., Biosynthesis and catabolism of monoterpenoids, FISCHER, N. H., On the biogenesis of pseudoguaianolides, Rev.
Chern. Rev., 87, 929-954 (1987). Latinoarner. Quim. 9, 41 (1978).
CROTEAU, R. and A. OUNDY, Cyclization of farnesyl pyrophosphate FIsCHER, N. H., Sesquiterpene lactones: Biogenesis and biomimetic
to the sesquiterpene olefins humulene and caryophyllene by an transfonnations, in Biochemistry of the Mevalonic Acid Path-
enzyme system from sage (Salvia officinalis), Arch. Biochem. way to Terpenoids (0. H. N. Towers and H. A. Stafford, eds.)
Biophys., 233, 838-841 (1984). Recent Advances in Phytochemistry Vol. 24, 161-201, Plenum
CROTEAU, R. and M. A. JOHNSON, Biosynthesis of terpenoids in Press, New York, 1990.
glandular trichomes, in Biology and Chemistry of Plant Tri- FISCHER, N. H., Plant terpenoids as allelopathic agents, in Ecologi-
chomes (E. Rodriguez, P. L. Healey, and I. Mehta, eds.), cal Chemistry and Biochemistry of Plant Terpenoids (J. B. Har-
133-186, Plenum Press, New York, 1984. borne and F. A. Tomas-Barberan, eds.), Phytochemical Society
CROTEAU, R. and M. A. JOHNSON, Biosynthesis of terpenoid wood of Europe Vol. 31, 377-398, Oxford University Press, Oxford,
extractives, in Biosynthesis and Biodegradation of Wood Com- 1991a.
ponents (T. Higuchi, ed.), 379-439, Academic Press, Orlando, FISCHER, N. H., Sesquiterpene lactones, in Terpenoids (B. V. Charl-
FL,1985. wood and D. V. Banthorpe, eds.), Vol. 7 of Modern Methods
CROTEAU, R., S. L. MUNCK, C. C. AKOH, H. J. FISK, and D. M. in Plant Biochemistry (J. B. Harborne and P. M. Dey, eds.),
SATI'BRWHITE, Biosynthesis of the sesquiterpene patchoulol 187-211, Academic Press, London, 1991b.
from farnesyl pyrophosphate in lesf extracts of Pogostemon FISCHER, N. H., E. J. OLIVIER, and R. D. FISCHER, The biogenesis
cahlin (patchouli): Mechanistic considerations, Arch. Biochem. and chemistry of sesquiterpene lactones, Fortschr. Chern. Org.
Biophys. 256, 56-68 (1987). Naturst., 38, 47-390 (1979).
Cun..ER, H. G., An historical perspective of ancient poisons, in FISCHER, N. H., J. D. WEIDBNHAMER, and J. M. BRADOW, Inhibition
Phytochemical Resoorees for Medicine and Agriculture (H. N. and promotion of germination by several sesquiterpenes. J.
Nigg and D. S. Seigler, eds.), 1-13, Plenum, Press, New York, Chern. Ecol.,15, 1785-1793 (1989a).
1992. FISCHER, N. H., J. D. WEIDENHAMER, and J. M. BRADOW, Dihydro-
DAHLGREN, R. M. T., S. R. JENSEN, and R. J. NIELSEN, A revised parthenolide and other sesquiterpene lactones stimulate
classification of the Angiospenns with comments on correlation witchweed germination, Phytochemistry, 28, 2315-2317
between chemical and other characters, in Phytochemistry and (1989b).
Angiosperm Phylogeny (D. A. Young and D. S. Seigler, eds.), FIsCHER, N. H., J. D. WBJDBNHAMER, J. L. RIOPEL, L. QUUANO, and
149-204, Praeger, New York, 1981. M. A. MENELAou, Stimulation of witchweed gennination by
sesquiterpene lactones: A structure-activity study, Phytochem-
DAWSON, O. W., D. C. ORIFFITHS, J. A. PICKEIT, L. J. WADHAMS,
istry, 29, 2479-2483 (1990).
and C. M. WOODCOCK, Plant-derived synergists of alarm phero-
mone from turnip aphid, Lipaphis (Hyadaphis) erysimi (Homo- FRAGA, B. M., Sesquiterpenes, in Terpenoids (B. V. Charlwood
ptera, Aphididae), J. Chern. Ecol., 13, 1663-1671 (1987). and D. V. Banthorpe, eds.), Vol. 7 of Modern Methods in Plant
Biochemistry (J. B. Harborne and P. M. Dey, eds.), 145-185,
DEHAL, S. S. and R. CROTEAU, Partial purification and characteriza-
Academic Press, London, 1991.
tion of two sesquiterpene cycIases from sage (Salvia officinalis)
GERSHENZON, J. and R. CROTEAU. Regulation of monoterpene bio-
which catalyze the respective conversion of famesyl pyrophos-
phate to humulene and caryophyllene, Arch. Biochem. Bio- synthesis in higher plants, in Biochemistry of the Mevalonic
phys., 261, 346-356 (1988). Acid Pathway to Terpenoids (0. H. N. Towers and H. A. Staf-
ford, eds.), Recent Advances in Phytochemistry Vol. 24,
DE ROSA, S., Terpenoids of marine plants, in Ecological Chemistry 99-160, Plenum Press, New York, 1990.
and Biochemistry of Plant Terpenoids O. B. Harborne and F.
GERSHBNZON, J. and R. CROTEAU, Terpenoids, in Herbivores: Their
A. Tomas-Barbenin, eds.), Phytochemistry Society of Europe,
Interactions with Secondary Plant Metabolites, Vol. I (0. A.
Vat.·31, 28-62, Oxford University Press, Oxford, 1991.
Rosenthal and M. R. Berenbaum, eds.), 165-219, Academic
DOSKOTCH, R. W., H. Y. CHENG, T. M. ODELL, and L. OIRARD, Press, San Diego, CA, 1991.
Nerolidol: An antifeeding sesquiterpene alcohol for gypsy moth Gn.aERT. B., Natural product derivatives in tropical insect and para-
larvae from Melaleuca leucadewon, J. Chern. Ecol., 6, site control, in Natural Products and the Protection of Plants (0.
845-851 (1980). B. Marini-Bellnlo, ed.), 225-252, Elsevier, Amsterdam, 1977.
DoUGLASS, S. K., J. A. JUVIK, H. PYUN, and R. M. COATES, Struc- OROSS, D., Phytoalexine und verwandte Pfianzenstoffe, Fortschr.
ture-activity relationship for analogs of ( + )-(E)-endo-j3-berga- Chern. Org. Naturst., 34, 188-247 (1977).
moten-12-oic acid, an oviposition stimulant of Helicoverpa zea
HALL,!. R., K. LEE, E. C. MAR, C. O. STARNES, and T. O. WADDELL,
(Boddie), J. Chern. Ecol., 19, 11-27 (1993).
Tenulin and helenalin and related cyclopentenones, J. Med.
ELus, B. E., Natural products from plant tissue culture, Nat. Prod. Chern., 20, 333-337 (1977).
Rep., 5, 581-612 (1988). HARBORNB, J. B., Introduction to Ecological Biochemistry, 2nd ed.,
EVANS, F. J. and R. J. SCHMIDT, Plants and plant products that Academic Press, New York, 1982.
induce contact dermatitis, Planta Medica, 38,289-316 (1980). HARBORNB, J. B., The role on phytoalexins in natural plant resis-
FARNSWORTH, N. R., T. O. HENDERSON, and D. D. SOB/ARTO, Plants taoce, in Natural Resistaoce of Plants to Pests (M. B. Green
with potential molluscicidal activity, in Plant Molluscicides, and P. A. Hedin, eds.), ACS Symposium Serres 296, 22-35,
131-204, John Wiley & Sons, Ltd., Chichester, 1987. American Chemical Society, Washington, DC, 1986a.
FENWICK, R. and M. MORGAN, Natural toxicants in plant foods, HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod.
Chern. Br., 1027-1029 (1991). Rep., 3, 323-344 (l986b).
 Sesquiterpenes 395

HARBORNE, J. B., Recent advances in chemical ecology. Nat. Prod. and N. I. MATZENBACHER, Trichothecene mycotoxins from Bra-
Rep., 7, 85-109 (1989). zilian Baccharis species, Phytochemistry, 30, 789-797 (1991).
HAY. M. E. and W. FENICAL, Marine plant-hervivore interactions: JARVIS, B. B .. N. B. PENA, M. M. RAO, N. S. COMEZOGLU, T.
The ecology of chemical defense, Annu. Rev. Beol. Syst., 19, COMEZOGLU, and N. B. MANOAVA, Allelopathic agents from
111-145 (1988). Parthenium hysteropherus and Baccharis megapotamica, in
HAY, M. E., P. E. RENAUD, and W. FENICAL, Large mobile versus The Chemistry of Allelopathy (A. C. Thompson, ed.), ACS
small sedentary herbivores and their resistance to seaweed Symposium Series, 268, 149-159, American Chemical Society,
chemical defenses, Oecologia, 75, 246-252 (1988). Washington, DC, 1985.
HEDIN, P. A., F. G. MAxwELL, and J. N. JENKINS, Jnsect plant JENNINGS, R. C .. K. J. JUDY, and D. A. SCHOOLEY, Biosynthesis of
attractants, feeding stimulants, repellents, deterrents, and other the homosesquiterpenoid juvenile hormone JH n [Methyl (2£,
related factors affecting insect behavior, in Proc. Summer Jnst. 6E, IOZ)-IO, ll-epoxy-3,7,ll-trimethyltridecadienoate] from
BioI. Control of Plant Jnsects and Diseases (p. G. Maswell and [5-3H]homomevalonate in Manduca sexta, J. Chern. Soc.,
F. A. Harris, eds.), 494-527, University of Mississippi Press, Chern. Commun. 21-22 (1975).
Jackson, 1974. JOHNSON, A. W., The interface of academic and industrial research.
HEGNAUER, R., The chemistry of the Compositae, in Biology and Chern. Br., 14, 332-337 (1978).
Chemistry of the Compositae, Vol. 1. (Y. H. Heywood, J. B. KELsEY, R. L., G. W. RBYNOLDS, and E. RODRIGUEZ, The chemistry
Harbome, and B. L. Tumer, eds.), 283-335, Academic Press, of biologically active constituents secreted and stored in plant
London, 1977. glandular trichomes, in Biology and Chemistry of Plant Tri-
chomes (E. Rodriguez, P. L. Healey, and I. Mehta, eds.),
IIENoERSON, T. 0., N. R. FARNSWORTH, and T. C. MYERS, Biochem-
187-241, Plenum Press, New York, 1984.
istry of recognized molluscicidal compounds of plant origin,
in Plant Molluscicides, 109-130, John Wiley & Sons, Ltd., KLAYMAN, D. L., Qinghaosu (artemisinin): An antimalarial drug
Chichester, 1987. from China, Science, 228, 1049-1054 (1985).
lIENDRYCH, R., Quantitive Ubersicht rezenter Cormobionten, Pre- KLAYMAN, D. L., Weeding out malaria, Nat. Hist., 10, 118-26
slia, 57, 359-370 (1985). (1989).
HERoUT, V., Some relations between plants, insects, and their iso- KUBO, I. and K. NAKANISHI, Insect antifeedants and repellents from
prenoids, in Progress in Phytochemistry Vol. 2 (1.. Reinhold African plants, in Host Plant Resistance to Pests (P. A. Hedin,
and Y. Liwschitz, eds.), 143-202, Jnterscience, London, 1970. ed.), ACS Symposium Series 62, 165-178, American Chemical
Society, Washington, DC, 1977.
HEROUT, V., Biochemistry of sesquiterpenoids, in Aspects of Ter-
penoid Chemistry and Biochemistry (T. W. Goodwin, ed.), Kut, J., Phytoalexins from the Solanaceae, in Phytoalexins (J. A.
53-94, Academic Press, London, 1973. Bailey and J. W. Mansfield, eds.), Wiley, New York, 1982.
Kut, J., Antifungal compounds from plants, in Phytochemical Re-
HERZ, W., Sesquiterpene lactones in the Compositae, in Biology
sources for Medicine and Agticulture (H. N. Nigg and D. S.
and Chemistry of the Compositae, Vol. 2. (Y. H. Heywood, J.
Seigler, eds.), 159-184, Plenum Press, New York, 1992.
B. Harbome, andB. L. Tumer,eds.), 337-357,AcademicPress,
London, 1977. KULLENBERG, B. and G. BERGSTROM, Chemical communication be-
tween living organisms, Endeavour, 34, 59-66 (1975).
HIKINO, H.. Recent research on oriental medicinal plants, in Eco-
nomic and Medicinal Plant Research, Vol. I (H. Wagner, H. KULLENBERG, B. and G. BERGSTROM, Hymenoptera aculeata males
Hikino, and N. R. Farnsworth, eds.), 53-85, Academic Press, as pollinators of Ophrys orchids, Zool. Scr., 5, 13-23 (1976).
London, 1985. KUTI, J. 0., B. B. JARVrs, N. MOKHTARI-RE.rAU, and G. A. BEAN,
HOLUB, M., J. TOMAN, and V. HEROUT, The phylogenetic relation- Allelochemical regUlation of reproduction and seed germination
ships of the Asteraceae and Apiaceae based on phytochemical of two Brazilian Baccharis species by phytotoxic trlchothe-
characters, Biochem. Syst. Beol., 15, 321-326 (1987). cenes, J. Chern. Ecol., 16, 3441-3453 (1990).

HOWARD, J. J., J. CAZIN JR., and D. F. WIEMER, Toxicity of terpe-

LAw, J. H., Biosynthesis of juvenile hormones in insects, in Biosyn-
thesis of Isoprenoid Compounds, Vol. 2 (J. W. Porter and S.
noid deterrents to the leafcutting ant Atta cephaloides and its
L. Spurgeon, eds.), 507-534, Wiley, New York, 1983.
mutualistic fungus, J. Chern. Ecol., 14, 59-69 (1988).
LEWIS. W. H., Plants used medicinally by indigenous peoples, in
ISMAN, M. B. and E. RODRiGUEZ, Larval growth inhibitors from
Phytochemical Resources for Medicine and Agticulture (H. N.
species of Parthenium (Asteraceae), Phytochemistry, 22,
Nigg and D. S. Seigler, eds.), 33-74, Plenum Press, New York,
2709-2713 (1983).
1992.
JACOBSON, M., Plants, insects, and man-Their interrelationships,
LoO:MIS, W. D. and R. CROTEAU, Biochemistry of terpenoids, in
Beon. Bot., 36, 346-354 (1982).
Lipids, Structure and Function (P. K. Stumpf, ed.), Vol. 4 of
JAENICKE, L., VI. Development: Signals in the development of The Biochemistry of Plants (P. K. Stumpf and E. E. Conn, eds.),
cryptogams, Prog. Bot., 52,138-198 (1991). 363-418, Academic Press, New York, 1980.
JARVlS, B. B., J. O. MrDIwo, D. Turnru., and G. A. BEAN, Interac- MABRY. T. J., Infraspecific variation of sesquiterpene lactones in
tion between the antibiotic trichothecenes and the higher plant Ambrosia (Compositae): Applications to evolutionary problems
Baccharis megapotamica, Science, 214, 460-461 (1981). at the populational level, in Phytochemical Phylogeny (J. B.
JARVIS, B. B., J. O. Mmrwo, G. A. BEAN, M. B. ABOUL-NASR, Harborne, ed.), 269-300, Academic Press, London, 1970.
and C. S. BARROS, The mystery of trichothecene antibiotics in MABRY, T. J., Major frontiers in phytochemistry, Recent Advances
Baccharis species, J. Nat. Prod., 51,736-744 (1988). in Phytochemistry Vol. 4, 273-306, Appleton Century Crofts,
JARVIS, B. B., N. MOKHfARI-RE.rAU, E. P. SCHENKEl., C. S. BARROS, New York, 1972.
396 Sesqui/erpenes

MABRY, T. 1., The chemistry of geographical races, in Pure App!. Einbaurersuche mit eH und '4C),-dopplemarkiertem Farnesol
Chern .• 34.377-400 (1973). in Cantharidin, Helv. Chim. Acta, 60, 1262-1272 (1977).
MABRY, T. 1. and J. E. GILL, Sesquiterpene lactones and other PICMAN, A. K., Biological activities of sesquiterpene lactones, Bio-
terpenoids, in Herbivores (G. A Rosenthal and D. H. Janzen, chern. Syst. Ecol., 14, 255-281 (1986a).
eds.), 502-537, Academic Press, New York, 1979. PICMAN, A. K., Aqueous solubility of isoalantolactone and its effect
MAcr, M. E., R. D. STlPANOVIC, and A A. BELL, Toxicity and on gennination and biomass production of redroot pigweed and
role of terpenoid phytoalexins in verticillium wilt resistance in late flowering goosefoot, Biochem. Syst. Bcol., 14, 361-364
cotton, Physiol. Plant Path., 26, 209-218 (1985). (1986b).
MACKIE, H. and K. H. OVERTON, Hydrolysis and isomerization PLATTNER, R. D., M. N. BEREMAND, and R. G. POWELL, Analysis
of trans, trans-farnesyl diphosphate by Andrographis tissue- of trichothecene mycotoxins by mass spectrometry and tandem
culture enzymes, Eur. J. Biochem., 77, 101-106 (1977). mass spectrometry, Tetrahedron, 45, 2251-2262 (1989).
MACKO, V., W. ACKLIN, C. HINDENBRAND, F. WEIBEL, and D. ARI- POMMERVILLE, J. C., J. B. STRlCKLAND, and K. E. HARDING, Phero-
GONI, Structure of the three isomeric host-specific toxins from mone interactions and ionic communication in gametes of
Helminthosporium sacchari, Experientia, 39, 343, 566 (1983). aquatic fungus Allomyces macrogynus, J. Chern. Ecol., 16,
MANN, J., Secondary Metabolism, Oxford University Press, Ox- 121-129 (1990).
ford, 1978; 2nd edition, 1987. PORTER, J, W. and S. L. SPURGEON (eds,), Biosynthesis of Isopren-
MARCHANT, Y. Y., F. BALZA, B. F. ABEYSEKERA, and G. H. N. oid Compounds, Vol. I, p. 177, Wiley, New York, 1981.
TOWERS, Molluscicidal activity of sesquiterpene lactones, Bio- POULTER, C. D. and H. C. RILLING, The prenyl transfer reaction.
chern. Syst. Ecol., 12, 285 (1984). Enzymatic and mechanistic studies of the 1'-4 coupling reaction
MASCARENHAS, 1. P., Sexual chemotaxis and chemotropism in in the terpene biosynthetic pathway, Ace. Chern. Res" 11,
plants, in Taxis and Behaviour (G. L. Hazelbauer, ed,), 307-313 (1978).
169-203, Chapman & Hall, London, 1978. POULlER, C. D., 1. C. ARGYLE, and E. A. MASH, Famesyl pyrophos-
MASH, E. A., G. M. GURRIA, and C. D. POULTER, Famesylpyrophos- phate synthetase, 1. BioI. Chern., 253, 7227-7233 (1978).
phate synthetase. Evidence for a rigid geranyl cation-pyrophos- POULTER, C. D., P. L. WIGGINS, and A. T. LE, Famesylpyrophos-
phate anion pair, J. Am. Chern. Soc., 103, 3927-3929 (1981). phate synthetase. A stepwise mechanism for the 1'-4 condensa-
MCCORMICK, J. p, and J, E. CARREL, Cantharidin biosynthesis and tion reaction, J. Am. Chern. Soc., 103, 3926-3927 (1981).
function in meloid beetles, in Pheromone Biochemistry (G. D, PREsTWICH, O. D. and G. J. BLOMQUIST, Pheromone Biochemistry,
Prestwich andG. J. Blomquist, eds.), 307-350, Academic Press, Academic Press, Orlando, FL, 1987.
Orlando, FL, 1987. RODRiGUEZ, E., G. H, N. TOWERS, and 1. C. MITCHELL, Biological
MENN, 1. J. and M. BEROZA, Insect Juvenile Honnones, Academic activities of sesquiterpene lactones, Phytochemistry, 15,
Press, New York, 1972. 1573-1580 (1976).
MUNAKATA, K., Insect antifeedants of Spodoptera litura, in Host RUCKER, G., SESQUITERPENES, Angew. Chern. Int. Ed., 12, 793-806
Plant Resistance to Pests (P. A. Hedin, ed.), ACS Symposium (1973).
Series 62,185-196, American Chemical Society, Washington, SAITO, K., T. NAGAO, S. TAKATSUKI, K. KOYAMA, and S. NATORI,
DC, 1977. The sesquiterpenoid carcinogen of bracken fern, and some ana-
O'DAY, D. H. and P. A. HORGEN, Sexual Interactions in Eucaryotic logues, from the Pteridaceae, Phytochemistry, 29, 1475-1479
Microbes, Academic Press, New York, 1981. (1990).
NEWMAN, A. A., (ed.), Chemistry of Terpenes and Terpenoids, SEAMAN, F. C., Sesquiterpene lactones as taxonomic characters in
Academic Press, London, 1972, the Asteraceae, Bot. Rev., 48, 121-595 (1982).
NAHRSTEDT, A., Chemische Waffen bei hoheren Pflanzen, Pharma- SEIGLER, D, S., The role of lipids in plant resistance to insects, in
zie in unserer Zeit, 8, 129-138 (1979). Plant Resistance to Insects (P. A Hedin, ed.), ACS Symposium
PAUL, V. 1. and W. FENICAL, Natural products chemistry and chemi- Series 208, 303-327, American Chemical Society Washington,
cal defense in tropical marine algae of the phylum Chlorophyta, DC, 1983.
in Bioorganic Marine Chemistry, Vol. I (P. 1. Scheuer, ed.), SLAMA, K. and C. M. WILLIAMS, Juvenile honnone activity for
1-29, Springer-Verlag, Berlin, 1987. the bug Pyrrhocoris apterus, Proe. Natl, Acad, Sci. USA, 54,
PAUL, V. 1., M, M. Lrrn.ER, D. S, LITTLER, and W. FENICAL, Evi- 411-414 (1965).
dence for chemical defense in tropical green alga Caulerpa ash- SPENCER, G. F., R. B. WOLF" and D, WEISLEDER, Gennination and
meadii (Caulerpaceae: Chlorophyta): Isolation of new bioactive growth inhibitory sesquiterpenes from Iva axil/aris seeds, J,
sesquiterpenoids, J. Chern. Ecol., 13, 1171-1185 (1987). Nat. Prod., 47, 730-732 (1984).
PEPPERMAN, A. B., JR, and E. ], BLANCHARD, Improvements in STADLER, E., Contact Chemoreception, in Chemical Ecology of
the synthesis of strigol and its analogs, in The Chemistry of Insects (W. J. Ben and R. T. Card", eds.), 3-35, Chapman &
Allelopathy (A. C. Thompson, ed.), ACS Symposium Series Han, London, 1984.
268,415-425, American Chemical Society, Washington, DC, STEVENS, K. L., Biological activity and chemistry of sesquiterpene
1985. lactones, in Isopentenoids in Plants (W. D. Nes, G. Fuller, and
PETER, M. G., W. D. WOGGON, C. SCHLA1TER, and H. SCHMID, L. Tsai, eds.), 65-80, Marcel Dekker, Inc., New York, 1984.
Einbaurersuche mit Geraniol und Faroeso} in Cantharidin, Helv. STOESSL, A, Structure and biogenetic relations: Fungal nonhost~
Chim. Acta, 60, 844-866 (1977). specific, in Toxins in Plant Disease (R. D, Durbin, ed,),
PETER, M, G., W. D. WOGGON, C. SCHLATI'ER, and H. SCHMID, 109-219, Academic Press, New York, 1981.
 Sesqu;terpenes 397

STOESSL, A., Stress compound formation in the interaction of potato WAGNER, H., Non-steroid, cardioactive plant constituents, in Eco-
tubers with Alternaria solani and A. solani metabolites, Physiol. nomic and Medicinal Plant Research, Vol. 2 (H. Wagner, H.
Plant Path., 20, 263-273 (1982). Hikino, and N. R. Farnsworth, eds.), 17-38, Academic Press,
SUTHBRLAND, O. R. W., C. H. WEARING, and R. F. N. HUTCHlNS, London, 1988.
Production of a-famesene, an attractant and oviposition stimu- WAGNER, H. and A. I'ROKSCH, Immunostimulatory drugs of fungi
lant for codling moth, developing fruit of ten varieties of apple, and higher plants, in Econontic and Medicinal Plant Research,
J. Chern. Beo!., 3, 625-631 (1977). Vol. 1 (H. Wagner, H. Hikino, and N. R. Farnsworth, eds.),
TBRANlSHI, R., R. G. BUTTERV, K. E. MATSUMOTO, D. J. STERN, 113-153, Academic Press, London, 1985.
R. T. CUNNINGHAM, and S. GOTHILF, Recent developments in WATERMAN, P. G. and A. I. GRAY, Chemical systematics, Nat.
chentical attractants for tephritid fruit flies, in Allelochenticals: Prod. Rep., 4, 175-203 (1987).
Role in Agriculture and Forestry (G. R. Waller, ed.), ACS Sym- WIEMER, D. F. and D. C. ALEs, Lasidiol angelale: Ant repellent
posium Series 330, 431-438, American Chentical Society, sesquiterpenoid from Lasianthaeafruticosa, J. Org. Chern., 46,
Washington, DC, 1987. 5449-5450 (1981).
THRELFAlL, D. R. and I. M. WHITEHEAD, Terpenoid phytoalexins: WONG, W., R. KAsAl, W. CHosm, Y. NAKAGAWA, K. MJzurANI, K.
Aspects of biosynthesis, catabolism, and regulation, in Ecologi- OHTANI, and O. TANAKA, Acyclic sesquiterpene oligoglycosides
cal Chentistry and Biochentistry of Plant Terpenoids (J. B. Har- frum pericarps of Sapindus delavayi, Phytochentistry, 30,
home and F. A. Tomas-Barberan, eds.), Phytochemistry Society 2699-2702 (1991).
of Europe Vo!. 31, 159-208, Oxford University Press, Oxford, YOSHIDA, Y., J. NOBUHARA, M. UCHIDA, and T. OKUDA, Buddledin
1991. A, B and C. Piscidical sesquiterpenes from Buddleja davidii
VANDER MEER, R., Semiochenticals and the red imported fire ant Franch, Tetrahedron Lett., 3717 (1976).
(Solenopsis invicta Buren) (Hymenoptera: Fornticidae), Fla. YosmoKA, H., T. J. MAaRY,andB. N. TiMMERMANN, Sesquiterpene
Entomo!.,66, 139-161 (1983). Lactones, University of Tokyo Press, Tokyo, 1973.
22
Diterpenes and Sesterterpenes

Introduction INI'KODVcnON
Biosynthesis
Diterpenes Derived from Monocyclic Precursors At least 2500 diterpenes are found in a large number of plant
Cembrene families (Banthorpe and Charlwood, 1980; Croteau and
Taxanes Johnson, 1981; Devon and Scott, 1972; Gershenzon and Cro-
teau, 1991; Hanson, 1991). Most of these compounds fit into
Cocarcinogenic Diterpenes from the Euphorbiaceae and
about 20 major groups (Fig. 22.1). Because diterpenes are
Thymelaeaceae
chemically complex and difficult to isolate, purify, and char-
Bicyclic Diterpenes
acterize, they probably have not been studied as thoroughly
Manool Derivatives
as monoterpenes and sesquiterpenes. Gibberellins and phy-
Labdane Derivatives
tol, the acyclic side chain of chlorophyll, occur in all plants.
Clerodane Derivatives
Diterpenes usually are components of plant resins and
Tricyclic Diterpenes
sometitnes are encountered as by-products from the isolation
Diterpenes related to ent-kaurene (5p,10a-configuration).
of essential oils (e.g., rosin or naval stores from turpentine
Other Structural Types Derived from the ent-Kaurane production). The most commonly encountered diterpenes are
Skeleton nonvolatile acids from conifers and legumes (Croteau and
Gibberellins Johnson, 1985; Langenheim, 1990).
Grayanotoxins Oleoresins characteristically contain 60% or more diter-
Diterpenes with a 5a, lOp-Configuration penes by weight. Because of the resinous nature of this mate-
Resins in Plants rial and its tendency to polymerize in the presence of oxygen,
Diterpenes in Chemosystematic Studies diterpenes are major components of physical barriers to in-
Biological Activity fection at wound sites (Croteau and Johnson, 1985).
Phytoalexins Methods for the isolation, purification, and characteriza-
Phytotoxins tion of diterpenes have been reviewed (Hanson, 1991). Most
Antifeedant Compounds diterpenes are isolated by extraction and purified by chroma-
Insecticidal and Nematicidal Activity tography. Thin-layer chromatography (TLC) and high-per-
Diterpenes with Juvenile Hormone Activity formance liquid chromatography (HPLC) have proven espe-
Diterpenes That Prevent Reproduction in Insects cially useful. The chromatography of diterpenes has been
Trail-Marking Pheromones reviewed (Coscia, 1984).
Diterpenes from Marine Plants A number of alkaloids based on diterpenoid structures
Sweeteners are discussed under Diterpene Alkaloids in Chapter 36.
Medicinal Properties
Antitumor Compounds BIOSYNTHESIS
Sesterterpenes
Biosynthesis The progenitor of C20 isoprenoids is geranylgeranyl pyro-
Biological Activity phosphate (GOPP) (1) (Fig. 22.2). The mechanism of con-
References densation probably is the same as that for sesquiterpenes

398
 Diterpenes and Sesterterpenes 399

levopimaric acid palustricacid abietic aeid (52)

sugiol

neoabietic acid pimaric acid (31) isopimaric acid (69) sandaracoplmaric acid (32)

<a) manool (30) torulosic acid larixol strobicacld

nagilactone G clerodadienoic acid trachylobanic acid incensole

o o
oyO';;{j,
HO~
~ ~o
otJ o 0

ginkgolide B (55) baccatinm

OH
phorbol (14) ryanodol R1=H, R1=OH
cinnzeylanol R,=OH, R,=H (82)
(b)

Fig. 22.1 (a & b). Some representative diterpenes (Croteau and lohnson. 1985; modified and used with permission of the copyright owner, Academic
Press. Orlando. FL).

(see Chapter 21). Geranyllinalyl pyrophosphate (2) also has ponent of the chlorophyll molecule and is found in all photo-
been suggested as a intermediate in the fonnation of some synthetic organisms. Phytol apparently is derived directly
diterpenes. from GGPP after reaction with chlorophyllide a (West,
Acyclic diterpenes are not common. Most known acyclic 1981).
diterpenes are derived from hydrolysis of GGPP and rela- Geranylgeranyl pyrophosphate synthetase activity has
tively simple modifications (Figs. 22.3 and 22.4). The fra- been found in carrot, pumpkin, and castor bean endospenn.
grant compound geranylgeraniol (3) is accumulated in the In preparations from pumpkin and castor bean endospenn,
wood of Cedrela toona (Meliaceae) (Croteau and Johnson, it was possible to separate famesyl pyrophosphate synthetase
1985). The monounsaturated diterpene, phytol (4), is a com- and geranylgeranyl pyrophosphate synthetase activity.
400 Diterpenes and Sesterterpenes

geranyIgeranyl pyrophosphate
farnesyl pyrophosphate
FPP GGPP(I)

OPP

~(( ~ct
lyU 6H ~u
geranylgeraniol (3) geranyJlinalyl pyrophosphate (GLPP) (2)

Fig. 22.2. Biogenesis of geranylgeranyl pyrophosphate (GGPP).

geranylgeraniol (3)
Cedrela toona wood, linseed oil
OR Phytophthora cactorum, bumblebees

geranyllinalool

jasmine oil, Norwegian spruce

isophytol
OH
# jasmine oil

9·hydroxygeranylgeraniol
(crinitol)
"" OH Cystoseira ennita, a brown alga

geranylcitronellol (89)
OH bumble bees

E-phylol (4)

Fig. 22.3.
"" OH all plants

Naturally occurring acyclic diterpenes (modified from West, 1981; used with permission of the copyright owner, Wiley, New York).
 Diterpenes and Sesterterpenes 401

acyclicditerpenes ~
opp

1 ~~ _ _ Sa,IOp·polycyclic diterpenes

#i
/
geranylgeraoyl OPP (1)
Sa,IOp·blcyclic dilerpenes
,.....
. . . .:
I labdanes
c1erodanes
=:::nes
and romedanes
rosanes

~ H
OPP 000=:"
-----+- Sp,10a-polycyclic diterpenes
abletanes
H

enantto-series
Sp,lOa-bicyclic diterpenes

cocarclnogenic diterpenes
----+ casbene
taxanes
cembrenes
macrocyclic diterpenes

Fig. 22.4. Geranylgeranyl pyrophosphate as a branch metabolite.

Geranylgeranyl pyrophosphate synthase (detected in pro- DITERPEl'IES DERIVED FROM MOI'lOCYCLIC

plastids of infected cells of Ricinus communis) had a molecu- PREClJRSORS
lar weight of 72,000 and required Mgz+ (Beale and MacMil-
lan, 1988). The enzymes with geranylgeranyl pyrophosphate Several monocyclic diterpenes occur in nature. Cembrene
synthetase activity also would accept pyrophosphates (5), found in the resinous material from several gymno-
smaller than famesyl pyrophosphate, indicating that famesyl sperms, is derived from the same intermediate leading to
pyrophosphate probably does not lie at the branch between taxanes (such as 6 and 7) that are only found in the gymno-
sterol and carotene metabolism in these organisms (Porter spermous family, Taxaceae (Sukh Dev and Misra, 1985)
and Spurgeon, 1981); that is, famesyl pyrophosphate is syn- (Fig. 22.5).
thesized by enzymes leading to the two series of compounds Other series of monocyclic diterpenes occur in angio-
independently. sperms (Fig. 22.6). Most important among these are duva-
As is the case in monoterpene and sesquiterpene biosyn- trienediol (8) (from tobacco), casbene (9) (from Ricinus
thesis, overall incorporation of mevalonic acid (MYA) and communis, Euphorbiaceae), and complex derivatives of cas-
other terpenoid precursors is low (about 1%) (Loomis and bene (e.g., 10-13) (pig. 22.6) from the Euphorbiaceae and
Croteau, 1980). Thymelaeaceae. Casbene is formed by the action of casbene
Many different mono-, bi-, trio, and tetracyclic diterpenes synthetase on geranylgeranyl pyrophosphate, an enzyme
are found and their formation may be rationalized by acid- with a molecular weight of 53,000 which requires Mg++ for
catalyzed, stereospecific cyclizations of geranylgeraniol and catalysis. The enzyme occurs in proplastids of seedlings.
analogs to yield a variety of diterpene skeleta. Similar pro- Casbene synthesis peaks about 12 hours after fungal attack
cesses are assumed to occur in vivo. (West et al., 1990). The biosynthesis oftltis diterpene phyto-
Although most bacteria do not make large quantities of alexin appears to involve transcriptional contol (West et al.,
terpenes, the Archaebacteria produce lipids that are Czo iso- 1990). Although conversion of mevalonate into isopentenyl
prenyl ethers of glycerol and its derivatives (in addition to pyrophosphate was not altered by fungal infection (Rhizopus
some C zo , C25 , and C30 acyclic compounds). These bacteria stolonifer), the activity of famesyl pyrosphate and geranyl-
do not synthesize fatty acids; the lipids in their membranes geranyl pyrophosphate synthase and casbene synthase were
are exclusively polyisoprenoid (Porter and Spurgeon, 1981). increased in infected plants (Beale and MacMillan, 1988).
402 Diterpenes and Sesterterpenes

---+ --
/
GGPP
cembrene (5)

HO

0 0

OH

(7) OCOCH,

Fig. 22.5. Biogenesis of cembrene and taxinine-K.

Pectic substances derived from the cell wall by the action properties. The latex of Euphorbia tirucalli contains a series
of fungal endopolygalacturonase have been identified as el- of highly unsaturated esters of phorbol (14) and 4-
icitors (Croteau and Johnson, 1985). deoxyphorbol along with some 3-0-acylingenol derivatives
esterified with the same unsaturated fatty acids (Fig. 22.7).
Cembrene In South Africa, honey from several cactiform species of
Euphorbia called "nors doring" produces a strong burning
The monocyclic diterpene, cembrene (5), has been sug-
sensation in the mouth or throat that persists for several hours
gested to be formed from GGPP with a Z-olefin linkage (Sosath et al., 1988). Tung oil and meal from the fruits of
(Newman, 1972) (Fig. 22.5). In view of other information, Aleurites is toxic to mammals. The meal can cause irritation
this compound is probably derived from a normal geranylg- to the skin and internal organs. The oil of Croton tiglium is
eranyl-OPP precursor. an extremely potent cathartic. A number of diterpenes, which
may comprise as much as 5% of the seeds have been isolated
Taxanes from this oil. One of these diterpenes (15) was toxic to the
Taxanes are found only in the gymnospermous family, larvae of Culex pipiens at 0.6 ppm (Harborne, 1986). The
the Taxaceae. Taxane alkaloids (derived from these ter- seeds of Jatropha curcas, J. gossypifolia, and related species
penes) are responsible for the extremely poisonous proper- also contain bioactive diterpenes (Hirota et al.; 1988). A
ties of the genus Taxus (Fig. 22.5). number of the diterpenes from these plants are tumor pro-
moting or cocarcinogenic; that is, they do not induce tumors
Cocarcinogenic Diterpenes from tbe themselves, but enhance the activity of carcinogens. When
Eupborbiaceae and TbymeJaeaeae cocarcinogenic diterpenes were applied after exposure to
subcarcinogenic doses of a carcinogenic compound, tumors
Many plants of the Euphorbiaceae and Thymelaeaceae were produced (Hecker, 1971, 1987a, 1987b). The calcium-
have long been known to be intensely irritating to the skin. and phospholipid-dependent protein, kinase C, is the recep-
Some produce severe skin lesions in humans and in livestock tive site of phorbol esters; the specific mechanisms for tumor
(Evans and Schmidt, 1980). Members of the genus Euphor- promotion are unknown, but they are related to protein ki-
bia are especially well known for their irritant and poisonous nase C activation (Alcaraz and Rios, 1991).lronically, many
properties. Polyfimctional diterpene esters of the tigliane, of these same compounds have proven antitumor effects.
ingenane, and daphnene types are responsible for the acrid Some diterpene esters are inflammatory agents but not pro-
 Y<'
Diterpenes and Sesterterpenes 403

tfjpp~, ~I
- -+ +-+
~ ~ ~

geranylgeranyl opp (1) ~ ~ cemhrane type

~~~ ingenane type jatropbane type casbene (9)

-dst~~
daphnane type (12) t1gllane type (11) lathyrane type (10)
20

19

17

2,7,1l-duvatriene-4,6-diol (8) jatrophone (13)

Fig. 22.6. Diterpenes derived from a monocyclic precursor (modified from Adolf and Hecker, 1977; used with pemrission of the copyright owner,
Laser Pages Publishing, Ltd., Israel).

moters (Alcaraz and Rios, 1991). The toxic effects and distri- mally occurs as a monoester or diester of aliphatic acids or
bution of!bese compounds have been reviewed (Alcaraz and occasionally as !be ester of an aromatic acid.
Rios, 1991; Evans and Kinghorn, 1977; Evans and Soper, Ingenol derivatives are restricted to several species of !be
1978; Hecker, 1971, 1977, 1987a, 1987b; Kinghorn, 1979). genus Euphorbia. These compounds exhibit toxicological
Many of !bese diterpenes occur as monoesters or diesters. and cocarcinogenic properties similar to !bose of !be phorbol
The most abundant croton oil diester, 12-0-tetradecanoylph- compounds.
orbol-13-acetate, is now widely used as a standard tumor- A number of plants from !be Thymelaeaceae are known
promoting agent (Marshall and Kinghorn, 1984). to be responsible for livestock poisoning. The active princi-
The major structural types known are !be phorbol esters ples of many of !bese plants are cocarcinogenic diterpenes
[e.g., a tigliane from Sapiumjaponicum (17)] (Croton, Sap- (Adolf et al., 1988). Among !be poisonous plants of !bis
ium, Aleurites, Mancinella, Baliospermum, and Euphorbia), family are Pimelia prostrata and Daphne mezereum. Intra-
ingenol esters [e.g., ingenane (18)] (Euphorbia), and daph- peritoneal injections of !be toxin of Pimelia (from New
nanes [e.g., daphnetoxin (19)] (Euphorbia, Hum, Hip- Zealand) of 3 fLglg of body weight were toxic to mice in 2
pornane, Excoecaria, Baliospermum, Thyrnelaeaceae) (Fig. h. The fruit and bark of Daphne species contain daphnetoxin
22.7); most of !be plants !bat contain !bese diterpenes are (19). The seeds contain mezerein (21). Bo!b of !bese com-
in subfamilies Crotonoideae and Euphorbioideae (Kinghorn, pounds can produce irritant dermatitis in humans, and mezer-
1979; Webster, 1975). ein (21) possesses cocarcinogenic activity as well (Alcaraz
Similar mixtures of compounds are found in many species and Rios, 1991). Odoracin (22), a structurally similar diter-
of !be genus Euphorbia as well as in o!ber species of !be pene from Daphne, possesses nematicidal activity in tests
Euphorbiaceae. 12-Deoxyphorbol (20) is one of !be most against the nematode, Aphelenchoides beseyi. At 5 ppm, !bis
common toxic diterpenoids in latex. This compound nor- diterpene is 100% nematicidal (Mabry and Gill, 1979). The
404 Diterpenes and Sesterterpenes

fl.
:dd
HO HOH CHOH
H
CH.OH
phorbol ester from phorbol ester from
ingenoJ (18) • Croton ligNum Saplumjaponicum (17)

CH.OH OH CH.OH
huratoxin from S-deoxyingenol from daphnetoxin (19)
Hum crepitalls and Euphorbia bigia1U/ulosa
Hippomane 1IUl1lcinella
OCO(CHv 12CH, C.ns OCOCH=CHCH-u=CH(CHV'~3 OCOCH,

~ 0 HO
~ H

HO ,9
o CH.OH
CH.OH

12·deoxyphorhnJ (20) mezereln (21)

goidiglaucin (27) R, = ·(CHv,CH" R. = ·O.CPh, R, = ·O.C(CHV14CH,
(a) gnidilatidin (lS) R, = ·(CH=CH),(CHv,CH" R. = ·O.CPh, R, = ·O.(CHV14CH,

Fig. 22.7 (a & b). CQcarcinogenic diterpenes from the Euphorbiaceae and Thymelaeaceae.

diterpenes ofyuan-hua, Daphne genkwa, have been reported phorbiaceae and Thymelaeaceae (Waterman and Gray,
to be abortifacient (Bingel and Fong, 1988). Four active 1987).
compounds have been isolated: yuanhuacine (23), yuanhua-
dine (24), yuanhuaf'me (25), and yuanhuatine (26). Yuanhua·
dine is administered intra-amnionically and yuanhuacine BICYCLIC DITERPEl'IES
either intra- or extra-amniotically to induce second-trimester
abortion (Bingel and Fong, 1988). GGPP is cyclized via two paths to produce the phosphate
Several compounds of this series also have antitumor ac- esters of the antipodal bicyclic labdadienols, thought to serve
tivity. Among these are gnidilatidin (27) and gnidiglaucin as precursors of the large majority of known bicyclic diter-
(28) from Gnidia lati/olia, Gnidia glaucus, and Diarthron penes (Hanson, 1971, 1973) (Fig. 22.8). Little direct mecha-
vesiculosurn (Thymelaeaceae) (Evans and Soper, 1978; nistic evidence concerning these cyclizations is available
Powell and Smith, 1980; Powell et al., 1985). (West, 1981). The initial cyclization steps yield labdane de-
The complexity of these structures and their limited dis- rivatives with either a 5a,10[3-confignration or a 5[3,lOa-
tribution suggest a close relationship between the Eu- configuration (Hanson, 1991).
 Dilerpenes and Sesterterpenes 405

°
~
°

CHpH CH,OH

odoracin (22)

:~

CH,OH

yuanbuacine (23) yuanbuadine (24)

CH,OH
(b) yua.hUaRD. (25) yuanhuatine (26)

Fig. 22.7. (continued)

Although these modes of condensation have been consid- ciano and LOpez, 1991). Most other diterpenes are thought
ered to be of systematic and phylogenetic significance, to be derived from one of the hypothetical cationic species
plants sometimes contain diterpenes with both configura- leading to compounds with these two configurations.
tions (e.g., Agathis australis, Araucariaceae). In other in-
stances, different species of the same genus possess diter- lIIanoolDerlvatives
penes with opposite configurations [e.g., Podocarpus
macrophyllus and P. !errugineus, Podocarpaceae Sclareol (29) and manool (30) may be derived from a
(Oehlschlaeger and Ourisson, 1967)]. Of these two types of geranyllinalyl pyrophosphate precursor (Fig. 22.9).
diterpenes, the 5J3,IOo<-configuration is almost certainly the
most widespread. 5o<,I0J3-Enantiomers are commonly re- Labdane Derivatives
ported in the literature, partly because several plants rich in
these compounds are of economic importance. Coniferous The bicyclic labdanoid diterpenes are characteristic of the
trees, in which this type of diterpene is particularly common, bark and needle resins of the Pinaceae, whereas tricyclic
cover about 8% of the land surface of the Earth (San Feli- types (especially the abietanes and pimaranes) dominate in
ciano and LOpez, 1991). Diterpenes with the 5o<,IOJ3-con- the wood resin (Croteau and Johnson, 1985) (Fig. 22.10).
fignration are common in gymnosperms, but are also known Highly oxygenated derivatives of the labdane series are
from the Asteraceae, Fabaceae, Lamiaceae, Rubiaceae, and widespread in the Asteraceae, Lamiaceae, Verbenaceae, and
Verbenaceae (de Mayo, 1959; MacMillan, 1971; San Feli- in the genus Croton of the Euphorbiaceae.
406 Diterpenes and Sesterterpenes

H'

5a,100...series

labda--8(17),13-dien-15-yl pyrophosphate

--+ OPP
Opp

W
~H;H'" OPP SO,IOu-,eries

Fig. 22.8. Cyclization of GGPP to Iabda-8(17),13-dien-15-yl pyrophosphate (modified from West, 1981; used with permission of the copyright owner,
Wiley, New York).

Clerodane Derivatives phylogeny of the Asteraceae, Lamiaceae, Verbenaceae, and

possibly Euphorbiaceae (Seigler, 1981).
Diterpenes of this series are fonned by rearrangement of
the labdane skeleton during cyclization (Hanson, 1991;
West, 1981) (Fig. 22.11). Many highly oxygenated diter-
penes of this series are found in the Lamiaceae and Verbena- TRICYCLIC DITERPEl'IES
ceae. There are over 400 known clerodanes; many of these
are from Teucrium species (Lamiaceae) (Hanson, 1991). Tricyclic diterpenes fall into four main classes: those with
Furan fonnation between C-15 and C-16 is a common the pimarane, abietane, rosane, or cassaine skeleta. Epimers
feature" in several groups of plants, notably the Asteraceae, at C-l3 exist among many groups of tricyclic diterpenes
Euphorbiaceae, Lamiaceae, and Verbenaceae. [e.g., pirnaric acid (31) and sandaracopimaric acid (32) (Fig.
As the structures of these compounds are quite complex 22.1)].

#.-.~# #
and they have restricted distributions, it is probable that their Subsequent modification of the precursor to both 5",,1013-
study will provide useful characters for understanding the and 5J3,10",-diterpenes may lead to diterpenes that differ in

OH

OH OH H

sclareol (29) manool (30) epl,dareol (33)

Fig. 22.9. Sclareol, 13-epi-scIareol, manooi.

 Diterpenes and Sesterterpenes 407

~
"'" opp

H' (I ~ ~H -

geranylgeranyl OPP (1) copalyl OPP (35) labdane

~b ~

cif
~-~
! beyerane

~~~
#v9#$abietane strobane kaurane atisirane
Fig. 21.10. Fonnation of labdane and tricyclic diterpenes (Croteau and Jolmson, 1985; modified and used with pennission of the copyright owner,
Academic Press, Orlando, FL).

configuration at position 13 [e.g., sclareol (29) and 13-epis- Most tetracyclic diterpenes possess the 5p,lOa-conftgu-
clareol (33) from Salvia sclarea (Fig. 22.9)]. ration, such as occurs in ent-kaurene (37) (Fig. 22.13), and
The cyclization of a labdadienyl pyrophosphate to the intermediate in the biosynthesis of gibberellins. These com-
tricyclic diterpene, pimara-8(9),15-diene (34) appears to be pounds appear to be widely distributed, but are common in
straightforward (West, 1981) (Fig. 22.12). The precursor the Euphorbiaceae (especially in Beyeria), the Asteraceae
role of labdadienyl pyrophosphates for tricyclic diterpenes (e.g., Stevia) and gymnosperms.
has been confirmed by the conversion of labda-8(l7),13- Incorporation of S_[1_2H] geranylgeranyl pyrophosphate
dien-15-yl pyrophosphate (35) to ent-sandaracopirnara- (1) into sandaracopimaradiene proceeds with overall anti-
8(l4),15-diene (36) in cell-free extracts from castor bean stereochemistry (Drengler and Coates, 1980). GGPP is ftrst
seedlings. converted to labda-8(l7),13-dien-15-yl (copalyl) pyrophos-
phate (35) and this intermediate compound is cyclized to a
tricyclic pimarenyl cation in the same manner shown above
DITERPEl'IES RELATED TO ent-KAVRENE (Fig. 22.12). However, instead of generating a tricyclic pi-
CSp.IO..·COl'lFlGVRADOI'l) maradiene by loss of a proton, cation 38 may further cyclize
to produce the tetracyclic cation. This cation has two fates.
The most widespread diterpenes are not the hydrocarbons Elimination produces ent-beyerene (39). Cation 38 also can
that generally are found in small quantities, but oxygenated form a carbonium ion that can react in three different ways.
polycyclic derivatives. Most of these compounds seem to be Cyclization of the C and D ring of diterpenes may also be
based on a relatively small number of hydrocarbon precur- viewed as taking place via nonclassical carbonium ion inter-
sors. Probably best known are those envolving ent-kaurene mediates (Geissman and Crout, 1969).
as the parent (West, 1981). In a cell-free system from Ricinus communis, GGPP is
408 Diterpenes and Sesterterpenes

lTanS-clerodanes kolavenyl OPP

(this one from Solidago arguta (Asteraceae»

clerodanes ~ R1 Rz
cis·clerodanes
R1=CH3, Rz=H
R1=CHzOAc, Rz=H
R1=CHzOH, Rz=H
R1=CHzOH, Rz=OH
R1=CH zOAc, Rz=OH
<a>
Fig. 22.11 (8 & b). Proposed pathway from GGPP to the clerodane skeleton of kolavenyl pyrophosphate and some typical clerodanes.

converted into four major products: d-sandaracopimara- plants) (MacMillan, 1971), grayanotoxins (42) (Ericaceae),
8(l4)-diene (36), ent-beyerene (d-beyerene) (39), ent-tra- and enmeins (43) [lsodon (syn. Rabdosia), Lamiaceae] arise
chylobane (I-trachylobane) (40), and ent-kaurene (1- by migration of bonds in ent-kaurene precursors (Newman,
kaurene) (37). The co-occurring phytoalexin casbene (9) is 1972). Feeding of suspected intermediates has demonstrated
derived directly from GGPP (Fig. 22.13). Each of these com- that the enmeins, steviol (44) (see the subsection sweete-
pounds is synthesized through the intermediacy of ent-l- ners), and other diterpenes are indeed derived from kaurene
labda-8(l7),13-dien-15-yl pyrophosphate (copalyl pyro- (Fig. 22.14) (Croteau and Johnson, 1985).
phosphate) (35) that is generated from GGPP by the enzyme A modified type of ent-kaurene system is found in the
copalyl pyrophosphate synthetase. ent-Beyerene, ent-trachy- diteepene alkaloids of the Garryaceae and the Ranunculaceae
lobane, and ent-kaurene are closely related and it is assumed (see Diteepene Alkaloids in Chapter 36).
that they are produced by closely related cyclases (Croteau
and Johnson, 1985). d-Sandaracopimara-8(14), 16-diene (36) GibbereJIins
has the opposite configuration at C-13 and would be ex-
pected to arise by another enzyme. Enzymes converting co- The gibberellins (41) are a large family of tetracyclic
palyl pyrophosphate to d-sandaracopimara-8(14),16-diene diteepenes apparently found in all plants (Beale and Willis,
and to ent-kaurene (I-kaurene) have been isolated (Croteau 1991). These diterpenoid acids were first isolated from the
and Johnson, 1985). fungus Gibberella fujikuroi, a pathogen that causes over-
growth of rice seedlings. The fungi may use these com-
pounds to soften cell walls and promote fungal invasion.
OTIIER STRUCTURAL TYPES DERIVED FROM
The gibberellins were shown to produce this effect and later
TIlE ent-KAURAl'IE SKELETON
were isolated from healthy plants as well.
The ent-kaurene skeleton is rearranged and subsequently These diteepenes now are known to have a number of
modified to produce a nnmber of important types of diter- hormonal effects in plants. Gibberellic acid activity has been
penes (Fig. 22.14). The gibberellins (41) (ubiquitous in found in all parts of higher plants. In general, the reproduc-
 Diterpenes and Sesterterpenes 409

o
J o
OH
H
1 H
0
o

columbin tloribundie acid tloridiolic acid

g§
a bitter principle from EuodiaJloribunda (Rutaceae)

~O
"...

~
o o
0

......6 0
~O
H
H -ko

o > OH Hoi OH
o o 0
teucrin B

Kg,
teucrin E teucrin F

CO
W
~O

O~OH
! 0

'"
o 0 CH30,C

teucrin G methyl barbascoate

(b) from Teucrium chamaedrys (Lamlaceae)

Fig. 22.11. (continued)

-=

labda-8(17),13-dien.15-yl pyrophosphate
copalyl OPP (35) pimara·8(9),lS·diene (34)

Fig. 22.12. Cyclization of labda-8(l4),13-dien-15-yl pyrophosphate to produce pimara-8(9),15-diene and related diterpenes (West, 1981; modified and
used with permission of the copyright owner, Wiley, New York).
410 Diterpenes and Sesterterpenes

~/
-'~~I (~~~~~ 8(14),16·dlene(36) /

1\1~7~' ~
~
i~-~=xb \ (l)·ent·t.achylobane(40)

+ ~
~g9r~$ ent-atisirene
ent~kaurene «l)~kaurene) (37)

Fig. 22.13. Proposed mechanism for the cyclization of GOPP to polycyclic diteIpenes of 5~,lOCl-configuration (modified from Robinson and West,
1970 and Schechter and West, 1969; used with pennission of the copyright owners, American Chemical Society and American Society for
Biochemistry & Molecular Biology).

.'
~ .
~.
12

OH
H __ H

Cg,H Cg,H
18 19

1 H f ~o OH
HO --- 0 HO OCOCH
HOi H J
gTayanotoxin I (42)

gibberellic acid AJ (41) kaurMI6·ene-15~one HO H 0

OH
enmein (43)

Fig. 22.14. Conversion of kaurene to several other skeletal types of diterpenes.

 Diterpenes and Sesterterpenes 411

tive parts contain more of these compounds than vegetative pathway to ent-kaurene (37) has been examined in cell-free
parts. Most gibberellins occur in plants as glycosides. Imma- cultures from Marah macrocarpus (Cucurbitaceae) and a
ture seeds are a particularly good source of these compounds number of other plants. The biosynthetic system in fungi
(Goodwin and Mercer, 1983). Giberellic acid (GA) synthesis and plants is identical (Loomis and Croteau, 1980).
occurs mainly in plastids of shoot and root tips, young
leaves, flower parts, immature seeds, and germinating em- Grayanotoxins
bryos. A number of plants in the Ericaceae contain andromedane
Giberellins stimulate stem elongation in rosettes and cer- diterpenes that cause poisoning in mammals (Fig. 22.16).
tain mutant plants. GA-stimulated cell elongation is due to For example, Rhododendron japonicum produces a series of
cell elongation, not cell division. These diterpene acids also four closely related toxic diterpenoids in its flowers (45-48).
stimulate bolting and flowering in many plants. Exogenously The leaves of another ericaceous shrub, Leucathoe grayana,
applied GA supplants the need for vernalization of many contain several andromedane diterpenes called grayanotox-
plants and stratification in many seeds. Numerous other ef- ins (42). The toxic principle of Pieris japonica also has been
fects have been observed (Goodwin and Mercer, 1983). found to be of this type of diterpenes. The leaves of Agauria
Methods for the isolation, purification and characteriza- salicifolia and other East African ericaceous plants are toxic
tion of gibberellins have been reviewed (Beale and Willis, to sheep and goats (Mabry and Gill, 1979). Several species
1991). of Kalmia in North America are highly toxic (Kingsbury,
The biosynthesis of gibberellins has been studied exten- 1964). Ten grayanoid diterpenes from Kalmia latifolia were
sively. These compounds are derived from ent-kaurene (37), found to be antifeedant for the gypsy moth, Lymantria dispar
with 5j3,lOa-stereochemislry (Fig. 22.15) (Phinney and (EI-Naggar et aI., 1980) and are probably responsible for the
Spray, 1990; Spray and Phinney, 1987; West, 1973). The toxicity of this species.

~
.
~I
'" I • OPP-
W·I~OPP
. H
H • ---

. g9.I: .
GGPP(l) labda·8,13·dlen·15·yl

gEl
pyrophosphate

. l·U . v. H.) _ _ ~
. I !
H~....... H-..
• H •
H H
• (. ).kaurene (37) • CO,H

12° OR

17

gibberellic acId A3 (41)

Fig. 22.15. Proposed biogenesis of gibberellins (modified from Beale and MacMillan. 1988 and Birch et aI., 1959; used with permission of the
copyright owners. the Royal Society of Chemistry, Cambridge, and Elsevier Science Ltd., The Boulevard. Langford Lane, Kidlington, OX5 1GB, UK)
412 Dilerpenes and Seslerlerpimes

~ ~
HOH o
H H
HO HO '7
¥O OH ~O oil
HO OCOCH, HO OCOCHzCH,
OCOCH,
grayanotoxin I (42) rhodojaponin I
asebotoxin I
Leucothoe Pieris RhotWdendro.

~ HO~
~O.O

O~ HO
.~
,tto \
~
OH
OCOCH,OH
OCOCHzCH, HO OCOCHzCH,
HO OH OH
(45) (46) (47)

~
HOH
o H
HO

HO OH
HO OH

(48) rhodojaponin In (49)

Fig. 12.16. Grayanotoxins.

Three grayanoid diterpenes are the most active constitu- mannia (Hepaticae). These compounds occur in the leaf-
ents from dried flowers of the Chinese insecticidal plant, surface resins of members of the tribe Ricinocarpoideae of
Rhododendron molle. Rhodojaponin III (49), the major com- the Euphorbiaceae (Croteau and Johuson, 1985). Com-
pound, has antifeedant, growth inhibitory, and insecticidal pounds of the rosane type occur in the wood of Erythroxylum
activity against the larvae of Leptinotarsa decemlineata and species (Erythroxylaceae) (Fig. 22.18) and those of the cas-
Spodoptera Jrugiperda (Klocke et al., 1991). saine type in the Fabaceae (subfamily Caesalpinioideae).
Erythrophleum alkaloids possess the last type of skeleton
(Nakanishi et al., 1974).
DITEKPEl'IES WITH A 5a.IOp·COl'lFlOURATlOI'l A complex series of (probably concerted?) migrations
must be invoked in order to explain the origin of some types
A large series of diterpenes possess a 50l, 1O~-configuration. of diterpenes such as rosenonolactone (54).
These are commonly accumulated in gymnosperms and in
members of the Fabaceae subfamily Caesalpinioideae. These
diterpenes have been isolated as by-products of turpentine RESIl'IS Il'I FLAl'ITS
manufacture and by collection of the resin from plants of
this group. Crude mixtures of many of these compounds Resins are mixtures of diterpenes (along with monoterpenes
have been called "naval stores." and sesquiterpenes in many instances) that are found in
ScJareol (29) and manool (30) (Fig. 22.9) are possibly plants of many different groups (Langenheim, 1990; Thomas
derived via the intermediacy of a geranyllinalyl pyrophos- 1970). Plants from about 10% of plant families produce res-
phate (2) precursor. Formation of the allylic ion (SO) permits ins. Most of the plants that produce large amounts of resins
further cyclization to rimuene (51) and abietic acid (52). The are tropical; all gymnosperms produce resins, but most only
former cation (SO) can again cycJize to compounds such as in small amounts (Langenheim, 1990). Resins also are com-
phyllocladene (53) and isophyllocladene (Fig. 22.17) monly found in the Anacardiaceae, Burseraceae, Clusiaceae
(Oehlschlager and Ourisson, 1967). (syo. Gutriferae or Hypericaceae), Dipterocarpaceae, Eu-
Many pimarane and abietane derivatives [mostly with phorbiaceae, Fabaceae, Rubiaceae, and Styraceae. In these
5a,IO~-configuration, e.g., abietic acid (52)] are found in families, compounds of both 50l,1O~- and 5~,IOIX-configura­
gymnosperms, but also are known from the Asteraceae, Eu- tions are encountered (Langenheim, 1973, 1981, 1990). Res-
phorbiaceae, Lamiaceae, and the liverwort genus Junger- ins from gymnosperms, mostly from the Araucariaceae and
 Dite/penes and Sestertelpenes 413

#-#' +

¢f'w gf/;
(50)
geranylgeranyl pyrophosphate (1) /

~~ CO,H
pbyUocladene (53)
abietic acid (52) rimuene (51)

Fig. 22.17. Biogenesis of phyllocladene and isophyllocladene (modified from Oehlschlager and Ourisson, 1967).

Pinaceae, usually are diterpenes with a 5a,IO[3-configura- notoxins of the Ericaceae, cembrene and its relatives in gym-
tion. Many plants serve as commerical sources of resins; the nosperms, and taxanes of the Taxaceae.
genera Hymenaea and Copaifera (Fabaceae) and Pinus and The bicyciic diterpenes of the Asteraceae, Euphorbiaceae,
Agatha (gymnosperms) are probably the best known produc- Lamiaceae, and Verbenaceae appear to have exceptional
ers. Resins are used as feedstocks for products such as insec- promise at the familial and generic level, but more study is
ticides, incense, varnishes, rosin, and adhesives, and as com- needed (Seigler, 1981).
ponents of drugs and polishes. Fossilized deposits of A series of seco-diterpenes, the ginkgolides (55), are only
diterpenes are known as amber (Langenbeim, 1990). known to occur in the leaves and root bark of Ginkgo biloba
(Fig. 22.19) (Nakanishi et aI., 1974).
Many diterpenes isolated from the genus Salvia (about
900 species) (Lamiaceae or Labiatae) possess quinonoid
DITERPENES Il'I CHEMOSYSTEJllATIC STUDIES structures (Luis, 1991). Callus cultures of Salvia miltiorrhiza
established from sterile seedlings produced diterpenes, and
Diterpenes appear to be useful for phylogenetic studies of one line accumulated both cryptotanshinone (56) and ferrug-
several groups of plants. Among these are cocarcinogenic inol (57) (0.8% and 1.3%, respectively, on a dry weight
diterpenes of the Euphorbiaceae and Thymelaeaceae, graya- basis). The red pigment, cryptotanshinone, is a potent inhibi-

r; ~ g5t~
__ b H
__

b •
H

~~
~: o
rosenonolactone (54) abietic acid (52)

Fig. 22.18. Biosynthesis of rosenonoIactone (modified from Torssell. 1983; used with pennission of the copyright owner, John Wiley & Sons, Ltd,
Chichester),
 90
414 Diterpenes and Sesterterpenes

~H °
0H
1 1 ° ::7

'"
1.& ° '"
\ Ii

cryptotanshinone (56) rerruginol (57) ginkgolide (55)

(58) a-4,8,13-duvatrien-l,3-diol (59) 8-4,8,13-duvatrien-l,3-dioI (60)

Fig. 22.19. Biologically active diterpenes.

tor of platelet aggregation (Charlwood and Charlwood, tities in dark-grown seedlings maintained under sterile con-
1991). ditions. When infected with potentially pathogenic fungi,
this enzyme and a synthetase that converts GGPP to casbene
(9, Fig. 22.6), a mactocyclic diterpene, are greatly increased
BIOLOGICAL ACTIVITY (West, 1981). It has been suggested that casbene is a phyto-
alexin as it appears to increase in amount in the presence of
fungi such as Rhizopus stolonifer, Aspergillus niger, and
Diterpenes are known to play diverse roles in plants. The
Fusarium moniliforme.
ubiquitous acyclic compound, phytol (4), is a part of the
Oryzalexins A (61), B (62), and C (63) are phytoalexins
chlorophyll molecule and is essential to photosynthesis. The
isolated from rice infected with Pyricularia oryzae (Fig.
biological activity of gibberellins (such as 41) as plant hor-
22.20) (Akatsnka et a1., 1985; Brooks and Watson, 1985;
monal substance is discussed above.
Kono et a1., 1985). These compounds appear to be deriva-
A series of diterpenes with antheridium-deve10ping prop-
tives of sandaracopimaradiene.
erties (antheridiogens) in several ferns is derived from path-
Another series of closely related phytoalexins have also
ways related to those that produce gibberellins. Compounds
been reported from rice (Threlfall and Whitehead, 1991).
with this activity have been isolated from Pteridium aquili-
These compounds, the momilactones (64 and 65), were re-
num, bracken fern, and well as several other species. Because
most ferns produce only nanogram quantities of these highly ported to arise in response to the same organism as oryzalex-
ins above or in response to ultraviolet light (Cartwright et
active substances, the discovery that ferns of the family
Schizaeaceae contained larger amounts has been of value
a1., 1980, 1981; ThrelfaII and Whitehead, 1991).
for continued work with these developmentally important Momilactones arise from GGPP via 9,(3H-labdadienyl
compounds. A new antheridiogen, ent-1a,10(3-dihydroxy- pyrophosphate (66) and 9-(3H-pimara-7,15-diene (67),
9a, 15a-cycI0-20-norgibberell-16-ene-7,9-dioic acid 10,19- whereas oryzalexins arise via copalyl pyrophosphate (35)
lactone (58), has been isolated from the fern Anemia mexi- and sandaracopimaridiene (36). Both groups of diterpenes
cana (Furber and Mandel, 1988; Furber et a1., 1989). appear to be synthesized in the cytosol. Only kaurene is
Duvatriendiols, primarily a- and (3-4,8,13-duvatrien-1,3- found in tissue not attacked by fungi. Chitin proved to be
diols (59 and 60), are synthesized in glandular trichomes of the best elicitor of momilactones in cell culture (West et aI.,
the leaves of tobacco. These preformed compounds have 1990). Chitinase probably breaks down the chitin to active
antifungal properties. Duvatriendiols inhibit spore germina- fragments (West et a1., 1990). Both types of phytoalexins
tion of Peronaspora tabacino with an ECso of 20 ",giml from rice are probably important in disease resistance, but
(Kuc, 1992). the relative importance of each has not been assessed (West
et a1., 1990).
An epimeric mixture of sclareol (29) and 13-epi-sclareol
FhytoaIexins
(33) has been shown to inhibit germination of spores of sev-
The famesyl transferase of germinating castor bean (Ri- eral rust diseases on tobacco, broad bean, dwarf bean, and
cinus communis, Euphorbiaceae) seeds occurs in small quan- wheat plants at 100 ppm (Wain, 1977).
 Dite/penes and Sestenerpenes 415

Phytotoxins Although fusicoccin is a C-25 compound, GGPP (1) is

Several well-known phytotoxins are diterpenes. Among first converted to a cyclic diterpene and then modified to
these are the colletotrichins, fusicoccin, and the cotylenins. produce fusicoccin (West, 1981). This compound is similar
The colletotrochins are produced by members of the fungal in some regards to the ophiobolanes (see the section Sester-
genus Colletotrichum. These fungi are pathogens on many terpenes, below) (Stoessl, 1981).
plants including sugar cane, peppers, tobacco and maize The cotylenins are leaf growth-promoting substances iso-
(Ballio, 1981; Stoessl, 1981). The fungal antibiotic, fusicoc- lated from a Cladosporium species (Fig. 22.22) (Stoessl,
cin (68), also is derived from diterpene metabolism (Fig. 1981).
22.21). This compound occurs in the fungus Fusarium amyg-
dati. In addition to the phytotoxic activity (wilting in the Antifeedant Compounds
hosts peaches and almonds), it was later discovered that this
group of compounds has plant growth-promoting activity. A number of diterpenes are known to have pronounced
Fusicoccin itself is the most active member of the group. antifeedant activity against herbivores (Mabry and Gill,
This compound readily induces wilt in tomato seedlings. 1979; Munakata, 1977). Abietic (52), dehydroabietic, 12-
Fusicoccins and the cotylenins affect a number of physiolog- methoxyabietic, sandaracopimaric (32), and isopimaric (69)
ical processes in plants. Among these are induction of water acid serve as feeding deterrents for the larch sawfly, Pristi-
uptake, proton extrusion, promotion of growth, stimulation phora erichsonii in single needles from new shoots of tama-
of seed gennination, opening of stomata, and abscission of rack (Larix laricina) (Ohigashi et al, 1981). The larvae of
leaves (Ballio, 1981; Ballio etal., 1991; Stoessl, 1981). Fusi- this insect do eat tofted needles on short shoots of the same
coccin (68) binds specifically and with high affinity to plas- trees.
malemma-enriched membrane preparations from maize Varieties of sunflower (Hetianthus annuus, Asteraceae)
(Ballio, 1981). that are resistant to attack by larvae of the sunflower moth

GGPP
.......

#~
W
98H·labdadienyl pyrophospbate (66) copalyl pyropbosphate (35)

i .....

HI

str 1
H

~
;:98;_Pimara·W7'15.diene(67) (+).,and CO Pimaradi:ne(36)

o ~ HO~ H I 0 HH ~ HO~ i H ""0

OH 0 H
o 0
momilactone A (64) momiladone B (65) oryzalexin A (61)

oryzalexin B (62) oryzalexin C (63)

Fig. 22.20. Oryzalexins and momilactones (Threlfall and Whitehead, 1991; modified and used with pennission of the copyright owner, Oxford
University Press. Oxford).
416 Dile/penes and Sesterterpenes

CHO OH
a colletotrichin

Fig. 22.21.Biogenesis of fusicoccin and colletotrichins (modified from Stoessl, 1981 and West, 1981; used with permission of the copyright owners,
Academic Press, Orlando, FL, and Wiley, New York).

(Homeosoma eleetellum) contain high concentrations of tra- in the Lamiaceae (Labiatae) and Verbenaceae but are found
chyloban-19-oic acid (70) and (- )-16-kauren-19-oic acid in other families as well (such as the Asteraceae).
(71) in their florets. As sunflower florets that contain only Species from the Lamiaceae contain a large variety of
small amounts of these compounds are a major portion of terpenes. Among these are a number of physiologically ac-
the diet of first ins tar larvae of this insect, it is likely that tive diterpenes (Fig. 22.24). One example is the type of com-
these acids serve as feeding inhibitors. At the 1% level, both pound found in the genus Isodon (syn. Rabdosia). These
kaurenoic and trachylobanoic acids decreased the growth ent-kaurenoid diterpenes possess feeding-deterrent activity.
of sunflower moth larvae and tobacco budworm (Heliothis Inflexin (73), isodomedin (74), and kamebanin (from I. in-
vireseens) by about 50%. At the 0.5% level, both reduced flexus, I. shikokianus, var. intermedius and I. kameba) all
larval growth of the cotton bollworm and the pink bollworm exhibit specific toxicity toward lepidopteran larvae, includ-
to less than 5% (Fig. 22.23) (Mabry and Gill, 1979). The Z- ing the African armyworm, Spodoptera exempta. These dit-
and E-isomers of ( - )-ozic acid (72) have been isolated from erpenes also are cytotoxic and have LDso values in the range
Helianthus oeeidentalis and may be associated with resis- of 4-5 J.'g/ml. All three have an exocyclic Ci,f3-unsaturated
tance to insect attack (Stipanovic et aI., 1979). carbonyl group and can undergo Michael additions in a simi-
Several series of complex oxygenated diterpenes are lar manner to sesquiterpene lactones (see Chapter 21) (Kubo
known to have antifeedant properties. Many of these occur and Nakanishi, 1977).
 Ditelpenes and Sesterterpenes 417

P P
HO CH,OCH3
OH
HO CH,OCH3

Ho,r0':('o
{)l0)"CH,OCH3

°
HO

HO'A--0H HO,A--0H
{)l0)"CH,OCH3 {)l0)"CH,OCH3
Fig. 22.22. Cotylenins (Sroessi, 1981; modified and used with pennission of the copyright owner, Academic Press. Orlando, F1..).

Clerodendrin A (75) and B, from verbenaceous plants, dendron infortunatum was considerably more active. Many
possess potent antifeeding activity against some insects (e.g., of these compounds have a bitter taste but no direct correla-
Spodoptera litura), but less against others (e.g., Calospilos tion between bitterness to humans, and feeding-deterrent ac-
miranda). Several genera of plants from this family contain tivity of insects could be established. Both clerodendrin A
similar compounds (Clerodendron, Caryopteris, and Calli- and B were active against the oriental tussock moth (Eu-
earpa). ent-Clerodane is the same compound as neo-clero- proetis subf/ava) at 1000 ppm and the European com borer
dane. Clerodendrin A (75) and B, from Clerodendron trieho- (Ostrinia nubialis) at 5000 ppm (Mabry and Gill, 1979; Mu-
tomum, gave 100% feeding inhibition against Spodoptera nakata, 1977).
litura at about 250 ppm, whereas clerodin (76) from Clero- Although clerodanes are feeding deterrents to many in-

~
~ (-)-ozic acid (72)
trachyloban-19-oic acid (70)

{-}-16-kauren-19-oic acid (71) levopimaric acid

Fig. 22.23. Diterpenes from Helianthus.

418 Diterpenes and Sesterterpenes

~
H ..•,,"
.../ 0 0

I
OH .
CH,COO ·····OH O 0;::: bCOCH,
OCOCH,

inflexin (73) ajugarin I (80)

16

OCOCH,
OCOCH, cinnzeylanol (82), R=H

"~_""'£oc.'
H

~"""'OH
CO,H
HO ......
CH, 0 15o:-hydroxy-(-)-kaur-16-en-19-oic acid (79)

s-r-~
C,H' 0 ..,

~
0"
OCOCH,
IbCOCHW'
OCOCH, ; .., /

cJerodendrin A (75)

CO,H

kaur-16-en-19-oic acid (77) (·)·kauran·16«·ol (78)

(a)

Fig. 22.24 (a & b). Diterpenes with biological activity.

sects. they are feeding stimulants for the turnip sawfly, Ath· miaceae, the ajugarins (such as 80) (a type related to the
alia rosae. The larvae of this insect feed on members of the ent-derodanes), exhibited antifeedant activity against
Brassicaceae; however, adults feed on Clerodendron tricho· Spodoptera exempta at 100 ppm and S. littoralis at 300 ppm.
tomum (Verbenaceae) and sequester derodanes in their bod·
ies (Gershenzon and Croteau, 1991). Insecticidal and Nematicidal Activity
Many genera of the Asteraceae contain highly oxygen- Many of the antifeedant compounds discussed above
ated compounds similar to those of the Lamiaceae and Ver- have insecticidal activity as welL Moreover, other diterpenes
benaceae discussed above. A series of these substances was are toxic to insects and nematodes when consumed. Ryanod-
found in the widespread, weedy genus Solidago (Asteraceae) ine (81), from Ryania speciosa (Flacourtiaceae) and several
(Cooper-Driver and Le Quesne, 1987). These indude four related species, is unique in that its insecticidal activity was
ent-kauranes: kaur-16-en-19-oic acid (77), ( - )-kauran-16o:- discovered as a part of a search for new insecticides (Crosby,
01 (78), ISo:-hydroxy-( - )-kaur-16-en-19-oic acid (71), and 1971). The toxic principle, ryanodine, is a diterpene esteri-
17-hydroxy-( - )-kaur-1S-en-19-oic acid (79). These ent- fied to a pyrrole-2-carboxylic acid (Crosby, 1971). Ryanod-
kauranes had antifeedant activity against Trirhabda cana- ine inhibits cardiomuscular calcium transport (Croteau and
densis (Cooper-Driver and Le Quesne, 1987). Johnson, 1985).
Feeding-deterrent compounds in Ajuga remota, La- Two diterpenes, cinnzeylanine (82) and cinnzeylanol
 Diterpenes and Sesterterpenes 419

0 O·glyc:osyl·glyeosyl

~ ~ ~
OH HO

I I

CO,CH3 CO,·glyeosyl
~ HO

(85) stevloside (94) (102)

4W
o ; !
J. i! H

• 0
HO H
OH

Isodonal (86) enmein(43) oridonin (87) nodosin(88)

o
(Y-o
NH

(b) ryanodlne (81) zoopolanol (99)

Fig. 22.24. (conlinued)

(83), from Cinnamomum zeylanicum (Lauraceae), have been ants (Lasius carniolicus) and bumblebees (Bombus ter-
demonstrated to possess insecticidal activity. At 2-4 ppm, restris and at least eight other species). These compounds
these compounds inhibited larval ecdysis in Bombyx mori are used by male bumblebees to mark their flight routes.
(Isogai et al., 1977; Seigler, 1983). The markings are species specific. The secretions of the ants
Diterpenes from the roots of Daphne odora (Thyme- setve as recognition marks in an alarm·defense system (Ahl-
laeaceae) are nematicidal against Aphelenchoides besseyi. quist et al., 1978; Bergstrom et al., 1970; Svensson and Berg-
Odoracin (22) caused 100% mortality at 5 J.Ioglml, whereas strom, 1977). It is not known if these compounds are synthe-
the related compound odoratrin (84) possessed greater activ- sized by the insects or if they are taken from plants.
ity (Fig. 22.7) (Chitwood, 1992). Cembrene derivatives also are found in some termites of
the genus Nasutitermes, where they may playa role as trail-
Diterpenes with Juvenile Hormone Activity marking pheromones (Nahrstedt, 1982).
A diterpene (85), from Persea (Lauraceae, avocado)
leaves, has been reported to possess juvenile hormone activ- Diterpenes from Marine Plants
ity (Mann, 1987). A number of diterpenes (as well as a variety of other
terpenes) have been isolated from marine algae. These differ
Dlterpenes That Prevent Reproduction
in large part from those of terrestrial plants (De Rosa, 1991;
in Insects
Hay and Fenical, 1988). The diterpene trialdehyde, halirned-
The diterpenes from Rabdosia (syn./sodon) species (La- atrial (90) produced by many species of Halimeda (a calcar-
miaceae) exhibit an inhibitory effect on the mitochondrial eous green alga of the Chlorophyta) is one of the most active
oxidative phosphorylation of the ovary of a termite queen compounds from this group of plants. Halirneditrial (90) has
of Macrotermes subhyalinus. Among the compounds tested structural similarities to warburganal and inhibits feeding by
were isodonal (86), enmein (43), oridonin (87), and nodosin reef fishes (Hay and Fenical, 1988). Both sesquiterpenes,
(88) (Kubo et al., 1981). such as flexilin, and diterpenes, such as trifarin (91), occur
in Caulerpa species. These two metabolites contain the 1,4-
Trail.Marking Pheromones diacetoxy-1,3-diene (bis-enol acetate) moiety, which is now
Several acyclic diterpenes such as geranylgeranial and known to be widespread among compounds derived from
geranylcitronellol (89), Fig. 22.3} have been isolated from green algae (De Rosa, 1991; Paul and Fenical, 1987). Algae
420 Diterpenes and Sesterlerpenes

of the family Udoteaceae have been shown to contain similar The fruit oil of the leguminous tree Pterodon pubescens
sesquiterpenoid compounds such as rhipocephalin and rhi- contains geranylgeraniol (3) and its terminal epoxide (98).
pocephanal (Paul and Fenical, 1987). Several of the sesqui- This terminal epoxide is highly lethal to the cercariae of
terpenes and diterpenes of these algae contain acetylenic Schistosoma mansoni, the causative agent of schistosomiasis
linkages. Many of these terpenoid compounds are active in (Gilbert, 1977). Other unsaturated diterpenes such as dehy-
a variety of bioassays and are quite likely responsible for droabietic acid, agathic acid, and copalic acid prevent cercar-
the antiherbivore properties of many green algae toward fish ial penetration.
and sea urchins (Paul and Fenical, 1987). A series of diterpenes from Sideritis (Lamiaceae) species
Brown algae from the order Dictyotales produce diter- have anti-inflammatory activity. These plants also contain
penes such as pachydictyol A (92). This compound deters anti-inflammatory flavonoids (Alcaraz and Rios, 1991).
feeding by tropical parrotfishes, the tropical sea urchin Dia- Gingkolides from Ginkgo bi/oba have anti-inflammatory ac-
dema antillarum, and the temperate omnivorous fishes La- tivity and antagonize platelet aggregation factor (Walton,
godon rhomboides and Diplodus holbrooki, but not by the 1992).
temperate sea urchin Arbacia punctulata, or the temperate Diterpene acids from Ponderosa pine needles when in-
amphipod, Ampithoe longimana, or the cosmopolitan poly- gested by cattle lead to reproductive failure. This is thought
chaete Piatyneris dumerilii. A related compound, dictyol-E to occur by interference with normal steroid metabolism
(93), deters feeding by the temperate fishes Logodon and (Croteau and Johnson, 1985).
Diplodus and by the temperate sea urchin Arbacia; however, Diterpenes from zoapatle (Montanoa tomentosa, As-
it is not effective against the temperate amphipod Ampithoe teraceae) are the active components of an aqueous extract
longimana or the polychaete Platyneris. These data suggest of the plant used for inducing labor, for increasing the tone
that diterpenes of this type limit herbivory by large mobile and frequency of uterine contractions during labor, for de-
herbivores, but not against the small and generally sedentary creasing postpartum bleeding, and potentially as a fertility-
polychaetes and amphipods which may be protected indi- regulating agent (Bingel and Fong, 1988). Zoapatanol (99)
rectly by the compounds (De Rosa, 1991; Hay et al., 1988). is one of the abortifacient diterpenes from this plant (Hanson,
1991).
Sweeteners
Antitumor Compounds
Stevioside (94, Fig. 22.25), a diterpene from Stevia re-
baudiana (Asteraceae), a plant from Paraguay, is intensely A number of antitumor norditerpenes, such as nagilactone
sweet. Steviol, the parent diterpene, is an oxygenated ent- C (100) and nagilactone F (101) from the gymnospermous
kaurene derivative and the biosynthesis involves ent-kaur- genus Podocarpus (Podocarpaceae), have activity against
human non-small-celliung tumor and colon tumor cells in
16-en-19-oic acid (West, 1981). Stevioside is about 300
culture (Alcaraz and Rios, 1991; Cassady et al., 1990). An-
times as sweet as sucrose on a molar basis (Harborne, 1982;
other interesting tumor-inhibiting diterpene is compound
Hodge and Inglett, 1974; Pezzuto et al., 1985; Soejarto et
(102) isolated from Taxodium distichum (Newman, 1972).
al., 1982). Steviol and related glycosides occur in amounts as
A casbane-type diterpene from Agrostistachys hookeri
high as 8% by weight of the plant leaves (Kinghorn, 1992).
(Euphorbiaceae) is active in the P-388lytuphocytic leukemic
A sweet labdane diterpene arabinoside, gaudichaudioside
cell culture system (Choi et aI., 1988).
A (95), has been isolated from aerial parts of Baccharis
Triptolide (103) and tripdiolide (104) from the celastra-
gaudichaudiana (Fullas et al., 1991).
ceous plant, Tripterygium wilfordii, exhibit antiturltor activ-
JIIedicinai Properties ity in the L-121O (lytuphoid leukemia), P-388, and KB test
systems (Cordell, 1978).
The 7-hydroxymethyl derivative of 6-Z-geranylgeraniol
A series of cytotoxic kaurene and B-secokaurene diter-
from Croton (Euphorbiaceae) stems appears to serve as a
penoids from Rhabdosia species (Lamiaceae) have potential
prostaglandin analog. This compound protects the digestive antitumor activity. Oridonin (87), lasiokaurin, emnein (43),
tract from the effects of ulceration and in promoting wound and nodosin (88) are some of the early bio.ctive compounds
healing (Croteau and Johnson, 1985). isolated from members of this genus (Alcaraz and Rios,
An unusual glycoside, wedeloside (96), from Wedelia 1991).
asperrima (Asteraceae), inhibits mitochondrial ADP/ATP Certain cocarcinogenic diterpenes also possess antitumor
carrier protein and has the ability to protect animals against activity (see above).
the toxic effects of aflatoxin B, (Croteau and Johnson, 1985;
Node et aI., 1982). The diterpene forskolin (97), from Coleus
SESTERTEKPEl'IES
barbatus and C. forskohlii (Lamiaceae), is an activator of
adenylate cyclase, and is an active inhibitor of the action of Sesterterpenes, C25 terpenoids, are relatively uncommon,
brefeldin A (a fungal metabolite with pronounced effects primarily fungal metabolites, although some compounds of
on protein trafficking in cells) (De Sousa and Shah, 1988; this series are known from higher plants and others from
Schreiber, 1992; Valdes et al., 1987; Wagner, 1988). This insect protective waxes (Cordell, 1974; Crews and Naylor,
compound also is of interest as an antihypertensive agent 1985; Hanson, 1972). The principal type of sesterterpenes
(Alcaraz and Rios, 1991; Hanson, 1991). includes the fungal metabolites known as ophiobolanes. Two
 Dite!penes and Sesterterpenes 421

OH

& 1

\
HO

OH
,
......' ' ' -
0

OCOCH3
OH

1
" OH

wedeloside (96) forskolin (97) HO

OH

HO~
~
CHO
OH
~ ~ .._"CHO
gaudichaudioside A (95)
H""
# CHO

halimedatrial (90)

pachydictyol A (92)

dictyol-E (93)

~ OH geranylgeraniol (3)

~
CH3COO

'" I ~
I I~ I~ I~ I
~OH (98)
OCOCH3
(a) trifarin (91)

OH

o o

triptolide (103) tripdiolide (104)

HO

(b) nagilactone F (101) nagilactone C (100)

Fig. 22.25 (a & b). Medically interesting ditetpenes.

422 Dilerpenes and Seslerterpenes

ophioboli. A (105) ophiobolin B (106)

cbeilanthatriol (107) gascardic acid (110) C01H

~~ r!fO~'"CO'H
notholaenic acid (109)

Fig. 22.26. Some representative sesterterpenes.

of the best known compounds of this series are ophiobolin BIOSYI'ITHESIS

A (105) and B (106) (Fig. 22.26).
A second type of sesterterpenes including cheilanthatrlol Sesterterpenes are derived from geranyJfarnesyl pyrophos·
(107) has been isolated from the fern Cheilanthes farinosa. phate (108) (Fig. 22.27) (Hanson, 1972). In the synthesis of
This compound appears similar in certain structural aspects ophiobolins and gascardic acid, the pyrophosphate leaving
to some trlterpenes (Khan et aI., 1971). group acts as the leaving·group initiating cyclization to an
A third type, including gascardic acid (110), appears to intermediate containing an II·membered ring. A I: 5 hy·
only be known from the insect Gascardia madagascariensis dride shift leads to the unusual 8-membered ring system of
(Hanson, 1972). the ophiobolins (Hanson, 1972).

! !

HO
opbiobolins
gascardic acid

Fig. 22.27. Proposed biosynthesis of opbiobolins and gascardic acid (Hanson, 1972; modified and used with pennission of the copyright owner,
Academic Press, London).
 Diterpenes and Sesterterpenes 423

BIOLOGICAL ACTIVITY BIRCH, A. J., R. W. RICHARDS, H. SMITH, A. HARRIS, and W. B.

WHALLEY, Studies in relation to biosynthesis. Rosenonolactone
A number of sesterterpenes from the genera Helminthospor- and gibberellic acid, Tetrahedron, 7. 241-251 (1959).
ium and Cochliobolus are phytoxins for their plant hosts. BROOKS, C. J. W. and D. G. WATSON, Phytoalexins. Nat. Prod.
Ophiobolin A (105) is found in many species of these two Rep., 2, 427-459 (1985).
genera. Ophiobolins induce chlorosis and necrosis of rice CARTWRIGHT, D. W., P. LANGCAKE, and J. P. RIDE, Phytoalexin
shoots and induce leakage of betacyanins from beet roots production in rice and its enhancement by a dichlorocyc1opro-
(Ballio, 1981). pane fungicide, Physiol. Plant Path., 17. 259-267 (1980).
Notholaenic acid (109) from Notholaena standleyi af- CARTWRIGHT. D. W., P. LANGCAKE, R. J. PaYCE, D. P. LawORTHY,
forded anti-HSV-I activity at a relatively high concentration and J. P. RIDs, Isolation and characterization of two phytoalex-
ins from rice as momilactones A and B, Phytochemistry, 20,
(25 f.Lgldisk) (Rinehart et aI., 1990).
535-537 (1981).
CASSADY, J. M., W. M. BAIRD, and C. CHANG, Natural products
as a source of potential cancer chemotherapeutic and chemo-
REFERENCES preventative agents, J. Nat. Prod., 53, 23-41 (1990).
CHARLWOOD, B. V. and K. A. CHARLWOOD, Terpenoid production
ADOLF, W. and E. HEcKER, Diterpene initants and cocarcinogens in plant cell cultures, in Ecological Chemistry and Biochemistry
in Euphorbiaceae and Thymelaeaceae: Structural relationships ofPIant Terpenoids (J. B. Harbome and F. A. Tomas-Barbenln,
in view of their biogenesis, Israel J. Chern., 16, 75-83 (1977). cds.), Phytochemistry Society of Europe Vol. 31, 95-132, Clar-
AnoLF, W., E. H. SEIP, E. HIlcKER, and S. F. DOSSAJI, Irritant endon, Oxford, 1991.
principles of the mezereon family (Thymelaeaceae), B. New CHITWOOD, D. J., Nematicidal compounds from plants, in Phyto-
skin irritants and tumor promoters of the daphnane and la- chemical Resources for Medicine and Agriculture (H. N. Nigg
aIkyldaphnane type from Synaptolepis kirkii and Synaptolepis and D. S. Seigler, cds.), 185-204, Plenum Press, New York,
re!usa, J. Nat. Prod., 51, 662-674 (1988).
1992.
AHLQUIST, L., G. BERGSTROM, and C. LlLlBNBERO, Acyclic diter-
CHOl, Y., J. M. PBZZUTO, A. D. KINGHORN, and N. R. FARNSWORTII,
pene alcohols: Occurrence and synthesis of geranylcitronellol,
Plant anticancer agents, XLVI. Cytotoxic casbane-type constitu-
phytol and geranylgeraniol, Prog. Chern. Fats Other Lipids, 16,
ents of Agrostistachys hookeri, J. Nat. Prod., 51, 110-116
231-255 (1978).
(1988).
AKATSUKA, T., O. KODAMA, H. SEKIDO, Y. KONO, and S. TAKEucm,
COOPER-DRIVER, O. A. and P. W. La QUBSNE, Diterpenoids as
Novel phytoalexins (oryzaIexins A, B, and C) isolated from rice
insect antifeedants and growth inhibitors: Role in Solidago spe-
blast leaves infected with Pyricularia oryzae. Part I: Isolation,
cies, in Allelochemicals: Role in agriculture and forestry (G. R.
characterization and biological activities of oryzalexins, Agric.
Waller, ed.), ACS Symposium Series 330, 534-550, American
BioI. Cbern., 49, 1689-1694 (1985).
Chemical Society, Washington, DC, 1987.
ALcARAz, M. J. and J. L. Rios, Pharmacology of diterpenoids, in
CORDELL, G. A., The occwrence, structure elucidation and biosyn-
Ecological Chemistry and Biochemistry of PIant Terpenoids (J.
thesis of the sesterterpenes, Phytochemistry, 13, 2343-2364
B. Harbome and F. A. Tomas-Barbenln, eds.), Phytochemistry
(1974).
Society of Europe Vol. 31, 230-263, Clarendon, Oxford, 1991.
CORDELL, G. A., Anticancer agents from plants, in Progress in
BAllIo, A., Structure-activity relationships, in Toxins in Plant Dis-
ease (R. D. Durbin, ed.), 395-441, Academic Press, New York, Phytocbemistry Vol. 5 (L. Reinhold, J. B. Harbome, and T.
Swain, cds.), 273-316, Pergamon, London, 1978.
1981.
BAllIo, A., S. CASTELLANO, S. CERRINI, A. EvroENTE, G. RAN- COSCIA, C. J., TrlRPBNOlDS. Vol. 1 of Handbook of Chromatography
DAZZO, and A. L. SEaRE, 'H-NMR conformational study of fusi- (0. Zweig and 1. Sherma, cds.), CRC Press, Boca Raton, FL,
cocein and related compoWlds: Molecular confonnation and 1984.
biological activity, Phytochemistry, 30. 1137-146 (1991). CREWS, P. and S. NAYLOR, Sesterterpenes: an emerging group of
BANTHORPE, D. V. and B. V. CHARLWOOD, The terpenoids, in Sec- metabolites from marine and terrestrial organisms, Fartschr.
ondary Plant Products (E. A. Bell and B. V. Charlwood, eds.), Chern. Org. Naturst., 48. 203-269 (1985).
185-220, Springer-Verlag, Berlin, 1980. CROSBY, D. G., Minor insecticides of plant origin, in Naturally

BEALE, M. H. and 1. MAcMn.LAN, The biosynthesis of C,-Cw Occurring Insecticides (M. Jacobson and D. O. Crosby, eds.),
terpenoid compounds, Nat. Prod. Rep., 5, 247-264 (1988). 177-239, Marcel Dekker, Inc., New York, 1971.
BEALE, M. H. and C. L. WlLLlS, Gibberellins, in Terpenoids (B. CROTEAU, R. and M. A. JOHNSON, Biosynthesis of terpenoids in
V. Charlwood and D. V. Banthrope, eds.), Vol. 7 of Modem glandular trichomes, in Biology and Cbemistry of Plant Tri-
Methods of Plant Biochemistry (J. B. Harbome and P. M. Dey, chomes (E. Rodriguez, P. L. Healey, and I. Mehta, eds.),
eds.), 289-330, Academic Press, London, 1991. 133-185, Plenum Press, New York, 1981.
BERGSTROM, G., S. S. STENHAGBN, and B. KULl.BNBERG, Volatile CROTEAU, R. and M. A. JOHNSON, Biosynthesis of terpenoid wood
components of the cephalic marking secretion of male bumble- extractives. in Biosynthesis and Biodegradation of Wood Com-
bees, Acta Chern. Scand., 24, 1481-1483 (1970). ponents (T. Higuchi, cd.), 379-439, Academic Press, Orlando,
BINGEL, A. S. and H. H. S. FONG, Potential fertility-regulating FL,1985.
agents from plants, in Economic and Medicine PIant Research, DE MAyo, P., The Higher Terpenoids. Interscience. New York,
Vol. 2 (H. Waguer, H. Hikino, and N. R. Farnsworth, eds.), 1959
73-118, Academic Press, London, 1988. DE ROSA, S., Terpenoids of marine plants, in Ecological Chemistry
424 Diterpenes and Sesterterpenes

and Biochemistry of Plant Terpenoids (J. B. Harborne and F. HANSON, J. R, The bicyclic diterpenes, in Progress in Phytochemis-
A. Tomas-Barbenin, eds.), Phytochemistry Society of Europe, try Vol. 3 (L. Reinhold and Y. Liwschitz, eds.), 231-285,
Vol. 31, 28-62, Clarendon, Oxford, 1991. Wiley-Interscience, New York, 1973.
DE SOUSA, N. 1. and V. SHAH, Forskolin-An adenylate cyclase HANSON, J. R., Diterpenoids, in Terpenoids (B. V. Charlwood and
activating drug from an Indian herb, in Economic and Medicinal D. V. Banthorpe, eds.), Vol. 7 of Modem Methods of Plant
Plant Research, Vol. 2 (H. Wagner, H. Hikino, and N. R. Farn- Biochemistry (P. M. Dey and J. B. Harborne, eds.), 263-287,
sworth, eds.), 1-16, Academic Press, London, 1988. Academic Press, London, 1991.
DEVON, T. K. and A. J. SCOTf, Handbook of Naturally Occurring HARBORNE, J. B., Introduction to Ecological Biochemistry, 2nd ed.,
Compounds, VoL 2, Academic Press, New York, 1972. Academic Press, New York, 1982.
DRENGEL, K. A. and R M. COATES, Stereochemistry of the SN 1 HARBoRNE,1. B., Recent advances in chemical ecology, Nat. Prod.
cyclization in the biosynthesis of ent-sandarocopimaradiene Rep., 3,323-344 (1986).
with enzyme extracts from seedlings of Ricinus communis L., HAY, M. E. and W. FENICAL, Marine plant-herbivore interactions:
J. Chern. Soc., Chern. Commun., 856-857 (1980). The ecology of chemical defense, Annu. Rev. EcoL Syst., 19,
EL-NAGGAR, S. F., R. W. DOSKOTCH, T. M. O'DELL, andL. GIRARD, 111-145 (1988).
Antifeedant diterpenes for the gypsy moth larvae from Kalmia HAy, M. E., P. E. RENAUD, and W. FENICAL, Large mobile versus
latifolia: Isolation and characterization of 10 grayanoids, 1. Nat. small sedentary herbivores and their resistance to seaweed
Prod., 43, 617-631 (1980). chemical defenses, Oecologia, 75, 246-252 (1988).
EVANS, F. J. and A. D. KINGHORN, A comparative phytochemical HECKER, E., Cocarcinogens from Euphorbiaceae and Thyme-
study of the diterpenes of some species of the genera Euphorbia laeaceae, in Pharmacognosy and Phytochemistry (R. Wagner
and Elaeophorbia (Euphorbiaceae), Bot. J. Linn. Soc., 74, and L. Hiirhammer, eds.), 147-165, Springer-Verlag, Berlin,
23-35 (1977). 1971.
EVANS, F. J. and R J. SCHMIDT, Plants and plant products that HECKER, E., New toxic, irritant and cocarcinogenic diterpene esters
induce contact dermatitis, Planta Medica, 38, 289-316 (1980). from Euphorbiaceae and from Thymelaeaceae, Pure AppL
EVANS, F. 1. and C. J. SOPER, The tigliane, daphnane, and ingenane Chern., 49, 1423-1431 (1977).
diterpenes. Their chemistry, distribution, and biological activi- HECKER, E., Cocarcinogens of the diterpene ester type as principal
ties, Lloydia, 41, 193-233 (1978). risk factors of cancer in Curacao and possibly in South China,
FULLAS, E, R A HUSSAIN, E. BORDAS, J. M. PeZZUTO, D. D. in Cancer of the Liver, Esophagus and Nasopharynx (G. Wagner
SOEJARTO, and A. D. KINGHORN, Gaudishaudiosides A-E, five and Z. You-Hui, eds.), 101-113, Springer-Verlag, Berlin,
novel diterpene glycoside constituents from the sweet-tasting 1987a.
plant, Baccharis gaudichaudianum, Tetrahedron, 47, HECKER, E., Tumour promoters of the irritant diterpene ester type
8515-8522 (1991). as risk factors of cancer in man, Bot. J. Linn. Soc., 94, 197-219
FURBER, M. and L. N. MANDER, Synthesis and confinuation of (1987b).
structure of the antheridium-inducing factor from the fern Ane- HIROTA, M., M. SUTTAJIT, H. SUGURl, Y. ENDO, K. SHUDO, V.
mia mexicana, J. Am. Chern. Soc., 110, 4084-4085 (1988). WONGCHAI, E. HECKER, and H. FUnKI, A new tumor promoter
FURBER, M., L. N. MANDER, 1. E. NESTER, N. TAKAHASHI, and H. from the seed oil of Jatropha curcas L., an intramolecular dies-
YAMANE, Structure of an antheridiogen from the fern Anemia ter of 12-deoxy-16-hydroxyphorpol, Cancer Res., 48,
mexicana, Phytochemistry, 28, 63-66 (1989). 5800-5804 (1988).
GEISSMAN, T. A. and D. H. G. CROUT, The Organic Chemistry of HODGE, 1. E. and G. E. !NOLEIT, Structural aspects of glycosidic
Secondary Plant Metabolism, Freeman Cooper. San Francisco, sweeteners containing (1'-2)-linked disaccharides, in Sweet-
1969. eners (G. E. Inglett, ed.), 216-234, A VI, Westport, CT, 1974.
GERSHENZON, J. and R CROTEAU, Regulation of monoterpene bio~ ISOGAI, A., S. MVRAKOSHI, A. SUZUKI, and S. TAMURA, Chemistry
synthesis in higher plants, in Biochemistry of the Mevalonic and biological activities of cinnzeylanine and cinnzeylanol, new
Acid Pathway to Terpenoids, G.H.N., Towers and H. A. Staf- insecticidal substances from Cinnamomum zeylanicum Nees.,
ford, eds., Recent Advances Phytochemistry, 24, 99-160, Agric. BioI. Chern., 41, 1779-1784 (1977).
Plenum Press, New York, 1990. KHAN, H., A. ZAMAN, G. L. CHETTY, A. S. GUPTA, and SUIm DEV,
GERSHENZON, J. and R CROTEAU, Terpenoids, in Herbivores: Their Cheilanthatriol-A new fundamental type in sesterterpenes, Te-
Interactions with Secondary Plant Metabolites, vol 1., G. A trahedron Lett., 4443-4446 (1971).
ROSENTHAL and M. R BERENBAUM, eds., 165-219, Academic KINGHORN, A. D., Cocarcinogenic mitant Euphorbiaceae, in Toxic
Press, San Diego, 1991. Plants (A. D. Kinghorn, ed.), 137-159, Columbia University
GILBERT, B., Natural product derivatives in tropical insect and para- Press, New York, 1979.
site control, in Natural Products and the Protection of Plants (G. KINGHORN, A. D., Plants as sources of medicinally and pharmaceut-
B. Marini-Bettolo, ed.), 155-199, Elsevier, Amsterdam, 1977. ically important compounds, in Phytochemical Resources for
GOODWIN, T. W. and E. L MERCER, Plant Biochemistry, 2nd ed., Medicine and Agriculture (H. N. Nigg and D. S. Seigler, eds.),
Pergamon, Oxford, 1983. 75-95, Plenum Press, New York, 1992.
HANSON, J. R., The biosynthesis of the diterpenes, Fortschr. Chern. KINGSBURY, 1., Poisonous Plants of the United States and Canada,
Org. Naturst., 29,395-416 (1971) Prentice-Hall, Englewood Cliffs, NJ, 1964.
HANSON, J. R, The di- and sesterterpenes. Part 1: The diterpenes, KLOCKE, J. A, M. Hu, S. CHIU, and L KUBO, Grayanoid diterpene
in, Chemistry of Terpenes and Terpenoids (A A Newman, ed.), insect antifeedants and insecticides from Rhododendron mol/e,
55-199, Academic Press, London, 1972. Phytochemistry, 30, 1797-1800 (1991).
 Diterpenes and Sesterterpenes 425

KONO, Y., S. TAKEUCHI, O. KODAMA, H. SEKlDO, and T. AKATSUKA, in Plants O. B. Prldharn, ed.), 83-109, Academic Press, London,
Novel phytoalexins (oryzalexins A, B, and C) isolated from rice 1967.
blast leaves with Pyricularia oryzae. Part II: Structural studies OHIGASm, H., M. R. WAGNER, F. MATSUMURA, and D. M. BENJA-
of oryzalexins, Agric. BioI. Chem., 49, 1695-1701 (1985). MIN, Chemical basis of differential feeding behavior of the larch
KUBO, l. and K. NAKANISHI, Insect antifeedants and repellents from sawfly, Pristiphora erichson;; (Hartig), J. Chern. Bcol., 7,
African plants, in Host Plant Resistance to Pests (P. A. Hedin, 599-614 (1981).
ed.), ACS Symposium Series 62, 165-178, American Chemical PAUL, V. and W. FENICAL, Natural products chemistry and chemical
Society, Washington, DC, 1977. defense in tropical marine algae of the phylum Chlorophyta, in
KUBO, I., N. ABO-KHATWA, T. KUBOTA, and M. TANIGucm, inhibi- Bioorganic Marine Chemistry, Vol. I O. W. Blunt, ed.), 1-29,
tion of mitochondial function in tennite queen ovary by Rab- Springer-Verlag, Berlin, 1987.
dosia diterpenoids, J. Nat. Prod., 44, 593-595 (1981). PBzzuro,J. M., C. M. COMPADRE,S. M. SWANSON, N. P. D. NANAY-
Kuc, J., Antifungal compounds in plants, in Phytochemical Re- AKKARA, and A. D. KlNGHORN, Metabolically activated steviol,
sources for Medicine and Agriculture (H. N. Nigg and D. S. the aglycone of stevioside, is mutagenic, Proc. Natl. Acad. Sci.
Seigler, eds.), 159-184, Plenum, New York, 1992. USA, 82, 2478-2482 (1985).
LANGBNHEIM, 1. H., Leguminous resin-producing trees in Africa PlDNNBy, B. O. and C. R. SPRAY, Plant hormones and the biosyn-
and South America, in Tropical Forest Ecosystems in Aftica thesis of gibberellins: The early-l3-hydroxylation pathway
and South America: A Comparative Review (B. J. Meggers, E. leading to GAl, in Biochemistry of the Mevalonic Acid Pathway
S. Ayensu, and W. D. Dockworth, ed.), 89-104, Smithsonian to Terpenoids (G. H. N. Towers and H. A. Stafford, eds.), Recent
Press, Washington, DC, 1973. Advances in Phytochemistry Vol. 24, 203-218, Plenum Press,
LANGBNHElM, J. H .. Terpenoids in the Leguminosae (R. M. Polhill New York, 1990.
and P. H. Raven, eds.) Royal Botanical Gardens, Kew, UK, PORTER, J. W. and S. L. SPURGEON (ods.), Biosynthesis of isopren-
1981. oid compounds, Vols. 1 and 2, Wiley, New York, 1981.
LANGBNHElM, J. H., Plant resins, Amer. Sci. 78, 16-24 (1990). POWELL, R. G. and C. R. SMITH, JR., Antitumor agents from higher
LoOMIS, W. D. and R. CROTEAU, Biochemistry of terpenoids, in plants, in The Resource Potential in Phytochemistry, (T. Swain
Lipids, Structure and Function, (p. K. Stumpf, ed.), Vol. 4 of and R. Kleiman, eds.), Recent Advances in Phytochemistry,
The Biochemistry of Plants (P. K. Stumpf and E. E. Conn, eds.), Vol. 14,23-51, Plenum Press, New York, 1980.
363-418, Academic Press, New York, 1980. POWELL, R. G., D. WBlSLEDBR, and C. R. SMITH, JR., Daphnane
LUIS, J. G., Chemistry, biogenesis, and chemotaxonomy of the dit- diterpenes from Diarthron vesiculosum: Vesiculosin and
erpenes of Salvia, in Ecological Chemistry and Biochemistry isovesiculosin, J. Nat. Prod., 48, 102-107 (1985).
of Plant Terpenoids (J. B. Harbome and F. A. Tomas-Barbenin,
RINEHART, K. L., T. G. HOLT, N. L. FREGEAU, P. A. KmFER, G. R.
eds.), Phytochemistry Society of Europe Vol. 31,63-82, Clar- Wn.sON, T. J. PERUN JR., R. SAKAl, A. G. THOMPSON, J. G.
endon, Oxford, 1991.
STROH, L. S. SHlEW, D. S. SBIGLER, L. H. Lt, D. G. MARTIN, C.
MADRY, T. J. and J. E. Gll.L, Sesquiterpene lactones and other J. P. GRlMMBLlKHUJ1ZEN, and G. G. GXoE, Bioactive compounds
terpenoids, in Herbivores (G. A. Rosenthal and D. H. Janzen, from aquatic and terresllial sources, J. Nat. Prod., 53, 771-792
eds.), 502-537, Academic Press, New York, 1979. (1990).
MAcMn.LAN, J., Diterpenes: The Gibberellins, in Aspects ofTerpe- ROBINSON, D. R. and C. A. WEST, Biosynthesis of cyclic diterpenes
noid Chemistry and Biochemistry (T. W. Goodwin, ed.), in extracts from seedlings of Ricinus communis L. ll. Conver-
153-180, Academic Press, New York, 1971. sion of geranylgeranyJ pyrophosphate into diterpene hydrocar-
MANN, J., Secondary Metabolism, Oxford University Press, Ox- bons and partial purification of the cyclization enzymes, Bio-
ford, 1978; 2nd edition 1987. chemistry, 9, 80-89 (1970).
MADSHALL, G. T. and A. D. KlNGHORN, Short-chain phorbol ester SAN FEUCIANO, A. and 1. LOPEZ, Recent chemistry of conifer terpe-
constituents of croton oil, J. Am. Oil Chem. Soc., 61, noids, in Ecological Chemistry and Biochemistry of Plant Ter-
1220-1225 (1984). penoids (J. B. Harbome and F. A. Tomas-Barbenin, eds.), Phy-
MUNAKATA, K., Insect antifeedants of Spodoptera litura in plants, tochemistry Society of Europe, Vol. 31, 1-27, Clarendon,
in Host Phmt Resistance' to Pests (P. A. Hedin, ed.), ACS Sym- Oxford, 1991.
posium Series 62, 185-196, American Chemical Society, SCHECHTER, I. and C. A. WEST, Biosynthesis of gibberellins. IV.
Wasbington, DC, 1977. Biosynthesis of cyclic diterpenes from trans-geranylgeranyl py-
NAHRSTEDT, A" Strukturelle Beziehungen zwischen pflanzlichen rophosphate, J. BioI. Chem., 244, 3200-3209 (1969).
und tierischen Se'kundiirstoffe, Planta Medica, 44, 2-14 (1982). SCHREIBER, s. L., Using the principles of orgarlic chemistry to
NAKANISHI, K., T. GOTO, S. 1m, S. NATORl, and S. NOzaB (ods.), explore cell biology, Chern. Eng. News, 22-32 (Oct. 24,1992).
Natural Products Chemistry, Vol. 1, Academic Press, New SBlGLBR, D. S., Terpenes and plant phylogeny, in Phytochemistry
York,1974. and Angiosperm Phylogeny (D. A. Young and D. S. Seigler,
NEWMAN, A. A., Chemistry of Terpenes and Terpenoids, Academic eds.), 117-148, Praeger, New York, 1981.
Press, New York, 1972. SEIGLER, D. S., The role of lipids in plant resistance to insects, in
NODB, M., M. SAl, K. FUJI, E. fuJITA, T. SIDNGU, W. H. WATSON, Plant Resistance to Insects (P. A. Hedin, ed.), ACS Symposium
and G. GRossm, Three new antitumor diterpenoids, llichorab- Series 208, 303-327, American Chemical Society, Wasbington,
dais A, C and D, Chern. Lett., 2023-2026 (1982). DC, 1983.
OEHLSCHLAGER, A. C. and G. OuruSSON, A comparison of in vivo SOBIARTO, D. D., A. D. KINGHORN, and N. R. FARNSWORTIf, Poten-
and in vitro skeletal transformations of diterpenes, in Terpenoids tial sweetening agents of plant origin in organoleptic evaluation
426 Diterpenes and Sesterterpenes

of Stevia leaf herbarium samples for sweetness, 1. Nat. Prod., borne and F. A. Tomas-Barberan, eds.), Phytochemistry Society
45, 590-599 (1982). of Europe, Vol. 31, 159-208, Clarendon, Oxford, 1991.
SOSATH, S., H. H. Orr, and E. HECKER, Irritant principles of the TORSSELL, K. B. G., Natural Products Chemistry, John Wiley &
spurge family (Euphorbiaceae). XIII. Oligocyc1ic and macrocy- Sons, Ltd., Chichester, 1983.
dic diterpene esters from latices of some Euphorbia species VALDEs L. 1., III, S. G. MISLANKAR, and A. G. PAUL, Coleus barba-
utilized as source plants of honey, J. Nat. Prod., 51, 1062-1074 tus (C.forskohlii) (Lamiaceae) and the potential new drug for-
(1988). skolin (coleonol), Econ. Bot., 41, 474-483 (1987).
SPRAY, C. R. and B. O. PHINNEY, Gibberellins in higher plants: WAIN, R. L., Chemical aspects of plant disease resistance, in Natu-
The biosynthetic pathway leading to GAl in Ecology and Me- ral Products and the Protection of Plants (G. B. Marini-Bettolo
tabolism of Lipids (G. Fuller and W. D. Nes, eds.), ACS Sympo- ed.), 483-508, Elsevier, Amsterdam, 1977.
sium Series 325, 25-43, American Chemical Society, Washing-
ton, DC, 1987.
WAGNER, H., Non-steroid cardioactive plant constituents, in Eco-
nomic and Medicinal Plant Research, Vol. 2 (H. Wagner, H.
STIPANOYIC, R. D., D. H. O'BRIEN, C. E. ROGERS, and T. E. THOMP-
Hikino, and N. R. Farnsworth, eds.), 17-38, Academic Press,
SON, Diterpenoid acids, (- )-cis- and (- )-trans-ozic acid, in
London, 1988.
wild sunflower, Helianthus occidentalis, J. Agric. Food Chern.,
27, 458-459 (1979). WALTON, N. 1., A fine chemical harvest, Chern. Br., 525-529
(1992).
STOESSL, A., Structure and biogenetic relations: Fungal nonhost-
specific, in Toxins in Plant Disease (R. D. Durbin, ed.), WATERMAN, P. O. and A. I. GRAY, Chemical systematics, Nat.
109-219, Academic Press, New York, 1981. Prod. Rep. 4, 175-203 (1987).
SUKH DEY and R. MISRA, CRC Handbook of Terpenoids. Diterpen- WEBSTER, G. L.. Conspectus of a new classification of the Euphor-
oids. Vol. I. Acyclic and Monocyclic Diterpenoids, 1-70, CRC biaceae, Taxon, 24, 593-601 (1975).
Press, Boca Raton, FL, 1985. WEST, c. A., Biosynthesis of gibberellins, in Biosynthesis and Its
SVENSSON, B. G. and G. BERGSTROM, Volatile marking secretions Control in Plants (B. V. Milborrow, ed.), 473-482, Academic
from the labial gland of north European Pyrobombus D. T. Press, London, 1973.
males (Hymenoptera, Apidae), Insectes Sociaux, 24, 213-224 WEST, C. A., Biosynthesis of diterpenes, in Biosynthesis of Isopren-
(1977). oid Compounds, Vol. I (1. W. Porter and S. L. Spurgeon, eds.),
THOMAS, B. R., Modern and fossil plant resins, in Phytochemical 375-411, Wiley, New York, 1981.
Phylogeny (J. B. Harbome, ed.), 59-79, Academic Press, Lon- WEST, C. A. t A. F. LOIS, K. WICKHAM, and Y. REN, Diterpene
don, 1970. phytoalexins: Biosynthesis and regulation, in Biochemistry of
THRELFALL, D. R. and I. M. WHITEHEAD, Terpenoid phytoalexins: the Mevalonic Acid Pathway to Terpenoids (G. H. N. Towers
Aspects of biosynthesis, catabolism, and regulation, in Ecologi- and H. A. Stafford, eds.), Recent Advances in Phytochemistry,
cal Chemistry and Biochemistry of Plant Terpenoids (1. B. Har- Vol. 24, 219-248, Plenum Press, New York, 1990.
23
Triterpenes and Steroids

Introduction major types (Fig. 23.1) (Connolly and Hill, 1991; Devon
Squalene biosynthesis and Scott, 1972).
Compounds Derived by Direct Cyclization of Squalene In contrast to the previously discussed mono-, sesqui-,
Squalene 2,3-Epoxide and diterpenes that are biosynthesized by linear condensa-
Triterpenes Derived via a Chair-Boat-Chair-Boat tion of five carbon units, triterpenes arise via dimerization of
Transition State. two farnesyl pyrophosphate units to produce an intennediate
Fonnation of Cycloartenol compound, squalene (1) (Fig. 23.2). Squalene (usually as
Phytosterols the 2,3-epoxide) may then be cyclized by several mecha-
Fonnation of Cholesterol nisms from different confonnations on an enzyme surface
Fonnation of Other Plant Sterols to produce the parent skeletal types of many different kinds
Biological Activity of Plants Sterols of triterpenes that undergo subsequent modification (Banth-
Progestans in Plants and from Animals orpe and Charlwood, 1980; Harrison, 1985). The structures
Progestagens from Insects of many triterpenes and steroids may be explained by the
Ecdysteroids "biogenetic isoprene rule" (see Chapter 18) (Connolly and
Phytoecdysteroids Hill, 1991).
Chemotaxis in Fungi When squalene 2,3-epoxide (2) is cyclized via a
Cucurbitacins chair-boat-chair-boat sequence, the fIrst isolable cyclic in-
Triterpenes Derived via a Chair-Chair-Chair-Boat tennediate (in plants) is cycloartenol (3) (Fig. 23.6). Cycloar-
Transition State tenol proves to be a key intennediate in the synthesis of
Biological Activity of Triterpenes certain other triterpenes and plant sterols. Many steroids are
Iridals and Irones from the Iridaceae common [if not ubiquitous, e.g., fl-sitosterol (4), stigmas-
terol (5), and campesterol (6) (Fig. 23.6)] and it is probable
References
that these pathways exist in all plants (Goad and Goodwin,
1973; Grunwald, 1975; 1980). Cholesterol (7) occurs in
small quantities, but appears to be a universal constituent of
INTRODUCTION plants. This sterol serves as a key intennediate in the synthe-
sis of a number of other steroidal compounds. A new system
of numbering sterols was recommended in 1989 (Goad,
Triterpenes constitute a significant portion of the lipid sub- 1991a).
stances of all plants; more than 4000 triterpenoids have been The analytical procedures and methodology for isolation,
isolated. These compounds are precursors to steroids in both purification, and characterization of plant sterols (Connolly
plants and animals. Steroids are components of membranes and Hill, 1991; Goad, 1991a; Patterson, 1984), as well as
in plants. Both triterpenes and steroids occur free, as glyco- general chromatographic techniques for triterpenes (Con-
sides (Boar and Allen, 1973), or in other combined fonns. nolly and Hill, 1991; Coscia, 1984), the application of C-13
Correct structures for many were not forthcoming until about nuclear magnetic resonance (NMR) spectroscopy to charac-
1940 (Connolly and Overton, 1972). The structures of terization oftriterpenoids and steroids (Goad, 1991a; Knight,
triterpenes and steroids may be subdivided into about 40 1974; Wehrli and Nishida, 1979), and the application of mass

427
428 Triterpenes and Steroids

HO

6-sitosterol (4) stigmasterol (5) campesteroI (6)

22 24 26
21 20

1623~":"
,"
I '
HO HO 0

cholesterol (7) lanosterol (15) estrone (21)

OH

HO o

parkeol (53) malabaricol

squalene (1)

Fig. 23.1. Some representative triterpenes and steroids.

spectrometry to structure elucidation of triterpenes (Goad, to yield presqualene pyrophosphate (9) is effected by the
1991a; Ogunkoya, 1981) have been reviewed. enzyme squalene synthase (EC 2.5.1.21). This synthetase is
A number of alkaloids containing steroid and triterpene a microsomal enzyme in every source that has been exam-
skeleta are discussed under Triterpenoid and Steroid Alka- ined and requires a divalent cation such as Mg2+ or Mn 2+
loids in Chapter 36. (Goodwin, 1985). Squalene synthetase can be solubilized
from yeast microsomal membranes with detergents such as
deoxycholate and has been purified to homogeneity. The
SQUALEI'IB BIOSYNTHESIS enzyme is a single polypeptide with a molecular weight of
47,000. This enzyme catalyzes both portions of the biosyn-
Squalene is derived from two farnesyl pyrophosphate (8) thesis of squalene (1) from farnesyl pyrophosphate (9)
units joined in a head-to-head (1'-1) condensation (Fig. (Agnew and Popja!<, 1978; Sasiak and Rilling, 1988; Zhang
23.2). The stereochemistry of this process is known from et aI., 1993). The first step, a 1'-2,3 prenyltransfer, yields the
tracer studies and the pathway includes presqualene pyro- intermediate presqualene pyrophosphate (9). The reaction
phosphate (9) as an intermediate. Mechanisms have been producing presqualene pyrophosphate (9) is substantially
proposed that explain most stereochemical features (Poulter, more rapid than the overall synthesis of squalene (1) (Poulter
1990); other evidence supports an ion-pair mechanism for and Rilling, 1981). In in vitro studies, this intermediate
the rearrangement (Fig. 23.2) (Croteau and Johnson, 1985; seems free to leave the enzyme, whereas, in contrast, under
Poulter, 1990). in vivo conditions, presqualene pyrophosphate is accumu-
The condensation of farnesyl pyrophosphate (FPP) units lated only in small amounts (Sasink and Rilling, 1988).
 Triterpenes and Steroids 429

5 H4 2 H 2 acceptor
PPOCH~"~ 1~3
3 CH3 I I PPOCH,.....- \.~
'-7 CM3 I I rotation
- - ~PPO.
{ppo, CH) -
P ~ SHR
.--:; h 0
CH3
H
around bond
•... ~... .,,; ,9
5' H 4' 3' 2' S .' H 2' 3' 44 donor

famesyl pyrophosphate (8) (numbers correspond to those of farnesyl pyrophosphate)

(numbers correspond to those of mevalonic acid)

PPOCH2 1 2
H

4'

(R)·presqualene pyrophosphate (9)

(a) (numbers correspond to those offarnesyl pyrophosphate)

PPO'

presqualene pyrophosphate (9) /

~~
'H'~~
H
~,~
OPP'
")'. +... /
',

'H'

NADPH'\

~~
HR~
t Q ,9

4(2)

squalene (1)
HS HR
(b) (numbel'S in parentheses are those of mevalonic acid) (other numbers are those for farnesyl pyrophosphate)

Fig. 23.2 (a & b). Squalene biosynthesis.

430 Triterpenes and Steroids

One protein is required for squalene synthesis, but two loss of the pro-S hydrogen at that center. The pro-R-hydro-
catalytic sites may be involved: one for the synthesis of pre- gen atom at C-I' of the second famesyl pyrophosphate mole-
squalene pyrophosphate and one for the reduction of this cule appears at C-I' of the cyclopropane ring (based on fam-
compound to squalene (Sasiak and Rilling, 1988). Although esyl pyrophosphate numbering) anti to the vinyJic
the enzyme is membrane bound by the carboxyl terminal substituent (Popjak et al., 1975). The prenylation offamesyl
domain, the catalytic domain apparently is free in the cytosol pyrophosphate (8) occurs at the (2-si-3-re) face of the double
(Zhang et al., 1993). Squalene synthase isolated by detergent bond of the prenyl acceptor, and C-I' of the donor is inverted
treatment from daffodil microsomal membranes was acti- during the process (Poulter and Rilling, 1981). In experi-
vated after the fIrst step of purifIcation by liposomes of phos- ments with [5- 2H2 1mevalonate (10), the squalene produced
phatidylcholine (Beriingheri et al., 1991). had three deuterium atoms and one hydrogen atom on the
Nerolidyl pyrophosphate does not serve as a precursor two central carbons. The proton furnished by NADPH dur-
for squalene (poulter and Rilling, 1981). The condensation ing the fmal conversion of presqualene pyrophosphate to
requires NADPH in yeasts and, if this reductant is not pres- squalene (4) occupied the pro-S-position. C-I of the acceptor
ent, famesyl pyrophosphate (8) accumulates. During the famesyl pyrophosphate (8) is undistorbed during the initial
course of the reaction, two pyrophosphate groups are re- condensation, but suffers inversion in later steps of the bio-
leased, one hydrogen lost and one hydrogen replaced by a synthesis. In the subsequent rearrangement to squalene, both
hydrogen from NADPH. One (lS)-hydrogen atom is re- C(l) and C(l') are inverted (poulter, 1990). The apparent
moved from famesyl pyrophosphate. net retention of the center that accepts the proton from
The fIrst reaction involves a prenyl transfer step in which NADPH (C-I' in Fig. 23.2) may be explained by two inver-
C(l') of one of the allyJic substrates (the donor) is bonded sion steps.
to the C(2)-C(3) double bonds of the other (the receptor) The mechanisms of the various steps of the biosynthesis
to produce a cyc1opropylcarbinyl diphosphate with a CI'-2- of squalene have been reviewed (Coates, 1976; Poulter,
3 structore (Poulter, 1990). In this manner, famesyl pyro- 1990; Poulter and Rilling, 1981).
phosphate (8) yields presqualene pyrophosphate (9). Only The structore of presqualene resembles that of chrysan-
(R)-presqualene pyrophosphate has been found in nature. thentic acid (11), a monoterpene acid found in pyrethrins
In studies of the biosynthesis of presqualene alcohol with (Fig. 23.3) that may be biosynthesized from two moieties
[l-0 181famesyl pyrophosphate, exchange of oxygen did not of dimethylallyl pyrophosphate (DMAPP) in a fashion anal-
occur. These observations indicate that the pyrophosphate ogous to the formation of presqualene pyrophosphate
group of one famesyl pyrophosphate molecule occurs in the (Poulter, 1990; Poulter and Rilling 1981).
product without change of confIguration in presqualene py- The steric and electronic factors regulating the complex
rophosphate (Popjak et al., 1975). series of steps have been explored using substrate analogs.
In the second reaction, the latter compound rearranges to 2-Methyl FPP (12) and 3-demethyl FPP (13) (Fig. 23.3) act
ai'-I structore with a saturated linkage between the two as efficient substrates for the second site on the enzyme
original famesyl residues in a series of steps that involve (i.e., the analogs serve as an acceptor) but do not undergo
loss of inorganic pyrophosphate and incorporation of hy- pyrophosphate expulsion or proton loss at the fIrst site (they
dride from NADPH to yield squalene (4) (poulter, 1990). cannot serve as donors). Ouly products of condensation with
Presqualene pyrophosphate (9) is formed from famesyl endogenous FPP (as the donor) are observed (Poulter and
pyrophosphate (8) with inversion at C-I' and concomitant Rilling, 1981).

H02~

H~
chrysanthemic acid (11) presqualene pyrophosphate (9)

o OH Opp

HO~OH 2

OPP
mevalonic add (10)
(13)

Fig. 23.3. Presqualene pyrophosphate and chrysanthemic acid and inhibitors for squalene biosynthesis.
 Triterpenes and Steroids 431

The mechanism probably resembles that for prenylation lished by deuterium labeling experiments and NMR spec-
of dimethylallyl pyrophosphate, geranyl pyrophosphate, and troscopy.
farnesyl pyrophosphate to yield mono-, sesqui-, and diter- Several pentacyclic triterpenes found in ferns also appear
penes, respectively (Hanison, 1985). Mechanisms in which to be formed from squalene (I), but not via the intermediacy
an "X-group" is involved, followed by a trans elimination of the 2,3-epoxide (2) (Fig. 23.4). Compounds, such as fer- .
and similar reactions, appear to be ruled out (Hanison, 1985; nane (16), are found in several ferns (Goodwin, 1973). 24-
Poulter, 1990; Poulter et al., 1979). Bthoxyhopane (17) has been reported from the fern Onoclea
The biosynthesis of squalene is similar to that of phytoene neriifolia. Although these triterpenes apparently are distrib-
in tetraterpene biosynthesis, except that in the fmal step, a uted widely in ferns, triterpenes derived from condensation
double bond is formed by elimination in tetraterpenes, of squalene 2,3-oxide frequently co-occur with them.
whereas a hydride from NADPH is added in triterpene bio-
synthesis (Poulter, 1990).
SQVALEl'IE 2.3-EPOXIDE

COMPOUi'lDS DERIVED BY DIRECT Squalene (1) is converted to (3S)-squalene 2,3-epoxide (2)

CYCLIZATlOI'l OF SQVALEl'IE by the action of microsomal squalene epoxidase in the pres-
ence of O2 and NADPH (BC 1.14.99.7) (Goad, 1991b;
A few simple eukaryotic organisms can cyclize squalene but Goodwin, 1985). Bpoxidation can occur at either end of the
do not produce sterols. For example, Tetrahymena pyri- squalene molecule, suggesting that an intermediate form is
formis (a protozoan) produces tetrahymanol (14), a pen- released from the synthetase enzyme complex prior to epoxi-
tacyclic analog of lanosterol (15) (Fig. 23.1), from squalene. dation and cyclization. Squalene epoxidase (squalene mo-
The mechanism of condensation of squalene (1) to tetrahy- nooxygenase; BC 1.14.99.7) is a two-component system.
manol in Tetrahymena pyriformis does not involve squalene The natore of the terminal oxidase has not been examined
2,3-oxide (2) and includes a protonation step, cyclization, in detail. The second component, a flavoprotein, has now
and, finally, addition of hydroxyl group from the medium. been fully characterized as an NADPH-cytochrome c re-
The stereochemistry of the proton predicted to be incorpo- ductase. The monooxygenase from rat liver has a molecular
rated at the 2113-position of the compound has been estab- weight of - 47,000, and the protein requires NADPH, cyto-

femane(l6) fllicane

CHzOCzHs

24-ethoxyhopane (17) diplotene

Fig. 23.4. Triterpenes derived by direct cyclization of squalene.

432 Triterpenes and Steroids

chrome P-450 reductase, FAD, and a detergent Triton X- epoxide and may be a completely concerted process or, in
100 for reconstitution of monooxygenase activity. The mo- other cases, may be interrupted at an intermediate stage to
nooxygenase from Candida albicans was destroyed when allow rearrangement to occur. Evidence from a wide variety
mixed with Triton X-IOO (Harrison, 1985, 1988). of biosynthetic systems indicates that only the (3S)-stereo-
The degree to which cyclization of (3S)-squalene 2,3- isomer is utilized. The initially formed cationic species are
epoxide (16) to the tetracyclic state is a concerted process purely hypothetical, but if these exist, they could presumably
has been the subject of much speculation. Because squalene be stabilized by enzymic interaction (Fig. 23.6).
2,3-epoxide cyclizes chemically more rapidly than similar
epoxides, it has been suggested that epoxide opening in this
Formation of Cycloartenol
precursor receives anchimeric assistance from the adjacent
double bonds (VIIJ;! Tamelen et al., 1966, 1968). Cycloartenol (3) is the major product of condensation of
squalene 2,3-epoxide (2) by a chair-boat-chair-boat transi-
tion state in plants. This condensation is effected by squa-
TRITERFEl'IES DERIVED VIA A lene-2,3-oxide-cycloartenol cyclase (Fig. 23.6). Micro-
C~OAT-CH~BOATTKANSn10N somes from Rabdosia japonica (Acanthaceae) showed
STATE activity for cyclizing·squalene 2,3-epoxide into cycloartenol,
~-amyrin, and a-amyrin. Purified cycloartenol cyclase had
A large number of triterpenes and all steroids in plants are a molecular weight of 54,000 (Abe et al., 1989a, 1989b).
derived by cyclization of squalene (as the 2,3-epoxide) via The enzyme squalene-2,3,-oxide-cycloartenol cyclase is
a chair-boat-chair-boat transition state (Fig. 23.5). The inhibited by a series of compounds designed to mimic the
cyclic structures formed can all be rationalized in terms of carbocationic intermediates that are postulated to participate
the ways that the squalene 2,3-epoxide (2) may be folded in the reaction process (Goad, 199Ib).
(pseudo-chair-and-boat conformations) on the enzyme sur- Squalene 2,3-epoxide (2) was converted into cycloartenol
face, with due consideration given to the stereoelectronic (3) in 22% yield by a microsomal fraction from Ochromonas
requirements for cyclization. Cyclization usually is initiated malhamensis (an alga). Retention of configuration at C-19
by acid-catalyzed opening of the ring of (3S)-2,3-squalene (the cyclopropyl carbon) is observed.

gammacerane cycloartane wcurbitane

#
H H

euphane lanostane dammarane

Fig. 23.S. Major structural types of triterpenes that arise from cyclization of squalene (as the 2.3-epoxide) via a chair-boat-chair-boat confonnation.
 Triterpenes and Steroids 433

H+

H !
~j0
H0V1--Y ~"''''.''''~
H I H
H H t
HO

12

23 cycloarteool (3)

HO

Fig. 23.6. Cyclization of squalene 2,3-epoxide via a chair-boat-chair-boat confonnation (modified from Luckner, 1972; used with the pennission of
the copyright owner, Academic Press, Orlando, Fl.).

FHYTOSTBKOLS of which are now considered erroneous), lanosterol has not

been found in plants. When labeled lanosterol is fed to
Many steroids (compounds with a tetracyclic cyclopentanop- plants, no label ends up in cycloartenol, but labeled cycloar-
erhydrophenanthrene ring system that have suffered the bio- tenol is incorporated readily into other phytosterols. Some
synthetic loss of methyl groups) are found in plants. Sterols fungi and certain other organisms appear to utilize either
playa major structural role in membranes of plant cells; the or both pathways. 2,3-0xidosqualene:lanosterol cyclase (BC
greatest sterol content is in the plasma membrane (Goad, 5.4.99.7) has been isolated from several plants (Goodwin,
1977, 1991a; Goodwin 1980, 1981; Heftrnan, 1973). All eu- 1985).
caryotic organisms synthesize sterols or have an absolute
requirement for them in the diet.
Formation of Cholesterol
In plants, sterols arise through the intermediacy of Cholesterol (7), the major sterol in most animals, also is
cycloartenol (3) and 2,3-oxidosqualene:cycloartenol cyclase found in small amounts in many plants, where it serves as
(EC 5.4.99.8), whereas, in animals, sterols arise from lanost- a precursor for other steroid derivatives, such as the proges-
erol (IS) and 2,3-oxidosqualene:lanosterol cyclase (EC tagens and cardiac glycosides (Heftman, 1984). In plants,
5.4.99.7) (Figs. 23.7 and 23.8). Despite several reports (all algae, and fungi, a variety of other phytosterols may serve
434 Triterpenes and Steroids

squalene (1)

(cbair·boat·cbair·boat)

HO

norlanosterol obtusifoliol (18)

HO
progestagens

t
dehydrolophenol desmosterol cholesterol (7) cardiac glyoosides

Fig. 23.7. Cholesterol biogenesis in plants (modified from Grunwald, 1980; modified and used with pennission of the copyright owner, Sprioger-
Verlag, Berlin),

in
animals
squalene (1) --+- --+-

HO
HO

lanosterol (15) cholesterol (7)

Fig. 23.8. Cholesterol biosynthesis in animals and some fungi.

 Triterpenes and Steroids 435

80 80

cycloeucalenol (19) obtuslfoliol (18)

Fig. 23.9. Cycloeucalenol and obtusifoliol.

as substrates for further metabolism, whereas in animals, methyl group at C-14 (Fig. 23.10). The order of removal of
cholesterol is the source of all steroid metabolites (Grun- methyl groups appears to vary from system to system.
wald, 1980; Harrison, 1988).
Experiments in which com, Zea mays, seeds produce ob- Formation of Other Plant sterols
tusifoliol (18) from cycIoeucalenol (19) upon gennination
in 2H20, the IH_NMR spectra indicate significant incorpora- In most plants, (3-sitosterol (4), campesterol (6), and stig-
tion of deuterium into the C-19 methyl group (Fig. 23.9). masterol (5) are the most abundant phytosterols. These com-
Experiments of this type provide some direct evidence for pounds arise from cycIoartenol (3) by a series of reactions
the intennediacy of these compounds in the biosynthesis of including modification of the side chain (Fig. 23.11) (Goad,
cholesterol in plants. 1991 b; Grunwald, 1980; Heftman, 1973). A series of olefinic
The three methyl groups that are lost in the fonnation of intennediates has been postulated and several metabolites
cholesterol (one at C-14 and two at C-4) probably are re- of this type have been isolated. Several basic skeletons of
moved via the oxidative sequence -Me to -CH2-OH, to phytosterols are encountered; these, together with some of
-CHO, to -C02H and decarboxylation, as indicated for the the biogenetic pathways by which they might arise, are given

24,15-dihydrolanosterol

80

80
o
cholesterol (7)

Fig. 23.10. Demethylation to produce cholesterol (Harrison, 1985; modified and used with penmssion of the copyright owner, The Royal Society of
Chemistry, London).
436 Triterpenes and Steroids

cycloartenol (3) 24·methylenecycloartenol cycloeucalenol (19)

obtusifoliol (18) 4a·methylergosta· 4a-methylergosta-

8,14,24(28).trien·3B·ol (20) 8,24(28)·dien·3B·ol

HO ~~~
•
H
I

24·methylenelophenol
--.
H0 1H
.
I

24-ethylidenelophenol
--
HO
H
I

stigmasta-7,24(28)·dien-30-ol
--+

eampesterol (6) stigmasterol (5)

Fig. 23.11. Proposed biosynthetic origin of plant sterols (modified from Grunwald. 1980; used with permission of the copyright owner, Springer-
Verlag, Berlin).

below (Fig. 23.11); again, a series of different pathways may Alkylation of the side chain may occur at C-24 with suc-
be involved (Mann, 1987). cessive methyl groups derived from S-adenosyl methionine.
Microsomal extracts from maize seedlings are capable of The first addition is catalyzed by a sterol-S-adenosylmethio-
catalyzing the alkylation of cycloartenol (3) by S-adenosyl- nine methyl transferase (Goad, 1991b; Goodwin, 1985). The
methionine to produce 24-methylenecycloartenol (Fig. side chain of (3-sitosterol (4) may be derived as in Fig. 23.12.
23.11) (Harrison, 1988). The enzymes involved in transfor-
mations of cycloeucalenol (19) to obtusifoliol (18) (cycloeu- Biological Activity of Plant Sterols
calenol : obtusifoliol isomerase), obtusifoliol to 4a-methyl-
ergosta-8,14,24(28)-trien-3(3-01 (20) (14a-methylsterol A number of plant sterols have been reported to possess
14a-demethylase), 4a-methylergosta-8,14,24(28)·trien-3(3- growth-regulating activity and developmental modification
01 to 4a-methylergosta·8,24(28)-dien-3(3-01 (L~14-sterol re- properties in plants (Heftman, 1975a, 1975b, Mandava,
ductase), 4a·methylergosta-8,24(28)-dien-3(3-01 to 24- 1979). For example, (3-sitosterol (4) initiates flower buds
methylenelophenol (1:>.8 .... 1:>.7-sterol isomerase), and sitos- in Chrysanthemum species; exogenously applied lanosterol
terol to stigmasterol (sterol-I:>.22-desaturase) have been stud- (15) (not a plant product) also stimulates flowering in these
ied (Fig. 23.11) (Goad, 1991b; Goodwin, 1985). plants. Estrone (oestrone) (21) and related compounds are
 Trite'penes and Steroids 437

reported to increase the number of flowers in Ecballium ela- calcium ions from the diet, stimulates transport of calcium
terium and to influence sex expression. None of these pro- ions in vivo, and, in conjunction with parathyroid honnones,
ceSses are well understood (Mandava, 1979). Despite the mobilizes calcium ions from bone (Williams et aI., 1989).
reports of in vitro growth-regulating activity of progestagens A South American plant, Solanum malacoxylon, produces
in plants, there is no evidence for in vivo activity. a lu,25-dihydroxy-vitamin D3 (24) glycoside with similar
Probably the most effective compound of this type for biological activity. Cattle that consume leaves of this plant
the modification of plant growth discovered to date is brass- exhibit all the symptoms of hypervitaminosis D (Williams
inolide (22), 2u,3u,22u,23u-tetrahydroxy-24u-methyl-f3- et aI., 1989).
homo-7-oxa-5u-cholestan-6-one, a steroidal lactone first The polyphagous insect Helicoverpa zea exhibited re-
isolated from the pollen of rape, Brassica napus (Brassica- duced growth on plants of Petunia hybrida. Steroidal com-
ceae) (Fig. 23.13) (Grove et aI., 1979; Mandava, 1988). Al- pounds, such as petuniasterone A (25), proved responsible
though the mechanism(s) by which this compound functions for this activity (Fig. 23.13) (Elliger and Waiss, 1989; Elliger
are not known, the effects are rapid (Meudt, 1987). Brassino- et aI., 1988).
lide affects specific target tissues that are sensitive to the A mixture of free sterols, steryl ferulates, and diglycerides
plant hormone indole-3-acetic acid and tissues that are gravi- from rice grain were found to induce oviposition of the rice
perceptive. Picomole quantities of brassinolide applied to grain weevil, Sitophilus zeamais. When the individual com-
young bean seedlings cause cell elongation, cell division, ponents of the mixture were tested separately, however, they
and splitting of the treated internode. The symptoms are repelled the female from egg laying (Harborne, 1987).
confined to the treated area, suggesting that brassinolide is 5f3-Cholestan-3,24-dione (26) serves as a trail-marking
not translocated. A series of related compounds have now pheromone for the gregarious caterpillars of Malacosoma
been isolated from several different plants (Meudt, 1987). neustria andM. americanum (Lepidoptera: Lasiocampidae).
Phytosterols are of particular importance to insects, nem- Larvae in the vanguard offoraging columns establish chemi-
atodes, and certain crustaceans, because they cannot synthe- cal trails as they explore new territory. These exploratory
size cholesterol de novo. These organisms degrade dietary trails are overmarked by fed larvae returning to the tent (Pe-
C 28 and C29 -phytosterols to C27 -sterols (usually cholesterol) terson, 1988). Successful foragers recruit colony mates to
or obtain C2?,sterols directly from other organisms (Ikek- feeding sites (Peterson, 1988). The caterpillars have a thresh-
awa, 1983). These steroidal products are then used in the old sensitivity of 10- 11 g/mm of trail (Harborne, 1989).
synthesis of biologically active sterols that the organisms
require. There is evidence that dealkylation of phytosterols
proceeds as indicated in (Fig. 23.14) (Harrison, 1985; Ikek- PROGESTAGENS
awa, 1983).
Progestanes in Plants and from Animals
Many animals convert steroidal precursors to vitamin D
(23) and related compounds (DeLuca and Schooes, 1983) Progestagens are steroids with 21 carbon atoms. Many
(Fig. 23.13). Vitamin D is involved in the assimilation of of these compounds have been isolated from animals (Fig.

Fig. 23.12. Proposed biogenesis of ~-sitosterol (Knights, 1973; modified and used with pennission of the copyright owner, The Royal Society of
Chemistry. London).
438 Triterpenes and Steroids

HO

la,lS·dihydrocholeealclferol (24)

o
petuniasterone A (25) brasslnolide (22) 58·cholestan·3,24-dione (26)

Fig. 23.13. Biologically active sterols (modified in part from DeLuca and Schnoes, 1983).

Fig. 23.14. Dealkylation of phytosterols (modified from Hanison, 1985; used with pennission of the copyright owner, The Royal Society of Chemistry,
London).
 Triterpenes and Steroids 439

HO

cholesterol (7)
'
~
.
~usP
HO::""

pregnenolone (27)
!
H-
0 b
-
H

progesterone (28)
* cytochrome P-4S0, 02,
!
r(i
OH OH

HO~
~Irr --"00
Oay.-
.
o
estradiol (31) cortisone (synthetic) testosterone (34)

androst-S-en-3p-ol-17-one aldosterone estrone (21)

Fig. 23.15. Progestagens from animals.

23.15), but a number of these steroids have been isolated have been shown to have pronounced effects on plant growth
from various plants as well (Farnsworthet al., 1975a, 1975b; and development (Geuns, 1978). Several cause increases in
Heftman, 1975a, 1975b). Among the plant-derived progesta- growth of various plant parts [e.g., estrone (21) increases
gens are pregnenolone (27) and progesterone (28) (which growth of root tips]. This compound also increases growth of
also occur as animal hormones). These compounds are pre- the pollen tube of Rumex tenuifolius. Application of estradiol
cursors of other plant progestagens (Fig. 23.15) (Grunwald, (31) to plants of Callistephus chinensis (Asteraceae) and
1980). A number of these compounds are identical to those Lemna minor (Lemnaceae) induces flowering. Application
in animals and other plant progestagens closely resemble of 171l-estradiol can replace the inductive cold period neces-
them (Geuns, 1978). sary for flowering in Cichorium intybus. Further effects on
In animals, the corticosteroids are syuthesized from pro- sex determination on plants and flowers have been observed
gesterone in the cortex of the adrenal gland. These very (Geuns, 1978). In the past, many of these treatments have
important 21-carbon steroids also may occur in plants. been difficult to repeat. The poor solubility of many of these
The C2rsteroids are derived from sterols by oxidative compounds and the necessity for an appropriate cosolvent
cleavage of the C-17 side chain. Cholesterol (7) is converted may be a major reason for these difficulties (Geuns, 1978).
to pregnenolone (27) (by C-17 side-chain cleavage) in a As previously mentioned (see Chapter 11), a number of
number of plants, although C2.-sterols [e.g.,Il-sitosterol (4)] isoflavones and their derivatives also possess estrogenic ac-
also may be converted (Grunwald, 1980). The conversion tivity, possibly because the overall shape of the molecules is
of cholesterol to pregnenolone has been demonstrated in a similar to that of steroids (Farnsworth et al., 1975a, 1975b).
number of plants (Fig. 23.16). Intermediates in the process
are 221l-hydroxycholesterol (29) and 20,22-dihydroxycho- Progestagens from Insects
lesterol (30). The conversion of pregnenolone (27) to proges-
terone (28) requires rearrangement of a double bond [A5(6) Several arthropods are rich sources of steroidal defensive
- A4 (5)]. The reverse reaction also has been observed in compounds. These progestagens are particularly common in
plants (Grunwald, 1980). water beetles of the subfamilies Colymbetinae and Dytisci-
Compounds with estrogenic activity occur in plants and nae. Beetles of these families accumulate steroidal toxins as
440 Triterpenes and Steroids

cholesterol (7) 22~-hydroxycholesterol (29) 20,22-dlhydroxycholesterol (30)

• cytochrome P-4S0, 02

pregnenolone (27) progesterone (28) 11 ~deoxycorticosteroDe

12-hydroxy-4,6-pregnadien-3,20-dione (32) cortexone (33)

Fig. 23.16. Proposed progestagen biogenesis in plants (modified from Grunwald, 1980; used with pennission of the copyright owner, Springer-Verlag.
Berlin) and progestagens from insects.

a poisonous milky liquid in the prothoracic glands. The ac- mg), now named ecdysone, was isolated in 1954 from 500
tivity of the toxin is not detected by the predator, until it bas kg (one-half ton) of silkworm pupae after about 10 years of
actually swallowed a beetle. Within a few minutes, it sickens work (Butenandt and Karlson, 1954; Karlson, 1956a,
and disgorges its prey. Predatory fish, feeding on the same 1956b)_ These hormones, produced by the insect larva, act
beetles, fall into a narcotic state and, subsequently, do not eat upon the epidermal cells and cause initiation of molting. The
the beetles. Most of these water beetle toxins are pregnane overall balance of juvenile hormone and the ecdysteroids
derivatives; at least 15 structures have been identified from controls the process of larval development and metamorpho-
a similar number of beetles. An individual Mexican water sis (Bowers, 1991; Camps, 1991; Rees, 1983). It is generally
beetle, Cybister tripunctatus, contains as much as 1 mg of agreed that 20-hydroxyecdysone «(3-ecdysone) (35) is the
12-hydroxy-4,6-pregnadien-3,20-dione (32). Each C.limba- active hormone that contruls different events during molting
Ius beetle contains the same amount of cortexone (33) (Fig. (Camps, 1991). Crustaceans also have an ecdysteroid re-
23.16). In addition to the pregnance derivatives, one beetle, qnirement. For example, callinecdysone-B (36) is found in
llybius, contains testosterone (34), dehydrotestosterone, es- the soft-shell crab and 20-hydroxyecdysone «(3-ecdysone)
tradiol (31), and estrune (21) (Harbome, 1982; Herout, 1970; (35) in crayfish.
Schildknecht, 1970, 1971). Although phytosterols are converted into other sterols
(such as cholesterol) that serve as components of membranes
and in other functions, one of the principal reasons that in-
ECDYSTEROmS sects require phytosterols is for conversion to ecdysteroids.
Insects differ in their ability to utilize different phytosterols
Some of the most interesting insect phytosterol derivatives (Svoboda and Thompson, 1987). Although cholesterol (7)
are the so-called moulting hormones or ecdysteroids (Fig. will satisfy this demand in most insects, most plants contain
23.17) (Hirino and Hirino, 1970). The first ecdysteroid (25 only traces of this sterol (Svoboda and Thompson, 1987).
 Triterpenes and Steroids 441

In some instances, the symbionts of insects may synthesize with material from necrotic tissues of saguaro and cardon.
or modify steroids used by the host insect (Herout, 1970). This species can tolerate concentrations of secondary com-
An interesting example of insect-host plant specificity pounds greater than the other fly species (Frank and
that involves phytosterols and ecdysterones is found in the Fogleman, 1992).
Sonoran Desert area of North America. Four Drosophila Cytochrome P-4S0 oxidases are involved in host-plant
species that are not especially closely related exhibit remark- utilization by these Sonoran Desert Drosophila species
able specificity in choosing to feed variously on rotting limbs (Frank and Fogleman, 1992).
of four cactus species from the Sonoran Desert (Frank and Phytosterols in the various cactus species exist as glyco-
Fogleman, 1992). Each species feeds on a specific cactus sides or saponins. Yeasts found in the rotting cactus plants
with very few exceptions. hydrolyze these saponins and make the phytosterol agly-
Drosophila pachea is limited to the senita cactus (Lopho- cones available to the insects (Spencer, 1987). Both 24R-
cereus schottii) by a biochemical dependency on sterols and 24S-methyl groups occur in phytosterols. Individuals of
found in that cactus. Unlike most insects that can utilize such Drosophila me/anogaster utilize both 24R- and 24S-meth-
compounds as l3-sitosterol (4) to synthesize ecdysones, flies ylcholest-S,7-dien-313-01 (37), whereas those of D. pachea
of this Drosophila species must have the compound schot- prefer the 24S-isomers and those of D. mojavensis prefer the
tenol (37) in which a particular double-bond position is 24R-isomers (Rees, 1983).
found (Fig. 23.18) (Harborne, 1982; Herout, 1970; Kircher, The co-occurring yeasts are an additional part of the co-
1982). This cactus contains 3-lS% dry weight of complex adapted system, as they are capable of hydrolyzing steroid
isoquinoline alkaloids that are toxic to both adults and larvae glycosides, and produce volatile compounds attractive to the
of Drosophila nigrospiracula, D. mojavensis, and D. mela- flies. Steroidal aglycones selectively inhibit Drosophila mo-
nogaster. javensis at levels that occur naturally in the plants (Fogleman
Individuals of Drosophila nigrospiracu/a use saguaro and Armstrong, 1989). The array of cactus alkaloids present
(Carnegiea gigantea) on the mainland and cardon (Pachy- was not poisonous to this species of fly. Nonadapted and
cereus pringlei) in Baja California. The two cacti are mor- nonspecialized insects do not exhibit selective tolerance to-
phologically and chemically similar (Frank and Fogleman, ward any given compound arrays and are susceptible to tox-
1992). Both have low concentrations of isoquinoline alka- icity by either steroidal aglycones or alkaloids or both. The
loids. aglycones may be relatively specific toxins for species of
The host plants for Drosophila mojavensis are agria Drosophila. Thus, the alkaloids appear to be a general deter-
(Sfenocereus gummosus) and organ pipe cactus (Stenocer- rent to herbivory, and the triterpenoid glycosides, and associ-
eus thurberi). The insect uses agria on the peninsula and ated hydrolytic glycosidases provide a specific toxification
organ pipe cactus on the mainland. These cacti do not contain against Drosophila species (Spencer, 1987).
alkaloids, but instead possess medium-chain fatty acids The complement of both alkaloids and phytosteroids now
(C.-C l2 ) and triterpene glycosides (Frank and Fogleman, is known to be more complex than previously thought, and
1992). a complex relationship of saponins, aglycones, alkaloids, and
Females of Drosophila mettleri oviposit in soil soaked yeasts determine on which of the cacti a particular Droso-

OH

OH

HO

HO HO
o
ergosterol (39)

OH OH

HO HO

HO HO
0 0
p-ecdysone (35) callinecdysone B (36)

Fig. 23.17. Ecdysteroids.

442 Triterpenes and Steroids

OH

OH

HO
2. dealkylation
atC-24
HO HO
3. oxidation o
P-sitosterol (7) a-ecdysone (34)

tI 1. dealkylation at C-24

oo?
H 2. oxidation

HO

24-R-methylcholest-5,7-dien-3B-ol (38) schottenol (37)

Fig. 23.18. Proposed path of synthesis of molting hormone from f3-sitosterol or schottenol (modified from Harhorne, 1982).

phila species will be successful (Frank and Fogleman, 1992; (40). Cholesterol (7) has been shown to be a precursor of 20-
Spencer, 1987). hydroxyecdysone (35) in Podoearpus elata and Polypodium
In another example, metamorphosis of the beetle Xyleb- vulgare (Goodwin et aI., 1970; Heftman et al., 1968). It is
orus ferrugineus will occur only if the larva receives the not clear, however, that cholesterol is an obligatory precur-
metabolic assistance of the fungus Fusarium solani. The sor. When [4<x-3Hl, [4f3- 3Hl and [3a- 3Hlcholesterol were
latter organism has the ability to introduce a b. 7 double bond administered to Polypodium vulgare,label from the 3a-posi-
into the nucleus of dietary sterols; such functionality is an tion was found to have migrated to C-4, that from 4f3 to C-
essential feature of all ecdysteroids. Larvae without the sym- 5, and that from 4a remained at C-4 (Davies et al., 1980).
biotic fungus will metamorphose only if provided with er- This leads to the proposal of the following mechanism in-
gosterol (39) (Fig. 23.17), a sterol that has this double bond cluding cholest-7-en-3f3-01-5,6-epoxide as an intermediate
(Kircher, 1982). (Fig. 23.20). Hydroxylation of the side chain occurs late in
An important group of plant and animal pathogens, the the biosynthetic sequence and introduction of a hydroxyl
nematodes, also possess a requirement for dietary sterols function at C-20 seems to be the last step in the synthesis.
(Chitwood et aI., 1987). Although complex in appearance, the pathways that lead to
this group of compounds are widely distributed, presumably
arose early in evolution, and have been maintained in many
PHYTOECDYSTEROlDS extant groups of plants.
Approximately 100 phytoecdysteroids have been re-
A number of plants synthesize ecdysteroids and sequester ported to occur in plants of about 80 plant families. Results
them in much greater concentrations than do insects (Fig. of a sensitive bioassay capable of detecting 5 X 10-9 g of
23.19). It is probable that these compounds serve as defen- 20-hydroxyecdysone (35) suggest that phytoecdysteroids are
sive systems in the plant much in the same way as juvenile found in 111 families (Camps, 1991). These compounds are
hormone mimics. For activity, a cis-A,B-ring fusion, a 6- most common in the ferns, Podocarpaceae (Gymno-
keto-7-ene group, a full sterol side chain with a 22R-oxygen spermae), and the Amaranthaceae. Among the angiosperms
function, an oxygen function at C-3, and oxygen groups at in which phytoecdysteroids are known to occur are Ajuga
C-14 and 2f3 are necessary. However, not all insects may (Lamiaceae), Aehryanthes, Spinaeea, and Chenopodium
respond in an identical manner (Camps, 1991). (Amaranthaceae), Vitex (Verbenaceae), Stachyurus (Stachy-
Because plants can synthesize ecdysteroids from meva- uraceae), and Diploelisia and Abuta (Menispermaceae)
lonic acid (10) and acetate, it is clear that they (in contrast (Harbome, 1986; Hikino and Takemoto, 1974; Miller et aI.,
to insects) possess the entire pathway leading to these impor- 1985). Phytoecdysteroids are not found in most fern allies
tant compounds. (Camps, 1991). A series of ecdysones were discovered even
The pathway leading to ecdysteroids in plants is not en- in the marine red alga Laurencia pinnata (Harbome, 1989).
tirely understood, but probably proceeds via intermediate The bracken fern, Pteridium aquilinum, has been shown
 Triterpenes and Steroids 443

OH

HO

HO

q5P
ponasterone A (42) inokosterone ponasterone B

Vo
(callinecdysone A)

OH
"",,,OHr

HO HO i
I (lH
HO HO H
o
pterosterone ponasterone C cyasterone

Fig. 23.19. Some representative phytoecdysteroids.

22 21 24
27
25

HO
cholesterol (7) 7-deoxycholesterol (40)
OH

OH

HO

o
o
Fig. 23.20. Proposed pathway leading to the fonnation of 20-hydroxyedcysone from cholesterol (modified from Davies et aI., 1980; used with
pennission of the copyright owner, the Biochemical Society. London).
444 Triterpenes and Steroids

to contain ecdysone (a-ecdysone) (41) and 20-hydroxyecdy- Labeling studies with [3- 3Hl-fucosterol (46) (the major
sone (35), as well as compounds that appear to be juvenile sterol of Aehlya) indicate that antheridiol and oogoniols 1
honnone mimics (Kaplanis et al., 1967). Although the com- and 2 are derived from this compound.
pounds had activity (including death without molting) when
injected into the nymphs of Sehistoeerca gregaria (the desert
locust), they were inactive when fed to the insects. However, CVCURBITACIl'IS
to the larvae of the cecropia silkwonn, Hyalophora cecropia,
ponasterone A (42) was highly active when incorporated
The most bitter and distasteful triterpenoids in plants proba-
into artificial diets. Toxicity (interfering with development
bly are the cucurbitacins, a series of about 20 tetracyclic
and reproduction) has been shown in other insects as well
triterpenes occurring in a number of plants of the Cucurbita-
(Camps, 1991; Robbins et al., 1970).
ceae (Fig. 23.22) (Ferguson and Metcalf, 1985; Gershenzon
Although there is evidence that phytoecdysteroids do af-
and Croteau, 1991; Lavie and Glotter, 1971; Mabry and Gill,
fect insect growth and development upon ingestion, this role
1979). Cucurbitacin B (47) and E (48) are the most common
should be evaluated in the context of multifaceted chemi-
of these compounds. Plant materials containing cucurbitac-
cally based resistance in plants (Camps, 1991).
ins are toxic to mammals, have strung purgative action, and
Brassinolates (see above) are similar to phytoecdysteroids
play a role in the defense against mammalian herbivores.
in structure (Camps, 1991).
Many have LCso values in the range 1-7 mg/kg (interperito-
neally) and 5-40 mglkg orally in mice. Humans can detect
the bitter taste of certain cucurbitacins as low as 1 ppb (Met-
CHEMMAXIS Il'I fUNGI calf, 1986), although not all cucurbitacins are bitter. For
example, the rhizomes of I(emsleya earnosijlora (Cucurbita-
In the phycomycete genus AchJya (a coenocytic water mold), ceae) contain six cucurbitacins: one is tasteless, two are
sexual reproduction is controlled by a set of two steroidal sweet, and three are bitter (Harborne, 1989). Cucurbitacin
derivatives: antheridiol and oogoniol. None of the strains B (47) (from Jpomopsis aggregata, Polemoniaceae) is cyto-
of Aehlya ambisexualis and A. heterosexualis fonn sexual toxic and has a EDso of 0.032 I1g/ml. Cucurbitacin F (49)
organs when grown alone, but when cultures of anyone type (from Elaeoearpus doliehostylus, EJaeocarpaceae) 0.52
are grown with an opposite type, sexual organs fonn and I1g/ml in the KB cell cultures system and lI-deoxocucurbi-
gametes are produced and fertilized. A complex series of tacin (50) had an EDso of 0.002 I1g/ml in the P-388 cell
events is involved in this process. Antheridiol (43) is re- culture system (Arisawa et al., 1984; Fang et al., 1984;
leased by female cells and induces the development of the Reddy et al., 1988).
male sex organs (the antheridia) as well as the synthesis of These highly oxygenated triterpenes have been recently
oogoniols (44 and 45) from fucosterol (46) (Jaenicke, 1991). shown to be interrelated chemically to the lanostane series.
A specific set of proteins is induced when the antheridia are They also are products of a chair-boat-chair-boat squalene
exposed to antheridiol. The antheridia are attracted to the condensation. A biogenetic sequence for cucurbitacins based
oogonia (an antheridium is a structure responsible for pro- on Coates (1976) is proposed (Fig. 23.23).
ducing male gametes and an oogonium is a structure respon- Seedlings of cucumber, Cueumis sativus, converted the
sible for producing female gametes). Fertilization tubes orig- triterpene (51) into cucurbitacin C (52); microsomal frac-
inating in the antheridia penetrate the oogonia and reach the tions from seedlings of Cueurbita maxima converted squa-
eggs. The nuclei from the male gametes migrate through the lene 2,3-epoxide into lOa-cucurbita-5,24-dien-3j3-01. Nei-
tubes and to the eggs. ther parkeol (53) (Fig. 23.1), cycloartenol (3), 11-
The presence of these honnones was demonstrated when ketocycloartenol, or 24,25-dihydro-9a,lIa-epoxyparkeol
male and female fungi were separated by membranes. An- were incorporated significantly, and no products with a cu-
theridiol (43) was first extracted from female hyphae and curbitane skeleton were isolated (Balliano et aI., 1983; Harri-
later synthesized. When this compound was put on small son, 1985).
polystyrene beads, antheridial hyphae were attracted to the Cucurbitacins originally were isolated from the Cucurbi-
beads. Antheridiol (a C 29 steroid) initiates antheridial taceae (they are found in most members of this family that
branches at 5-10 pg/ml. The structural requirements are have been examined), but are now known to occur in other
highly specific (Fig. 23.21). families. Among these are the Begoniaceae (Begonia),
A second honnone, produced by the male hyphae, has Brassicaceae (Jberis), Datiscaceae, Desfontainiaceae (Des-
been shown to be a mixture of oogoniols 1 and 2 (44 and Jontainia), Elaeocarpaceae (Elaeocarpus), Euphorbiaceae,
45). These steroidal compounds are active at 620 and 460 Liliaceae (Phormium), Polemoniaceae (/pomopsis), Primu-
ng/ml, respectively. Cultures of Achlya ambisexualis pro- laceae (Anagallis), Rosaceae (Purshia), Rubiaceae (Hin-
duced oogoniols only afrer treatment with antheridiol. An- tonia), Scrophulariaceae (Gratiola, Picrorhiza), Sterculia-
theridiol appears to induce the fonnation of an enzyme nec- ceae (Helieteres), and Thymelaeaceae (Gyrinops) (Arisawa
essary for the production of oogoniols (Mascarenbas, 1978; et aI., 1984; Blasko and Cordell, 1988; Dreyer and Trousdale
O'Day and Horgen, 1981). 1978; Fang et aI., 1984; Hegnauer, 1989; Kupchan et aI.,
 Triterpenes and Steroids 445

antheridlol (43)

OH

(CH,),CHCOO

<JOgoDiol-! (44) oogoniol-2 (45)

HO

ruCOlilerol (46)

Fig. 23.21. Structure of antheridiol and oogoniois.

1978; Mata et at, 1990; Paris and Tessier, 1972; Reddy et allomonal effects of cucurbitacins. The beetle, Epilachna
at, 1988; Reguero et al., 1987; Stuppner et al., 1991). In borealis, has developed a system of "trenching" or cutting
cucurbits, cucurbitacins are known from all plant parts (Met- a circular area of tissue off from the rest of the leaf and
calf, 1986). apparently preventing movement of cucurbitacins into the
Cucurbitacins are reported to possess growth-regulating area. The beetle eats this tissue and then relocates. The
activity in plants (Mandava, 1979). In animals, they exert amount of cucurbitacins in wounded plant tissue has been
cytotoxic activity and are purgatives. Cucurbitacin E gluco- shown to increase, and beetles fed this tissue show a reduc-
side, from Gratiola officinalis (Scrophulariaceae) has tion in fituess (Carrol and Hoffman, 1980; Tallamy, 1985).
cardiotonic activity (Wagner, 1988). Cucurbitacins from Despite the fact that transport of cucurbitacins into the
several plants have cytotoxic activity and antitumor proper- wounded area is a possible explanation, there is currently
ties (Blasko and Cordell, 1988). Cucurbitacin F (49) is cyto- little evidence for the transport of cucurbitacins in plants.
toxic in the KB and P-388 in vitro cell culture lines. Although cucurbitacins serve as allomones for most other
The presence of cucurbitacins is correlated with the inhi- insects, these compounds are kairomones, or attractants, for
bition of feeding by the polyphagous mite Tetranychus urti- many insects associated with the Cucurbitaceae (Metcalf,
cae, which is often damaging to nonbitter cucurbits. These 1985). For example, when cucumber beetles are offered a
compounds also inhibit feeding by a number of leaf beetles; choice of nonbitter and bitter (cucurbitacin containing)
among them are Phyllotreta undulata, P. tetrastigma, and fruits, they feed almost exclusively on the bitter fruits (11
Phaedon cochleariae. In/beris amara (Brassicaceae), cucur- : 1). When given the same choice, the honeybee, Apis mellif-
bitacins I and E (48) from the seeds and the green parts are era, will select food materials lacking the cucurbitacins (Har-
potent inhibitors for the flea beetle Phyllotreta nemorum borne, 1982).
(Mabry and Gill, 1979). A scenario for the evolutionary and behavioral interac-
Other insects seem to have evolved systems to avoid the tions between plants of the Cucurbitaceae and their herbi-
446 Triterpenes and Steroids

HO 0

OAc OAc

HO

o HO'"''

OH HO 0

OH OH

HO

o 0

OH

OH
OAc
HO

OH

ll·deoxocucurbitacin (50)
22.deoxocucurbitacin D

Fig. 23.22. Representative cucurbitacins.

lanosterol (in animals)

HO

parkeol
squalene 2,3-epoxide
H

OAc
3'oxidation
ll-oxidation
• side chain
oxidation HO
acetylation

Fig. 23.23. Biogenetic pathway for cucurbitacins (Hanison et aI., 1985; modified and used with pennission of the copyright owner, The Royal Society
of Chemistry, London).
 Triterpenes and Steroids 447

vores has been proposed (Andersen and Metcalf. 1987; Met- 1989b; Croteau and Johnson. 1985; Goodwin. 1985). 13-
calf. 1986; Metcalf and Lampman. 1989). Plants that possess Amyrin cyclase from Rabdosia japonica had a molecular
the ability to produce these compounds do not suffer attacks weight of 28.000. whereas that from peas had a molecular
by generalist herbivores. When changes io ability of co- weight of 35.000 (Abe et al .• 1989a. I 989b).
adapted herbivores to handle the cucurbitacins present occur. In the oleanane-ursane group of triterpenes. postulated
some of the iosects develop detoxification and excretion hydride shifts [as proposed by Eschenmoser and co-workers
pathways to neutralize the harmful effects of cucurbitacios. (1955)1 have now been demonstrated. Incorporation of [4-
As these herbivores expand ioto a new ecological niche. a 13C]MVA into oleaoolic acid (56). ursolic acid (63). and
concommitant change io the ability to detect these com- several other metabolites was accomplished with tissue cul-
pounds is favored. and. ultimately. the herbivore develops tures of Isodon japonicus (Lamiaceae). Followiog introduc-
high blood and tissue levels of conjugates of cucurbitacios as tion of [4- 13C]mevalonate and [1.2- 13C2 1acetate. the pen-
defensive substances (Andersen and Metcalf. 1987; Metcalf. tacyclic triterpene compounds were examined by 13C_NMR
1986). spectroscopy. Rings 0 and E were formed by the routes
There seems to have been an ancestral association of leaf predicted by Eschenmoser et al. (1955). The labeling pattern
beetles of the tribe Luperioi (Coleoptera: Chrysomelidae: at carbons 4. 23. and 24 of 3-epi-maslioic acid (64) indicates
Galerucioae) and plants of the family Cucurbitaceae. The that this series is formed from (3S)-squalene epoxide. rather
scope of this association presently is worldwide. In the New than from the (3R)-epimer.
World. the subtribe Diabroticioa contaios some of the most The 13C_NMR spectrum showed enrichment of carbons
important insects of the world. io an economic sense. Among 13. 18. and 19 io oleanolic acid (56) and confirmed the se-
these are cucumber beetles and com root worms. These io- quence 65-68 in the formation of oleananes (Fig. 23.26)
sects are capable of recogniziog cucurbitacios. which serve (Harrison. 1988).
as kairomones for most (if not all) members of this group of Incorporation of [3.5- 13C21mevalonate into ursolic acid
beetles. at extremely low levels. A number of these diabrotid by cell cultures of Perillafrutescens var. acuta (Lamiaceae)
beetles appear to accumulate cucurbitacios as allomones for has been examioed. In another series of experiments with
protection against predators (Ferguson and Metcalf. 1985). [2- 13C,4,4-2H2 1mevalonate. the postulated 1.2 migration of
Although primarily adults feed on plants with these com- the hydride ion from C-19 to C-20 (ursane numbering) was
pounds. both the eggs and larvae also contain cucurbitacios. confirmed (Harrison. 1988).
which may play a role io discouragiog egg predators such Thus. the origioal proposals of Eschenmoser and co-
as ants (Ferguson and Metcalf. 1985; Metcalf. 1986). workers (1955) have proven largely accurate in predicting
the routes of biosynthesis of many pentacyclic triterpenes.
Compounds such as (69) (from gum mastic. Pistacia len-
TRITEKPBI'IES DERIVED VIA A liseus. Anacardiaceae) and (70) (Fig. 23.24) may be regarded
CIIA1K-CHAIR-CHAIK-BOAT TRAl'ISITION as products of trapping of a cationic intermediate of the cycli-
STATE zation of squalene 2.3-epoxide (2) to tetracyclic and pen-
tacyclic triterpenes (Harrison. 1988).
Cyclization of squalene 2.3-epoxide (2) via a
chair-chair-chair-boat transition state gives rise to many of
the most common plant triterpenes. Pentacyclic triterpenoid BIOLOGICAL ACTIVITY OF TRlTEKPBI'IES
compounds such as a- and l3-amyrin (54 and 55). oleaoolic
acid (56). taraxerol (57). taraxasterol (58). lupeol (59). be- Although triterpenes have been considered to be relatively
tulin (60). euphol (61). friedelio (62). and ursolic acid (63) ionocuous plant constituents. several have been established
may constitute several percent of the dry weight of many recently to have pronounced physiological activity. Many
plants (Fig. 23.24). The bark of trees such as the cork oak. triterpenes have antiherbivore activity. Although phytoster-
Quercus suber. and the white birch. Betula alba. contaio ols are required for iosect and fungal growth. triterpenoid
large amounts oftriterpenes. Friedelio (62) and cerio (71) are compounds may interfere with this process and exhibit anti-
the maio compounds io cork (Croteau and Johnson. 1985). herbivore or antifungal effects (Croteau and Johnson. 1985).
Biogenetic schemes for the formation of most of these In general. those that are highly oxygenated seem to be more
groups of triterpenes have been proposed (Figs. 23.25 and active io this regard (Seigler. 1983). Triterpenes and steroids
23.26) (Coates. 1976). Most of these iovolve shifts of methyl with sugar moieties attached have soap-like properties.
and hydride groups along with skeletal rearrangements to These compounds. called saponios. are discussed io Chapter
achieve the final structures. In practice. most of these ioter- 24. Another group of triterpenes with highly modified struc-
mediates have not been substantiated. tures. restricted to a group of closely related plant families.
The cyclization of 2.3-epoxysqualene to l3-amyrin (55) are discussed io chapter 25.
by microsomal fractions from pea cotyledons. which io- Toxic triterpenoids are major constitueuts of plants from
volves the enzyme 2.3-oxidosqualene: l3-amyrin cyclase popUlations of Lantana camara (Verbenaceae) that produce
(EC 5.4.99). has been demonstrated (Abe et al.. 1989a. poisoning of livestock. The active compounds often are 1an-
448 Triterpenes and Steroids

o no

friedelin (62) p·arnyrin (55) oleanolic acid (56)

no no
a·amyrin (54) ursolic acid (63) taraxerol (57)
(.)

no
no
# #

"'~
""
!H~~,
euphol (61)
no
Ii

lupeol (59)
dammaradienol

no no
no
betulic acid germanicol
(b) betulin (60)

fi fi ~

no
0
(70)
(69)
no
(0)

Fig. 23.24 (a-c). Representative pentacyclic triterpenes.

 Triterpenes and Steroids 449

-
[1.2- 13 Cl-acetate

HO HO
lupeol other pentacyclic triterpenes a-amyrin (54)
(a)

#
""

HO
HO
I
'II
'.
II

(66)
i
-- HO

,. /
21
/
HO
HO

ursolic acid (63) (68) oleanolic acid (56)

(b)

Fig. 23.25. Biogenesis of pentacyclic triterpenes from chair-chair-chair-boat confonnation of squalene (modified from Harrison, 1985 and Herbert,
1981; used with permission of the copyright owners, the Royal Society of Chemistry, London and Chapman & HalJ, London), respectively.
450 Triterpenes and Steroids

HO

ursolic acid (63)

3-epi-maslinic acid (64)

Fig. 23_26. Incorporation of labeled mevalonic acid into oleanolic acid (Harrison, 1988; modified and used with pennission of the copyright owner,
The Royal Society of Chemistry, London).

jacquinonic acid (75) Iantadene A (72) n·hydroxytingenone (76) OH

o 0
HO
HO~O
soyasapogenol B (74) tingenone (maitenine) (77) papyriferic acid (73)

Fig. 23.27. Biologically active triterpenes.

 Triterpenes and Steroids 451

tadenes A (72) and B (Fig. 23.27). These triterpenes are dry weight). This is more than 25 times the level in mature
toxic intraruminally to sheep at doses of 80 and 200 mg/kg internodes. Furthermore, the acid is deterrent to hares at the
of body weight, respectively. A related compound, the C4 - 2% level in oatmeal (Harborne, 1986).
hydroxy derivative of lantadene A, has been isolated from Ursolic acid (63) plays a role in the a1lelopathic effects
a species Lippia rehmanni and found to be toxic to rabbits of several other types of secondary metabolites in sandhill
(Mabry and Gill, 1979). vegetation. Dihydrocinnamic acid (derived from ceratiolin)
Detailed investigations of the browsing behavior of the from Ceratiola ericoides (Empetraceae), a series ofmonoter-
snowshoe hare (Lepus americanus) on the Alaska paper penes including camphor, myrtenal, borneol, and carvone
birch (Betula resinifera) and the green alder (Alnus crispa), from Conradina canescens (Lamiaceae), and the monoter-
particularly during winter months, suggest that certain deter- penes menthofuran, epievodone, and calarninthone from
rent chemicals provide protection to particular stages in the Calamintha ashei (Lamiaceae) all displayed biological ac-
life cycle, at certain seasons, or in some tissues, and not tivity. However, when these compounds were dissolved in
others (Harborne, 1986). Juvenile growth-phase internodes a saturated aqueous solution of ursolic acid, allelopathic ac-
are made unpalatable to the hare by extremely high concen- tivity was greatly increased. Later work suggests that monot-
trations of the triterpene papyriferic acid (73) (up to 30% erpenes may be sufficiently soluble in water to produce toxic

~R:".
'" I -.
~R:::,., ~R!\
1'....H~
....
O H c ' F RH
"'"
He-;
h
'" '"

-
h h h
HOH,C
HOH,C

OH
/
HOH,C
OHC~ OH H
H

~
/HCh

HOH,C H
OHCH OH 0

(+)-22-me~I-Y'CYclOiridal '-....... Q~+ ~

......... .........

iridal
-
three isomers of irone from natural Iris oil

Fig. 23.28. Iridals (modified from Jaenicke and Marner. 1986; used with pennission of the copyright owner, Springer-Verlag, Vienna),
452 Triterpenes and Steroids

effects in any case (Fischer et al., 1994; Weidenheimer et al., REFERENCES

1993). Ursolic acid (63) appears to act as anatoral surfactant
(Fischer et al., 1988). ABE, I, U. SANKAWA, and Y. EBIZUKA, Purification of 2,3-oxido-
As the roots of parasitic plants attach to those of the host squalene: Il-amyrin cyclase from pea seedlings, Chern. Phann.
plant, a special structore, the haustorium, is formed. The Bull., 37, 536-538 (1989a).
formation of haustoria in induced by exudates from the host ABE, I., Y. EBIZUKA, S. SEO, and U. SANKAWA, Purification of
plant. When the roots of the hemiparasite Agalinis purpurea squalene-2,3-epoxide cyclases from cell suspension cultures of
(Scrophulariaceae) approach those of the host, Lespedeza Rabdosiajaponica Hara, FEBS Lett., 249,100-104 (1989b).
sericea, soyasapogenol B (74) induces haustorial formation AGNEW, W. S. and G. popiK, Squalene synthetase. Solubilization
(Fig. 23.27) (Lynn, 1985). from yeast microsomes of a phospholipid-requiring enzyme, J.
Jacquinonic acid (75) from Jacquinia pungens BioI. Chern., 253, 4574-4583 (1978).
(Theopbrastaceae) inhibits feeding by the leafcutter ant Alta ANDERSEN, J. F. and R. L. METCALF, Factors influencing disnibu-
cephalotes. As this compound is nonvolatile, tactile contact tion of Diabrotica spp. in blossoms of cultivated Cucurbita spp.,
by the ant is probably necessary for sensing the presence J. Chern. Beol., 13, 681-699 (1987).
of this triterpene. A mixture of triterpenoids from Cordia AR.!SAWA, M., J. M. PEzzUTO, A. D. KlNOHORN, G. A. CORDELL,
alliodora (Boraginaceae) similarly prevents feeding by this and N. R. FARNSWOR1H, Plant anticancer agents. XXX. Cucurbi-
ant (Harborne, 1987). tacins from Ipomopsis aggregata, J. Phann. Sci., 73, 411-413
(1984).
Several pentacyclic nortriterpene quinonemethides occur
in plants of the Celastraceae (Catha, Euonymus, Glyptopeta- BALLIANo, G., O. CAPUTO, F. VIOLA, L. DELPRINo, and L. CATfEL,
lum, May tenus, Peritassa, Plenckia, and Salacia) (Bavovada Cyclization of squalene-2,3-epoxide to lOa-cucurbita-5,24-
dien-3~-ol by microsomes from Cucurbita maxima seedlings,
et al., 1990). These compounds are cytutoxic in several sys-
Phytochemistry, 22, 915-921 (1983).
tems. 22-Hydroxytingenone (76) had an EDso of 0.073
BANTHORPE, D. V. and B. V. CHARLWOOD, The terpenoids, in Sec-
""g/ml in P-388 and 0.62 ""giml in a KB cell cultore system
ondary Plant Products (E. A. Ben and B. V. Chatlwood, eds.),
(Bavovada et al., 1990). Maitenine (or tingenone) (77) ex- 185-220, Springer-Verlag, Berlin, 1980.
hibits in vivo antitumor activity, cytotoxicity, and a pro-
BAVOVADA, R., G. BLASKO, H. SHIEH, G. M. PEzzUTO, and G. A.
nounced inhibitoty effect on protein and RNA synthesis CORDELL, Spectral assignment and cytoxicity of 22-hydroxy
(Cordell, 1978). Tinger,Jne completely inhibits Trypano- lingotone from Glyptopetalum sc/erocarpum, Plant. Medica,
soma cruzi in vitro at 30 ""giml (Borris and Schaeffer, 1992). 56, 380-382 (1990).
BELINGHHRI, L., P. BEYER, H. KulINIG, and M. GLEIZES, Solubiliza-
tion and panial purification of squalene synthase from daffodil
IRIDALS AI'ID IRONES FROM THE IRIDACEAE
microsomal membranes, FEBS Lett., 292, 34-36 (1991).
Extracts of iris (Iris, lridaceae) rhizomes have a pungent and BLASKO, G. and G. A. CORDELL, Chemistry of plant-derived anti-
bitter taste. Tissue of the yellow flag iris, Iris pseudacorus, cancer agents, in Economic and Medicinal Plant Research, Vol.
2 (H. Wagner, H. Hikino, andN. R. Farnsworth, eds.),119-191,
of Europe, is poisonous to man and animals, either fresh
Academic Press, London, 1988.
or dried, and produces unpleasant irritations of the mucous
BOAR, R. B. and J. Au.EN,Il-Amyrin triterpenoids, Phytochemistry,
membranes, vomiting, and bloody diarrhea. Rhizoma iris
12,2571-2578 (1973).
was formerly a constituent of folk medicine and was consid-
ered a potent purgative. An essential oil is prepared from BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from
plants, in Phytochemical Resoutces for Medicine and Agricul-
the roots of Iris pa/lida and I. germanica. Mter harvesting
ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum
and sun-drying, the roots assume a pleasant violet-like scent Press, New York, 1992.
which increases with time. The oil obtained by steam distilla-
BOWERS, W. S., Insect hormones and antihonnones in plants, in
tion is . 'essence d'iris" or . 'orris root oiL" The odoriferous Herbivores: Their Interactions with Secondary Plant Metabo-
principle is called "irone." Three isomers are the major lites (G. A. Rosenthal and M. R. Berenbaurn, eds.), 431-456,
component of this essential oil (Fig. 23.28). In contrast to Academic Press, San Diego, CA, 1991.
the ionones that are widespread in nature, irones are found BUTENANDT, A. and P. KARLsON, Ueber die Isolierung eines Meta-
primarily in orris root oil. Freshly harvested rhizomes do morphosehormone der Insekten in kristallisierten Form, Z. Na-
not contain irones; the scent develops gradually over 3-4 turforsch., 9b, 389-391 (1954).
years of storage of the root stocks. It seems unlikely that CAMPS, F., Plant ecdysteroids and their interaction with insects, in
irones arise by enzymatic processes; oxidizing agents accel- Ecological Chemistry and Biochemistry of Plant Terpenoids (J.
erate formation of these compounds. The precursor of these B. Harbome and F. A. Tomas-Barbenin, eds.), Phytochemistry
compounds is iridal (Jaenicke and Marner, 1986), which is Society of Europe Vol. 31, 331-376, Clarendon, Oxford,1991.
derived from squalene. A biogenetic pathway has been pro- CARROLL, C. R. and C. A. HOFFMAN, Chemical feeding deterrent
posed which rationalizes the formation of iridal and related mobilized in response to insect herbivory and counteradaption
compounds (Jaenicke and Marner, 1986). Methylation at C- by Epilachna tredecimnotata, Science, 209, 414-416 (1980).
22 appears to be involved in formation of irones possibly CHITWOOD, D. J., R. LoZANO, W. R. LUSBY, M. J. THOMPSON, and
via 22-methyl-'Y-cycloiridal. 1. A. SVOBODA, Metabolism and function of sterols in nema-
 Triterpenes and Steroids 453

todes, in Ecology and Metabolism of Plant Lipids (G. Fuller FARNSWORTH, N. R., A. S. BINGEL, G. A. CORDELL, F. A. CRANE,
and W. D. Nes, eds.), ACS Symposium Series 325, 200-217, and H. H. S. FONG, Potential value of plants as sources of new
American Chemical Society, Washington, DC, 1987. antifertility agents II, J. Pharm. Sci., 64, 717-754 (1975b).
COATES, R. M., Biogenetic-type rearrangements of terpenes, FERGUSON, J. F. and R. L. METCALF, Cucurbitacins. Plant-derived
Fortschr. Chem. Org. Naturst., 33, 73-230 (1976). defense compounds for Diabroticites (Coleoptera: Chrysomel-
CONNOLLY, J. D. and R. A. HiLL, Triterpenoids, Nat. Prod. Rep., idae), J. Chern. Ecol., 11, 311-318 (1985).
6,475-501 (1991). FISCHER, N. H., N. TANRISEVER, and G. B. Wll.LlAMSON, Allelopa-
CONNOLLY, J. D. and K. H. OVERTON, The triterpenoids, in Chemis- thy in the Florida scrub community as a model for natural herbi-
try of Terpenes and Terpenoids (A. A. Newman, ed.), 207-287, cide actions, in Biologically Active Natural Products: Potential
Academic Press, London, 1972. Use in Agriculture (H. G. Cutler, ed.), ACS Symposium Series
380,233-249, American Chemical Society, Washington, DC,
CORDELL, G. A., Anticancer agents from plants, in Progress in
1988.
Phytochemistry Vol. 5 (L. Reinhold, J. B. Harborne, and T.
Swain, eds.), 273-316, Pergamon, London, 1978. FISCHER, N. H., G. B. Wll.LIAMSON, J. D. WElDENIlAMER, and D.
COSCIA, C. J., Terpenoids, Vol. 1 of Handhook of Chromatography
R. RICHARDSON, In search of allelopathy in the Florida scrub:
The role of terpenoids, J. Chem. Eco!., 20, 1355-1380 (1994).
(G. Zweig and 1. Sherma, eds.), CRC Press, Boca Raton, FL,
1984. FOGELMAN, J. C. and L. ARMs'rn.ONG, Ecological aspects of cactus
triterpene glycosides I. Their effect on fitness components of
CROTBAU, R. and M. A. JOHNSON, Biosynthesis of terpenoid wood
Drosophila mojavensis, J. Chern. Ecol., 15, 663-676 (1989).
extractives, in Biosynthesis and Biodegradation afWood Com-
pounds (T. Higuchi, ed.), 379-439, Academic Press, Orlando, FRANK, M. R. and J. C. FOGLEMAN, Involvement of cytochrome P-
FL, 1985. 450 in host-plant utilization by Sonoran Desert Drosophila,
Proc. Natl. Acad. Sci. USA, 89, 11998-12002 (1992).
DAVIES, T. G., W., J. S. LoCKLEY, R. BOlD, H. H. REBs, and T.
W. GOODWIN, Mechanism of formation of the AlB cis ring junc- GERSHENZON.1. and R. CROTEAU, Terpenoids, in Herbivores: Their
tion of ecdysteroids in Polypodium vulgare, Biochem. J., 190, Interactions with Secondary Plant Metaholites (G. A. Rosenthal
537-544 (1980). and M. R. Berenbaum, eds.), 165-249, Academic Press, San
Diego, CA, 1991.
DE LUCA. H. and H. K. SCHNOES, Vitamin D: Recent advances,
Annu. Rev. Biochem.. 52, 411-439 (1983). GEUNS, 1. M. C., Steroid hormones and plant growth and develop-
ment, Phytochemistry, 17, 1-14 (1978).
DEVON, T. K. and A. I. Scon, Handhook of Naturally Occurring
Compounds, Academic Press, New York, 1972. GoAD, L. J., The biosynthesis of plant sterols, in Lipids and Lipid
Polymers in Higher Plants (M. Tevini and H. K. Lichtenthaler,
DREYER, D. L. and E. K. TROUSDALE, Cncurbitacins in Purshia
eds.), 146-168, Springer-Verlag, Berlin, 1977.
tridentata, Phytochemistry, 17, 325-326 (1978).
GoAD, L. 1., Phytosterols, in Terpenoids (B. V. Charlwood and D.
EICHENBERGER, W., Steryl glycosides and acylated steryl glyco-
V. Banthorpe, eds.), Vol. 7 of Methods in Plant Biochemistry
sides, in Lipids and Lipid Polymers in Higher Plants (M. Tevini
(P. M. Dey andJ. B. Harhorne, eds.), 369-434, Academic Press,
and H. K. Lichtenthaler eds.), 169-182, Springer-Verlag, Ber-
London, 1991 a.
lin, 1977.
GOAD, L. J., Inhibition of phytosterol biosynthesis and the conse-
ELLIGER, C. A. and A. C. WAlSS, JR., Insect growth inhibitors from
quences for plant growth, in Ecological Chemistry ~ Bio-
Petunia and other solanaceous plants, in Insecticides of Plant
chemistry of Plant Terpenoids (J. B. Harborne and F. A. Tomas-
Origin (J. T. Amason, B. J. R. Philogene, and P. Morand, ed.),
Barberan, eds.), Phytochemistry Society of Europe Vo!' 31,
ACS Symposium Series 387, 188-206, American Chemical
209-229, Clarendon, Oxford, 1991b.
Society, Washington, DC, 1989.
GOAD, L. J. and T. W. GOODWIN, The biosynthesis of plant sterols,
ELLIGER, C. A., M. E. BENSON, W. F. HADDON, R. E. LUNDIN, A.
in Progress in Phytochemistry Vol. 3 (L. Reinhold and Y.
C. WAISS, JR., and R. Y. WONG, Petuniasterones, novel ergos-
Liwschitz, eds.), 357-362, Wiley-Interscience, 1973.
tane-type steroids of Petunia hybrida Vilm. (Solanaceae) having
insect-inhibitory activity. X-ray molecular structure of the GoODWIN, T. W., Recent developments in the biosynthesis of plant
22,24,25-[(methoxycarbonyl)orthoacetatel of 7o<,22,24,25-te- triterpenoids, in Terpenoids: Structure, Biogenesis and Distribu-
trahydroxyergosta-1,4-dien-3-one and of 1o<-acetoxy-24,25- tion, Vol. 6 (Y. C. Runeckles and T. J. Mabry, eds.), 97-115,
epoxy -70< - hydroxy - 22 - (methylthiocarhonyl)acetoxyergost - 4- Academic Press, London, 1973.
en-3-one,1. Chern. Soc., Perkin I, 711-717 (1988). GOODWIN, T. W., Biosynthesis of sterols, in Lipids: Structure and
EsCHBNMOSER, A., L. RUZICKA, O. JEGER, and D. AIuooNl, Zur Function (P. K. Stumpf, ed.), Vol. 4 of Biochemistry of Plants
Kenntnis der Triterpene. Eine stereochemische Interpretation (P. K. Stumpf and E. E. Conn, eds.), 485-507, Academic Press,
der biogenetischen Isoprenregel bei den Triterpenen, Helv. New York, 1980.
Chim. Acta, 38, 1890-1904 (1955). GOODWIN, T. W., Biosynthesis of plant sterols and other triterpen-
FANG, X., C. H. PHOEBE JR., J. M. i'EzzUTO, H. H. H. FONG, N. R. oids, in Biosynthesis of Isoprenoid Compounds, Vol. 1 (J. W.
FARNSWORTH, B. YELLIN, and S. M. HEcIIT, Plant anticancer Porter and S. L. Spurgeon, eds.), 443-480, Wiley, New York,
agents, XXXIV. Cucurbitacins from Elaeocarpus dolichostylus, 1981.
1. Nat. Prod., 47, 988-993 (1984). GOODWIN, T. W., Membrane-bound enzymes in plant sterol biosyn-
FARNSWORTH, N. R., A. S. BINGEL, G. A. CORDELL, F. A. CRANE, thesis, in The Enzymes of Biological Membranes, 2nd ed. (A.
and H. H. S. FONG, Potential value of plants as sources of new N. Martonosi, ed.), 205-226 Plenum Press, New York (1985).
antifertility agents I, J. Pharm. Sci., 64, 535-598 (1975a). GoODWIN, T. W., N. J. DESOUZA, E. L. GHISALBERTI, and H. H.
454 Triterpenes and Steroids

REEs. Studies on insect moulting hormones: Biosynthesis of Insect hormones: Alpha ecdysone and 20-hydroxyecdysone in
ecdysone, ecdysterone, and 5j3-hydroxyecdysterone in Polypo- bracken fern, Science, 157, 1436-1437 (1967).
dium vulgare, Phytochemistry, 9, 1247-1252 (1970). KARLsON, P., Biochemical studies on insect hormones, Vitam.
GROVE. M. D., G. F. SPENCER. W. K. ROHWEDDER, N. MANDAVA, Horm., 14, 227-266 (1956a).
J. F. WOOLEY, J. D. WARTHEN, JR .• G. L. STEFFBNCE, J. L. KARLsON, P., Chemische Untersuchungen tiber die Metamor-
FLIPPEN-ANDERSON, and J. C. COOK, JR., Brassinolide, a potent phosehormone der Insekten, Ann. Sci. Nat. Zool. BioI. Anim.,
growth-promoting steroid isolated from Brassica napus pollen, 18, 125-138 (1956b).
Nature, 225, 1065-1066 (1979). KIRCHER, H. W., Chemical composition of cacti and its relationship
GRUNWALD, c., Plant sterols, Annu. Rev. Plant Phys., 26, 209-236 to Sonoran desert Drosophila, in Ecological Genetics and Evo-
(1975). lution (J. S. F. Barker, and W. T. Starmer, ed.), 143-158, Aca-
GRUNWALD, C., Steroids, in Secondary Plant Products (E. A. Bell demic Press, Sydney, 1982.
and B. V. Charlwood, eds.), 221-256, Springer-Verlag, Berlin, KIRCHER, W. C., Sterols in the leaves of Cheirodendron gaudichau-
1980. dU tree and their relation to Hawaiian Drospohila ecology. J.
HARBORNE, I. B., Introduction to Ecological Biochemistry, Aca- Insect Physiol., 15,1167-1173 (1969).
demic Press, New York, 1982. KNIGHT, S. A., Carbon-13 NMR spectra of some tetra- and pen-

HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod. tacyclic triterpenoids, Org. Magn. Res., 6, 603-611 (1974).
Rep., 4, 323-344 (1986). KNIGHTS, B. A., Sterol metabolism in plants, Chern. Br., 9, 106-111
HARsORNE, J. B., Chemical signals in the ecosystem, Ann. Bot., (1973).
60 (Suppl. 4), 39-57 (1987). KUPCHAN, S. M., H. MBSHULAM, and A. T. SNEDEN, New cucurbi-
tacins from Phormium lenax and Marah oreganus, Phytochem-
HARBORNE, I. B., Recent advances in chemical ecology, Nat. Prod.
istry,17, 767-769 (1978).
Rep., 7, 85-109 (1989).
LA VIE, D. and E. GWITER, The cucurbitanes, a group of tetracyclic
HAruusON, D. M., The biosynthesis oftriterpenes and steroids, Nat.
triterpenes, Fortschr. Chern. Org. Naturst., 29,307-362 (1971).
Prod. Rep., 2, 525-560 (1985).
LYNN, D., The involvement of allelochemicals in the host selection
HARRISON, D. M., The biosynthesis of triterpenes, steroids, and
of parasitic angiosperms, in The Chemistry of AUelopathy (A.
carotenoids, Nat. Prod. Rep., 5, 387-415 (1988).
C. Thompson, ed.), ACS Symposium Series 268, 55-81, Ameri-
HEFTMANN, E., Steroids, in Phytochemistry Vol. 2 (L. P. Miller, can Chemical Society, Washington, DC, 1985.
ed.), 171-226, Van No"trand Reinhold, New York, 1973. MABRY, T. J. and J. E. GILL, Sesquiterpene lactones and other
H8FrMAN, E., Functions of steroids in plants, Phytochemistry, 14, terpenoids, in Herbivores (G. A. Rosenthal and D. H. Janzen,
891-901 (1975a). eds.), 502-537, Academic Press, New York, 1979.
HBFl'MAN, E., Steroid hormones in plants, Lloydia, 38, 195-209 MANDAVA, N. B., Natural products in plant growth regulation, in
(1975b). Plant Growth Substances (N. B. Mandava, ed.), ACS Sympo-
IiEFrMAN, E., Metabolism of cholesterol in plants, in Isopentenoids sium Series 111, 135-213, American Chemical Society, Wash-
in Plants (W. D. Nes, G. Fuller, and L. Tsai, eds.), 487-518, ington, DC, 1979.
Marcel Dekker, Inc., New York, 1984. MANDAVA, N. B., Plant growth-promoting hrassinosteroids, Annu.
HEFl'MAN, E., H. H. SAVER, and R. D. BBNNEIT, Ecdysterone bio- Rev. Plant Phys. Plant Mol. BioI., 39, 23-52 (1988).
synthesis in Podocarpus elata, Phytochemistry, 7, 2027-2030 MANN, J., Secondary Metabolism, Oxford University Press, Ox-
(1968). ford, 1978; 2nd edition, 1987.
HEGNAUER, R, Euphorbiaceae. in Chemotaxonomie der Pflanzen, MASCARENHAS, J. P., Sexual chemotaxis and chemotropism in
Vol. 8,440-474, Birkhiiuser Verlag, Basel, 1989. plants, in Taxis and Behaviour (G. L. Hazelbauer, ed.),
HERBERT, R. B., The Biosynthesis of Secondary Metabolites, Chap- 169-203, Chapman & Hall, London, 1978.
man & Hall, London, 1981. MATA, R., M. DEL RAyo C., E. CERVERA, R. BYE, and E. LINARES,
Secondary metabolites from Hintonia latif/ora, Phytochemistry,
HEROOT, V., Some relations between plants, insects and their iso-
29, 2037-2040 (1990).
prenoids, in Progress in Phytochemistry Vol. 2 (L. Reinhold
and Y. Liwschitz, eds.), 143-202, Interscience, London, 1970. MBTcALF, R. L., Plant kairomones and insect pest control, m. Nat.
Hist. Surv. Bull. 33, 175-198 (1985).
HIKINO, H. and Y. HIKINo, Arthropod moulting hormones, Fortschr.
Chern. Org. Naturst., 28, 256-312 (1970). MaTcALF, R. L., Coevolutionary adaptations of rootworm beetles
(Coleoptera: Chrysomelidae) to cucurbitacins, J. Chern. Ecol.,
HIKINo, H. and T. TAKEMOTO, Ecdysones of plant origin, in Inverte-
12, 1109-1124 (1986).
brate Endrinocrinology and Hormonal Heterophylly (W. J. Bur-
dette, ed.), 185-203, Springer-Verlag New York, 1974. MaTCALF, R. L. and R. L. LAMPMAN, The chemical ecology of
Diabroticites and Cucurbitaceae, Experientia, 45, 240-247
IKEKAWA, N., Sterol metabolism in insects and biosynthesis of ec- (1989).
dysone in the silkworm, Experientia, 39, 466-472 (1983).
MBUDT, W. J., Chemical and biological aspects of brassinolide, in
JAENICKE, L., VI. Development: Signals in the development of Ecology and Metabolism of Plant Lipids (G. Fuller and W.
cryptogams, Prog. Bot., 52, 138-189 (1991). D. Nes, eds.), ACS Symposium Series 325, 53-75, American
JAENICKE, L. and F. MARNER, The irones and their precursors, Chemical Society, Washington, DC, 1987.
Fortschr. Chern. Org. Naturst., 50, 1-25 (1986). MILLER, R. W., J. CLARDY, J. KOZLOWSKI, K. L. MIKOLAJCZAK, R.
KAPLAN!S, J. N., M. J. THOMPSON, W. E. ROBBINS, andB. M. BRYCE, D. i'LAITNER, R. G. POWELL, C. R. SMITH, D. WElSLBDER, and
 Triterpenes and Steroids 455

Z. QI-TAI, Phytoecdysteroids of Diploc/isia glaucescens seed, ROBBINS, W. E., J. N. KAPLANIs, M. J. THOMPSON, T. J. SHORTINO,
Planta Medica, 51, 40-42 (1985). and S. C. JOYNER, Ecdysones and synthetic aoalogs: Molting
MORGAN, E. D. and N. B. MANoAVA, Handbook of Natural Pesti- hormone activity and inhibitive effects on insect growth, meta-
cides: Insect Growth Regulators, Part. A., CRC Press, Boca morphosis, and reproduction, Steroids, 16, 105-125 (1970).
Raton, FL, 1987. SASIAK, K. and H. C. RILLING, Purification to homogeneity and
NEs, W. D., The biochemistry of plant sterols, Adv. Lipid Res., some properties of squalene synthase, Arch. Biochem. Biophys.,
15,232-324 (1977). 260, 622-627 (1988).
NEs, W. D. and G. W. PATIERSON, Effects of tetracyclic and pen- SCHrLOKNECHT, H., Die Wehrchemie von Land- und Wasserkiifern,
tacyclic triterpenoids on growth of Phytophthora cactorum, J. Angew. Chern., 82,17-25 (1970).
Nat. Prod., 44, 215-220 (1981). SCHrLOKNECHT, H., Evolutionary peaks in the defensive chemistry
O'DAY, D. H. and P. A. HOROEN. Sexual interactions, in Eucaryotic of insects, Endeavour, 30,135-141 (1971).
Microbes, Academic Press, New York, 1981. SEIGLER, D. S., The role of lipids in plant resistance to insects, in
Plant Resistance to Insects (p. A. Hedin, ed.), ACS Symposium
OOUNKOYA, L' t Application of mass spectrometry in structural
Series 208, 303-327, American Chemical Society, Washington,
problems in triterpenes, Phytochemistry, 20, 121-126 (1981).
DC,1983.
PARIS, R. and A. 'fEsSlER, Chimie Vegetal. Presence de substances
SPENCER, K. C., Specificity of action of allelochemicals: Diversifi-
du groupe des cucurbitacines chez diverses Euphorhiacees tox-
cation of glycosides, in Allelochemicals: Role in Agriculture
iques africaines, notamment chez Maprounea mebranacea Pax
and Forestry, ACS Symposium Series 330, 275-288, American
et K. Hofm., C. R. Acad. Sci. Paris, 274, 321-323 (1972).
Chemical Society, Washington, DC, 1987.
PATIERSON, G. W., Chemical and physical methods in the analysis STUPPNBR, H., E. P. MiiLLBR, and H. WAGNER, Cucurbitacins from
of plant sterols, in Isopentenoids in Plants (W. D. Nes, G. Fuller,
Picrorhiza kurrooa, Phytochemistry, 3D, 305-310 (1991).
and L. Tsai, eds.), 307-312, Marcel Dekker, Inc., New York,
SVOBODA, 1. A. and M. J. THOMPSON, Variability in steroid metabo-
1984.
lism aroong phytophagous insects, in Ecology and Metabolism
PETERSON, S. C., Chemical trail marking and following by caterpil- of Plant Lipids (G. Fuller and W. D. Nes, eds.), ACS Sympo-
lars of Malacosoma neustria, J. Chern. Ecol., 14, 815-824 siura Series 325, 176-186, American Chemical Society, Wash-
(1988). ington, DC, 1987.
POPIAK, G., H. NGAN. and W. AONEW, Stereochemistry of the bio- TALLAMY, D. W., Squash beetle feeding behavior: an adaptation
synthesis of presqualene alcohol, Bioorg. Chern., 4, 279-289 agaiost induced cucurbit defenses, Ecology, 66, 1574-1579
(1975). (1985).
POULTER, C. D., Biosynthesis of non-head-to-tail terpenes. Forma- VAN TAMELEN, E. E., J. WILLET, M. SCHWARlZ, and R. NADEAU,
tion of 1'-1 and 1'-3 linkages, Acc. Chern. Res. 23, 70-77 Nonenzymic laboratory cyclization of squalene 2,3-oxide, J.
(1990). Am. Chern. Soc., 88, 5937-5938 (1966).
POULTER, C. D. and H. C. Rn.uNG, Conversion of farnesyl pyro- VAN TAlWELEN, E. E., Bioorganic chemistry: Sterols and cyclic ter-
phosphate to squalene, in Biosynthesis of Isoprenoid Com- pene terntinal epoxides, Acc. Chern. Res., 1, 111-120 (1968).
pounds, Vo!'1 (J. W.PorterandS.L.Spurgeon,eds.),413-441, WAGNER, H. t Non-steroid, cardioactive plant constituents, in Eco-
Wiley, New York, 1981. nomic and Medicinal Plant Research, Vol. 2 (H. Wagner, H.
POULTER, C. D., E. A. MASH, 1. C. ARGYLE, O. J. MUSClO, and Hikino, and N. R. Farnsworth, eds.), 17-38, Academic Press,
H. C. Rn.uNG, Famesyl pyrophosphate synthetase. Mechanistic London, 1988.
studies of the 1'-4 coupling reaction in the terpene biosynthetic WEHRLI, F. W. and T. NISHIDA, The use of carbon-13 nuclear mag-
pathway, J. Am. Chern. Soc., 101, 6761-6763 (1979). netic resonance spectroscopy in natural products chemistry,
REDDY, K. S., A. J. AMoNKAR, T. G. MCCLOUD, C. CHANG, and Fortschr. Chern. Org. Natorst., 36, 1-229 (1979).
J. M. CASSADY. Spinosides A and B. Two cytotoxic cucurbitacin WEIDENHAMER, J. D., F. A. MACIAS, N. H. FISCHER, and G. B.
glycosides from Desfontainia spinosa, Phytochemistry, 27, WILUAMSON, Just how insoluble are monoterpenes?, J. Chern.
3781-3785 (1988). Ecol.,19, 1799-1807 (1993).
REES, H. H., Ecdysteroids. in Biosynthesis of Isoprenoid Com- WILLIAMS, D. H., M. J. STONE, P. R. HAUCK, and S. K. RAHMAN,
pounds, Vol. 2 (J. W. Porter and S. L. Spurgeon, eds.), 463-505, Why are secondary metabolites (natural products) biosynthes-
Wiley, New York, 1983. ized?, J. Nat. Prod., 52,1189-1208 (1989).
REGUERo, M. T., R. MATA, R. BYE, E. LINARES, aod G. DeLGADO, ZHANG, D., S. M. JENNINGS, G. W. ROBINSON, and C. D. POULTER,
Chemical studies on Mexican plants used in traditional medi- Yeast squalene synthase: Expression, purification, and charac-
cine. n. Cucurbitacins from Hintonia latiflora, J. Nat. Prod., terization of soluble recombinant enzyme, Arch. Biochem. Bio-
50,315-316 (1987). phys., 304, 1133-143 (1993).
24
Saponins and Cardenolides

Introduction elucidation of many saponins has been possible (Hostett-

Steroidal saponins mann et al., 1991).
Triterpenoid Saponins The most common sugars of triterpenoid saponins are
Biological Activity of Saponins hexoses (mainly o-glucose and o-galactose), pentoses
Saponins in Food and Forage Plants (mainly 0- and L-arabinose and o-xylose), methyl pentoses
Saponins and Insects (L-rhamnose and quinovose), and uronic acids (o-glucuronic
Medicinal Uses of Saponins acid and n-galacturonic acid). Different sugars, mainly glu-
Alteration of Taste Properties cose, galactose, rhamnose, xylose, and arabinose have been
Sweeteners identified as components of the carbohydrate moiety of ste-
Cardenolides roid saponins (see Chapter 15).
Biosynthesis The complexity of saponins resnlts from the variety of
Distribution of Cardiac Glycosides combinations of the different sapogenins and numerous sug-
Biological Activity of Cardiac Glycosides ars. For example, the saponins of alfalfa, Medicago sativa,
Interactions with Insects and Other Animals (Fabaceae) vary in composition as well as in absolute
Medicinal Properties amount. Some alfalfa varieties contain as many as 30 differ-
Arrow Poisons ent saponins (Applebaum and Birk, 1979).
References Many of the biological properties for which saponins are
known are due to this hydrophobic/hydrophilic asymmetry
and consequent ability to lower surface tension (Gershenzon
and Croteau, 1991; Hostettmann et al., 1991). In general,
Il'ITKODVCTIOI'l both the sugar and the aglycone are necessary for actiVity.
Saponins have soap-like properties in aqueous solutions.
Saponins are compounds that possess a polycyclic aglycone This is a major reason for acute toxicity when they are in-
moiety with either a steroid (typically C27 ) or triterpenoid jected intravenously. These compounds have been used to
(C30 ) structure attached to a carbohydrate unit (a monosac- captore fish (as piscicides) since ancient times (Howes,
charide or oligosaccharide chain) (Fig. 24.1). These sugar 1930). Saponins in the diet are more toxic to cold-blooded
units are composed variously of pentoses, hexoses, or uronic than to warm-blooded animals. Most saponins hemolyze red
acids. The existence of uridine-5'-diphosphate-glucose blood cells (Marston and Hostettmann, 1991).
(UDPG):sterol transglucosylase actiVity in particulate prepa- When macerated plant material or plant extracts contain-
rations from higher plants is well established (Goodwin, ing saponins is shaken with water, a thick stable foam usually
1985). results (Hostettmann et aI., 1991). This is the basis of a
Saponins are classified into two major groups based on simple, reasonably specific test for the general presence of
the structore of the aglycones that are known collectively as these compounds. Saponins also are detected by the Lieber-
sapogenins. Steroidal sapogenins usually contain 27 carbon mann-Burchard test, which is useful for distinguishing be-
atoms; those of triterpenoid sapogenins contain 30. Approxi- tween triterpenoid and steroidal saponins (Chandel and Ras-
mately 360 sapogenins and 750 triterpene glycoside struc- togi, 1980).
tures are known, but it is only recently that full structore Saponins are widespread among plants, having been re-

456
 Saponins and Cardenolides 457

OH
oH~OH
OH

P-D-glucosyl H~~~O
OH 0 0
o HO H

a_L-rhamnosy~o#.o-.,....<.OH
HO~HO
~O. ;0 P-D-glucosyl

OH
sarsaporilloside

Co,?~O
H

~OH
OH OH W OH
0

OH
disoc:ln

Co,?~~l
~H~ OH OH
HO
0
asparanin B (9)

HO OH

FIg. 24.1. Typical sapunins.

ported from more than 500 plants from at least 90 different been reviewed (Hostettrnan, 1985, 1986; Hostettrnan et aI.,
families (Bondi et aI., 1973), these substances have heen 1991; Mahato et aI., 1982; Marston and Hostettrnan, 1991).
isolated from all parts of plants: leaves, stems, roots bulbs, Among the most useful techniques are high-performance liq-
flowers, and froits, although they tend to be concentrated in nid chromatography (HPLC), centrifugal thin-layer chroma-
the roots of many species. There is considerable seasonal tography (CTLC), column chtomatography, and Qroplet
variation in the amounts of saponins present, and often sub- countercurrent chromatography (DCCC) (Hostettrnan, 1986;
stantial variation within plant populations in the amount of Hostettrnan et aI., 1991; Marston and Hosteltmann, 1991).
compound is present. Low saponin lines of many crop plants, Mass-spectrometry techniques such as field ionization de-
for example, alfalfa, have been selected. However, as much sorption (FlO), chemical ionization (CI), and fast atom bom-
as 30% by weight of some plants is saponins (Hostettrnann bardment (FAB) have proven useful for determination of
et aI., 1991). structures of intact saponins (Hostettrnann et aI., 1991; Mar-
Several plants produce saponins in plant tissue culture. ston and Hostettrnan, 1991).
Among these are Dioscorea species, Trigonella foenum- A number of saponic alkaloids are discussed in Chapter
graecum, Agave wightii, and Tribulus terre.tris (EIlis, 36.
1988). Because of its economic importance, much research
has been directed toward producing diosgenin in culture. The Steroidal SaponiDs
highest level of diosgenin-like saponins has been obtained in
suspension cultures of Dio.corea deltoidea, where 7-8% of Steroidal saponins (those based on C27-steroidal sapoge-
the dry weight was obtained (Ellis, 1988). nins) are found in a large number of plant families, but are
Only in the last few years has isolation of single compo- particularly common in the monocotyledonous families
nents from mixtures of saponins been feasible. In many pre- Agavaceae, AmaryIlidaceae, Dioscoreaceae, and Liliaceae
vious studies, the sugar portion was removed by hydrolysis (Dahlgren et aI., 1981). The vast majority of steroidal sapo-
and the mixture of sapogenins fractionated. Many of the nins are based on furostan- or spirostan-type precursors
techniques needed for isolation of the intact saponins have (Hostettrnan et aI., 1991). A series of steroidal saponins from
458 Saponins and Cardenolides

Lilium pardarinum has been isolated and characterized by and Rastogi, 1974; Chandel and Rastogi, 1980; de Mayo,
a variety of high-field nuclear magnetic resonance (NMR) 1959). These glycosides may possess from I to 10 sugar
techniques (Shimomuraet aI., 1989). In many of these com- units (Agarwal and Rastogi, 1974). Triterpene glycosides
pounds, there is an AlB cis relationship (i.e., rings A and are especially common in the Aceraceae, Amaranthaceae,
B are joined in a cis manner). These compounds also are Asteraceae, Campanulaceae, Caryophyllaceae, Chenopodia-
commonly encountered in the dicotyledonous families Scro- ceae, Hippocastanceae, Lamiaceae, Oleaceae, Papaveraceae,
phulariaceae and Solanaceae. Other types of steroidal sapo- Phytolaccaceae, Polygalaceae, Rubiaceae, Rutaceae, Sapin-
nins have been reported from the Arecaceae, Fabaceae, Fou- daceae, Sapotaceae, Verbenaceae, and Zygophyllaceae. Sa-
quieriaceae, and Zygophyllaceae. ponins involving f3-amyrin, or other biosynthetically related
In most plants in which these steroidal saponins occur, triterpene moieties, are widespread among plants (Boar and
the sapogenins are combined with a number of sugars to Allen, 1973). Some representative triterpenoid sapogenins
form saponins. Generally, as many as five sugar units may from the Fabaceae are illustrated (Fig. 24.2) (Applebaum
form a chain that comprises the glycoside linkage. The link- and Birk, 1979).
age to the sapogenin is always f3 and always in the C-3
position (Grunwald, 1980; Takeda, 1972).
As is true for most phytosterols, cholesterol is the usual
precursor (see Chapter 23), but in some instances f3-sitos- BIOLOGICAL ACTIVITY OF SAPOl'llNS
terol, demosterol, or other phytosterols can serve in this role.
Several steroidal saponin sulfates have been reported Many saponins have generalized antibiotic properties, such
from starfish (Hostettmann, 1985). Sea cucumbers secrete as antifungal and antimicrobial effects. Phytopathogenic
water-soluble glycosides, holothurins, that possess greater fungi often are sensitive to the presence of saponins. The
hemolytic ability than saponlns of plant origin. These com- effects of steroidal and triterpenoid saponins are sometimes
pounds are mixtures of glycoside snlfates (Agarwal and Ras- quite different (Agarwal and Rastogi, 1974).
togi, 1974). Water-soluble bidesmosidic saponlns (those with sugars
Alkaloids such as solanidine and tomatidine are related atrached to two different parts of the aglycone) lack the typi-
to diosgenin in terms of structure. These cholesterol-derived cal properties of saponlns and, generally, are inactive as anti-
metabolites usually occur as the 3-0-glycosides (e.g., sola- biotics. It has been proposed that these bidesmosidic com-
nine and tomatine) (Mann, 1987) (see Chapter 36). pounds are transport forms from leaves to other parts of
the plant (Hostettmann et al., 1991). Once the tissues are
Trlterpenoid Saponins damaged, enzymes convert bidesmosidic saponins into mo-
Triterpenoid saponins also are widespread in nature, nodesmosidic derivatives that have defensive functions. The
being known from at least 90 families of plants (Agarwal conversion of ot-hederin (1) from hederasaponln C (2) in

HO

soyasapogenol A soyasapogenol B soyasapogenol C

HO
~ HH

CHzOH
•

soyasapogenol D
0
HO

HO

medicagenic acid (3) hederagenin (4)

Fig. 24.2. Several triterpenoid sapogenins from the Fabaceae (modified from Applebaum and Birk, 1979; used with permission of the copyright owner,
Academic Press, New York),
 Saponins and Cardenolides 459

g1c
I
ara-O
I
glc

n
(6)
(5)

CHO

o-(l/
o NHCH,

glc glc
I I
ara-Q ara-O
I I
glc g1c (8)
(7)

Fig. 24.3. Saponic phytotoxins from oats.

H edera helix (Araliaceae) is thought to represent such an Larvae of the leek-moth, Acrolepiopsis assectella, feed
example (Fig. 24.4) (Hostettmann et aI., 1991). on the leaves of various Allium species, but usually avoid
Because saponins are surface-active agents and act as the flowers. The saponin aginoside was isolated from these
detergents, they can inactivate a number of enzymes. Many flowers and shown to be toxic to the larvae. The level of
saponins also cause disintegration of membranes and, in the toxicity was reduced by addition of cholesterol or sitosterol
case of erythrocytes, they cause hemolysis. This action is to the diet of the larvae (Harmatha et aI., 1987).
related to the presence of cholesterol, as addition of choles- Saponins, such as asparanin B (9) (Fig. 24.1) from the
terol counters some of the deleterious effects of saponins seed of Asparagus adescendens (Liliaceae), are active
(Agarwal and Rastogi, 1974). Addition of cholesterol also against the nematode Meloidogyne incognita at concentra-
causes precipitation of certain saponins from aqueous solu- tions as low as 200 IlgiL (Chitwood, 1992).
tions (Bondi et aI., 1973). Many active molluscicides are saponins (Farnsworth et
Saponins can cause changes in the microstrucmre of cell aI., 1987; Henderson et aI., 1987; Marston and Hostettrnann,
membranes. An attempt has been made to relate the hemo- 1991). These compounds are important because of the poten-
lytic activity to the struclnre and composition of the saponin tial for controlling the snail vectors for schistosomiasis (bil-
molecule. Saponins that contain medicagenic acid (3) and
harzia), which affects about 300 million people worldwide.
hederagenin (4) (Fig. 24.2), as their aglycones can be precip-
Four triterpenoid saponins with strung molluscicidal activity
itated by cholesterol and have strong hemolytic activity.
are found in the fruits of ivy (Hedera helix Araliacene), and
Blocking of the carboxylic acid groups in medicagenic acid
others in the fruits of Lonicera japonica (Caprlfoliaceae)
results in complete loss of hemolytic activity. The sapogenin
(Hostettmann, 1986).
to sugar ratio is also important in this activity.
Alfalfa saponins are known to inhibit the germination of The saponins from Phytolacca dodecandra (Phytolacca-
letmce seeds, as are those of certain other species. They also ceae) contain a mixture of triterpene glycosides (mainly
inhibit growth of the soil fungus Trichoderma viride. based on oleanolic acid) that are highly active against several
The resistance of oats (Avena sativa, Poaceae) to the take- of the intermediate snail hosts of schistosomiasis. This was
all fungus Ophiobolus graminis is due to a fluorescent "wot fIrst discovered by the observation of dead snails along
tip glycoside." This resistance factor, "avenacin," has streams in Ethiopia where the fruits of Phytolacca are used
proven to be a mixmre of four pentacyclic triterpene glyco- for laundry purposes (Hostettmann, 1985; Hostettmann et
sides (5-8). Compounds (5 and 7), which possess anthranilic aI., 1991). To date, one of the.most effective sources of
acid residues as a part of the molecule, have the greatest molluscicidal saponins is the fruit of Phytolacca dodecandra
fungicidal properties (Fig. 24.3) (Harborne, 1986). (Phytolaccaceae), These fruits contain up to 25% glycosides,
460 Saponins and Cardenolides

primarily those of oleanolic acid. The most active compound Forage legumes are unsuitable for the development of
in the mixture of saponins from these fruits is lemmatoxin, several locust species. A diet of alfalfa increases mortality
[4'-O-(j3-o-glucopyranosyl-3'-O-(j3-o-galactopyranosyl)-j3- and extends the duration of larval development of Mela-
o-glucopyranosyllolean-12-ene- 28-oic acid (10) (Fig. 24.4) noplus sanguinipes and M. differentialis. Adults derived
(Henderson et ai., 1987). Lemmatoxin is toxic in the range from these larvae have small, and usually short, forewings.
1.5-3.0 mg/L (Marston and Hostettmann, 1991). Similar results are obtained with Schistocerca gregaria.
An emulsion from Balanites aegyptiea (Zygophyllacene) Feeding of Locusta on Trifolium pratense also results in high
containing three molluscicidal saponins killed not only the mortality. Fecal pellets of saponin-fed locusts are wet, in
snail vectors but also the free-living stages of the schisto- contrast to the dry pellets of locusts on a saponin-free diet.
some (Henderson et al., 1987). This emulsion is also lethal to Water resorption is impaired in the hindgut and the hemo-
fish and tadpoles (Marston and Hostettmann, 1991). Extracts lymph volume decreases as a consequence. Studies with
from Solanum nodiflorum and from S. mammosum contain other surface active agents indicate, however, that the sapo-
effective molluscicides (LD.5 25 ppm, 24 h). The active nins have a specific effect in addition to the surface-active
components are steroidal saponic alkaloids (Henderson et properties. Inclusion of saponins from alfalfa in diets of in-
al., 1987). One of the most promising molluscicidal plants sects that normally eat this plant had little affect on them.
to date is Swartzia madagaseariensis (Fabaceae). The fruits The presence of saponins in woods of Ternstroemia ja-
of this plant, common in many regions of Africa, are effec- ponica (Temstroemiaceae or Theaceae) is reputed to be re-
tive against both common species of schistosomiasis-trans- sponsible for the antitermite effects observed in ancient
mitting snails, and are a fish poison (Marston and Hostett- woods of temples (Agarwal and Rastogi, 1974).
mann, 1991).
Medicinal Uses of saponins
saponins in Food and Forage Plants
Certain saponins have antitumor, piscicidal, mollusci-
Saponins generally are not absorbed readily in the intes- cidal, spermicidal, sedative, expectorant, and analgesic prop-
tinal tract. Often these compounds must be accompanied by erties (Hostettman et al., 1991). Preparations such as glycyrr-
an irritant in the intestines to have much activity. Despite hizae radix (from Glycyrrhiza glabra, Fabaceae), are useful
the fact that they are not absorbed, saponins do produce as expectorants and antitussive agents. The major saponic
severe gastroenteritis in the intestines. compound, glycyrrhizin (glycyrrhizic acid) (11) (Fig. 24.6)
Saponins are present in many pastore plants (Bondi et is responsible for the sweetuess (50 times sweeter than su-
al., 1973). Appreciable quantities accumulate in some, espe- crose), antiulcer, and glucocorticoid-like properties of the
cially in certain forage legumes. These compounds are prob- drug (Hikino, 1985). Glycyrrhizin preparations are used in
ably involved in the condition known as ruminant bloat. By Japan to treat chronic hepatitis and cirrhosis (Hikino and
altering the surface tension of the ruminal contents, saponins Kiso, 1988). The saponins of Zizyphus jujuba (Rhamnaceae)
cause gas formed in the rumen by anaerobic bacterial fer- exert sedative activity in mice (Hikino, 1985).
mentation to be entrapped as a froth. When fed to cattle, Some saponins have anti-inflammatory properties. The
saponins elicit characteristic symptoms of bloat. In mono- mixtore of saponins from Bupleurum falcatum (Apiaceae)
gastric animals, saponins are responsible for reduced growth (a crude drug used in oriental medicine) is effective (400
rate and decrease in food consumption. Poultry are particu- mg/kg) for the treatment of edema in rats and possesses anti-
larly sensitive to alfalfa that contains saponins. Swine are inflammatory activity (Hikino and Kiso, 1988). The sapo-
less sensitive (Applebaum and Birk, 1979). nins saikosaponin a and d appear to be responsible for most
Soybeans and sugar beets also contain saponins that have of the activity.
been shown to be deleterious under some circumstances (Ap- The crude saponins of Thea sinensis (Theaceae) also have
plebaum and Birk, 1979; Bondi et al., 1973). pronounced antiexudative and anti-inflammatory properties.
A pseudocereal of the Andes of South America, Cheno- Phytolaeea americana roots are reputed to possess anti-
podium quinoa, owes its bitter taste to the presence of tri- inflammatory properties in Korean medicine (Chandel and
terpenoid saponins. These must be removed before the seeds Rastogi, 1980; Hostettmann et al., 1991). Similar properties
are eaten. A number of the most important saponins have have been demonstrated for a number of other saponins
been characterized (Bumouf-Radosevich et al., 1985). (Agarwal and Rastogi, 1974).
Ginseng, Panax ginseng and P. quinquefolium (Aralia-
saponins and Insects ceae), contains a series of triterpenoid saponins. Over
Soybean saponins have been shown to be detrimental to 350,000 pounds of ginseng, worth more than $25,000,000,
the larvae of Callosobruehus chinensis, an oligophagous in- were shipped from the United States in 1978 (Shibata et al.,
sect that lives on several legumes but that cannot live on 1985; Tyler et al., 1981). Ginseng is a favorite remedy in
soy beans, peanuts, and certain other legumes. Chinese and Korean medicine and is used primarily as a
In studies with the rust-red flour beetle Tribolium casta- means of maintaining balanced homoestasis or tu restore
neum, alfalfa saponins inhibited growth, but this effect was Yang (Shibata et al., 1985). A number of physiological ef-
reduced by addition of cholesterol to the diet. fects have been observed (Chandel and Rastogi, 1980);
 Saponins and Cardenolides 461

non·molluscicidal
OR saponin from PhYto Iocca dodecandra
o

RO"'-~ _0 RO
R'O~O
RO RO~ _'''0, ,
RO~O
OR

OR
RO"'-~ _0 RO
R~~O
RO RO~ _"'0, ,
RO~O
OR CR,OR

moJiuscicidal saponin from Phytolacca dodecandra

OR
RO~ ~O
RO~~ .~ ____ 0,
RO
,

RO~
lR(:: ;t~0 lemma!oxi" (10)

(a) OR

l
! ~~~OR
O~.~ORO~OR
HO~OR 0

~
OR
OR
RO~O, '
RO~O OR OR
CR,OR hederasaponin C (2)

RO --r---...oJ
~ RO
OR

RO~O, '
RO~O

RO ---r----..oJ
(b)
~ RO

Fig. 24.4. Molluscicidal saponins.

462 Saponins and Cardenolides

OR

RO

diosgenin (12) (15)

progesterone (14) cortisone (13)

Fig. 24.5. Diosgenin. progesterone, and an intennediate in the biosynthesis of diosgenin.

whether any of these are sufficient to explain the purported fied in the leaves of an asclepiadaceous vine Gymnema sylv-
medicinal value of ginseng is open to question. Both the estre. These saponins have been called taste-distortion
Oriental and American species contain a similar complement agents because of their ability to suppress the perceived
of saponins; the chemistry and techniques for analysis of sweetness of sugars and, to a lesser extent, the taste of amino
these compounds has been reviewed (Shibata et a1.. 1985). acids. Chewing the root of this plant blocks perception of
The same array of ginseng saponins are produced in root the sweet taste of sugars, saccharin, and cyclamates for hours
cultures as in the intact plants; furthermore. the saponin con- (Hodge and Inglett, 1974). These compounds may protect
tent of the cultures exceeded that of the plants (CharI wood plants against mammalian and insect herbivores by dimin-
and Charlwood, 1991; Ellis, 1988; Shibata et aI., 1985). ishing the animals ability to sense the presence of sugars
A related plant, Eleutherococcus senticosus or Siberian (Harborne, 1982).
ginseng, has been used for medicinal purposes as well (Farn-
sworth et al., 1985).
The sapogenin, diosgenin (12), which occurs as the 3-0- Sweeteners
glycoside in several species of Dioscorea (Dioscoreaceae),
Several saponins are known to taste sweet to humans.
is important as a precursor for the chemical synthesis of
Glycyrrhizin or glycyrrhizic acid (11), a triterpenoid glucu-
cortisone (13) and a number of contraceptive drugs, such as
ronide from Glycyrrhiza glabra (licorice, Fabaceae), is about
progesterone (14). The sapogenin is found in the roots of
50 times as sweet as sucrose (Fig. 24.5). Licorice is added
these plants in quantities as large as 5-6% of the dry root
weight. Diosgenin is biosynthesized from cholesterol via in- to chewing gum, chocolate candy, cigarettes, smoking mix-
termediate (15), but the complete pathway has not been elu- tures, chewing tobaccos, and snuff. Pharmaceutically, lico-
cidated (Fig. 24.5). rice is used to mask the flavor of many distasteful drugs.
Several saponins have been found to have cardiotonic Licorice root extract also is used to treat peptic ulcers. Large
activity, but usually less than that observed for the cardenol- amounts of glycyrrhizic acid can modify heart action (Hodge
ides (see the following section) (Mahato et aI., 1982). and Inglett, 1974; Tyler et aI., 1981).
Glycyrrhizin (11) appears to inhibit HIV-l cell binding Another sweet-tasting saponin, osladin (17), occurs at
(Kinghorn, 1992). rather low concentrations (about 0.03%) in the roots of the
fern Polypodium vulgare (Hodge and Inglett, 1974). Similar
steroidal glycosides have been isolated from the rhizomes
Alteration of Taste Properties
of Polypodium glycyrrhiza (Kim et aI., 1988; Kim and Kin-
A class of triterpenoid saponins called gymnemic acids ghorn, 1989). The major intensely sweet compound of that
[based on gymnemagenin (16) (Fig. 24.6)] has been identi- plant is polypodoside A, a 26-0-a-L-rhamnopyranosyl-pol-
 Saponins and Cardenolides 463

ypodogenin-3-0- a-L-rhamnopyranosyl-(I - 2)-/3-o-gluco- CARDEI'IOLIDES

pyranoside (18). Podogenin, (22S,25R,26R)-3/3,26-dihy-
droxy-22,26-e poxy-6-oxo-5a-cholest-7-ene, serves as the Cardiac glycosides or cardenolides (mostly CZ3 compounds)
aglycone (Kim and Kinghorn, 1989). The glycoside is ap- are an important group of secondary metabolites that are
proximately 600 times sweeter than a 6% solution of sucrose. used medicinally and, under other circumstances, for arrow
Polypodogenin is the A7 •8 -isomer of osladin (Kim et aI., poisons and for ordeal ceremonies (Connolly and Hill,
1988). A flavonoid glycoside, (+ )-afzelechin-7-0-/3-o-api- 1991). All cardenolides have a 3/3-oxygen function, a 14/3-
oside, from the same plant, has a pronounced bitter taste hydroxyl group, and an a,/3-unsaturated-"y-lactone attached
(Kim and Kinghorn, 1989). at the 17/3 position (Connolly and Hill, 1991). These bioac-
The sweet-tasting compounds from Abrus fruticulosus tive glycosides are especially well known in plant-insect
andA. precatorius (Fabaceae) also are a mixture of triterpene interactions. Several examples of cardiac-active compounds
glycosides related to cycloartenol (Choi et aI., 1989; Fullas are known, but most plant-derived cardenolides are sapo-
et aI., 1990). Abrusosides A-D (19-22) are 30-100 times nins. The sapogenin usually is derived from CZ1 progesta-
sweeter than a 2% solution of sucrose. gens and modified by an additional acetate (from malonate)
The triterpene glycoside, mogroside V (23), from the unit to form a 5- or 6-membered lactone ring (Miller, 1973).
dried fruits of Thladiantha grosvenorii (Cucurbitaceae) is Large numbers of individual glycosides are found in various
approximately 250 times sweeter than sucrose (Hussain et plant tissues. For example, at least 24 cardiac glycosides
aI., 1990; Matsumoto et aI., 1990). are found in the leaves of Strophanthus boivinii. In general,

O'HO~
'V--'v --;t(

HO~O, !>
HO~

~O osladin (17)

OH

HO

(a) gymnemagenln (16) glycyrrhetic acid

Fig. 24.6 (a-c). Saponins with sweet or taste~modifying properties.

464 Saponins and Cardenolides

·....'0 ...·''''0

~
polypodoside A (18)
OH

HO OH OH
HO~ _0 . o
HO~O
o
HO ---r--.. o )
~ HO

HO (19)

HO~ _0 0
HO~
OH

HO
HOH~O (20)
HO~ __ O, ,0
O
HHO~
OCH)

HO
HO~ __ O, ,0
HO HO~
HO~ ~O. ? (21)
HO~
(b) OH

HO
OHO~ ..--0. ,0
HO HO~
HO~ -~ :0
Iio~
OH OH

HO
HO~P ~..--O. ~
HO~-----O~
OH t?o.~.--"~~,/
OH
(0)

Fig. 24.6. (continued)

 Saponins and Cardenolides 46S

cardenolides are accumulated in epidermal cells, laticifers, BIOSYNTHESIS

and phloem elements of both external and internal phloem
of leaves and stems. In tissue culture of Digitalis, cardenol- Cardenolides typically have 23 carbon atoms, possess a 14-
ides are accumulated in the vacuoles (Malcolm, 1991). l3-hydroxyl group, an a,l3-unsaturated "V-lactone ring, and
Many of the sugars found in cardenolides are restricted various sugars attached at the 313-position (Harbone, 1991).
in distribution (see Chapter 15). These include D-glucose, Biosynthesis of cardenolides in Digitalis involves three dis-
D-fucose, D-digitoxose, D-digitalose, D-diginose, D-sarmen- tinct phases: the formation of a mevalonate-derived steroid
tose, D-cymarose, D-thevetose, and L-thevetose (Connolly nucleus, the formation of a butenolide ring by addition of
and Hill, 1991). an additional unit of acetate (malonate), and glycosidation
The procedures for isolation, purification, and characteri- with up to four sugars (Groeneveld et al., 1991). Cardenol-
zation of cardiac glycosides have been reviewed (Connolly ides in most plants are formed by condensation of a C21 -
and Hill, 1991; Malcolm, 1991). Several effective methods pregnane derivative [pregnenolone (3-I3-hydroxypregn-5-
for the isolation and purification of cardenolides by HPLC en-20-one) or progesterone] with acetate (malonate) (Fig.
(Groeneveld et al., 1990, 1991) and TLC (Malcolm et al., 24.7). Labeling work with Strophanthus kombe demon-
1989; Martin and Lynch, 1988) have been reported. strated that [4- 14C]3-I3-hydroxy-5a-pregnane-20-one (24)
The 13C_NMR spectra of several cardenolides are of value and progesterone (14) are incorporated into cardiac glyco-
for characterization of this group of compounds (Tori et al., sides. Incorporation experiments with stem disks and slices
1973). demonstrated an acetate-derived butenolide ring in calo-

HO

cholesterol
RO~O~
(24) ,tropbanthldln
o 0

HO,!
~CO>H_
HO •

scillaren A

glucosyl-O-rhamnosyl- 0

Fig. 24.7. Some intennediates in the biogenesis of cardenolides.

466 Saponins and Cardenolides

tropin (25) and uscharidin (26) (Fig. 24.8) in Asclepias cura- tasting and poisonous to higher animals (Fig. 24.8) (Har-
ssavica (Lee and Seiber, 1983); however, feeding of [1- borne, 1991; Seiber et al., 1984). Nonetheless, insects from
l3C]acetate at lower levels to Asclepias curassavica stem about eight orders feed on these plants. Most are coadapted
disks produced uscharidin in which there was no significant species capable of consuming plants that contain cardiac
introduction of l3C into the portion of the butenolide ring glycosides. Many of the compounds are metabolized by the
containing C-22 and C-23 (Groeneveld et aI., 1990, 1991). insects, but others are taken up and sequestered unchanged
More than 20 sugars, most of these uncommon from other (Harbome, 1989, 1991). Many ofthese insects are aposemat-
sources, have been identified from cardiac glycosides (see ically labeled (Rothschild, 1973).
Chapter 15) (Miller, 1973; Seiber et aI., 1984). The sugar Among these are the monarch butterflies, Danaus plexip-
moieties of cardenolides are normal sugars or unbranched pus. When the larvae of these butterflies feed on Asclepias
aldohexoses, mostly of the 6-deoxy or 2-deoxy type. species such as Asclepias curassavica, they store the toxic
The cardenolides of the Asclepiadaceae differ from those compounds cal actin (27) and calotropin (25) within their
of the Apocynaceae and the Scrophulariaceae, in that the bodies. As few other insects can eat these milkweeds, the
former have a Sa(trans AlB) and the latter a S(3(cis AlB) insect has little competition for its food source. The larvae
ring junction. The latter type is useful clinically in humans, pass the compounds on to the adult stage of the insect which
whereas the former normally is not (Seiber et al., 1984). is brightly colored and distasteful and/or toxic to many pred-
At least 30% of the cardiac glycosides of young plants of ators. This warning coloration is called aposematic marking
Asclepias curassavica occurs in the latex (Groeneveld et al., or labeling. The larvae of monarch butterflies have potent
1991). emetic properties when fed to blue jays (not a normal preda-
tor of the monarch butterfly). Birds that consume the larvae
quickly learn not to repeat the mistake. In central Mexico,
DISTRIBUTION OF CARDIAC GLYCOSIDES where the insects winter, other birds eat the adult butterflies
with no apparent ill effects (Brower, 1984; Harbome, 1982;
More than 400 cardiac glycosides are known. Most occur Herout, 1970; Malcolm, 1991). These birds remove the outer
in the Apocynaceae, Asclepiadaceae, Celastraceae, Brassi- portions of the insect that contain the emetic compounds,
caceae, Fabaceae, Euphorbiaceae, Moraceae, Ranuncula- and ouly eat the inner parts that lack the compound.
ceae, Scrophulariaceae, Tiliaceae, Sterculiaceae, and Lilia- Because the butterfly carries warning coloration
ceae. Related compounds with 6-membered lactone rings (aposematic labeling) that the bird learns to recognize
(bufadienolides) occur in the Liliaceae and Ranunculaceae quickly, only a fraction, about 50% of the insects, need to
(Malcolm, 1991). have the toxic compounds within their bodies for all of the
A number of frogs and toads and some plants make car- insects to be protected (Brower, 1969). Both the plants and
diac-active compounds that have steroidal structures but are the insects differ in cardenolide content. Monarch butterflies
not glycosides (Miller, 1973). do not accumulate each of the compounds in the plant
equally well. Some cardenolides are absorbed intact, others
are biochemically modified, and yet others are excreted.
BIOLOWCAL ACTIVITY OF CARDIAC Over its natural range in North America, the female mon-
GLYCOSIDES arch butterfly may lay her eggs on several Asclepias species
that differ in the complement and amount of cardenolides
Despite the fact that cardenolides usually are described as present. By TLC analysis of adult butterflies, it is possible
bitter tasting, milkweed poisoning in livestock is a persistent to determine the species upon which the larva feed. Depend-
problem. Normally, animals will not eat the apparently dis- ing on the complement and quantity of cardenolides present
tasteful plants, but in time of scarcity, many animals (espe- in the host plant, the larvae are able to adjust their diet to
cially sheep) succumb to this type of poisoning (Malcolm, modify the amount of cardenolide sequestered (Harborne,
1991; Seiberet al., 1984). As little as 0.8 oz. of dry Asclepias 1991).
labriformis leaf is fatal to a 100-lb sheep (Harbome, 1991). The cardenolides within the monarch butterfly's tissues
Although it is well known that cardiac glycosides are are tightly bound to the cuticle in the various organs. Presum-
sequestered by insects, it is less well known that parasitic ably, this is a mechanism to avoid self-poisoning and to keep
plants of the family Loranthaceae also sequester these com- these cellular toxins away from neuronal tissues (Harbome,
pounds when they grow on hosts containing cardenolides 1991).
(Boonsong and Wright, 1961; Rothschild, 1973). Certain other butterflies mimic the color pattern of the
Monarch butterflies and also are protected from the predators
Interactions with Insects and Other even though they do not have the cardiac glycosides in their
Animals
bodies.
Most members of the milkweed family (Asclepiadaceae) Other aspects of the chemical ecology of Danaus species
and many members of the related family Apocynaceae syn- involve pyrrolizidine alkaloids and their metabolites (see
thesize cardiac glycosides. These substances are both bitter Chapter 30).
 Saponins and Cardenolides 467

HO

HO
HO

ouabagenin scillarenin A
(a)

o o

~
OCH3

HO 0 OH

!HO~,o
o

••,,,....
d
4'
5'
0
2'
1'.,
H "''>'0
oleandrin (28)

o calotropin (25)

o
~
OCH3" 19
HO" 3' 0 OH,1

6' 5' l' 3, 7 OH

OH

o neriifnlin(3!) ~O ••",

,,,,,,,lo1"""'o
° 0
calactin (27)

)rO""
,lAA,,·,o
uscharidin (26)

""" 0 H
(b)

Fig, 24,8 (a-c), Represen t ative cardiac glycosides,

468 Saponins and Cardenolides

erysimoside (29)

uo~ ...--0. ~~o\

UOiio~O ~O OU
ou OU

erycbroside (30)

(0)
Ul'~
o
OU
0
~ OU
0
OU

Fig. 24.8. (continued)

Insects such as Iygaeid bugs fed on Nerium oleander also terrent fractions of Erysimum cheiranthoides and characteri-
accumulate cardiac glycosides, as does a bright yellow zation of the active compounds revealed that cardenolides
aphid, Aphis nerii (Rothschild, 1973). The sugar~rich secre~ are responsible for this avoidance. In particular, the strop-
tion of these aphids, called honeydew, contains cardenolides hanthidin glycosides, erysimoside (29) and erychroside (30),
found in the host plant (Molyneux et aI., 1990). The ability were strongly deterrent to the females of Pieris rapae (Fig.
of this aphid to acquire these compounds suggests that they 24.7) (Dimock et aI., 1991; Renwick et aI., 1990, 1991; Sach-
are found in the phloem of Nerium oleander plants (Moly- dev-Gupta et aI., 1991). A strophanthidin glycoside was iso-
neux et aI., 1990). lated from leaves of Cheiranthus X allionii which had simi-
The larvae of the bug, Caenocoris nerii, feed on ripening lar activity (Rothschild et aI., 1988). This cardenolide had
seed pods of Nerium oleander plants and also accumulate no effect on another crucifer pest, the diamondback moth,
cardenolides such as oleandrin (28). At night, when Plutella xylostelta (Renwick et aI., 1990).
aposematic colors are less useful, these insects abandon the When neriifolin (31) is applied to the roots of canteloupe
conspicuous pods and shelter in twos or threes on the under- plants, more than 70% mortality of neonate larvae of the
side of leaves of the plant (Rothschild, 1973). codling moth (Laspeyresia pomonelta) occurred (Reed et
After attacking and consuming milkweed bugs (Oncopel- ai, 1982). The toxin appears to have a systemic effect. The
tus fasciatus) raised on seeds of milkweed (Asclepias syr- development and molting of larvae of Acrolepiopsis assec-
iaca), the mantid Tenodera ardifolia sinensis regurgitates tella are inhibited when the larvae are fed digitonin. This
and shows signs of poisoning by cardenolides (Berenbaum insect normally feeds on species of Allium (Liliaceae) that
and Miliczky, 1984). do not have cardenolides. The addition of cholesterol to the
The North African aposematic grasshopper, Poekilocerus diet prevents these deliterious changes (Arnault and Mau-
bufonius, like the monarch butterfly, also feeds on milk- champ, 1985).
weeds and accumulates the cardenolides. Unlike the mon- An African mammal, the hyrax, eats oleander with im-
arch, the grasshopper ejects the cardenolide materials as a punity and seems to excrete the cardiac glycosides in its
noxious foam from specially located poison glands (Seiber urine, which is used by Cape Bushmen for tipping poison
et aI., 1984). arrows (Rothschild, 1973).
Gravid females of Pieris rapae oviposit on several mem- The larvae of a number of chrysomelid beetles synthesize
bers of the Brassicaceae and other families that possess glu- and accumulate cardiac glycosides (pasteels et aI., 1988,
cosinolates; however, they avoid plants of the genera Erysi- 1989). In this instance, the beetles do not take the compounds
mum and Cheiranthus (see Chapter 17) (Rothschild et aI., from the host plants (many of which lack cardenolides) but
1988; Sachdev-Gupta et aI., 1991). Fractionation of the de- biosynthesize them from cholesterol.
 Saponins and Cardenolides 469

l'Iedicinai Properties cardiovascular drug (Lewis, 1992). Digitoxin (33) is the

main saponic glycoside from digitalis (Tyler et a!., 1981).
Cardiac glycosides or cardenolides have a highly specific
This glycoside is absorbed readily from the intestinal tract;
and powerful effect on the cardiac muscle in mammals. Ou-
intravenous and oral toxic doses are almost equal. In con-
bain (32) and other cardiac glycosides inhibit vertebrate so-
trast, ouabain (32) is much more toxic when given intra-
dium pumps [(Na+ + K+)-ATPasel that play an essential role
venously (Malcolm, 1991). The toxicities of some common
in maintaining transmembrane ionic gradients (Malcolm,
cardenolides have been summarized (Malcolm, 1991).
1991). There are two principal types: the cardenolides and
Several cardiac glycosides have been reported to have
the bufadienolides (Tyler et a!., 1981). Cardenolides have a antiviral properties (Vanden Berghe et aI., 1985). Others
5-membered and bufadienolides have a 6-membered unsatu- have been reported to have antitumor or cytotoxic properties.
rated lactone ring. In some instances, these saponic com- Among these are calotropin (25) from Asclepias curassavica
pounds are emetic and they are frequently toxic. The (Kupchan et a!., 1964) and neriifolin (31) and 2'-acetylnerii-
(Na++ K+)-ATPase in cardiac muscle has been proposed as folin from Thevetia thevetioides (Apocynaceae) seeds
the pharmacological receptor for cardiac glycosides (Seiber (McLaughlin et aI., 1980).
et a!., 1984). The therapeutic level is often about 50% of The cardioactive compounds from Bryophyl/um species
the toxic dose (Tyler et a!., 1981). Cardiac glycosides are are bufadienolides with potent cytotoxic activity (Yamagishi
saponins and have most of the properties ascribed to these et aI., 1989). Bryophyllin B (35) (Fig. 24.9) from Bryophyl-
compounds as well. fum pinnatum possesses an unusual orthoacetate-containing
Many plants containing cardiac glycosides have been structure; this compound has an EDso of less than 80 ng/ml
used for medicinal purposes (Kinghorn, 1992). Digitalis in the KB cell culture system (Yamagishi et a!., 1989).
(Scrophulariaceae) is particularly important, but ouabain
(from the seeds of Strophanthus spp. or wood of Acokanth- Arrow Poisons
era schimperi, both Apocynaceae) also is widely used. Di- A number of plants containing cardiac glycosides [e.g.,
goxin (34) from Digitalis lanata is the most widely used seed extracts of Strophanthus gratis and S. kombe (Apocyna-

""~", o~~~/~"
o

~ 08
08

~o, 0
08
80
80
~
08
bryophyUin B (35) ouabain (32)

Fig. 24.9. Medically important cardiac glycosides.

470 Saponins and Cardenolides

o
o

allosyl-O-deoxyallosyl-O gulosyl-O-deoxytalosyl-o
OH OH

o o

allosyl-O-deoxyallosyl-O glucosyl-O-deoxytalosyl-O
OH
Fig. 24.10. Cardenolides from Lophopetalum toxicum.

ceae) species1 have been used as arrow poisons (Lewis, chromatography-mass spectrometry of oleanane- and ursane-
1992). In some cases, the active compounds have been iso- type triterpenes-Applications to Chenopodium quinoa
lated and confinned to be cardenolides. The active principles triterpenes, Phytochemistry, 24, 2063-2066 (1985).
of the arrow poison from Lophopetalum toxicum (Celastra- CHANDEL, R. S. and R. P. RASTOGI, Triterpenoid saponins and sapo-
ceae) consists of four cardiac glycosides (Fig. 24.10). genins: 1973-1978, Phytochemistry. 19. 1889-1908 (1980).
c.wu.WOOD, B. V. and K. A. c.wu.WOOD, Terpenoid production
in plant cell cultures, in Ecological Chemistry and Biochemistry
of Plant Terpenoids (J. B. Harbome and P. M. Dey, eds.), Phyto-
REFERENCES chemistry Society of Europe Vol. 31,95-132, Clarendon, Ox-
ford, 1991.
AGARWAL, S. K. and R. P. RASTOGI, Triterpenoid saponins and CHITWOOD, D. J., Nematicidal compounds from plants, in Phyto-
their genins, Phytochemistry, 13. 2623-2645 (1974).
chemical Resources for Medicine and Agriculture (H. N. Nigg
APPLEBAUM, S. W. and Y. BlRK, Saponins, in Herbivores (G. A. and D. S. Seigler, eds.), 185-204, Plenum Press, New York,
Rosenthal and D. H. Janzen, eds.), 539-566, Academic Press, 1992.
New York, 1979.
CHOl, Y., R. HUSSAIN, J. M. i'Ezurro, A. D. KINGHORN, and J. F.
ARNAULT, C. and B. MAUCHAMP, Ecdysis inhibition in Acrolepi- MORTON, Abrusosides A-D, four novel sweet-tasting triterpene
opsis assectella larvae by digitonin: Antagonistic effects of cho- glycosides from the leaves of Abrus precatorius, J. Nat. Prod.,
lesterol, Experientia, 41, 1074-1077 (1985). 52. 1118-1127 (1989).
BERENBAUM, M. R. and E. Mu.!CZKY, Mantids and milkweed bugs: CONNOU..Y, J. D. and R. A. HnL, Cardenolides, in Terpenoids (B.
Efficacy of aposematic coloration against invertebrate preda- V. Charlwood and D. V. Banthorpe, eds.), Vol. 7 of Methods
tors, Am. Midi. Nat., 111, 64-68 (1984). in Plant Biochemistry (J. B. Harbome and P. M. Dey, eds.),
BOAR, R. B. andJ. ALLEN,/l-Amyrin triterpenoids, Phytochemistry, 361-368, Academic Press, London, 1991.
12. 2571-2578 (1973). DAHLGREN, R. M. T., S. ROSENDAL-JBNSBN, and B. J. NIELSEN, A
BONDI, A., Y. BIRK, and B. GESTETNER, Forage saponins, in Chem- revised classification of the Angiospenns with comments on
istry and Biochemistry of Herbage, Vol. 1 (G. W. Butler and correlation between chemical and other characters, in Phyto-
R. W. Bailey, eds.), 511-528, Academic Press, London, 1973. chemistry and Angiosperm Phylogeny (D. A. Young and D. S.
BOONSONG, C. and S. E. WRIGHT, The cardiac glycosides present Seigler, eds.), 149-204, Praeger, New York, 1981.
in mistletoes growing on Nerium oleander, Aust. J. Chern., 14, DE MAyo, P., The Higher Terpenoids, Interscience, New York.
449-457 (1961). 1959.
BROWER, L., Ecological chemistry, Sci. Am., 220. 22-29 (1969). DIMOCK, M. B., J. A. A. RENwlCK, C. D. RADKE, and K. SACHDEV-
. BROWER, L., Chemical defence in buttedlies, in The Biology of GUPTA, Chemical constituents of an unacceptable crucifer. Ery-
Butterflies (R. I. Vane-Wright and P. R. Ackery, eds.), Sympo- simum cheiranthoides, deter feeding by Pieris rapae, J. Chern.
sium of the Royal Entomological Society, London 11. 109-134, Ecol., 17. 525-533 (1991).
Academic Press, London, 1984. EillS, B. E .• Natural products from plant tissue culture, Nat. Prod.
BURNOUF-RAnosEVICH, M., N. E. DELFEL, and R. ENGLAND, Gas Rep., 5. 581-612 (1988).
 Saponins and Cardenolides 471

FARNSWORTII, N. R., T. O. HENDERSON, and D. D. SOEIARTO, Plants Economic and Medicinal Plant Research, Vol. 2 (H. Wagner,
with potential molluscicidal activity. in Plant Molluscicides, H. Hikino and N. R. Farnsworth, eds.), 39-72, Academic Press,
131-204, John Wiley & Sons, Ltd., Chichester, 1987. London, 1988.
FARNSWORTII, N. R., A. D. KINGHORN, D. D. SOEIARTO, and D. P. HODGE, J. E. and G. E. lNGLB1T, Structural aspects of glycosidic
WALLER, Siberian ginseng (Eleutherococcus senticosus): Cur- sweeteners containing {l'-2)-linked disaccharides, in Sweet-
rent status as an adaptogen, in Economic and Medicinal Plant eners (G. E. Inglett), 216-234, AVI, Westport, CT, 1974.
Research, Vol. 1 (H. Wagner, H. Hikino and N. R. Farnsworth, HosTETI'MANN, K., Isolation and identification of saponins and re-
eds.), 155-210, Academic Press, London, 1985. lated terpenoid glycosides, in Advances in Medicinal Plant Re-
FULLAS. F., Y. CHOIt A. D. KINGHORN, and N. BUNYAPRAPHATSARA, search (A. J. VIietnck and R. A. Dommisse, eds.), 225-241,
Sweet-tasting triterpene glycoside constituents of Abrus fruticu- Wissenschaftliche Verlagsges. Stuttgart, 1985.
losus, Planta Medica, 56, 332-333 (1990). HOSTETrMANN, K., Current level of technology in the separation,
GBRSHBNZON, J. and R. CROTEAU, Terpenoids. in Herbivores: Their isolation, and structural analysis of natural products, in Ad-
Interactions with Sec~ndary Plant Metabolites, Vol. I (G. A. vances in Medicinal Phytochemistry (D. Barton and W. D. Ollis,
Rosenthal and M. R. Berenbaum, eds.), 165-219, Academic eds.), 67-76, Libbey, London, 1986.
Press, San Diego, CA, 1991. HOSTEITMANN, K., M. HOSTETIMANN, and A. MARSTON, Saponin.,
GOODWIN, T. W., Membrane-bound enzymes in plant sterol biosyn- in Terpenoids (B. V. Charlwood and D. V. Banthorpe, eds.),
thesis, in The Enzymes of Biological Membranes, 2nd ed. (A. Vol. 7 of Methods in Plant Biochemistry 0. B. Harbome and
N. Martonosi, ed.), 205-226, Plenum, New York, 1985. P. M. Dey, eds.), 435-471, Academic Press, London, 1991.
GRoENEVELD,H. W.,H. STBDL,B. VANDERBERG,andJ. C. ELiNGS, HOWES, F. N., Fish-poison plants, Bull. Misc. Inform. Kew, No.
Rapid, quantitative HPLC analysis of Asclepias fruticosa L. and 4, 129-153 (1930).
Danaus plexippus L. cardenolides, J. Chern. Ecol., 16, HUSSAIN, R. A., Y. LIN, L. 1. POVEDA, E. BORDAS, B. S. CHUNG,
3373-3382 (1990). J. M. i'EzzUTO, D. D. SOEIARTO, and A. D. KINGHORN, Plant-
GROENEVELD, H. W., B. VAN DEN BERG, J. C. ELlNGs, and D. derived sweetening agents: Saccharide and polyol constituents
SEYKENS, Cardenolide biosynthesis from malonate in Asclepias of some sweet-tasting plants, J. Ethnopharmacol., 28, 103-115
curassavica, Phytochemistry, 29, 3479-3486 (1990). (1990).
GROENEVELD, H. W., A. BINNEKAMP, and D. SEYKENS, Cardenolide KIM, J. and A. D. KINGHORN, Further steroidal and flavonoid con-
biosynthesis from acetate in Asclepias curassavica, Phytochem- stituents of the sweet plant, Polypodium giycyrrhiza, Phyto-
istry, 30, 2577-2585 (1991). chemistry, 28, 1255-1228 (1989).
GRUNWALD, C., Steroids, in Secondary Plant Products (E. A. Bell KIM, J., J. M. Pezzuro, D. D. SOEIARTO, F. A. LANG, and A. D.
and B. V. Char1wood, eds.), 221-256, Springer-Verlag, Berlin, KINOHORN, Polyporoside A, an intensely sweet constituent of
1980. the rhizomes of Polypodiwn glycyrrhiza, J. Nat. Prod., 51,
HAReORNB, J. B., Introduction to Ecological Biochemistry, 2nd ed., 1166-1172 (1988).
Academic Press, New York, 1982. KINGHORN, A. D., Plants as sources of medicinally and pharmaceut-
liARBORNE, J. B., Recent advances in chemical ecology. J. Chern. ically important compounds, in Phytochemical Resources for
Ecol., 3, 323-344 (1986). Medicine and Agriculture (H. N. Nigg and D. S. Seigler, eds.),
75-95, Plenum Press, New York, 1992.
HARBORNE, J. B., Recent advances in chemical ecology. Nat. Prod.
Rep., 7, 85-109 (1989). KupcHAN, S. M., J. R. KNox, J. E. KELsEY, and J. A. S. RENAULD,
HAReORNB, J. B., Recent advances in the ecological chemistry of Calotropin, a cytotoxic principle isolated from Asclepias syr-
plant terpenoids, in Ecological Chemistry and Biochemistry of iaca, Science, 146, 1685-1686 (1964).
Plant Terpenoids 0. B. Harbome and P. M. Dey, eds.), Phyto- LEE, S. M. and J. N. SEIBER, Biosynthetic preparation of cardenol-
chemistry Society of Europe Vol. 31, 399-426, Clarendon, Ox- ides from [lYCjacetic acid by stem discs of the milkweed
fotd, 1991. Asclepias curassavica, Phytochemistry, 22, 923-927 (1983).
HARMATHA, J., B. MAuCHAMP, C. ARNAULT, and K. SLAMA, Identi- LEWls, W. H., Plants used medicinally by indigenous peoples, in
fication of a spirostane-type saponin in the flowers of leek with Phytochemical Resources for Medicine and Agriculture (H. N.
inhibitory effects on growth of leek-moth larvae, Biochem. Syst. Nigg and D. S. Seigler, eds.), 33-74, Plenum Press, New York,
Ecol.,15, 113-116 (1987). 1992.
HENDERSON, T. 0., N. R. FARNSWORTII, and T. C. MYERS, Biochem~ MAHATO, S. B., A. N. GANGULY, and N. P. SAHU, Steroid saponins,
istry of recognized molluscicidal compounds of plant origin, Phytochemistry, 21, 959-978 (1982).
in Plant MoUuscicides, 109-130, John Wiley & Sons, Ltd., MALcOLM, S. B., Cardenolide-mediated interactions between plants
Chichester, 1987. and herbivores, in Herbivores: Their Interactions with Second-
HEROlIT, V., Some relations between plants, insects and their iso- ary Plant Metabolites, Vol. 1 (G. A. Rosenthal and M. R. Beren-
prenoids, in Progress in Phytochemistry, Vol. 2, L. Reinhold baum, eds.), 251-296, Academic Press, San Diego, CA, 1991.
and X. Liwschitz, eds., 143-202, Interscience, London, 1970. MALCOLM, S. B., B. J. CocKReLL, and L. P. BROWER, Cardenolide
HIKINo, H., Recent research on oriental medicinal plants, in Eco- fingerprint of monarch butterflies reared on common milkweed.
nomic and Medicinal Plant Research, Vol. I (H. Wagner, H. Asclepias syriaca L., J. Chern. Ecol., 15, 819-853 (1989).
Hikino, and N. R. Farnsworth, eds.), 53-85, Academic Press, MANN, J., Secondary Metabolism, Oxfotd University Press, Ox-
London, 1985. fotd, 1978; 2nd edition, 1987.
HIKINo. H. and Y. KISO, Natural products for liver diseases, in MARSTON, A. and K. HOSTETTMANN, Plant saponins: Chemistry
472 Saponins and Cardenolides

and molluscicidal action, in Ecological Chemistry and Bio- tion in insects, in Insect/Plant Relationships (H. F. van Emden,
chemistry of Plant Terpenoids (J. B. Harborne and P. M. Dey, ed.), 59-83, Royal Entomological Society, Symposium 6,
eds.), Phytochemistry Society of Europe Vol. 31, 264-286, Blackwell, London, 1973.
Clarendon, Oxford, 1991. ROTHSCHILD, M., H. ALBORN, G. STBNHAGEN, and L. M. SCHooN-
MARTIN, R. A. and S. P. LYNCH, Cardenolide content and thin- HOVEN, A strophanthidin glycoside in Siberian wallflower: A
layer chromatography profiles of monarch butterflies, Danaus contact deterrent for the large white butterfly, Phytochemistry,
plexippus 1.., and their larval host-plant milkweed, Asclepias 27, 101-108 (1988).
asperula subsp. capricornu (Woods.) Woods., in North Central SACHDBV-GUPl"A, K., J. A. A. RENwICK, and C. D. RADKE, Isolation
Texas, J. Chem. Ecol., 14, 295-318 (1988). and identification of oviposition deterrents to cabbage butterfly,
MATSUMOTO, K., R. KAsAl, K. OHTANT, and O. TANAKA, Minor Pieris rapae, from Erysimum cheiranthoides, J. Chern. Ecol.,
cucurbitane glycosides from fruits of Sirai/ia grosvenori (Cu- 16,1059-1067 (1990) [also see J. Chem. Ecol.,17, 247 (1991)].
curbitaceae), Chem. Pharm. Bull., 38, 2030-2032 (1990). SEIBER, J. N., S. M. LEE, and J. M. BENSON, Chemical characteris-
McLAUGLlN, J. L., B. FREEDMAN, R. G. POWELL, and C. R. SMITH, tics and ecological significance of cardenolides in Asclepias
JR., Neriifolin and 2'-acetylneriifolin: Insecticidal and cytotoxic (milkweed) species, in Isopentenoids in Plants (W. D. Nes, G.
agents of Thevetia thevetioides seeds, J. Econ. Entomol., 73, Fuller, and L. Tsai, eds.), 563-588, Marcel Dekker, Inc., New
398-402 (1980). York,1984.
MILLER, L. P., Glycosides, in Phytochemistry, Vol. 1 (L. P. Miller, SHIBATA, S., O. TANAKA, J. SHOO, and H. SAITO, Chemistry and
ed.), 297-375, Van Nostraod Reinhold, New York, 1973. pharmacology of Panax, in Economic and Medicinal Plant Re-
MOLYNEUX, R. J., B. C. CAMPBELL, and D. L. DREYER, Honeydew search, Vol. 1 (H. Wagner, H. Hikino, and N. R. Farnsworth,
analysis for detecting phloem traosport of plant natural products, eds.), 217-283, Academic Press, London, 1985.
J. Chem. Ecol., 16, 1899-1909 (1990). SHIMOMURA, H., Y. SACHIDA, and Y. MIMAKI, Steroidal saponins,
PASTEELS, J. M., M. ROWELL-RAHlER, T. RANDoux, J. C. BRAEK- pardarinoside A-G from the bulbs of Lilium pardarinum, Phy-
MAN, and D. DALOZE, Pyrrolizidine alkaloids of probable tochemistry, 28,3161-3170 (1989).
host-plant origin in the pronotal and elytral secretion of the TAKEDA, K., The steroidal sapogenins of the Dioscoreaceae, in
leafbeetle Oreina cacaliae, Ent. Exp. Appl., 49, 55-58 (1988). Progress in Phytochemistry Vol. 3 (L. Reinhold and Y.
PASTEBLS,1. M., M. ROWELL-RAHIER, J. C. BRAEKMAN, D. DALOZE Liwschitz, eds.), 287-333, Wiley-Interscience, New York,
and S. DUFFEY, Evolution of exocrine chemical defense in leaf- 1972.
beetles (Coleoptera: Chrysomelidae), Experientia, 45,295-300 TORI, K., H. ISHn, Z. W. WOl.KOWSKl, C. CHACHATY, M. SANGARE,
(1989). F. Pm.ou, and G. LUKACS, Carbon-13 nuclear magnetic reso-
REED, D. K., B. FREEDMAN, and T. L. LAoD JR., Insecticidal and nance spectra of cardenolides, Tetrahedron Lett. (13)
antifeedant activity of neriifolin against codling moth, striped 1077-1080 (1973).
cucumber beetle, and Japanese beetle, J. Econ. Entomol., 75, TYLER, V. E., L. R. BRADY, and 1. E. ROBBERS, Pharmacognosy,
1093-1097 (1982). 8th ed., Lea and Febiger, Philadelphia, 1981.
RENwICK, J. A. A., C. D. RADKE, and K. SACHDBV-GUPl"A, Toler- VANDBN BBRGHE, D. A., A. J. VLlBTINCK, and L. VAN HOOF, Present
ance of cardenolides of Erysimum cheiranthoides by the dia- status and prospects of plant products as antiviral agents, in
mondback moth, Plutella xylos/ella, Symp. BioI. Hung., 39, Advances in Medicinal Plant Research (A. J. Vlietinck and R. A.
527-528 (1990). Domisse, ods.), 47-99, Wissenschaftliche Verlagsges. Stuttgart,
RENwICK, J. A. A., C. D. RADKE, and K. SACHDBV-GUPl"A, Chemi- 1985.
cal constituents of Erysimum cheiranthoides deterring oviposi- YAMAGlSm, T., M. HARUNA, X. YAN, J. CHANG, and K. LEB, Antitu-
tion by the cabbage butterfly, Pieris rapae, J. Chem. Ecol., 15, mor Agents, 110. Bryophyllin B, a novel potent cytotoxic bufa-
2161-2169 (1989) [also see J. Chem. Ecol., 17, 247 (1991)]. dienolide frum Bryophyllum pinna/urn, J. Nat. Prod., 52,
ROTHSCHILD, M., Secondary plant substances and warning coloura- 1071-1079 (1989).
25
Limonoids, Quassinoids, and Related
Compounds

Introduction types of triterpenes are fonned. There is little direct evidence

Biosynthesis for many of the intennediates and enzymes involved in the
Limonoids (Tetranortriterpenoids) synthesis of limonoids, quassinoids, and other compounds
Quassinoids (Decanortriterpenoids) of this general derivation. Proposed biogenetic schemes are
Pentanortriterpenoids based largely on the occurrence of probable intennediate
Chemosystematic Studies compounds. Euphol (1) and tirucallol (2), which differ in
Biological Activity configuration at C-20, are both fonned from this pathway.
Antifeedant Activity Although most quassinoids and C30 triterpenes of the Rutales
Medicinal and Antitumor Properties have the C-20 (R) configuration, suggesting tirucallol as
Limonin in Orange Juice the precursor, C-20 (S)-euphol also was converted to the
References limonoid nimbolide (6) (Fig. 25.1) more efficiently than tiru-
callol inAzadirachta indica leaves (Champagne et aI., 1992).
The identity of the actual precursor of limonoids and quassi-
INTROovcnOI'l noids is not known with certainty. A7 -Euphol (8) and/or A 7 _
tirucallol (butyrospennol) (7) appear to be later intennedi-
ates in the biosynthetic sequence. The apo-rearrangement
Two major groups of metabolically altered triterpenes, the
occurs with introduction of a 14-unsaturation, migration of
limonoids (tetranortriterpenoids) and the quassinoids (dec-
the C-14 methyl group to C-8, and introduction of oxygen at
anortriterpenoids) are derived from euphol (1) (Fig. 25.1)
C-7 (Connolly, 1983), and may involve opening of a 70<,80<-
or tirucallol (2) (Fig. 25.2). To date, these compounds are
epoxide (9 and/or 10).
restricted in distribution to the Rutaceae, Meliaceae, Cneora-
Relatively simple C30 compounds, sometimes called pro-
ceae, Simaroubaceae, and, perhaps, the Burseraceae (Con-
tolimonoids, that appear to be derived from apo-euphol (11)
nolly and Hill, 1991), and occur variously in seeds, fruits,
and/or apo-tirucallol (12), have features suggesting their role
and wood of plants of these families. In general, the limonoid
as precursors and lack ring opening, are found in the Melia-
and quassinoid content of leaves has not been examined
ceae and Rutaceae (Connolly, 1983; Dreyer, 1983). It is not
(Taylor, 1983).
certain whether oxygenation precedes or follows the apo-
Small amounts (0.33%) of quassins are produced by cal-
rearrangement, as both melianone (13) (in which the C-14
lus and suspension cultures of Picrasma quassinoides
methyl group has not migrated) and grandifoliolenone (14)
(Charlwood and Charlwood, 1991).
(in which migration has taken place) have been isolated (Fig.
25.3).
A similar compound, turreanthin (5), from Turreanthus
BIOSYl"fl1lESIS africanus (Meliaceae), has a side-chain oxidation pattern that
suggests a potential furan ring (Connolly, 1983).
Both limonoids and quassinoids are derived from a cation
such as (3) (Fig. 25.2), produced by condensation of a Limonoids (Tetranortriterpenoids)
chair-chair-chair-boat configured squalene 2,3-epoxide The limonoids (more than 300 described compounds) are
(4) precursor. By subsequent rearrangement, many other a stereochemically homogeneous group of tetranortriter-

473
474 Limonoids. Quassinoids. and Related Compounds

P
~~ ySP';
H H~~
H6~ I _0

CH'C~O
O
HO" HO" CH,Co-O.yJOH

Co
eupbol (1) tUndlssol grandifollone

rc~
if o
0

'0
limonin (19)

®
kbivorin (23) mexicanolide (25)

CH~O'C
, OCH,

o I o "
CH,-O I

CH'C~ ~C:;H' o
! 0 0

nimbin (26) nimbolide (6) quassio (31)

Fig. 25.1. Some representative metabolically altered triterpenes.

~oo~- squalene 2,3-epoxlde (4) + (3) \

~ ;~--~~~
HO

! ':
,~ (A'-UrucaJlol)
butyro.permol(7)
HO

tirucallol (2) H
HOr·
eupbol (I) \
R
H

HO #
l""
~~:~#~
\ ,,_.
o

~
HO
0
(9)
--

~
HO
I

oo~~ ~
Amencan SocIety of Pbannacognosy, Downers Grove, Dlinois).
••
Fig. ~.2. Bi,osynthesis of apo-eupbol and apo-tirucallol (modified from . . . .
SiddiqUI et aI., 1988; used with pennission of the copyright owner, the
 Limonoids. Quassinoids. and Related Compounds 475

0
OH

!{r4!~ 00

~~o#o¥ o~
turreanthin (5) me1ianone (13) grandlfoliolenone (14)

(Y0H (Yo
,
o
°
(15) (16)

Fig. 25.3. Protolimonoids.

penes only found in the Meliaceae, Rutaceae, Cneoraceae, tabolites in various plants that synthesize and accumulate
and Harrisonia (Simaroubaceae) (Champagne et al., 1992; limonin supports this pathway. As members of the genus
Harborne, 1991). A report from Balsamodendron pubeseens Citrus (Rutaceae) synthesize only nomilin (20) in the leaves
(Bursemceae) warrants reconfirmation (Taylor, 1983). Li- and stems (the principal site of biosynthesis), this compound
monoids have a furan ring attached as a side chain at C- appears to be moved to other sites (e.g., the fruits) and subse-
17 to a cyclopentenophenanthrene nucleus which may have quently modified (Hasegawa et aI., 1984).
undergone oxidative opening of one or more rings (Taylor, Other compounds undergo A-ring cleavage as well; for
1983). example, tricoccin S22 (21), from the Cneoraceae (Connolly,
Oxidative modification eventually results in the removal 1983).
of the four terminal side-chain carbons and formation of a Cleavage of ring B of intermediate compounds similar to
l3-substituted furan ring. A hemiacetal (15) and a lactone khivorin (23) (Fig. 25.1) may give rise to compounds such
(16) from Chisoeheton panieulatus (Meliaceae) represent as andirobin (22) and methyl angolensate (24) (a B,D-seeo-
stages between the intact side chain and the furan ring. limonoid) (Fig. 25.6). This cleavage also may be concealed
Cedrelone (17) and a series of other compounds have
suffered cleavage of the side chain but retain the apo-euphol

c>
skeleton intact (Fig. 25.4) (Connolly and Overton, 1972).
Other classes of limonoids may have the A, B, C, or D
ring (or a combination of rings) cleaved. For example, gedu-
~O
I

o~
nin (18) has a cleaved D ring (D-seeo-lirnonoids) (Fig. 25.4),
whereas limonin (19) has both a cleaved A ring and D ring
(A,D-seeo-limonoids) (Fig. 25.5). The initial products of the
oxidation process are concealed by secondary cyclization. °
Both the epoxide and II-lactone systems are characteristic of
limonoids. OH
°
A plausible pathway for the formation of 1imonin from
cedrelone (17) gedUDin (18)
d '-tirucallol (7) has been proposed (Fig. 25.5) (Arigoni et
al., 1960; Dreyer, 1984). The presence of related minor me- Fig. 25.4. Limonoids with an intact apo-eupbol skeleton.
476 Limonoids, Quassinoids, and Related Compounds

HO

Ll7 -tirucallol (7)

k)
1'0 Q5P''0H~
~O ,~O

""~- O~OO
0- OH

0
HO - 0 0
o
deacetylnomilinate deacetylnomilin

nomilin (20) obucanone (47) obacunoate

o ~O C)
.nomilin ~-
~'Oo_O~lloo
acetyl-lyase 0 0 0 ~
** obacunone
A-ring lactone
hydrolase isoobacunoate Jimonin (19)

Fig. 25.5. Proposed biosynthetie pathway to limonin (modified from Dreyer, 1984; used with pennission of the copyright owner, Marcel Dekker, Inc.,
New York).

by other reactions (Connolly and Overton, 1972) such as to those of limonoids. Li?-Euphol (8) and/or Li? -tirucallol
those seen in the fDlmation of mexicanolide (25) which has (butyrospermol) (7) (Fig. 25.7) appear to be early intermedi-
a bicyclo[3.3.IJnonane system (Connolly, 1983). Cleavage ates in the biosynthetic sequence. The apo-rearrangement
of ring C yields compounds such as nimbin (26) (a C-seco- occurs with the introduction of a 14-unsaturation, migration
limonoid) (Fig. 25.6) (Connolly, 1983). Most compounds of the C-14 methyl group to C-8 and the introduction of
of this group occur in the genera Melia and Azadirachta oxygen at C-7 (Connolly, 1983) and may involve opening
(Connolly, 1983). of a 7a,8a-epoxide. As is true for many limonoids, the D
As previously mentioned, gedunin (18) (Fig. 25.4) has ring of an intermediate compound is expanded by the biolog-
undergone cleavage of the Dring. D-Ring cleavage is often ical equivalent of a Baeyer-Villiger reaction to produce a
associated with other modifications of the molecule as well S-lactone (Polonsky, 1983). Opening of that lactone, fol-
(Connolly, 1983). lowed by recyclization to the 7-hydroxyl group (and produc-
tion of another 8-lactone) (27) yields a possible intermediate
Quassinoids (Decanortriterpenoids) for the different routes required for subsequent transforma-
tions (Polonsky, 1983). Several C25 -quassinoids could logi-
Quassinoids are decanortriterpenes known to occur only cally be derived from this intermediate (Fig. 25.7). Among
in the family Simaroubaceae (Polonsky, 1983, 1986). More these are simarolide (28), picrasin A (29), and soulameolide
than 120 compounds and 5 major structural types have been (30). All possess a C-l7 and none possess a C-12 oxygen
described (Polonsky, 1986). function (Polonsky, 1983). Introduction of an oxygen at C-
The biosynthetic precursors of quassinoids are similar 12 could lead to cleavage of the C J3 -C l7 bond and formation
 Limonoids. Quassinoids. and Related Compounds 477

ceae (Dreyer, 1984; Mondon and Epe, 1983). Many tetranor-

triterpenoids (such as 21, Fig. 25.6) also have been isolated
from eneorum tricoccum (Epe and Mondon, 1979; Mondon
and Epe, 1983).

CHEl'IOSYSTEJllATIC STUOmS
nimbin(26)
This series of compounds and pathways provides one of
the best examples of the application of terpene chemistry to
phylogeny. Limonoids characterize the family Meliaceae,
where they are diverse and abundant; a more limited range
of structures is found in the Rutaceae and Cneoraceae
(Champagne et aI., 1992). Rearrangement of the A ring leads
to formation of the typical rutaceous limonoids (also found
in Harrisonia), whereas rearrangement of both the A and B
rings results in the highly derived limonoids characteristic
andirobin (22) mexlcanolide (25)
of the Cneoraceae. In the subfamily Sweetenoideae of the
Meliaceae, D-seco-limonoids are further oxidized to B,D-

c> C>
«ft
seco structures, which may then be extensively rearranged
tu yield more complex structures. in contrast, members of
the subfamily Melioideae produce limonoids through a vari-
fo ~i ::;;- ...·OH
ety of pathways leading tu B-, A-, AB-, and C-seco com-
pounds (Champagne et aI., 1992). Quassinoids are found
0 0 0 only in members of the Simaroubaceae. Based on the distri-
o 0 ~ bution of limonoids and quassinoids, it seems probable that
COZCHJ the Meliaceae and Simaroubaceae arose independently from
methyl angoJensate (24) tricoccln Su (21)
Rutaceae-like ancestors. The Simaroubaceae has the most
highly modified triterpenoid chemistry of the three families
Fig. 25.6. Limonoids that exhibit B-ring or C-ring cleavage. (Simao et aI., 1991; Waterman, 1983). Within the Rutaceae,
other chemical changes have occurred that presumably arose
after the separation of the Meliaceae and Simaroubaceae
of C20 quassinoids (Fig. 25.8). All members of this last group
from Rutaceous stock (da Silva et aI., 1984; Gershenzon and
contain C-12 oxygen functions (Polonsky, 1983).
Mabry, 1983; Hegnauer, 1983; Seigler, 1981).
Quassin (31), the parent compound of this group, shares
Data from triterpenes and chromosomes shed light on the
certain structural similarities with merogedunin (32) that can
he produced by treatment of gedunin (18) (Fig. 25.4) with placement of several enigmatic groups. Nymania capensis,
a small South African tree that previously has been placed
alkali. Gedunin suffers a fragmentation to a a-lactone and
in a number of families, contains limonoids that strongly
furan-3-aldehyde. The reaction is typical of limonoids hav-
ing the O-ring structure of gedunin and is called the "meroli- suggest affiliations with the Meliaceae. The compounds
monol rearrangement" (Connolly and Overton, 1972; Geiss- present are similar to those of the genera Guarea, Trichilia,
and Aphanamixis species and support placement of the genus
man and Crout, 1969). Although it has heen suggested that
in the subfamily Melioideae (MacLachlan and Taylor, 1982;
quassinoids could arise in this manner, such a synthesis re-
Taylor, 1983). Another such problem involves the genus
quires nonphysiological conditions.
inadequate data exist to define clearly the pathway of
Ptaeroxylon (Ptaeroxylaceae), which produces chromo-
somes similar to the genera Spathelia and Harrisonia but
biosynthesis.
lacks limonoids. The presence of limonoids in the genus
Pentanortrlterpenoids
Harrisonia suggests that genus may be misplaced within the
Simaroubaceae. Other studies suggests that Harrisonia is
A series of pentanortrlterpenoids has been isolated and related to the genus Spathelia (Taylor, 1983; Waterman,
studied from the family Cneoraceae (Fig. 25.9). This family 1983). Indeed, placement of the entire trihe Spathelioideae
has been placed in the Rutaceae by some (Thome, 1976) is somewhat questionable and, in many regards, is intermedi-
but differs (among other features) in the presence of this ate to the Rutaceae, Cneoraceae, and Ptaeroxylaceae (Water-
group of compounds. Although limonoids are found in the man, 1983). The separation of these taxa from proto-simar-
family, those closely related to limonin itself are lacking. A oubaceous ancestors must have come before the evolution
number of isomers of the cneorin-tricoccin series are of the quassinoid pathway. All (except Spathelia) have lost
known; these compounds are representative of the Cneora- the ability to synthesize alkaloids, Harrisonia and Spathelia
478 Limonoids, Quassinoids, and Related Compounds

HO
¥~y5P~¢0I~
A7••• phol (20~) (8)
I ---+-
HO
::0 ---..
HO

HO
....----+-
OH
21

20
23
COzH
.1.7·tirucallol (20a.) (7)
17

o
o

soulameolide (30) picrasin A (29) simarolide (28)

Fig.· 25.7. Possible scheme for the formation of quassinoids (modified from Polonsky, 1983; Academic Press, London).

have lost the ability to synthesize coumarins, and the Ptero· groups of plant metabolites. Some compounds of this group
xylaceae have lost the ability to synthesize limonoids (Wa· taste bitter to humans and, presumably, to other mammals
tennan, 1983). (Champagne et aI., 1992). Both vegetative parts and the
wood of many plants of the Rutaceae, Meliaceae, and Sima·
BIOLOGICAL ACTIVITY roubaceae are resistant to insect attack. In accord with these
observations, many limonoids, quassinoids, and their rela-
Many metabolically altered triterpenes contain epoxides, tives possess a variety of biological activities.
lactones, furans, and cyclopentanoid systems, functional

rttfu
units that are associated with biological activity in other Antifeedant Properties
A number of limonoids and quassinoids from the Ruta·
° CH3
ceae, Meliaceae, and Simaroubaceae are insect antifeedants.
o '" These compounds have been extensively investigated, as
CHJO they may provide relatively specific, biodegradable insecti·
--. I H cides and antifeedants.
I "0 0 Although extracts of seeds of many meliaceous plants
have many interesting bioactivities, the best known example
(27) quassin (31) is extracts of the neem tree (Anon., 1992; Klocke et aI.,
1989; Kraus et aI., 1987; Saxena, 1989; Smutterer et aI.,

~ ~
H 1980). The neem tree, Azadirachta indica (MeJiaceae), is
I OH I OH native to India, but grows commonly in the Near East and

o 17 0 0 HO~' 0 0
in certain regions of Africa. Extracts of the seed oil have
both insecticidal and antifeedant properties (Harbome, 1989;
Kraus et aI., 1987; Stone 1992); at least 200 species of insects
and mites are affected (Champagne et aI., 1992). In Israel,
merogedunin (32) merokhivorin
it has been noted that the tree is never eaten by the desert
Fig. 25.8. Czo and C Z5 quassinoids. locust, Schistocerca gregaria, a generalist insect known to
 Limonoids, Quassinoids, and Related Compounds 479

cneorin C cneorinK

o
-t--t--f-O
o
tricoccin C14

Fig. 25.9. Pentanortriterpenes and tetranortriterpenes from the Cneoraceae.

eat virtually all vegetation. Extracts of neem tree seeds are bug, Oncopeltus fasciatus (Champagne et al., 1989), but
repellent at low concentrations. The probable active com- other studies failed to reproduce this activity (Champagne
pound is the tetranortriterpene azadirachtin (33) (Fig. 25.10) et aI., 1992). Insect ecdysis inhibitors from plants have been
(Broughton et al., 1985; Kraus et al., 1985, 1987; Ley et aI., reviewed (Kubo and Klocke, 1986).
1987, 1992; Schroeder and Nakanishi, 1987). Azadirachtin Extracts and exudates of neem trees, Azadirachta indica
gives 100% feeding inhibition at 40 JLglL against Schistoc- (MeIiaceae), have biological activity against a variety of dif-
erca gregaria (desert locust) (Kubo and Nakanishi, 1977) ferent nematodes. For example, a neem seed extract applied
and against the fall armyworm (pC9S = 0.1 JLgldisk) (Kubo to tomato plants as a root drench at 125 JLg/ml inhibited
and Klocke, 1981). Azadirachtin has an ECso of 0.36 ppm reproduction of the root-rot nematode Meloidogyne javanica
against larvae of Peridroma saucia, the variegated cutwonn (Chitwood, 1992). Azadirachtin (33), at 10 JLg/ml, inhibits
(Champagne et al., 1989). In contrast, azadirachtin has no microfIlarial release in the animal-parasitic nematode,
antifeedant activity against nymphs of the migratory grass- Brugia pahangi. A similar mode of action may be involved
hopper, Melanoplus sanguinipes, although the topical EDso in phytoparasitic nematodes (Chitwood, 1992).
is 4.5 JLglg insect (Champagne et al., 1989). Feeding deterre- Curiously, when azadirachtin (33) is fed as a part of a
ncy involves interaction with one set of sensory receptors, blood meal to Rhodnius prolixus, a vector of Chagas disease,
whereas the toxic effects probably involve different systems subsequent parasitic infestation of the arthropod by the try-
in the insect. panosome Trypanosoma cruzi is prevented, but azadirachtin
The mode of toxicity of azadirachtin when consumed by is not directly toxic to the trypanosome (Borris and Schaef-
insects appears to involve the neuroendocrine regulation of fer, 1992).
juvenile and molting honnone titers (Rembold, 1989). Azad- Other compounds from Azadirachta indica, including Ii-
irachtin produces no observable effects on larvae between monin (19), nomilin (20), nimbin (24), nimbalide, azidirone,
molts. Only after the initiation of the insect molting cycle and salannin also are feeding deterrents. Nimbolide (6) has
(occurring simultaneously with the fonnation of new cuticle) cytotoxic activity (LDso of 0.25 JLg/ml against KB and 0.065
does the toxicity become evident. The larvae usualIy die as JLglml against P-388 celI culture systems (Kigodi et aI.,
a result of being encased in the unshed exuviae that prevents 1989).
feeding, excretory, and locomotory functions (Kubo and Another widely grown meliaceous tree, the chinaberry or
Klocke, 1981). Cedrelone (17) (Fig. 25.4) is the only other Persian lilac (Melia azadirachta), also contains a variety of
compound of this series known to inhibit ecdysis (Klocke, toxic limonoids. This tree is known for its cathartic, emetic,
1987). This compound inhibited molting of the milkweed and anthelminthic properties (Kraus et al., 1987). Although
480 Umonoids, Quassinoids, and Related Compounds

"Co
23

2O~

12
~ 21

",'
CH3CO-O

CH3CH,CH(CH3)CO· 0
azadirachtin (33)
meliatoxin At (34)

CH3COO""...

CH 3COO"·..··

CH,CH,CH(CH,)CO- 0
(CH3lzCHCO-O
meliatoxin B2 (37) (CH3hCHCO· 0 meliatoxin A2 (36)
meliatoxin 8 1 (35)

Fig. 25.10. Some antifeedant limonoids.

azadirachtin has been reported from this plant and might be noids [such as azadirachtin (33)], the most active compounds
expected to occur there, the presence of this compound from appear to be intact apo-euphollimonoids with a 14,15-epox-
Melia azadirachta requires confirmation (Kubo and Klocke, ide and either a 19/28 lactol bridge or a cyclohexenone A
1981). Furthermore, chemical variation in this widely dis- ring (Champagne et aI., 1992). Examples of the latter type
seminated species exists, complicating interpretation of phy- include anthothecol, cedrelone (17), sendanin (40), trichiro-
tochemical reports. kanin (41), toosendanin (39), and meliacin Az (36).
Meliatoxins Al (34), BI (35), Az (36), and B2 (37) are Certain other meliaceous seed extracts were equally ac-
the compounds responsible for most of the fruit toxicity of tive to azadirachtin in bioassays (Mikolajczak and Reed,
some variants of this plant (Oelrichs et a1., 1983), but other 1987). Those of Aglaia cordata were more potent than ex-
insect antifeedants also are present. These fruits are espe- tracts of neem toward larvae of Spodoptera jrugiperda (Mi-
cially toxic to swine (Oelrichs et a1., 1983). Other com- kolajczak et aI., 1989). A Iimonoid compound from Carapa
pounds isolated from Melia azedarach include ohchinolid procera had antifeedant activity comparable to that of azadi-
A and B, nimbolidin A and B, and nimbolinin B (Kraus et rachtin, but much weaker insecticidal properties. This com-
aI., 1982). pound, methyl 313-isobutyryloxy-l-oxomeliac-8(30)-enate
Melianone (13) (Fig. 25.3) from the leaves of Melia azad- (42), also was found in Khaya senegalensis and K. nyasica
irachta has antifeedant activity. Meliantriol (38) (Fig. (Mikolajczak et aI., 1988).
25.11), from the fruit, inhibits feeding of the desert locust Toonacilin (43) and 6-acetoxytoonacilin (44) from the
Schistocerca gregaria (Lavie et aI., 1967). bark of Toona ciliata (Meliaceae) (B-seco-limonoids) ex-
Toosendanin (39) from the Chinese species Melia toosen- hibit powerful antifeedant activity against the Mexican bean
danan displays activity as strong as azadirachtin against lar- beetle (Kraus et aI., 1978).
vae of Spodoptera litura and has better activity than azadi- Four additional bioactive compounds with intact apo-eu-
rachtin against the yellow stemborer, Scirpophaga phol skeletons were isolated from the meliaceous tree Trichi-
incertulas, and the Asiatic comborer, Ostrinia furnacalis lia emetica (syn. T. roka) of East Africa (Nakatani et aI.,
(Fig. 25.11) (Towers et aI., 1989). 1981). The Iimonoids of this tree had antifeedant properties,
Despite the fact that the neem tree is widely known, limo- but none were as potent as azadirachtin (Kubo and Klocke,
noids from other sources also have pronounced effects on 1981). Trichilin A (45) differs from meliatoxin Al (35) only
insect feeding and growth; many of these have been tabu- in the presence of a hydroxyl substituent at C-12 (Oehlrichs
lated (Champagne et aI., 1992). Apart from the C-seco limo- et aI., 1983).
 Limonoids. Quassinoids. and Related Compounds 481

A series of six quassinoids isolated from Simaba mul- activity of Spodoptera exempta at the 20-ppm level in leaf-
tiflora and Soulamea soulameoides were shown to be active disk tests and has antibiotic and cytotoxic activity (2.2
as growth inhibitors and as antifeedants against Heliothis fLg/ml, KB test). Although the East African variety of Harri-
virescens (cotton budworm) andSpodopteraJrugiperda (fall sonia abyssinica contains harrisonin, the West African vari-
armyworm) (Klocke et al., 1985). ety lacks this compound (personal communication, P. G.
A nomilin derivative (46) from the seeds of a Xylocarpus Waterman).
species [erroneously reported as being from Uncaria gambia
(Rubiaceae)] has been studied (Ahmed et al., 1978). The Medicinal and Antitumor Properties
identify of the Xylocarpus species has been suggested to be
X. mol/uscensis, but possibly should be Xylocarpus gra- Several Iimonoids have anticancer activity. Bioassay-
natum. guided fractionation led to the isolation of amoorastatin (49),
A species of enigmatic affinities, Harrisonia abyssinica, a cytotoxic limonoid from Aphanamixis grandifolia that has
usually placed in the Simaroubaceae (but see the discussion the same B-, C-, and D-ring substituents and the same rela-
above), contains both the widespread Iimonoid bitter princi- tive stereochemistry as meliatoxin A2 (Fig. 25.11) (Cham-
ple, obacunone (47) and a second Iimonoid, harrisonin (48). pagne et al., 1992; Oehlrichs et al., 1983). 14,15-Epoxy-
Harrisonin has insect antifeedant activity, is cytotoxic, and prieurianin (SO) from Guarea guidona also possessed
has antibacterial activity. This compound inhibits feeding antitumor activity. Aphanostatin (51), sendanin (40), and 12-

OH

HO~H OCOCH,CO

~
HO j'

.o~
I
o 0
CH,COO OH
HO H

r:
meliantriaol (38) toosendanin (39) sendanin (40)

OCOCHkO OH. o

~
OO "
CH,COO '
o 0 0
OH CH,CO O· OH
CH,CH,CH(CH,)COO H
OCOCH(CH,)CH,
methyl 38wisobutyryloxy- trichirokanin (41) trichilin A (45)
l-oxomeliac-8(30)-enoate (42)
OH 1: 0

CH'C~OO
CH,CH,CH(CH,)CHOHCo-O
i iii
HO I
..... 0 0
o CH,COO' OH
CH,CH,CH(CH,)COO H
CO,CH,

C>
4fiP
OCOCHJ 15,16-epoxyprieurlauin (SO) apbanostatin (51)

CH,COO 1-'0
~ CHCOO 6)--0 1: 0

~o
HO Ii
1i
o l
'" o 0 0
o CH,CO 0 OH
CO,CH, i OCOCH, HO
OHO OH H
toonacllin (43) 6-acetoxytoonacilin (44) harrisonin (48) amoorstatin (49)

Fig. 25.11. Some biologically active limonoids.

482 Limonoids, Quassinoids, and Related Compounds

hydroxyamoorastatin also had pronounced activity in tests tive against Plasmodium Jalciparum, one of the organisms
against a murine P-338 lymphocytic leukemia cell line responsible for malaria (Polonsky, 1983). Bruceantin (53)
(Champagne et a!., 1992), However, most limonoids lack in and simalikalactone D (55) had ICso of 0.8 ng/ml and 0.9
vivo activity. ng/ml values, respectively, against PlasmodiumJalciparum.
The limonoid nimbolide (6) (Fig, 25, I) from Azadarichta These compounds were the most potent of a series tested
indica inhibited Plasmodium Jalciparum in vitro (LDso 0.95 for antimalarial, antileukemic, and cytotoxic activity (Blasko
fJ-g/ml), but was inactive in vivo in mice. and Cordell, 1988; Borris and Schaeffer, 1992). Several
Several quassinoids are known to have antileukemic ac- other quassinoids have in vivo antimalarial activity, but tox-
tivity (Blasko and Cordell, 1988; Cordell, 1978; Polonsky, icity precludes use of most of these in man. Simalikalactone
1983). Only compounds with a Czo skeleton have antileuke- D also has powerful antileishmaniacal activity (Borris and
mic activity. Quassimarin (51) from Quassia amara and Schaeffer, 1992).
bruceoside A (52) from Brucea javanica and a series of other A mixture of quassinoids from Hannoa undulata (Sima-
quassinoids from Brucea antidysenterica (all Simarou- roubaceae) prevented penetration of tomato roots by infec-
baceae) (Fig. 25.12) are active in this regard. Bruceantin tive juveniles of the root-rot nematode, Meloidogyne javan-
(53) has a high degree of antitumor activity in P-388, LE, ica, at 1-5 fJ-g/mI. The major components of the mixture
LL and B 16 tissue culture systems (Cordell, 1978). Six quas- were chapparrinone (56), klaineanone (59), and glaucaruba-
sinoids from Simaba multiflora and Soulamea soulameoides lone (57) (Chitwood, 1992). The primary effect appeared to
were shown to be active against several tumor systems be on the motility of the nematodes.
(Klocke et a!., 1985).
Bruceantin binds tightly to ribosomes and this is the major Limonin in Orange Juice
mechanism by which it inhibits protein synthesis (Blasko
and Cordell, 1988). Fruits of the genus Citrus constitute one of the world's
Other quassinoids are known to have antiviral activity most important horticultural crops. Production is at least 45
(Polonsky, 1983). Among these are isobruceine A (54), si- million tons, of which about 45% is converted into a variety
malikalactone D (55), chaparrinone (56), glaucarubolone of processed products (Maier, 1983). After expression of
(57), and castelanone (58). citrus juices, especially orange juice, precursors that occur
Certain quassinoids [e.g., simalikalactone D (55)] are ac- in the juice are converted to bitter limonoids, primarily li-

HO ';lH_ OH OH

O~:~~'o~~~ _<-O~o·,co:ct(
yu. ~J. W'''O~O o~,. ~J.OH
quassimarin (51) isobrucein A (54) bruceoside A (52)

OH OH
OH

chaparrinone (56) glaucarubolone (57)

castelanone (58)

simaIikalactone D (55) bruceantin (53) klaineanone (59)

Fig. 25.12. Medicinally promising quassinoids.

 Limonoids, Quassinoids. and Related Compounds 483

o
W
o
o
i!

0
Co
OH
CO,H
o

oW o
i
. '

0
~O
0
CO,H

limonoie acid A-ring lactone (OO) 17·debydrolimoDoic acid A-ring lactone (61)

Fig. 25.13. Limonoic acid A-ring lactone and nonbitter debydro-derivative of limonin.

monin (19) (Fig. 25.5) and, to a lesser extent, nomilin (20). SUKH DBV, P. G. FERRINI, E. R. GLAZIER, A. MBLERA, S. K.
The bitter taste of limonin can be detected at concentrations i'RAnHAN, K. SCBAFFNER, S. STBRNHELL, J. F. TEMPLETON, and
ranging from 0.075 to 5 ppm (Champagne et al., 1992). S. TOBINGA, Constitution of limoDin, Experientia, 16, 41-49
Losses to the California citrus industry attributable to unac- (1960).
ceptable bitterness in juice products alone are estimated at BLASKO, G. and G. A. CORDELL, Recent developments in the chem-
$8,000,000 per year. istry of plant-derived anticancer agents, in Economic and Me-
No evidence of limonoid biosynthesis in fruit or seed dicinal Plant Research, Vol. 2 (H. Wagner, H. Hikino, and N.
R. Farnsworth, eds.), 119-191, Academic Press, London, 1988.
tissues exists, despite the fact that most of the limonins are
found in the seeds of mature fruits. Limonoid synthesis oc- BORRIS, R. P. and J. M. SCBABFFBR, Antiparasitic agents from
curs in the leaves and limonoids are transported into the plants, in Phytochemical Resources for Medicine and Agricul-
ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum
fruits (Maier, 1983). In citrus tissues, the natorally occurring
Press, New York, 1992.
precursor of limonin is a salt of limonoic acid A-ring lactone
(60) (Fig. 25.13) in which the A ring is closed and the D BROUGHTON, H. B., S. V. LEY, A. M. Z. SLAWIN, D. J. Wn.LIAMS,
and E. D. MORGAN, X-ray crystallographic structure determina-
ring is open. This tasteless compound is stable only in the
tion of detigloyldihydroazdirachtin and reassignment of the
salt form (Maier, 1983). In the presence of acid or the en- structure of the limonoid insect antifeedant azadirachtin, 1.
zyme citrus limonoate D-ring hydrolase, the D-ring lacton- Chern. Soc., Chern. Commun., 46-47 (1985).
izes to form limonin (19). The rate oflactonization is acceler-
CHAMPAONB, D. E., M. B.lsMAN, and G. H. N. TOWBRS, Insecticidal
ated by pasteurization of the juice. In the fruit, the precursor activity of phytochemicals and extracts of the Meliaceae, in
appears to be located in a compartment of the cell where Insecticides of Plant Origin (Amason, J. T., B. 1. R. Philogene,
the pH is neutral or alkaline, probably the cytoplasm (Maier, and P. Morand, eds.), ACS Symposium Series 387, 95-109,
1983). American Chemical Society, Washington, DC, 1989.
A number of methods have been developed to prevent CHAMPAGNB, D. E., O. KOUL, M. B. lsMAN, G. G. E. SCUDDER,
the formation of limonin in orange juice. Application of and G. H. N. TOWERS, Biological activity of limonoids from
auxin (and a number of synthetic compounds with auxin the Rutales, Phytochemistry, 3J, 377-394 (1992).
activity) in a preharvest treattnent prevents the synthesis of CHARLWOOD, B. V. and K. A. CHARLWOOD, Terpenoid production
nomilin (20) and hence the formation of limonin (19) (Ha- in plant cell cultures, in Ecological Chemistry and Biochemistry
segawa et al., 1986). Five species of bacteria have been re- of Plant Terpenoids (J. B. Harbome and F. A. Tomas-Barberan,
ported that have the ability to metabolize limonins. The use eds.), Phytochemistry Society of Europe Vol. 31,95-132, Clar-
of immobilized Arthrobacter globiformis cells at a favorable endon, Oxford, 1991.
pH (where both limonin D-ring lactone hydrolase and limo- CHITWOOD. Nematicidal compounds from plants, in Phytochemical
noate dehydrogenase of the bacteria are active) to convert Resources for Medicine and Agriculture (H. N. Nigg and D. S.
limonin to the nonbitter dehydro-derivative (61) is effective Seigler, eds.), 185-204, Plenum Press, New York, 1992.
in prevention of bitterness in orange jnice (Hasegawa et al, CONNOLLY, J. D., Chemistry of the limonoids of the Meliaceae
1982; Maier, 1983). and Cneoraceae, in Chemistry and Chemical Taxonomy of the
Rutale, (P. G. Waterman and M. F. Grundon, eds.), 175-213,
Academic Press, London, 1983.
REFERENCES CONNOLLY, J. D. and R. A. Hn.r., Triterpenoids, in Terpenoids (B.
V. Charlwood and D. V. Banthorpe, eds.), Vol. 7 of Methods
ANON., NEEM. A Tree for Solving Global Problems, National Acad- in Plant Biochemistry (1. B. Harbome and P. M. Dey, eds.),
emy Press, Washington, DC, 1992. 331-359, Academic Press, London, 1991.
AHMED, F. R., A. S. NG, and A. G. FALUS, 7a-Acetoxydihydronom- CONNOLLY, J. D. andK. H. OVERTON, The triterpenoids, in Chemis-
ilin: Isolation, spectra, and crystal structure, Can. J. Chern., 56, try of Terpenes and Terpenoids (A. A. Newman ed.), 207-287,
1020-1025 (1978). Academic Press, London, 1972.
AIuGGNI, D., D. H. R. BARTON, E. J. COREY, O. JEGER, L. CAGLIOTI, CORDEU., G. A., Anticancer agents from plants, in Progress in
484 Limonoids. Quassinoids. and Related Compounds

Phytochemistry Vol. 5 (L. Reinhold, 1. B. Harborne, and T. and P. Morand. eds.), ACS Symposium Series 387, 136-149,
Swain, eds.), 273-316, Pergamon, London, 1978. American Chemical Society, Washington, DC, 1989.
DA SILVA, M. F. D. G. F., O. R. GOITLIEB, and D. E. DREYER, KRAus, W .• M. BOKEL. A. KLENK. and H. POHNL, Structure of
Evolution of limonoids in the Meliaceae, Biochem. Syst. Beol., azadirachtin and 22.23-dihydro-23~-methoxyazadirachtin, Te-
12,299-310 (1984). trahedron Lett., 26, 6435-6446 (1985).
DREYER, D. L., Limonoids of the Rutaceae, in Chemistry and KRAus. W .• M. BOKEL, A. BRUHN. R. CRAMER. I. KLAIBER, A.
Chemical Taxonomy of the Rutales (P. G. Waterman and M. KLENK, G. NAGL. H. POHNL, H. SADLO, and B. VOGLER, Struc-
F. Grundon, eds.), 215-245, Academic Press, London, 1983. ture detennination by NMR of Azadirachta indica A. Juss.
DREYER, D. L., Biogenetic relationships of degraded triterpenes in (Meliaceae). Tetrahedron, 43, 2817-2830 (1987).
the Rutales, in Isopentenoids in Plants (W. D. Nes, G. Fuller, KRAus, W., W. GRIMMINGER, and G. SAWITSKI, Toonacilin and 6-
and L. Tsai, eds.), 267-290, Marcel Dekker, Inc., New York. acetoxytoonacilin, two novel ~-seco-tetranortriterpenoids with
1984. antifeeding activity. Angew. Chern. Int. Ed., 17, 452-453
(1978).
EPE, B. and A. MONDON, Zur Kenntnis der Bitterstoffe aus Cneora-
ceen, XII, Tetrahedron Lett., 2015-2018 (1979). KRAus. W .• R. KRAMER. M. BOKEL. and G. SAWITZKI. New insect
antifeedants from Azadirachta indica and Melia azedarach, in
GEISSMAN, T. A. and D. H. G. CROUT. Organic Chemistry of Sec-
Natural Pesticides from the Neem Tree (Azadirachta indica A.
ondary Plant Metabolism, Freeman Cooper, San Francisco,
Iuss.) (H. Schmutterer. K. R. S. Ascher. and H. Rembold. eds.).
1969.
Proceedings 1st International Neem Conferences. 53-62. 1980.
GERSHENZON, 1. and T. 1. MABRY, Secondary metabolites and the
KUBo, I. and J. A. KLoCKE, Limonoids as insect control agents,
higher classification of Angiosperms, Nordic J. Bot.. 3, 5-34
Les Mediateurs Chemiques. in Les CoUoques de I'INRA 7.
(1983).
206-219, 1981.
HARaORNE. J. B., Recent advances in chemical ecology, Nat. Prod. KUBO, I. and J. KLocKE, Insect ecdysis inhibitors, in Natural Resis-
Rep., 7, 85-109 (1989).
tance of Plants to Pests (M. B. Green and P. A. Hedin, eds.).
HAREORNE. 1. B., Recent advances in the ecological chemistry of ACS Symposium Series 296. 206-219. American Chemical
plant terpenoids. in Ecological Chemistry and Biochemistry of Society. Washington, DC, 1986.
Plant Terpenoids (1. B. Harborne and F. A. Tomas-Barbenin, KUBO, I. and K. NAKANISm, fusect antifeedants and repellents from
eds.). Phytochemistry Society of Europe Vol. 31, 399-426, African plants, in Host Plant Resistance to Pests (P. A. Hedin,
Clarendon, Oxford, 1991. ed.). ACS Symposium Series 62,165-178. American Chemical
HASEGAWA, S., M. N. PATEL, and R. C. SNYDER, Reduction of Society, Washington, DC, 1977.
limonin bitterness in navel orange juice serum with bacterial LAvm, D., M. K. JAIN. and S. R. SHPAN-GABRIELITH, A locust
cells immobilized in acrylamide gel. 1. Agric. Food Chern., 30, phagorepeUent from two Melia species, 1 Chern. Soc., Chern.
509-5II (1982). Common. 910-9II (1967).
HASEGAWA, S.• R. D. BENNETT, and V. P. MArnR, Biosynthesis of LEY, S. V., H. LoVELL, and D. 1. WILLIAMS, Chemistry of insect
limonoids in Citrus seedlings, Phytochemistry, 23,1601-1603 antifeedants from Azadirachta indica, Part 14. Absolute con-
(1984). figuration of azadirachtin, 1. Chern. Soc., Chern. Commun..
HASEGAWA, S .. V. P. MAIER, Z. HERMAN, and P. au. Phytohormone 1304-1306 (1992).
bioregulation of nomilin biosynthesis in Citrus limon seedlings, LEY, S. V .• D. SANTAFIANOS, W. M. BLANEY, and M. S. J. SIM-
Phytochemistry, 25,1323-1325 (1986). MONDS. Synthesis of a hydroxy dihydrofuran acetal related to
HEGNAUER, R., Chemical characters and the classification of the azadirachtin: A potent insect antifeedant. Tetrahedron Lett.• 28,
RutaIes, in Chemistry and Chemical Taxonomy of the RutaIes 221-224 (1987).
(P. G. Waterman and M. F. Grundon, eds.). Annual Proceedings MAcLACHLAN. L. K. and D. A. H. TAYLOR, Limonoids from Ny-
Phytochemistry Society of Europe 22). 401-440, Academic mania capensis, Phytochemistry, 21,1701-1703 (1982).
Press, London, 1983. MArnR, V. P., Chemistry and significance of selected Citrus limo-
KtOODI, P. G. K., G. BLASKO, Y. THEBTARANONTH, J. M. PEzzuro, noids and flavonoids, in Chemistry and Chemical Taxonomy
and G. A. CORDELL, Spectroscopic and biological investigation of the Rutales (P. G. Waterman and M. F. Grundon, eds.),
of nimbolide and 28-deoxonimbolide from Azadirachta indica, 319-342. Academic Press, London, 1983.
1. Nat. Prod.• 52, 1246-1251 (1989). MrnOLAJCZAK, K. L. and D. K. REED. Extractives of seeds of the
KLOCKE, A., Natural plant compounds useful in insect control, in Meliaceae: Effects on Spodopterafrugiperda (1. E. Smith),Aca-
Allelochemicals: Role in Agriculture and Forestry (G. R. Wal- lymma vitlatum (F.). and Artemia salina Leach, 1. Chern. Ecol.,
ler, ed.). ACS Symposium Series 330. 396-415. American 13, 99-III (1987).
Chemical Society. Washington. DC, 1987. MrnOLAJCZAK, K. L .• D. WEISLEDER, L. PARKANYI. and 1. CLARDY.
KLOCKE, 1. A.. M. AruSAWA. S. S. HANDA, A. D. KINGHORN, G. A limonoid antifeedant from seed of Carapa procera, 1. Nat.
A. CORDELL, and N. R. FARNSWORTH, Growth inhibitory, insec- Prod.• 51,606-610 (1988).
ticidal and antifeedant effects of some antileukemic and cyto- MrnOLAJCZAK, K. L., B. W. ZILKOWSKI. and R. 1. BARTELT. Effect
toxic quassinoids on two species of agricultural pests, Experien- of meliaceous seed extracts on growth and survival of Spodopt-
tia, 41,379-382 (1985). era frugiperda (J. E. Smith), J. Chern. Ecol., 15, 121-128
KLOCKE,1. A., Limonoids, phenolics and furanocoumarins as insect (1989).
antifeedants. repellents, and growth inhibitory compounds, in MONDON, A. and B. EPE, Bitter principles of Cneoraceae, Fortschr.
Insecticides of Plant Origin (1. T. Amason, B. 1. R. Philogene, Chern. Org. Naturst., 44, 101-188 (1983).
 Limonoids, Quassinoids. and Related Compounds 485

NAKATANI, M., 1. C. JAMES, and K. NAKANISKI,lsolation and struc- racyclic & riterpenoids and their derivatives from Azadirachta
tures of trichilins, antifeedants against the southern anny wonn, indica, J. Nat. Prod., 51,30-43 (1988).
J. Am. Chern. Soc., 103, 1228-1230 (1981). SIMAO, S. M., E. L. BARRElROS, M. F. D. G. F. DA SILVA, and
OELRICHS, P. B., M. W. HILL, P. J. VALLELY, J. K. MAcLEOD, O. R. GOTI'LIEB, Chemogeographic evolution of quassinoids in
and T. F. MOLINSKI, Toxic tetranortriterpenes Melia azedarach, Simaroubaceae, Phytochemistry, 30, 853-865 (1991).
Phytochemistry, 22, 531-534 (1983). SMUTfERER, H., K. R. S. ASCHER, and H. REMBOLD, Natural re-
POWNSKY, J., Chemistry and biological activity of the quassinoids, sources from the neem tree, in Proceedings 1st International
in Chemistry and Chemical Taxonomy of the Rutales (P. G. Neem Conference, 1980.
Watennan and M. F. Grundon, eds.), 247-266, Academic Press, STONE, R.o A biopesticidal tree begins to blossom, Science, 255,
London, 1983. 1070-1071 (1992).
POLONSKY, J., Recent advances in the quassinoid field, in Advances TAYLOR, D. A. H., Biogenesis. distribution, and systematic signifi-
in Medical Phytochemistry (D. Barton and W. D. Ollis, eds.), cance of limonoids in the Meliaceae, Cneoraceae. and allied
125-137, Libbey, London, 1986. taxa, in Chemistry and Chemical Taxonomy of the Rutales (P.
REMBOLD, H., Azadirachtins: Their structure and mode of action, G. Watennan and M. F. Grundon, eds.), 353-375, Academic
in Insecticides of Plant Origin (Amason, J. T., B. J. R. Philo- Press, London, 1983.
gene, and P. Morand, eds.), ACS Symposium Series 387, THORNE, R. F., A phylogenetic classification of the Angiospennae.
150-163, American Chemical Society, Washington, DC, 1989. in Evolutionary Biology, Vol. 9 (M. K. Hecht, W. C. Steere
SAXENA, R. C. o Insecticides from neem, in Insecticides of Plant and B. Wallace, eds.), 35-106, PlenumPress,NewYork,1976.
Origin (J. T. Amason, B. 1. R. Philogene, and P. Morand, eds.),
TOWERS, G. H. N., P. A. SPENCER, and E. RODIUGUEZ, Recent topics
ACS Symposium Series, 387, 1l0-135, American Chemical
in phytochemical ecology research, in Phytochemical Ecology:
Society, Washington, DC, 1989.
Allelochemicals, Mycotoxins, and Insect Pheromones and Allo-
SCHROEDER, D. R. aud K. NAKANISHI, A simplified isolation proce- mones (C. H. Chou and G. R. Waller, eds.), 1-17, Institute of
dure for azadirachtin, J. Nat. Prod., 50, 241-244 (1987). Botany, Academia Sinica Monograph Series 9, Taipei, 1989.
SEIGLER, D. S., Terpenes and plant phylogeny, in Phytochemistry Waterman, P. G. Phylogenetic implications of the distribution of
and Aogiospenn Phylogeny (D. A. Young and D. S. Seigler, secondary metabolites within the Rutales Chemistry and Chemi-
eds.), 117-148, Praeger, New York, 1981. cal Taxonomy of the Rutales; P. G. Watennan & M. F. Grundon,
SIDDIQUI, S., B. S. Siddiqui, S. Faizi, and T. Mainnood, Tet- eds., 377-400 Academic Press, London, 1983.
26
Tetraterpenes or Carotenoids

Introdnction Carotenoids are C40 polyolefinic metabolites derived

Biosynthesis from mevalonic acid. Almost all members of this group are
Fonnation of Geranylgeranyl Pyrophosphate acyclic or have chains that tenninate with one or two 6-
Fonnation of Prephytoene Pyrophosphate membered rings. All have the potential for a vast number of
Fonnation of Phytoenes geometric isomers, but, in practice, almost all have entirely E
Acyclic Carotenoids (trans) configurations; many carotenoids are coloted be-
Alicyclic Carotenoids cause of the extended conjugated systems they contain. The
Oxygenated Carotenoids stroctures for approximately 500 carotenoids are known, but
Site of Synthesis only about one-fourth of these are from plants (Britton,
Chemosystematic Studies 1983). The leaves of all green plants contain the same major
Carotenoids in Algae carotenoids: (3-carotene (usually 25-30% of the total) (1),
Cartenoids in Fungi lutein (about 45%) [(3R,3'R,6'R)-(3,E-carotene-3,3' -diol] (2),
Biological Activity violaxanthin [(3S,5R,6S,3'S,5'R, 6'S)-5,6,5',6'-diepoxy-
Carotenoids in Photosynthesis 5,6,5',6'-tetrahydro-(3,(3-carotene-3,3'-diol] (3), and neoxan-
Photoresponses in Plants thin [(3S,5R,6R,3'S,5'R, 6'S)-5',6'-epoxy-5,6,5',6'-tetra-
Carotenoids and Vision hydro-(3,(3-carotene-3,5,3'-triol] (4) (Young, 1993a, 1993b).
Development in animals Carotenoids also are found in fruits [e.g., Iycopene (5) (Fig.
Pigmentation in Animals 26.5) in tomatoes and capsanthin (6) in red pepper] and flow-
Pigmentation of Flowers and Fruits ers (often (3-carotene or Iycopene). (3-Carotene is the orange
Uses of Carotenoids pigment of carrots (Mann, 1987). Although usually in
Metabolites of Carotenoids smaller percentages, E-carotene [(6'R)-(3,E-carotene] (7) may
Plant Growth-Regulating Compounds occur in amounts up to 40% of the carotene fraction. The
Carotenoids Metabolites as Fungal Pheromones most widespread carotenoids are a- (7) and (3-carotene (1)
References (Fig. 26.1).
The properties and stroctures of tetraterpenes differ
markedly from those of the lower terpenes (Ramage, 1972).
Im'RODVcnOI'l Carotenoids (the major representatives of this class) are
mostly linear polyene systems. The high degree of unsatu-
Carotenoids, which occur in all plants, bacteria, and fungi, ration renders them both heat and light sensitive. These
are probably the most widely distributed of all natural pig- compounds are an experimentally demanding group of
ments; they are involved in many fundamental processes compounds with which to work (Britton, 1983; Ramage,
such as photosynthesis and mammalian vision. These highly 1972).. Methods for purification of carotenoids are similar
unsaturated lipids also serve as vegetative, floral, and fruit to those for other terpenoids, but, because of the highly
pigments in plants and as pigments in the feathers of birds, unsaturated nature of the compounds, special care must
outer parts of insects, as well as the skins of fish and other be taken (Britton, 1991). Care to avoid oxidation is most
animals (Britton, 1976, 1983; Buchecker, 1982; Kayser, important; extracts that contain chlorophyll must be han-
1982). dled with special care. The presence of chlorophyll often
 Tetraterpenes or Carotenoids 487

a-carotene (7)

J3~carotene (l)

HO
fucoxanthin (17)
~"'"""' OH

~4 o
HO capsanthin (6)

HO
""$0
..
C,.., ",

neoxanthin (4) OH
OH

lutein (2) OH

HO
violaxanthin (3)

Fig. 26.1. Common carotenoids.

leads to photoisomerization of carotenoids (Britton, 1991). tion or analysis of carotenoids is a common practice (Brit-
Chromatographic separations should be carried out under ton and Young, 1993).
reduced light. Most carotenoids also are sensitive to acid Ultraviolet and visible spectroscopy have been important
(Britton and Goodwin, 1982, Britton and Young, 1993), for characterization of this group of compounds. Circular
and most plant tissues are sufficiently acidic to isomerize dichroism also is of value for establishing the stereochemis-
labile tetraterpenes. Reversed-phase high-performance liq- try of many tetraterpenoids (Noack, 1982). Mass spectrome-
uid chromatography (HPLC) is a useful method that averts try (Budzikiewicz, 1982) has proven to be a useful technique
structural modification of most carotenoids. Particularly for elucidation of structures of carotenoids. However, nu-
common is the production of the furanoid derivatives (5,8- clear magnetic resonance (NMR) is undoubtedly the most
epoxides) from carotenoids that contain 5,6-epoxide powerful technique for the investigation of carotenoid struc-
groups. General chromatographic data for carotenoids have tures (Britton, 1991; Britton and Young, 1993; Englert,
been reviewed (Britton, 1991; Britton and Goodwin, 1982, 1982). In a high-field IH_ or I3C-NMR spectrum, signals
Britton and Young, 1993, Coscia, 1984; Young, 1993a, due to the olelfinic region of the carotenoid molecule are
1993b). The saponification of total extracts prior to isola- resolved well, can be assigned, and their coupling relation-
488 Tetraterpenes or Carotenoids

ships identified, and the geometric configuration of the ca- geranyl pyrophosphate (GPP), or farnesyl pyrophosphate
rotenoid determined (Britton and Young, 1993). (FPP). C IO or C l5 prenyl derivatives do not appear to be
products, in agreement with former studies on GGPP syn-
thase from Cucurbita (see Chapter 22). Although GPP and
BIOSY1'lTIlESIS FPP are involved as intermediates, GGPP probably is the
first product to leave the active site (Dogbo and Camara,
Tetraterpenes or carotenoids are synthesized from mevalo- 1987) Data from affinity-labeling studies confirm that
nate precursors. Those involved in photosynthesis are syn- DMAPP, GPP, and FPP are all bound at the same site on
thesized in the chloroplast, but the enzymes specific for the enzyme.
carotenoid biosynthesis are encoded in the nucleus, synthe-
sized in cytoplasmic ribosomes, and transported into the Formation of Frephytoene Pyrophosphate
chloroplast (Britton, 1993). Chloroplasts at different stages
in development seem to differ in their ability to synthesize The first step in the formation of prephytoene pyrophos-
carotenoids autonomously from CO2 or by importation of phate (10) and phytoene (8) is the prenyl-transfer step, dur-
isopentenyl pyrophosphate (isopentenyl diphosphate) (Brit- ing which C(I ') of one of the allylic substrates is bonded
ton, 1993). to the C(2)-C(3) double bond of the other to produce a
Isopentenyl pyrophosphate isomerase and geranylgeranyl cyclopropyIcarbinyl pyrophosphate with a Cl'-2-3 structure
pyrophosphate synthase are particularly important in the bio- (Fig. 26.2). In this manner, geranylgeranyl pyrophosphate
synthesis of plastid terpenoids (carotenoids, chlorophylls, (9) yieldS prephytoene pyrophosphate (10) (Poulter, 1990).
plastoquinones, tocopherols, and phylloquinones) (Dogbo In some instances, the ability to produce prephytoene py-
and Camara, 1987). Development of cell-free systems have rophosphate (10) is closely linked to the production of phy-
been important for the study of biosynthesis of carotenoids toene (8) and appears to involve one enzyme with two activi-
(Bramley, 1985). The general features of carotenoid biosyn- ties (Britton, 1993; Dogbo et al., 1988). With an enzyme
thesis are similar in higher plants, algae, fungi, and bacteria. isolated from Capsicum chromoplast preparations, biosyn-
Biosynthesis of the first carotenoid in the pathway, phytoene thesis procedes via condensation of two molecules of gera-
(8), may be divided into two basic steps: (I) formation of nylgeranyl pyrophosphate (9) directly to phytoene (8)
geranylgeranyl pyrophosphate (9) and (2) dimerization of (Dogbo et al., 1988). The enzyme phytoene synthase has
geranylgeranyl pyrophosphate to yield prephytoene pyro- been purified to homogeneity; phytoene synthase from this
phosphate (10) followed by concomitant rearrangement to source is a monomer with a molecular weight of 47,500
phytoene (Dogbo and Camara, 1987; Poulter, 1990). Forma- (Dogbo et al., 1988).
tion of further carotenoid products involves a series of reduc- Although the enzyme from Capsicum has both types of
tion, oxygenation, and ring-closure steps. activity, prephytoene pyrophosphate is an isolable product in
other situations. The conversion of GGPP (9) to prephytoene
Formation of Geranylgeranyl pyrophosphate (10) has been demonstrated with a soluble
Pyrophosphate tomato plastid enzyme system and chloroplasts isolated from
Phaseolus vulgaris by nonaqueous methods (Spurgeon and
The enzymes of carotenoid biosynthesis have been iso- Porter, 1983). The only cofactor that appears to be required
lated from chromoplast stroma after osmotic shock of iso- is Mg2+ or Mn2+. There is no requirement for a reduced
lated chromoplasts. Several steps of purification are required pyridine nucleotide as exists for the formation of squalene.
to separate the desired activities from nonspecific phospha- Prephytoene pyrophosphate (10) appears to be the only isola-
tases and prenyl pyrophosphate hydrolases. The initially iso- ble intermediate involved in the process. This compound has
lated enzyme complex has several other types of activity the same absolute stereochemistry as presqualene pyrophos-
(GPP synthase, FPP synthase, GGPP synthase, GGPp-gera- phate.
nyl transferase, and phytoene synthase), and additional chro-
matographic steps are needed to resolve IPP isomerase and
Formation of Phytoenes
GGPP synthase activity. Isopentenyl pyrophosphate iso-
merase (EC 5.3.3.2) from Capsicum chromoplasts is a mono- In the second step of phytoene formation, inorganic pyro-
meric protein with a molecular weight of 33,500. Geranyl- phosphate is expelled and a proton is eliminated. Pyridine
geranyl pyrophosphate (GGPP) synthase from the same nucleotide cofactors are not involved, as is the case in
source also displays dimethylallyl transferase (2.5 .1.1), gera- triterpene biosynthesis (Fig. 26.3) (Poulter, 1990).
nyl transferase (2.5.1.10), and farnesyl transferase (2.5.1.30) Both 15,15'-Z- (11) and 15,15'-E-phytoene (12) isomers
activity (Britton, 1993; Dogbo and Camara, 1987). This di- appear to be formed in vivo, with concomitant loss of one
meric enzyme has a molecular weight of 74,000. The close I-pro-(S)-hydrogen [originally the 5-pro-(S) of MVA] from
association of these two enzymes may provide a mechanism each molecnle of GGPP (I5,15'-Z-phytoene) or one I-pro-
for channeling IPP toward synthesis of GGPP. GGPP syn- (R)-hydrogen (15,15'-E-phytoene) (Fig. 26.4) (Britton,
thase from this source is capable of using isopentenyl pyro- 1993; Harrison, 1988; Poulter, 1990; Spurgeon and Porter,
phosphate (IPP), dimethylallyl pyrophosphate (DMAPP), 1983).
 Tetraterpenes or Carotenoids 489

H
geranylgeranyl phyrophosphate (9)

PPOCH
H'"

rotation around bond

prephytoene pyrophosphate (10)

Fig. 26.2. Proposed mechanism for the conversion of GGPP to prephytoene pyrophosphate.

NADPH'~

Iycopersene (13)

E-15,15'-phytoene (12)

Z-15,15'-phytoene (11)

Fig. 26.3. Proposed mechanism for the conversion of prephytoene pyrophosphate to lycopersene, Z~phytoene, and E-phytoene (Poulter, 1990; adapted
and used with permission of the copyright owner, the American Chemical Society, Washington, DC).
490 Tetraterpenes or Carotenoids

HO\/ H,s;H
H01~~ R -----110>
: 3 4 5 OH opp

geranylgeranyl pyrophosphate ~
(SR)·[2-'''c,s.3H,Jm.valonic acid
"':= 14C

.;)!Y .
HO,e ,
o
\
3
3H 3H

'" 4 OK

3H
i H

Z-15,15'·phytoe•• (11)

E-15,15'·phyt.... (11)
t

Iycopene (5)

Fig. 26.4. Labeling patterns of band E-15,15'-phytoene and lycopene (Goodwin, 1973 and Williams et aI., 1967b; modified and used with pennission
of the copyright owner, the Biochemical Society, London).

It does not appear that Iycopersene (13), a tetraterpene hydrogens alternatively to the left and right of the central
homologous to squalene, is an intennediate in tetraterpene triene system (Fig. 26.4) (Britton, 1993). The hydrogens re-
biosynthesis (Bramley, 1985). This compound does, how- moved were originally the 5-pro-(R) and 2-pro-(S) hydro-
ever, occur in a number of organisms including carrot roots. gens of MYA (Spurgeon and Porter, 1983). In higher plants,
In most plants that have been studied, 15-Z-phytoene evidence for the involvement of NADP+ and FAD has been
(7,8,1l,12,7',8',1l',12'-octabydro-t!J,t!J-carotene) (11) is the presented (Britton, 1983).
predominant form, although in some bacteria the 15-E form In tomatoes and other higher plants, formation of Iyco-
(12) is most common (Spurgeon and Porter, 1983). pene (5) from Z-phytoene (11) occurs via ,-carotene (16).
When (3R,S,5R)-[2- 14C,5- 3Htlmevalonate was incubated Z-phytoene (11) is converted to Z-phytufluene (14), E-
with slices of tomato, the phytoene isolated shOWed a 3HJ 14C phytofluene (15), ,-carotene (16), neurosporene (17), and,
ratio of 8: 8 (Fig. 26.4). These results indicate that there was fmally, Iycopene (5) by extracts of tomatu fruit plastids
no loss of the 5R hydrogen of mevalonate (based on Fig. (Fig. 26.5) (Porter et al., 1984; Spurgeon and Porter, 1983).
26.4, formation of Z-phytoene occurred). However, in exper- The desaturase (and following cyclase) enzymes are
iments with [2_ 14C,5-3H2 ]mevalonate, the ratio was closer integral membrane proteins that can be solubilized by the
to 14:8, instead of the 16:8 ratio expected, corresponding detergent CHAPS. Enzymatic activity is lost when the
to loss of two hydrogens. Thns, in the formation of Z-phy- enzymes are solubilized, but it is restored when reconstitu-
toene (11), both of the pro-(R) hydrogens of mevalonate are ted in proteoliposomes (Britton, 1993). In the first phase
retained, whereas the two pro-S hydrogens are lost (Fig. 26.3 of desaturation, the conversion of (15Z)-phytoene into
and 26.4) (Spurgeon and Porter, 1983). (15ZH-carotene occurred in the dark, with oxygen as an
essential requirement. The reaction was independent of
Acyclic carotenoids cofacturs such as NADP+ and NADPH. (15ZH-Carotene
could not be desaturated further, but when this compound
Once formed, 15,15'-Z-phytoene (11) is desaturated to was isomerized by light, the isomeric mixture formed
produce Iycopene (5) via stereospecific removal of pairs of underwent further desaturation to produce Iycopene, not
 Tetraterpenes or Carotenoids 491

as the all E fonn, but as the (7Z,9Z,7'Z,9'Z)-isomer, proly- other types of terpenoid closures now are known to occur
copene (14) (Fig. 26.6) (Britton, 1993). Part of the uncer- by ionic mechanisms, it is quite possible that a similar mech-
tainty of products in many studies may arise from the anism may prevail here as well.
inability to distinguish isomers by chromatographic and In higher plants and algae, these desaturation reactions
absorption spectral techniques. and the subsequent cyclization reactions appear to be under
In certain other plants (including other types of toma- direct nuclear control, although the carotenoids of photo-
toes), prolycopene (14) is the main carotenoid produced. synthetic tissues are fonned in the chloroplast (Britton,
The structures for a number of Z compounds related to 1993). In fruits, these changes occur in chromoplasts (Brit-
prolycopene have been resolved recently (Porter et aI., ton, 1983).
1984). The importance of these compounds has not been Several types of evidence indicate that the 13 ring (13-
resolved at present. ionone) does not arise by isomerization of the € ring (Il-
ionone) (Britton, 1993). Use of [2- 14C,2- 3H 2 1mevalonate
Alicyclic Carotenoids (Fig. 26.8) as substrate in carrot root slices indicates that
four hydrogens are lost frum carbon atoms that arise from
Lycopene (5), a major pigment of tomatoes, is the final C-2 of mevalonate in the desaturation of phytoene (8) to
acyclic carotenoid; subsequent cyclization gives Il- and 13- Iycopene (5). As the two types of rings (i.e., 13 and €)
carotenes (7 and 1). Three types of ring closure are shown arise independently, in subsequent steps one additional
(Fig. 26.7) (Spurgeon and Porter, 1983). However, as many tritium should be lost in the fonnation of the € ring and

HO,eP --
H H
O H H

OH
Opp

geranylgeranyl OPP (9)

mevalonic add

lopp

~;:H
~
I prephytoene OPP (10) ~

1
lycopersene(13)

15,15I~E·phytoene (12)
H H
IS,IS'-Z-phytoene (11)

(a) 15,lS'·Z-phytofluene (14)

Fig. 26.5. (continued)

492 Tetraterpenes or Carotenoids

IS,IS'-E-phytoOuene (15)

neurosporene (17)

!
'" "" '"
::,... ::,..
"" '" '"
lycopene (5)

j
"" "" '" ::,..
""
::,..

(b)

Fig. 26.5 (8 & b). Biogenesis of acyclic carotenoids (modified from Britton, 1993. used with pennission of the copyright owner. Chapman & Hall,
London).

none in the formation of the J3 ring. J3-Carotene should The cyclases responsible for formation of cyclic carot-
have a 3H/'4C ratio of 12:8 compared to a value of 16:8 enoids are integral membrane proteins, as are the desatura-
for phytoene. The expected value for a-carotene would tion enzymes described above. The desaturation reactions
be 11: 8. In experimental work, these ratios were closely require oxygen, whereas the cyclase reactions require strictly
approximated (Fig. 26.8) (Spurgeon and Porter, 1983). anaerobic conditions. All E-lycopene did not serve as a sub-
Proposed pathways for the formation of a number of strate despite the fact that all the products were exclusively
alicyclic carotenes are summarized in Fig. 26.9 (Britton, all E-isomers. Cyclization was shown to proceed only with
1993; Spurgeon and Porter, 1983). the (7Z, 9Z, 7'Z, 9'Z)- and (7Z, 7'Z)-isomers. However, in

11 15
:::,.., :::,.., :::,..,
1Z IJ 14 15'

7'

prolycopene (14)
[(7Z,9Z,7'Z,9'Z}-lycopene]

Fig. 26.6. Prolycopene.

 Tetraterpenes or Carotenoids 493

Fig. 26.7. Ring closure of carotenoids and types of 6·membered alicyclic end rings (modified from Herbert, 1981; used with pennission of the
copyright owner, Chapman & Hall, London).

o HsH.
T }
H01~ll4 ~ •
5
OH
HR Hs Hs lHR
T
(4R)~[2~14C,4.lH]~mevalonate

N o HsH
TW.T"",}_T
H
H

}
.
1 •
HOzC 1 3 4 , -+-
I •

TT
• OH
3H 3H H H
H

l2~14C,2-3Hl]-mevalonate
~T
}
_=14c

T T
FIg. 26.8. Labeling patterns for biosynthesis of alicyclic rings (Goodwin and Williams, 1965 and Williams et al., 1967a; modified and used with
permission of the copyright owner, the Biochemical Society, London).
494 Telralerpenes or Carolenoids

15,15 ·Z·phytonuene (14) _ _

;r~~~~~,,~~~~~~~~~~~~~~;r,,~~

r p.carotene (1)

Fig. 26.9. Summary of biogenesis or alicyclic carotenes (Britton, 1993; modified and used with pennission of the copyright owner, Chapman & Hall,
London),

the green alga Scenedesmus, cyclization only occurred after Site of Synthesis
the light-catalyzed isomerization of prolycopene to the all
E-isomer (Britton, 1993). Carotenoids are synthesized in the chloroplasts of plants,
but are under direct nuclear control (Britton, 1993; Young,
Oxygenated Carotenoids 1993a). These pigments also appear to be synthesized in
tissues such as flower petals, roots, and fruits. Extracts of
Oxygenated derivatives of carotenoids also are common tomato plastids have the ability to utilize IPP to produce
and widely distributed. These colored substances are known phyloene as well as other carotenoids.
collectively as xanthophylls. The formation of xanthophylls
involves an aerobic mechanism. (3-Carotene is converted ox-
idatively to lutein (2), zeaxanthin [(3R, 3'R)-(3,(3-carotene-
3,3'-diol] (15), and, finally, to violaxanthin (3) (Fig. 26.10) CHEIIIOSYSTEl'IATIC STUDIES
(Britton, 1993). Labeling studies have shown that hydroxyla-
tions such as those involved in the conversion of zeaxanthin Both carotenoids and xanthophylls have been found in all
and lutein are formed by the direct replacement of the hydro- leaf tissues examined. Cyclic carotenes and xanthophylls
gen by OR (Britton, 1993). from leaf tissues have both (3- and .-ring types. Only dicyclic
Rearrangement of these compounds leads to metabolites xanthophylls, and particularly those with C-3 or C-3', have
such as capsanthin [(3R, 3'S, 5'R)-3,3'-dihydroxy-(3,k-caro- been found in the biosynthetic tissues of higher plants. Acy-
ten-6'-one] (6), a red pigment from peppers (Capsicum an· clic carotenoids normally are not present (Young, 1993a).
nuum, Solanaceae). This compound may be derived from The same four m~or carotenoids, namely (3-carotene (I),
rearrangement of an epoxide intermediate as shown (Fig. lutein (2), violaxanthin (3), and neoxanthin (4), have been
26.10) (Riittimann, 1982). Capsanthin (6), capsorubrin (16), found as the major carotenoid constituents of leaf tissues of
and cryptocapsin are the major pigments of red peppers (Riit- all plants examined to date (Young, 1993a). Purple and green
timann, 1982). phototrophic bacteria rarely contain any of the monocyclic
 Tetraterpenes or Carotenoids 495

and dicyclic carotenoids found in plants, algae, and cyano- (Britton, 1983; Young, 1993a). Many members of the Chlo-
bacteria (Britton, 1993). rophyceae possess the ability to accumulate secondary carot-
enoids in response to stress conditions. The pigment compo-
Carotenoids in Algae sition of the Euglenophyceae more closely resembles that
of the chromophyte algae than those of the Chlorophyta
Although carotenoids have been of limited taxonomic (Young, 1993a). Red algae possess both j3-carotene (1), Ci-
value in higher plants, data concerning the distribution of carotene (7), and oxygenated derivates, such as lutein (2)
these compounds have proven useful in studies of algal and and zeaxanthin (15) (Young, 1993a).
bacterial groups (Britton, 1983; Goodwin, 1971, Young, In some instances, carotenoids are present outside the
1993a). Many algae differ from higher plants in the carot- chloroplast. The pigment of the light-sensitive "eye-spot"
enoids found in their chloroplasts. Acetyienic carotenoids of Euglena and of reproductive areas in certain algae (such as
are restricted to aquatic, mainly marine, animals and algae. Ulva) is comprised partially of carotenoids (Britton, 1983).
Fucoxanthin (17) (Fig. 26.1) (from Fucus) is a widespread
pigment in brown algae (Young, 1993a). j3-Carotene and Carotenoids in Fungi
violaxanthin also are present in high amounts. The chloro-
plast content of green algae (Chlorophyta), however, is quite Carotenoids are found widely, but not universally, aroong
similar to that of higher plants, suggesting a closer evolution- fungi. However, most fungi synthesize a very limited spec-
ary link than between angiosperms and other groups of algae trum of these compounds. The yellow pigment of the edible

OH

f3-carotene (1) lutein (2)

~ OH

HO
(3R,3'R).zeaxanthin (15)

OH

HO
violaxanthin (3)

OH

HO

V. ,
o :::::-...:::-.. ~ ~

(3S,5R,3'S,5'R)·capsorubin (1:
OH

-A-
OH

'" 0'
HO
capsanthin (6)

Fig. 26.10. Biogenesis of oxygenated carotenoids or xanthophylls modified from (Milborrow et al., 1982 and data from Camara and Moneger, 1981.
496 Tetraterpenes or Carotenoids

mushroom Cantharellus cibarius is an unusual carotenoid The light-harvesting complex of photosystem I has been iso-
(Britton, 1983). lated and shown to contain 80-120 molecules of chlorophyll
(Young, 1993b).
The light-harvesting complexes, particularly those associ-
BIOLOGICAL ACTIVITY ated with photosystem 11, are rich in both chlorophylls and
xanthophylls (Young, 1993b). Of the approximately 250
chlorophyll molecules per P-680 in photosystem II, it has
Most carotenoids are colored and absorb light in the visible
been estimated that about 80% are found in the chlorophyll
range of the spectrum. The specific wavelength of absorption
is a function of the structure, in particular, the number and
alb-xanthophyll complexes of the light-harvesting complex
II (Young, 1993b).
arrangement of double bonds and oxygenated substituents,
Carotenes are essential accessory pigments in photosyn-
but also the solvent and other factors (Britton, 1983). Most
thesis, but their exact role is not known. In in vitro experi-
carotenoids are yellow to orange (sometimes red) when iso-
ments, carotenoids do not fluoresce, but they are able to
lated.
sensitize the fluorescence of chlorophyll in green plants
(Britton, 1983).
Carotenoids in Photosynthesis
Carotenoids also protect the cell from damage due to pho-
As mentioned above, the leaves of all green plants contain tooxidation catalyzed by light-absorbing pigments such as
the same major carotenoids: J3-carotene (1), lutein (2), viola- chlorophylls. This effect was noted in comparisons of nor-
xanthin (3), and neoxanthin (4). Smaller amounts of other mal carotenoid-containing bacteria with mutants in which
compounds are often present. These compounds are located the carotenoids are replaced by the more saturated and color-
in the chloroplast grana as chromoproteins (Britton, 1983; less carotenoid, phytoene (8) (Ramage, 1972). Many bleach-
Britton et al., 1982). ing herbicides inbibit carotenoid biosynthesis; the plants
Each photosystem consists of two multiprotein binding usually die because of photooxidation (Britton, 1991).
complexes, namely the core complex, or reaction center, and Three carotenoids, violaxanthin (3), antheraxanthin (18),
the antenna pigments organized into light-harvesting com- and zeaxanthin (15), undergo rapid, light-induced changes
plexes. The pigment content and composition of photosys- in concentration. These interconversions sometimes are
tern 11 is not as well documented as that for photosystem I, called the "xanthophyll cycle" (Fig. 26.11) (Demmig-
largely because there are still difficulties in obtaining clean Adams and Adams, 1993). The content of violaxanthin in
and relatively uncontaminated preparations of the individual leaves can be reversibly altered in plants by exposure to
complexes. Photosystem I from higher plants contains a core light. In the presence of light, violaxanthin is deepoxidized
complex that is enriched in J3-carotene and chlorophyll a. to produce first antheraxanthin and then zeaxanthin. This

"'" "'" "'" "'" ""

violaxanlhln (3) Jr
OH

HO
""
anderaxanthln (18)

Jr
"'" "'" "'" "'" "" "'" "'"
(3R,3'R)·......lhin (15)

Fig. 26.11. The "xanthophyll cycle" (modified from Demmig-Adams and Adams, 1993; used with permission of the copyright owner, Chapman &
Hall. London).
 Tetraterpenes or Carotenoids 497

plant <arotenoids t
stored as palmitate and stearate
t
stored as palmitate and stearate

~~;:,...
U~'''CH'OH b~';:,...
~ ;:,...

I -. I
isomerase 17
r
! l1.z·RII"~
retinol (vitamin A) (19) ll-Z-retinol CRlOR

I"'·'·""no' ........""'" . "......""'..

~~~ all.E·retinai (20) CHO

~t
opsin + nerve impulse

.
protem·(CH,).·NH
.#
rhodopsin (21)

Fig. 26.12. Chemistry of visual pigments.

reaction is catalyzed by violaxanthin deepoxidase. In dark- and higher plants, or phototaxis in algae (Spurgeon and Por-
ness, violaxanthin again accumulates. This reaction is cata- ter, 1980).
lyzed by zeaxanthin epoxidase. The enzymes responsible are
found in the thylakoid membranes of the chloroplast. The Carotenoids and Vision
degree to which violaxanthin is converted into zeaxanthin Animals require vitamin A (retinol) (19) or a carotenoid
varies with the light level or photon flux density (Demmig- precursor for normal growth and vision (Liu, 1982). An early
Adams and Adams, 1993). However, zeaxanthin can ouly symptom of vitamin deficiency is a decreased ability to see
increase in excessive light until the violaxanthin pool is ex- in dim light. Vitamin A aldehyde, or lI-Z-retinal (20), has
hausted. The maximal degree of conversion of violaxanthin an essential role in vision (Fig. 26.12); this pigment is the
to zeaxanthin typically is reached at photon flux densities chromophore of the visual pigment rhodopsin (Buck et al.,
exceeding those required for normal growth of the plant. In 1991). lI-Z-Retinal is formed in the intestinal mucosa by
full sun at noon, violaxanthin (3) represented ouly about oxidative cleavage of (3-carotene, a process catalyzed by the
5-12% of the total xanthophyll cycle pool. Shade leaves enzyme (3-carotene dioxygenase (Britton, 1983). The lI-Z
tend to form and accumulate zeaxanthin (15) at photon flux isomer of retinal combines with a protein, opsin, via a Schiff-
densities much below those that would lead to formation of base linkage to form rhodopsin (21). Rhodopsin is located
zeaxanthin in a sun leaf. The deepoxidation reaction, in ef- in the rods and cones of the retina, and when exposed to
fect, leads to thermal dissipation of excess solar energy. The light, geometric isomerism occurs, producing all E-retinal
excess is removed in a safe and controllable fashion that with concomitant release of the protein moiety. This confor-
protects the photosynthetic apparatus against the adverse ef- mational change is somehow translated into a nerve impulse
fects of excessive light (Demmig-Adams and Adams, 1993). and transmitted to the brain. lI-Z-Retinal is regenerated in
The "xanthophyll cycle" has been found in higher plants, a further sequence of reactions and the net result is vision
ferns, mosses, and both green and brown algae. (Britton, 1983; Buck et al., 1991; Liu, 1982).

Photoresponses in Plants Development in Animals

Carotenoids also may be concerned with other functions, Carotenoids appear to playa fundamental role in develop-
for example, photoresponses such as phototropism in fungi ment in animals. For example, all E-retinoic acid (22) ap-
498 Tetraterpenes or Carotenoids

14-hydroxy-4,1l-retro-retinol (24) E-retinoic acid (22)

retinol (vitamin A) (23)

o

o
canthaxanthin (25)

Fig. 26.13. Biologically active carotenoids and carotenoid derivatives.

pears to be a natural signaling substance involved in limb imus (Fig. 26.14) (Goodwin, 1983). Complexes of this type
pattern formation in chicks (Thaller and Eichele, 1987)_ Reti- also are partly responsible for the colors of bird feathers and
nol (23) is essential for the growth of a variety of cell types in other animal pigments. These complexes are readily dena-
culture. A retinol derivative, 14-hydroxy-4,14-retro-retinol tured by heat. Many of the carotenoids of invertebrates ap-
(24), has growth-promoting properties in animal cells (Fig. pear to be rare.
26.13) (Buck et aI., 1991).
Pigmentation of flowers and Fruits
Pigmentation in Animals
The background pigmentation of the aboveground parts of
The colors of the yolks of eggs of birds, many bird feath- most plants is green. In the course of evolution, selection also
ers, the skin of fishes, and the pigments of many animals has occurred for colors other than green in more specialized
other than mammals may be directly attributed to the pres- plant parts, especially those involved in reproduction. At the
ence of carotenoids (Buchecker, 1982; Castillo, 1982; same time, selection for animal visual systems capable of vis-
Kayser, 1982; Yamaguchi, 1982). The pink color offlamin- ualizing these flowers, fruits, pollen, and other plant parts
gos is produced by ketocarotenoids, mainly canthaxanthin with modified colors has occurred. In general, the vision of
(25) (Britton, 1983). birds and mammals is especially sensitive to reds, yellows,
Many of the colors used for aposematic labeling by ani- and blues. The vision of many insects differs and, although
mals (bright reds, oranges, and yellows) are produced by insects often are sensitive to colors visible to other animals,
carotenoid pigments (Rothschild, 1975, 1978). These com- these organisms are also sensitive to portions of the ultraviolet
pounds ultimately arise from plants (or from symbiotic bac- spectrum (Britton, 1983). These pigments serve as syno-
teria?), as animals cannot synthesize carotenoids. mones or kairomones that are involved in attraction of the ani-
Complexes of carotenoids with proteins in animals often mals to plants. These coloration patterns also may serve as
are responsible for pigmentation, especially in instances warning signals for would-be hervivores or predators.
where colors other than yellow or orange are involved. This The major pigments of plants often consist of anthocya-
is especially true of purple, green, blue, and black pigments nins, other flavonoids (see Chapter 11), chlorophyll, and
(Britton, 1983; Britton et aI., 1982). Although the precise a complement of carotenoids. It is not unusual to observe
manner in which these interactions occurs is not understood, instances where all three types of pigments are implicated,
absorption maxima of the complexes are different from those even in the same plant or plant structure.
of the carotenoids themselves. The blue color of lobsters, The yellow coloration of flowers may be produced by a
for example, is composed of crustacyanin and a protein. Pec- number of substances, but carotenoids frequently are impor-
tonoxanthin (alloxanthin) (26) and a similar protein complex tant among them (Fig. 26.14). Almost all yellow and lemon-
is responsible for the color of the giant scallop, Pecten max- yellow flowers contain xanthophylls, such as zeaxanthin (15)
 Tetraterpenes or Carotenoids 499

and its 5,8-epoxides auroxanthin and f1avoxanthin (27), as natto, has been used as a food colorant and flavoring in Latin
major pigments. Deep-orange flowers may contain large America for generations, a practice now common in most
amounts of J3-carotene (1) or Iycopene (5). The carotenoids tropical areas of the world. The seeds of this plant contain
in petals are concentrated in the chromoplasts and may be bixin (31), the methyl ester of 6,6'·diapocarotenoic acid. In
present in bound form linked to protein or esterified with fatry the Iberian peninsula, the use of Crocus sativus (saffron,
acids (Britton, 1983, 1991; Harborne, 1982). These pigments, Iridaceae) as a food coloring material is widespread. Some
accompanying pigments of other chemical classes (especially of the yellow pigments of this plant, such as crocein (29),
f1avonoids), and the associated morphology of flowers are all also are carotenoids (Rothschild, 1975). Many carotenoids
important in the effective reproduction of plants. (e.g., J3-carotene) are antioxidants. This property has led to
The immature fruits of most plants are green. These green the suggestion that they are important in the diet and can
fruits usually contain the same pigments as leaves of the afford protection against some forms of cancer and other
plant on which they occur. As the fruits mature, the numher diseases (Britton, 1991).
of chloroplasts and the amount of pigmentation increases.
Chloroplasts are converted to chromoplasts and the chloro-
phylls gradually disappear. The major pigments of tomatoes
are lycopene (5) and J3-carotene (1); that of watermelon is JllETABOLITES OF CAKOTEI'IOmS
phytoene (8). Carotenoids constitute the most abundant pig-
ments of Pyracantha (Rosaceae), Sambucus, Citrullus vul- Many compounds with fewer than 40 carbon atoms, but with
garis, Citrus, Taxus, Rosa, Cotoneaster, Capsicum, and carotenoid·like structures, are found in nature. Synthesis of
many other fruits. these compounds, sometimes called apocarotenoids, appears
to occur mainly by catabolism of carotenoids (Parry and
Horgan, 1991). Apocarotenoids in the range of C,-C[3 are
USES OF CAKOTEI'IOmS
found in plant essential oils, but others, ranging up to C30
Carotenoids, especially J3-carotene (I), often have heen used compounds, are essentially nonvolatile. However, knowl-
for coloring food products. This is especially common in edge of the biochemistry of carotenoid metabolism is limited
such fatry foods as margarine. Bixa orellana, achiote or an- (parry and Horgan, 1991). In vitro, photuoxidation of carot-

~·carotene (1)

croeein (29)
CO,.gI....yl.()..gl.c..yl
gI.cosyl.O·gI.cosyl·O,C

Iycopen. (5)

DRvoxanthin (27)
HO

pectenoxanthin (26)
(alloxanthin)
HO
Fig. 26.14. Carotenoid pigments.
500 Tetraterpenes or Carotenoids

~'"~
trixagol (40) jhanic acid (41)

CPu (42)
o
~~
,
'::;0

(43)
o -- HO
~

OH

3-hydroxy-5,6-epoxy-5,6-dihydro-J}-ionol (30)

3-oxoactinidol (31) p-damascenone (33) 1-(2,3,6-trimethylphenyl)-but-2-ene-l-one (32)

cI
megastigma-5,8(E)-dien-4-one (34) megastigma-5.7(E)-9-trien-4-one (35) (36)

~
"".,..
H
~ '.
o ""'"'
(37) (38) (39)

Fig. 26.15. Metabolites of carotenoids.

enoids produces a range of C9 -C 15 products similar to those gal (40) from Bellardia trixago (Scrophulariaceae) and
found in plants, but it is not known how the process occurs jhanic acid (41) from Eupatorium jhanii, have been deter-
in vivo. mined. The defensive secretions of two species of termites
Some carotenoid metabolites apparently arise during the contain (42) and (43).
processing of plant material [e.g., (3S, SR, 6S)-3-hydroxy-
S,6-epoxy-S,6-dihydro-j3-ionol (30), 3-oxoactinidiol (31),
and 1-(2,3,6-trimethylphenyl)-but-2-ene-I-one (32)] are PLAl'IT GROWTH RBGULATING COMPOUNDS
formed during curing of tobacco (Fig. 26.IS). These com-
pounds are related to j3-darnascenone (33) and the aromatic (+ )-(S)-Abscisic acid (44) is involved in a number of hor-
carotenoids. monal roles in plants (Fig. 26.16) (Milborrow, 1983). This
Megastigma-S,8(E)-dien-4-one (34) has been obtained plant growth-regulating compound, probably found in all
from Virginia tobacco, from passion fruit, and, along with higher plants, acts as a growth inhibitor and is involved in
megastigma-S,7(E),9-trien-4-one (35), from Osmanthus ab- bud and seed dormancy. (+ )-(S)-Abscisic acid is a potent
solute (an oil used in perfumery). The passion fruit, Pas- effector of stomatal closure. Abscisic acid also is linked to
siflora edulis, has also afforded two other related compounds water stress resistance in plants. To date, the presence of
(36) and (37) and two further edulan derivatives (38) and abscisic acid has not been confirmed in bryophytes (Milbor-
(39). row and Netting, 1991).
The structures of two carotenoid-like diterpenoids, trixa- Although abscisic acid is synthesized as a sesquiterpene
 Tetraterpenes or Carotenoids 501

in several fungi (e.g., Cereospora rosicola), this apparent such as phaseic acid (48), xanthoxin (47), and E-farnesol,
sesquiterpene is derived by modification of carotenoids in are capable of initiating stomatal closure in plants, a process
higher plants (Willows and Milborrow, 1989). The syn- involved in drought resistance (Fig. 26.16) (Harborne, 1982;
thetic compound fluridone inhibits carotenoid metabolism Milborrow and Netting, 1991; Varner and Ho, 1976).
at the stage of phytoene dehydrogenase (Parry and Horgan,
1991); treatment of maize seedlings with this inhibitor carotenoids l'IetaboliUes as Fungal
strongly inhibits formation of abscisic acid (Moore and Pheromones
Smith, 1984). The stereochemistry of abscisic acid pro-
duced by either process is identical (Willows and Milbor- Compounds such as trisporic acids serve as pheromonal
row, 1989). substances in the reproduction of fungi of the order Mucor-
The most likely precursors of abscisic acid (44) are 9'- ales (Zygomycetes) (Fig. 26.17). The principal genera in-
Z-violaxanthin (45) and 9-Z-neoxanthin (46). E-Violaxan- volved are Mucor, Phyeomyees, and Blakeslea (Mascaren-
thin (3) is converted to 9'-Z-neoxanthin in fluridone-treated has, 1978). The same pheromones apparently initiate sexual
etiolated plants of Lyeopersieon and Phaseolus seedlings development in all mucoraceous fungi; furthermore, this pro-
following exposure to light (Parry and Horgan, 1991). Cleav- cess appears to be more complex than originally envisioned
age of these xanthophylls across the 11,12- (11',12'-) double (Jaenicke, 1991; Miller and Sutter, 1984; Sutter and Whi-
bond may produce xanthoxin, which is readily converted to taker, 1981).
abscisic acid by plant tissues. Xanthoxin (47) serves as a The mating type of a heterothallic strain of these fungi is
source of abscisic acid in tomato and wheat. The epoxide not apparent when the mycelium is growing vegetatively or
oxygen atom becomes the tertiary hydroxyl oxygen atom of during asexual reproduction. However, when adjacent colo-
abscisic acid. nies of opposite mating types grow together and nutrient lev-
Abscisic acid and similar sesquiterpenoid compounds, els are adjusted to modify purely vegetative growth, zygo-

~,
CO,HO

(+)·(S)-absclsie acid (44)

MM o
"'"
OH

phasele acid (48)

~
CO,HHO

xanthonin (47)
0"'" CHO

9'·Z-violaxanthin (46)

OH

~C~
~OH
9'·Z_neoxanthin (45)

/~
HO

X~~CHO ~O'H
OH
"'" ~- "'"
OH ~ -. "'" OH ~
HoU~l 0
Q CHO
0

xanthonin (50) abscisic acid (44)

Fig. 26.16. Probable biosynthesis of abscisic acid (modified from Parry and Horgan. 1991; used with pennission of the copyright owner, Elsevier
Science Ltd.. The Boulevard, Langard Lane. Kidlington OXS 1GB. UK).
502 Tetraterpenes or Carotenoids

phores are produced. These are well developed in Mucor, the ture of trisporate pheromones: the methyl esters of trisporic
organism usually studied. Zygophores are formed even when acid B (49), trisporic acid C (50), and trisporic acid E, as
the two mating types are of different species. The zygophores well as the corresponding 4-dihydro-derivatives of these
are recognizable because they show an increased level of ca- compounds (such as 51) (Sutter, 1986; Sutter and Whitaker,
rotenoid pigmentation. After contact occurs, septa form, and 1981). These mating-type-specific pheromones induced the
the nuclei from the ( + ) and ( - ) type undergo sexual combi- formation of zygophores in (- )-cultures of both Phyco-
nation (Mascarenhas, 1978). In addition to the accumulation myces blakesleeanus and Mucor mucedo. (- )-Cultures of
of j3-carotene, mixed cultures also produce trisporic acids Phycomyces blakesleeanus are at least 100-fold more effec-
(Fig. 26.18) that mediate sexual reproduction in the Mucor- tive than ( + )-cultures in converting the methyl esters into
ales. The necessary complement of trisporic acids is not pro- trisporic acid C (Miller and Sutter, 1984; Sutter, 1986; Sutter
duced by single strains of either mating type or by other heter- and Whitaker, 1981). However, trisporic acids are inactive in
othallic species. Some apparently cooperative mechanism is (- )-cultures. Trisporin (52) [with the pro-(S)-methyl intact]
involved (Bu'Lock, 1983; Mascarenhas, 1978). and trisporol (53) [with the pro-(S)-methyl converted to a
Trisporic acids are formed from j3-carotene (1) via retinal carbinol group] are ( - )-pheromones [i.e., produced by ( - )-
(20) with further loss of two carbon atoms. As these com- cultures] (Jaenicke, 1991; Sutter and Whitaker, 1981). Both
pounds induce additional carotenoid biosynthesis, biosyn- ( + )- and ( - )-cultures oxidize the 4-hydroxy to a carbonyl
thesis of these compounds is self-amplifying. Yields of group, apparently by the same enzyme. This reaction may
trisporic acids in mixed cultures (+ and -) are typically be important in (+ )-cultures to avoid self-stimulation by
200-1000 ",g/m!. compounds capable of inducing formation of zygospores
(+ )-Cultures of Phycomyces blakesleeanus make a mix- (Jaenicke, 1991).

~H/ CO,H
.0
:? ......" OH

trisporic acid B (49) trisporic acid C (SO)

o
o

trisporol B trisporin C

~'
trisporol C trisporin B
o H OH
H

OH OH

methyl trisporate C methyl4·dihydrotrisporate C (51)

Fig. 26.17. Trisporic acid and related metabolites.

 Tetraterpenes or Carotenoids 503

o
p·carotene (1) --+

4-dihydrotrisporin
~O
OR /

~"
OR
4·dlhydrotrisporol !
(+) OR
trisporol (53)

~" __------------~~~~O
metbyl4-dihydrotrisporate

~Co~O~--------+-----~
I'·'
o

o
trisporic acid
Fig. 26.18. Reactions in the fonnation of trisporic acids (Sutter and Whitaker, 1981; modified and used with pennission of the authors).

REFERENCES BRITTON, G., G. M. ARMrrr, S. Y. M. LAu, A. K. PATEL, and

C. C. SHONE, Carotenoproteins, in Carotenoid Chemistry and
BRAMLEY. P. M., The in vitro biosynthesis of carotenoids, Adv. Biochemistry (G. Britton and T. W. Goodwin, eds.), 237-251,
Lipid Res., 21, 243-279 (1985). Pergamon, Oxford, 1982.
BRI1TON. G., Biosynthesis of carotenoids, in Chemistry and Bio- BUCHECKER. R. t A chemist's view of animal carotenoids, in Carot-
chemistry of Plant Pigments, 2nd ed., Vol I (T. W. Goodwin, enoid Chemistry and Biochemistry (G. Britton and T. W. Good·
ed.), 262-327, Academic Press, London, 1976. win, eds.), 175-194, Pergamon, Oxford, 1982.
BRfITON, G., The Biochemistry of Natural Pigments, Cambridge BUCK, J., F. DERGUINI, E. LEVI, K. NAKANISHI, and U. HAAMMER-
University Press, Cambridge, 1983. UNO, Intracellular signaling by 14·hydroxy-4,I4-retro·retinol,
BRITTON, G., Carotenoids, in Terpenoids (B. V. Charlwood and D. Science, 254, 1654-1656 (1991).
B. Banthorpe, eds.), Vol. 7 of Methods in Plant Biochemistry (J. BUDZIKIEWIcz, H., Mass spectra of carotenoids-labeling studies, in
B. Harbome and D. M. Dey, eds.), 473-518, Academic Press, Carotenoid Chemistry and Biochemistry (G. Britton and T. W.
London, 1991. Goodwin, eds.), 155-165, Pergamon, Oxford, 1982.
BRmoN, G., Biosynthesis of carotenoids, in Carotenoids in Photo- Bu'LoCK, 1. D., Trisporic acids, in Biosynthesis of Isoprenoid
synthesis (A. Young and G. Britton, eds.), 96-125, Chapman & Compounds, Vol. 2 (1. W. Porter and S. L. Spurgeon, eds.),
Hall, London, 1993. 437-462, Wiley, New York, 1983.
BRfITON, G. and T. W. GOODWIN (eds.), Carotenoid Chemistry and CAMARA, B. and R. MONEGER, Carotenoid biosynthesis. In vitro
Biochemistry, Pergamon, Oxford, 1982. conversion of antheraxanthin to capsanthin by a chromoplast
BRITTON, G. and A. 1. YOUNG, Methods for the isolation and analy· enriched fraction of Capsicum fruits, Biochem. Biophys. Res.
sis of carotenoids, in Carotenoids in Photosynthesis (A. Young Commun., 99, 1117-1122 (1981).
and G. Britton, eds.), 409-457, Chapman & Hall, London, 1993. CASTILLO, R., General survey of the carotenoids in Crustacea, in
504 Tetraterpenes or Carotenoids

Carotenoid Chemistry and Biochemistry (G. Britton and T. W. chemistry of hydroxylation of the carotenoid lutein in Calen-
Goodwin, eds.), 211-224, Pergamon, Oxford, 1982. dula ojficinalis, Phytochemistry, 21, 2853-2857 (1982).
COSCIA, C. J., Terpenoids, Vol. 1 of Handbook of Chromatogra- MILLER, M. L. and R. P. SlITTER, Methyl trisporate E, a sex
phy (G. Zweig and J. Sherma, eds.), CRC Press, Boca Raton, pheromone in Phycomyces blakesleeanus, 1. BioI. Chern., 259,
FL, 1984. 6420-6422 (1984).
DEMMIG-ADAMS, B. and W. W. ADAMS ill, The xanthophyll MOORE, R. and 1. D. SMITH, Growth, graviresponsiveness and
cycle, in Carotenoids in Photosynthesis (A. Young and G. abscisic-acid content of Zea mays seedlings with Fluridone,
Britton, eds.), 206-251, Chapman & Hall, London, 1993. Planta, 162, 342-344 (1984).
DOGBO, O. and B. CAMARA, Purification of isopentenyl pyrophos- NOACK, K., Circular dichroism of carotenoids and its use in
phate isomerase and geranylgeranyl pyrophosphate synthase investigations of their structures, configurations, and confor-
from Capsicum chromoplasts by affinity chromatography, Bio- mations, in Carotenoid Chemistry and Biochemistry (G. Brit-
chim. Biophys. Acta, 920, 140-148 (1987). ton and T. W. Goodwin, eds.), 135-153, Pergamon, Oxford,
DOOBO, 0., A. LAFilRRIERE, A. D'iIARuNGUE, and B. CAMARA, 1982.
Carotenoid biosynthesis: Isolation and characterization of a PARRY, A. D. and R. HORGAN, Carotenoid metabolism and the
bifunctional enzyme catalyzing the synthesis of phytoene, biosynthesis of abscisic acid, Phytochemistry, 30, 815-821
Proc. Nat!. Acad. Sci. USA, 85, 7054-7058 (1988). (1991).
ENGLERT, G., N.M.R. of Carotenoids, in Carotenoid Chemistry PORTER, J. W., S. L. SPURGEON, and N. SATlIYAMOORTlIY, Biosyn-
and Biochemistry (G. Britton and T. W. Goodwin, eds.), thesis of carotenoids, in Isopentenoids in Plants (W. D. Nes,
107-134, Pergamon, Oxford, 1982. G. Fuller, and L. Tsai, eds.), 161-183, Marcel Dekker, Inc.,
GOODWIN, T. W., Algal carotenoids, in Aspects of Terpene Chem- New York, 1984.
istry and Biochemistry (T. W. Goodwin, ed.), 315-356, Aca- POULTER, C. D., Biosynthesis of non-head-to-tail terpenes. Fonna-
demic Press, London, 1971. tion of 1'-1 and 1'-3 linkages, Acc. Chern. Res., 23, 70-77
GooDWIN, T. W., Prochira1ity in biochemistry, Essays Biochem., (1990).
9, 103-160 (1973). RAMAGE, R., Carotenoid chemistry, in Chemistry of Terpenes
GoODWIN, T. W., The Biochemistry of the Carotenoids, Vol. 2 and Terpenoids (A. A. Newman, ed.), 288-336, Academic
Animals, 2nd ed., Chapman & Hall, London, 1983. Press, London, 1972).
GOODWIN, T. W. and R. J. H. Wn.LIAMS, A mechanism for the ROTHSCHILD, M., Remarks on carotenoids in the evolution of
cyclization of an acyclic precursor to fonn ~-carotene, Bio- signals, in Coevolution of Animals and Plants (L. E. Gilbert
chern. J., 94, 5c-6c (1965). and P. H. Raven, eds.), 20-51, University of Texas Press,
HARBORNE, J. B., Introduction to Ecological Biochemistry, 2nd Austin, 1975.
ed., Academic Press, New York, 1982. ROTHSCHILD, M., Carotenoids in the evolution of signals. Experi-
HARRISON, D. M., The biosynthesis of triterpenoids, steroids, and ments with insects (1974-1976), in Biochemical Aspects of
carotenoids, Nat. Prod. Rep., 5, 387-415 (1988). Plant and Animal Coevolution (J. B. Harborne, ed.), 259-276,
HERBERT, R. B., The Biosynthesis of Secondary Metabolites, Academic Press, New York, 1978.
Chapman & Hall, London, 1981. RfrmMANN, A., Synthesis and stereochemistry of red pepper
JAENICKE, L., VI. Development: Signals in the development of carotenoids, in Carotenoid Chemistry and Biochemistry (G.
Cryptogams, Prog. Bot., 52, 138-189 (1991). Britton and T. W. Goodwin, eds.), 71-86, Pergamon, Oxford,
KAYSER, H., Carotenoids in insects, in Carotenoid Chemistry and 1982.
Biochemistry (G. Britton and T. W. Goodwin, eds.), 195-210, SPURGEON, S. L. and J. W. PORTER, Carotenoids, in Lipids:
Pergamon, Oxford, 1982. Structure and Function (p. K. Stumpf, ed.), Vol. 4 of The
LID, R. S. H., Synthetic and structural studies of visual pigments, Biochemistry of Plants (P. K. Stumpf and E. E. Conn, eds.),
in Carotenoid Chemistry and Biochemistry (G. Britton and 420-484, Academic Press, New York, 1980.
T. W. Goodwin, ods.), 253-264, Pergamon, Oxford, 1982. SPURGBON, S. L. and J. W. PORTER, Biosynthesis of carotenoids,
MANN, J., Secondary Metabolism, Oxford University Press, Ox- in Biosynthesis of Isoprenoid Compounds, Vol. 2 (J. W.
ford, 1978; 2nd edition, 1987. Porter and S. L. Spurgeon, eds.), 1-122, Wiley, New York,
MASCARENHAS, J. P., Sexual chemotaxis and chemotropism in 1983.
plants, in Taxis and Behaviour (G. L. Hazelbauer, ed.), SUTIER, R. P., Apotrisporin-E: A new sesquiterpenoid isolated
169-203, Chapman & Hall, London, 1978. from Phycomyces blakesleeanus and Blakeslea trispora, Exp.
Mn..BoRRow, B. V., Biosynthesis of abscisic acid and related Mycol., 10, 256-258 (1986).
compounds, in Biosynthesis of Isoprenoid Compounds, Vol. SlITTER, R. P. and J. P. WHITTAKER, Zygophore-stimulating pre-
2 (J. W. Porter and S. L. Spurgeon, eds.), 279-295, Wiley, cursors (pheromones) of trisporic acids active in ( - )-Phyco-
New York, 1983. myces blakesleeanus, J. BioI. Chern., 256, 2334-2340 (1981).
MILBORROW, B. V. and A. G. NETI1NG, Abscisic acid and deriva- THALLER, C. and G. EICHELB, Identification and spatial distribu-
tives, in Terpenoids (B. V. Charlwood, and D. B. Banthorpe, tion of retinoids in the developing chick limb bud, Nature,
eds.), Vol. 7 of Methods in Plant Biochemistry (1. B. Hatborne 327, 625-628 (1987).
and D. M. Dey, eds.), 213-261, Academic Press, London, VARNER, J. E. and D. T. Ho, Hormones, in Plant Biochemistry,
1991. 3rd ed. (1. Bonner and J. E. Varner, eds.), 714-770, Academic
MILBORROW, B. V., I. E. SWlFf, and A. G. NETTING, The stereo- Press, New York, 1976.
 Tetraterpenes or Carotenoids 505

WILLIAMS, R. J. H., G. BRITTON, and T. W. GOODWIN, The YAMAGUCHI, M., Carotenoids in sponges, in Carotenoid Chemis-
biosynthesis of cyclic carotenes, Biochem. J., 105, 99-105 try and Biochemistry (G. Britton and T. W. Goodwin, eds.),
(1967a). 225-235, Pergamon, Oxford, 1982.
Wn..LIAMS, R. J. H., G. BRITION, 1. M. CHARLTON, and T. W. YOUNG, A. J. Occurrence and distribution of carotenoids in
GOODWIN, The stereospecific biosynthesis of phytoene and photosynthetic systems, in Carotenoids in Photosynthesis (A.
polynnsaturated carotenes, Biochem. J., 104, 767-777 Young and G. Britton, eds.), 16-71. Chapman & Hall, Lon-
(1967b). don, 1993a.
Wn..LOWS, R. D. and B. V. Mn..BORROW, The stereochemistry of YOUNG, A. 1. Carotenoids in pigment-protein complexes, in
the hydrogen atoms at C-5' of abscisic acid, Phytochemistry. Carotenoids in Photosynthesis (A. Young and G. Britton,
28, 2641-2642 (1989). eds.), 72-95, Chapman & Hall, London, 1993b.
27
Introduction to Alkaloids

Introduction dine and about 3600 kg of ajmalacine are produced annually.

What Is an Alkaloid? Many of these alkaloids come from trees that require several
Isolation of Alkaloids years for growth and production of alkaloids; some are ex-
Biosynthetic Origin of Alkaloids tremely expensive. For example, vincristine from Catharan-
Some General Reactions Involved in the Formation of thus roseus has a value of about $22,000 per gram (Ver-
Alkaloids poorte et aI., 1991). To date, however, attempts to produce
Distribution of Alkaloids alkaloids by biotechnological methods usually have not pro-
Biological Activity vided sufficient quantities of alkaloids to be economically
Biological Activity of Alkaloids in Plants useful. Berberine is the alkaloid closest to industrial-scale
Biological Activity of Alkaloids in Animals production, but uses for the pure compound are limited. The
Alkaloids as Allomones and Kairomones prospects for sanguinarine perhaps are better. Nonetheless,
Uses of Alkaloids by Man there is a need for studies leading to insight into the regula-
References tion of secondary metabolism in order to produce levels of
alkaloids necessary for industrial processes. On the other
hand, cell cultures provide an excellent system for studying
Il'ITKODVCTION biosynthesis (Verpoorte et aI., 1991).
The nuclear magnetic resonance (NMR) spectroscopy of
Although plants containing alkaloids have been used by man most major groups of alkaloids and the determination of
for at least 3000 years as medicines, teas, and potions (Cor- alkaloid structores by spectral methods such as NMR and
dell, 1978), the compounds responsible for activity were not mass spectroscopy (MS) have been reviewed (Crabb, 1982;
isolated and characterized until the 19th century. Many Highet and Wheeler, 1985). Many chromatographic tech-
proved to be challenging chemical problems and the struc- niques for alkaloids have been surveyed (Baerheim Svend-
tures were resolved only in recent years. At present, there sen and Verpoorte, 1983; Hashimoto et al., 1988; Verpoorte
are at least 10,000 characterized alkaloids (Dalton, 1991; and Baerheim Svendsen, 1984).
Wink, 1993). The exact number is difficult to estimate, be-
cause many alkaloids listed in compendia are not naturally Wbat Is an Alkaloid?
occurring. Based on the NAPRALERT Database, almost
16,000 alkaloids have been reported from plant, animal, and Probably the ouly single character that distinguishes all
marine sources (Verpoorte et aI., 1991). About 20-30% of alkaloids is that they have nitrogen. In general, this nitrogen
higher plants accumulate alkaloids. Perhaps 60-70% of spe- comes from amino acids, is incorporated into a heterocyclic
cies of the Solanaceae and Apocynaceae contain alkaloids ring, and is basic. Pelletier (1983) defined an alkaloid as a
(Hartmann, 1991; Verpoorte et al., 1991; Wink, 1993). cyclic compound containing nitrogen in a negative oxidation
Because of their importance, many attempts have been state, which is of limited distribution among living organ-
made to produce alkaloids in plant tissue culture. About 30 isms. Some authors accept a series of more narrowly defined
alkaloids account for most commercial interest, primarily as groups, based on biosynthetic origin (Hegnauer, 1988). Al-
medicines, flavorings, or poisons (Verpoorte et al., 1991). kaloids almost always have physiological activity in animals,
Approximately 300-500 metric tons of quinine and quini- although some have limited effects (e.g., betalain pigments).

506
 Introduction to Alkaloids 507

Alkaloids are derived from many biosynthetic pathways, in- dicting the general direction for approach to these nitrogen-
cluding those of amino acid, polyketide, shikimic acid, ace- ous secondary metabolites.
tate, and terpenoid metabolism. In this book, a broad per- Most alkaloids are derived biosynthetically from orni-
spective is adopted; simple amines, nitrogenous bases of thine, lysine, nicotinic acid, anthranilic acid, phenylalanine,
terpenoid origin, neutral compounds such as colchicine, and tyrosine, and tryptophan (Fig. 27.1) (Leete, 1983). In some
purines are all considered to be alkaloids. Many alkaloids are groups, however, the source of the nitrogen is not from an
poisonous to mammals and a large number are medicinally amino acid.
useful. Feeding studies of labeled precursors for the study of
Many of the topics briefly mentioned in this introductory alkaloids have been summarized and reviewed (Leete,
chapter will be discussed in more detail in subsequent chap- 1983).
ters.
Some General Reactions Involved in tbe
Isolation of Alkaloids Formation of Alkaloids
The basic properties of alkaloids simplify isolation of Certain chemical reactions are common to the formation
these compounds as a group, although separation of individ- of many types of alkaloids (Geissman and Crout, 1969).
ual compounds from mixtures of alkaloids may be difficult Among these are decarboxylation, Schiff-base formation,
(Hartmann, 1991; Waterman, 1993). Alkaloids generally are Mannich-base condensation, and imino-aldol-type conden-
extracted with nonpolar solvents such as chloroform, which sations.
is, in turn, extracted with an acidic solvent. This acidic liquid Schiff-base formation may occur between any amine and
is then made basic and reextracted with a nonpolar solvent carbonyl compound to yield an imine system (Fig. 27.2a).
to yield a mixture of free alkaloids. However, some alkaloids In the Manuich condensation, a C-C-N system is gener-
occur as nonbasic forms such as quaternary amines or as N- ated by the addition of a carbanion to the Schiff base formed
oxides. N-Oxides must be reduced to the corresponding by condensation of a ketone or aldehyde with an amine. Both
bases before they can be extracted in acid. There are many primary and secondary amines may participate (Fig. 27.2b).
color and precipitation reactions for the detection of alka- Charged intermediates produced by the involvement of sec-
loids (Waterman, 1993). Most of these are of value for the ondary amines facilitate the reaction by increasing the elec-
detection of alkaloids as a group, but they do not permit tron deficiency at the carbon atom which is subject to attack
identification of particular compounds. Some of these re- by the carbanion species. As Manuich condensations occur
agents, such as those that form "Reinecke salts," permit the best at about pH 6-7, appreciable amounts of the protonated
investigator to isolate the precipitate and regenerate alka- Schiff base and the attacking anionic species can exist
loids. The preparation of many alkaloid testing reagents and (Geissman and Crout, 1969).
wet chemical methods is outlined (Cordell, 1978, 1981; Aldol and aldol-like condensations between compounds
Cromwell, 1955). containing imino groups also occur frequently. This enamine
variation of the aldol-type reaction may be illustrated by the
Biosyntbetic Orlgln of Alkaloids
formation of tetrahydroanabasine, as in Fig. 27.2c.
Numerous biosynthetic studies of alkaloids have been In addition to these reactions, several other common or-
published since the seminal article by Robinson (1917) pre- ganic chemical conversions occur. Hydrogenation of imino

CH,NH,
I
fH,NH, ~CO'H
fH, fH,
~H, fH, V NH,
~HNH, fH, phenylalanine
CO,H
fHNH, nicotinic acid
~CO'H
ornithine
CO,H
lysine (X
7'1
CO'H

HO
~ NH,

(()X
I I
~
~ NH,
CO'H ~ NH,

anthranilic acid
tyrosine

r=rTCO,H

NH HN~N NH,

histidine
tryptophan

Fig. 27.1. Amino acids commonly involved in the fonnation of alkaloids (Geissman and Crout, 1969),
588 Introduction to Alkaloids

a) :::e=o
" ,
b) -CH +
+ HlN-R

C=O
--
H,O

/' /'

--
with 8 primary amine
/ R-Nlt-C-C
//
R-NH, + O=C"
\\

--
with a secondary amine

R"NH+ ./ OH·
R/ O=C,

H
-';I:(;H-CH,- _ _ -NH-cr-cH, -

o o
-CH"CH=N- -CH-CII=N-

~ cg:.....,B~
c)

N N+

0-0
4,l-piperldeine

G,oH NH
N
tetrahydroanabaslne

0 - -- 0
d)

-2H+
·20 0
N
-2B+
·20
N
-2H+
--+
·20
-2H+
0 N
e) ::::C=N- ~ 'CH-NH- """-
/CH-NH-
~ ./
'C=N-
+20 ./

I) -N=C-CH- -NH-C=C--
I I I I o
OH [0]" [0]
g) :::::~-CH- - /N-CH,- -
" N-C-II amide
./
carbinolamine

Fig. 27.2. Some general reactions involved in the fonnation of alkaloids (Geissman and Crout, 1969).

functions is found commonly (Fig. 27.2d). The reverse of in two biologically active forms, pyridoxal-5'-phosphate and
this reaction (i.e., dehydrogenation of saturated amines) is pyridoxamine-5-phosphate (Fig. 27.3a). Reactivation of the
also observed (Fig. 27.2d). This process may continue sev- coenzyme is effected by a simple reversal of the sequence
eral times and, in ring systems, may produce aromatic com- called transamination. In other instances, a-keto acids are
pounds (Fig. 27.2e). converted into a-amino acids (Geissman and Crout, 1969).
Isomerization of an imino to an a,f3-unsaturated amino The reaction of amino acids to produce keto acids with flavin
function also occurs (Fig. 27.2f). This isomerization occurs adenine dinucleotide (FAD), which occurs in animals and
in the mechanism postulated for the conversion of ~ '-piper- microorganisms, also may be important in the formation of
ideine into the tetrahydroanabasine above (Fig. 27.2c). alkaloids (Figure 27.3b).
Oxidation may occur at an activated position adjacent to Decarboxylation and transamination proceed via the in-
nitrogen. The products usually are carbinolamines or amides termediacy of an amino acid-pyridoxal phosphate complex.
(Fig. 27.2g). Pyridoxal phosphate is related to vitamin B6 and is of pivotal
a-Amino acids are converted to the corresponding a-keto importance in amino acid metabolism. This cofactor occurs
acids by reaction with one form of vitamin B6 , pyridoxal- in two forms: pyridoxal-5'-phosphate (aldehyde form) and
5'-phosphate, a coenzyme that is of fundamental importance pyridoxamine (amine form), which mediate a reversible in-
in amino acid metabolism (Fig. 27.3). This compound exists terconversion of a-amino acid and a-keto acids via a Schiff-
 Introduction 10 Alkaloids 509

a)

pyridoxal pyridoxamine

RCHC02H '--- RCOCO,H

I
NH,

pyridoxal pyridoxamine
HO,CCH,CH,COCO,H ~_=:::::::..-G== HO CCH CH CHCO H
.. ' "I '
NH,
",·ketoglutaric acid glutamic acid

HO,CCH,CH,COCO,H RCOCO,H + HO,CCH,CH,CHCO,H

I
NH,

b) RCHCO H FAD FADH, RCCO,H

I ''---=_/!:::..:.... II _
H,O
RCOCO,H+ NH,
NH, 0

Fig. 27.3. Transamination by pyridoxal phosphate (Geissman and Crout, 1969).

base intennediate. In cases where symmetrical labeling is phenylalanine and tyrosine, respectively, is another enzy·
observed, the free base may be fonned, but, in many cases, matic process of importance in the fonnation of alkaloids
the label of symmetrical molecules is incorporated nooran· (Fig. 27.4).
domly, suggesting that a cofactor·bound compound is in· Primary amines often are converted to the corresponding
valved. aldehydes by oxidation (Fig. 27.4). These aldehydes then
N·Methyl groups often are present. These usually are de· may participate in additional reactions such as the fonnation
rived from S·adenosyl methionine. The point at which the of Schiff bases. 'The enzymes that catalyze these reactions
methyl group is introduced varies from pathway to pathway. are known as monoamine oxidases and diamine oxidases.
The fonnation of cinnamic acid and p.coumaric acid from Monoamine oxidases act on monoamines, whereas diamine

o
~CO'H
~
CO'H
phenylalanine ammonia lyase
19'1
Nu, (PAL) "'" +NH,

~
COlH

~
"",CO'H
19'1 tyrosine ammonia lyase

HO "'" NH, (TAL)

"",I
HO +NH,

monoamine
oxidase
a) RCHO

b) CH,NH, CHO
diamine
I
(CH,).
oxidase I 0=2, putrescine
(CH,).
I
CH,NH,
I 0=3, cadaverine

CH2NH2

Fig.17.4. Phenylalanine ammonia lyase and tyrosine ammonia lyase; monoamine and diamine oxidases (Geissman and Crout, 1969).
510 Introduction to Alkaloids

oxidase enzymes act on diamines, such as putrescine and A number of alkaloids have been demonstrated to act as
cadaverine. phytoalexins (Brooks and Watson, 1985). Among these are
Examples of each of the above types of reactions will be lupanine in Lupinus species and rutacridone epoxide in Ruta
given in the chapters that follow. graveolens.
Many alkaloids are large molecules with pathways quite
distinct from those of primary metabolism, and the cost to
DlSTRlBUfION OF ALKALOIDS the plant may be related to production of the compounds
themselves rather than just allocation of nitrogen (Bernays,
At least 20% of all plant species contain alkaloids in quan- 1983).
tities of about 0.01 % of the dry weight or greater. Alkaloids
may occur in any plant part, but they are usually accumulated
Biological Activity of Alkaloids in Animals
in plant parts involved in biological interactions, for exam-
ple, in the plant epidermis (Hartmann, 1991; Wink, 1993). Although more than 10,000 alkaloids are known, fewer
Data from the distributional patterns of alkaloids often are than 2-5% have been studied for their biochemical proper-
taxonomically useful (Gomes and Gottlieb, Hegnauer, 1988; ties (Wink, 1993). The ecological interpretation of patterns
Mothes, 1966; Seigler, 1977; Waterman, 1993; Waterman of alkaloid distribution have been summarized (McKey,
and Gray, 1987). 1974). Despite the potent physiological effects of many alka-
The most important alkaloid containing families are the loids, there are many fewer articles dealing with the chemical
Amaryllidaceae, Apocynaceae, Asteraceae (Eupatorieae, Se- ecology of this group of compounds than might be predicted.
necioneae), Boraginaceae, Euphorbiaceae, Fabaceae, Laur- The activity of alkaloids against herbivores, toxicity in verte-
aceae, Liliaceae, Loganiaceae, Menispennaceae, Papavera- brates, cytotoxic activity, the molecular targets of alkaloids,
ceae, Ranunculaceae, Rubiaceae, Rutaceae, and Solanaceae mutagenic or carcinogenic activity, antibacterial, antifungal,
(Cordell, 1978; Dalton, 1991). antiviral, and allelopathic properties, and their possible roles
Alkaloids are found most commonly in dicotyledonous as phytoalexins have been tabulated (Wink, 1993). Many
angiosperms and are uncommon in algae, bryophytes, ferns, alkaloids are sufficiently toxic to animals to cause death
and monocotyledonous angiosperms. However, alkaloids are if eaten. Several (e.g., nicotine and anabasine) are used as
known from the lower plant Lycopodium, and from the mon- insecticides (Jacobson, 1982).
ocotyledonous families Amaryllidaceae, Dioscoreaceae, Lil- Many alkaloids act on the nervous system, one of two
iaceae, Orchidaceae, and Poaceae. Alkaloids are not com- important information systems in animals. The nervous sys-
mon in gymnosperms, although they are encountered in tem transmits perceived environmental changes rapidly to a
several conifers and occasionally in other gymnospermous specific processing center. The fundamental unit involved
groups. in this change is the neuron, with the information being trans-
Although alkaloids largely are plant products, a number ferred across a minute gap (2 x 10-5 mm) called a synapse
of these compounds are found in animals such as insects (Cordell, 1981). Although the information is passed electri-
and amphibians. Many of the alkaloids found in animals also cally within the neuron, it is passed chemically between neu-
occur in plants (Nahrstedt, 1982). rons by neurotransmitters. The four most commonly recog-
nized neurotransmitters are acetylcholine, noradrenaline,
dopamine, and serotonin. The reactive site for these chemi-
BIOLOGICAL ACTIVITY cals on the adjacent neuron is called a receptor (Cordell,
1981).
Biological Activity of Alkaloids in Plants There are two main parts to the nervous system: the cen-
Most alkaloids are subject to relatively rapid turnover in tral nervous system (CNS) and the autonomic nervous sys-
plants; diurnal variations in concentration have been ob- tem (ANS). The CNS is the major integrating system of the
served in many. The amounts of other alkaloids vary season- body. The ANS connects the spinal cord at each vertebra
ally. For example, the amount of pedoline in tall fescue is and connects the CNS to each muscle and to glands. The
less than 1000 ",gig dry weight, except in July and August, ANS is divided classically into two systems: the sympathetic
when levels exceeding 3000 ",gig dry weight are reached. and parasympathetic (Cordell, 1981).
The fate of the catabolized alkaloid molecules is not well Neurons that release acetylcholine, noradrenalin, dopa-
understood (Robinson, 1974; Waller and Nowacki, 1978). mine, and serotonin are called cholinergic, adrenergic, dopa-
Although few alkaloids have been tested for plant growth- minergic, and serotonergic, respectively (Cordell, 1981).
regulatory activity, several are known to inhibit growth of Cholinergic drugs elicit responses that mimic stimulation
plants. For example, tomatine, a steroidal alkaloid from to- of cholinergic nerves by direct action on effector cells sup-
matoes, and narciclassine and narciprimine from daffodils plied by cholinergic nerves (choline), by indirect action in-
have antimitotic activity in plants. Lycoricidinol and lycori- volving the inhibition of cholinesterase (which maintains
cidine from Lycoris radiata showed growth-inhibiting activ- high levels of acetylcholine postsynaptically) (e.g., physo-
ity on Avena coleoptile sections (Mandava, 1979). stigmine), and by direct stimnlation of cholinergic effector
 Introduction to Alkaloids Sl1

cells (pilocarpine, arecoline, nicotine, and muscarine) (Cor- synthetic diethyl amide, LSD), mescaline, morphine (often
dell,1981). as the synthetic diacetyl derivative heroin), and nicotine.
Cholinergic blocking agents do just the opposite. Three Alkaloids are particularly important in arrow poisons
main groups of these drugs are also recognized. Atropine- based on Chondrodendron (Menispermaceae) and Strychnos
like compounds occupy the postsynaptic receptor site and (Loganiaceae) species, but they are the active ingredients of
prevent normal neurotransmitter action. Curare and similar arrow poisons from many other species as well (Cordell,
drugs antagonize the nicotinic action of acetylcholine at the 1978).
motor end plates of skeletal muscles. Ganglionic blocking
agents depress autonomic ganglia by depolarizing the end
plate (Cordell, 1981). REFERENCES
Adrenergic drugs are those that elicit responses similar
to the stimulation of adrenergic nerves, that is, release of BAERHEIM SVENDSEN, A. and R. VERPOORTE, Chromatography of
noradrenaline. Epinephrine and ephedrine are examples. Ad- Alkaloids. Part A: Thin-layer Chromatography. (J. Chromatog-
renergic blocking agents prevent stimulation of sympathetic raphy Library Vol. 23A), Elsevier, Amsterdam, 1983.
nerves by antagonizing the effects of drugs such as epineph- BERNA YS, E. A., Nitrogen in defense against insects, in Nitrogen
rine. Many of the ergot alkaloids have this type activity (Cor- as an Ecological Factor (J. A. Lee, S. McNeill, and I. H. Rorison,
dell,1981). eds.), 321-344, Blackwell, Oxford, 1983.
BROOKS, C. J. W. and D. G. WATSON, Phytoalexins, Nat. Prod.
Alkaloids as Al10mones and Kairomones Rep., 2, 427-459 (1985).
CORDELL, G. A., ALKALOIDS, in Kirk-Othmer Eocyclopedia of
Many alkaloids play roles as allomones (Hedin et al.,
Chemical Technology, Vol. I, 3rd ed., 883-943, Wiley, New
1974; Levinson, 1976; McKey, 1974). A large number of York,1978.
these compounds limit hervivory by non-coadapted herbi-
CORDELL, G. A., Introduction to Alkaloids, Wiley, New York,
vores.
1981.
Alkaloids generally are not present in nonflowering
CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annu.
plants. It has been suggested that the rise and diversification
Rept. NMR Spectrosc., 13, 59-210 (1982).
of this group of compounds is a result of coevolution with
CROMWELL, B. T., The alkaloids, in Modeme Methoden der Pflan-
mammals. Alkaloids taste bitter to most mammals; many of
zenanalyze, Bd. 4 (K. Paech and M. V. Tracey, eds.), 367-516,
these animals avoid food plants with high alkaloid content Sprioger-Verlag, Berlin, 1955.
(Swain, 1977). As reptiles cannot detect alkaloids as well
DALTON, D. R, Alkaloids, in Kirk-Othmer Encyclopedia of Chem-
as mammals (they are about 30 times less sensitive), it was ical Technology, Vol. I, 4th ed., 1039-1987, Wiley, New York,
suggested that alkaloids may have had an important role in 1991.
the demise of dinosaurs (Swain, 1977). The status of this
GEISSMAN, T. A. and D. H. G. CROUT, Organic Chemistry of Sec-
idea must be revised, as dinosaurs now are considered more ondary Plant Metabolism, Freeman Cooper, San Francisco,
closely related to birds than reptiles, and the demise of dino- 1969.
saurs appears to be linked to the collision of a large meteor GoMES, C. M. R. and O. R GOTILIEB, Alkaloid evolution aod
with the earth. aogiosperm systematics, Biochem. Syst. Ecol., 8, 81-87 (1980).
In systems involving coadaption, alkaloids may play roles HARTMANN, T., Alkaloids, in Herbivores: Their Interactions with
as kairomones (Levinson, 1976). In other instances, animals Secondary Plant Metabolites, Vol. I, 2nd ed. (G. A. Rosenthal
even use alkaloids as pheromonal substances (Robinson, and M. R. Berenbaum, eds.), 79-121, Academic Press, San
1979). Diego, CA, 1991.
HASHIMOTO, Y., K. KAWANISHI, and M. MORIYASU, Forensic chem-
Uses of Alkaloids by Man istry of alkaloids by chromatographic analysis, in The Alka-
loids, Vol. 32 (A. Brossi, ed.), 1-77, Academic Press, New
Many of the most important therapeutic agents for man York,1988.
are alkaloids; among these are atropine, berberine, caffeine, HEolN, P. A., F. G. MAXWELL, and J. N. JENKINS, Insect plant
cocaine, codeine, colchicine, emetine, ephedrine, ergota- attractants, feeding stimulants, repellents, deterrents, and other
mine, hyoscyamine, morphine, physostigmine, pilocarpine, related factors affecting insect behavior, in Proceedings Sum-
quinidine, quinine, reserpene, scopolamine, strychnine, the- mer Institute for Biological Control Plant Insects and Diseases
ophylline, tubocurarine, and yohimbine. (F. G. Maxwell and F. A. Harris, eds.), 494-527, University of
Alkaloids are among the most important groups of antitu- Mississippi Press, Jackson, 1974.
mor compounds that have been studied (Cordell, 1978, 1981; HEONAUER, R., Biochemistry, distribution and taxonomic relevance
Suffness and Cordell, 1985). Vincristine and vinblastine, of higher plant alkaloids, Phytochemistry, 27, 2423-2427
from Catharanthus roseus (periwinkle, Apocynaceae), are (1988).
used in the treatment of lenkemia. HERBERT, R. B.. The biosynthesis of plant alkaloids and nitrogenous

Others alkaloids are drugs of abuse. These include areco- microbial metabolites, Nat. Prod. Rep., 3, 185-204 (1986).
line, cocaine, N ,N-dimethyltryptamine, lysergic acid (as the HIGHET, R. J. and J. W. WHEELER, The study of alkaloid structures
512 Introduction to Alkaloids

by spectral methods, in The Alkaloids, Vol. 24 (A. Brossi, ed.), (G. A. Rosenthal and D. H. Janzen, eds.), 414-448, Academic
287-348, Academic Press, New York, 1985. Press, New York, 1979.
JACOBSON, M., Plants, insects and man-Their interrelationships, SEIGLER, D. S., Plant systematics and alkaloids, in The Alkaloids,
Beon. Bot., 36, 346-354 (1982). Vol. 16 (R. H. F. Manske, ed.), 1-82, Academic Press, New
LEETE, E., The biosynthesis of alkaloids, Specialist Periodic Re- York,1977.
ports, Biosynthesis, Vol. 7 (R. B. Herbert and T. J. Simpson, SUFFNESS, M. and G, A. CORDELL, Antitumor alkaloids, in The
eds.), 102-223, Chemistry Society, London, 1983. Alkaloids, Vol. 25 (A. Brossi, ed.), 1-369, Academic Press,
LEVINSON, H. Z., The defensive role of alkaloids in insects and New York, 1985.
plants, Experientia, 32, 408-411 (1976). SWAIN, T., Secondary compounds as protective agents,'Annu. Rev,
MANDAVA, N. B., Natural products in plant growth regulation, in Plant Physiol., 28, 479-501 (1977).
Plant Growth Substances (N. B. Mandava, ed.), ACS Sympo- VERPOORTE, R. and A, BAERHEIM SVENDSEN, Chromatography of
sium Series Ill, 135-213, American Chemical Society, Wash- Alkaloids. Part B: Gas-liquid. Chromatography and High-Per-
ington, DC, 1979. formance Liquid Chromatography (Journal of Chromatography,
Library., Vol. 23B), 1-435, Elsevier, Amsterdam, 1984.
McKEy, D., Adaptive patterns in alkaloid physiology, Am. Nat.,
108, 305-320 (1974). VERPOORTE, R., R. VAN DER HEIJDEN, W. M. VAN GULIK, and H.
1. G. TEN HOOPEN, Plant Biotechnology for the Production of
MOTHES, K., Biogenesis of alkaloids and the problem of chemotax-
Alkaloids: Present Status and Prospects, Vol. 40 of The Alka-
onomy, Lloydia, 29, 156-171 (1966).
loids (A. Brossi, ed.), 1-187, Academic Press, New York, 1991.
NAHRSTEDT, A., Strukturelle Beziehungen zwischen pflanzIichen
WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and Metabo-
und tierischen Sekundarstoffe, Planta Medica, 44,2-14 (1982).
lism in Plants, Plenum Press, New York, 1978.
PELLETIER, S. W., (ed.), Alkaloids: Chemical and Biological Per- WATERMAN, P. G., Alkaloids: General observations, in Alkaloids
spectives, Vol. l~present, Wiley-Interscience, New York, and Sulphur Compounds (P. G. Waterman, ed.), Vol. 8ofMeth-
1983-19. ods in Plant Biochemistry (P. M. Dey and J. B. Harborne, eds.),
ROBINSON, R., A theory of the mechanism of the phytochemical 1-16, Academic Press, London, 1993.
synthesis of certain alkaloids, 1. Chern. Soc., 111, 876-899 WATERMAN, P. G. and A I. GRAY, Chemical systematics. Nat.
(1917). Prod. Rep., 5, 175-203 (1987).
ROBINSON, T., Metabolism and function of alkaloids in plants, Sci- WINK, M., Allelochemical Properties or the Raison d'Etre of Alka-
ence, 184, 430-435 (1974). loids, Vol. 43 of The Alkaloids (G. A. Cordell, ed.), 1-105,
ROBINSON, T" The evolutionary ecology of alkaloids, in Herbivores Ac~demic Press, New York, 1993.
28
Simple Amines, Simple Aromatic
and Pyridine Alkaloids

Introduction a large number of plants that are not necessarily closely

Simple Amines related.
Biogenesis of Simple Amines
Biological Activity of Amines in Plants
Biological Activity of Amines in Animals SIMPLE AJIIINES
Amines from Capsicum Species
Diamines and Polyamines in Plants Several simple aliphatic monoamines, formed by decarbox-
Polyamine Alkaloids ylation of amino acids or by transamination of aldehydes,
Simple Aromatic Alkaloids occur widely in plants (Fig. 28.1) (Geissman and Crout,
Hordenine 1969; Guggisberg and Hesse, 1983; Smith, 1977a,b). Many
Ephedrine involve subsequent N-methylation. These compounds are
Khat called "protoalkaloids" in some references (Hegnauer,
Mescaline and Related Alkaloids 1988); more than 1100 compounds of this type are known
Gramine (Verpoorte et aI., 1991).
Psilocybin The chemical and instrumental techniques for the isola-
Pyridine Alkaloids tion and characterization of amines have been reviewed
Nicotine (Crabb, 1982; Smith, 1993).
Ricinine and Related Alkaloids
Biogenesis of Simple Amines
Areca Alkaloids
Dioscorea and Related Alkaloids The biosynthesis of simple amines appears to involve a
Other Pyridine Alkaloids single enzyme with broad specificity. An enzyme with this
References activity, isolated from red algae (Rhodophyceae), was capa-
ble of decarboxylation of the L-forms of leucine, valine, iso-
leucine, norvaline, 2-aminobutyric acid, phenylalanine, me-
thionine, cysteine, and homocysteine. A number of other
INI'RODVCTlON amino acids were unable to serve as precursors (Smith,
1980).
Many alkaloids are formed by simple, usually short, series Although methylamine is widespread in plants, it is prob-
of reactions that are quite common in plants. For instance, ably produced as an artifact of isolation in many cases. Other
simple amines are derived in a one-step reaction from amino simple amines arise from transamination. An enzyme system
acids by decarboxylation. AIkaIoids of this type subse- from Mercurialis perennis (Euphorbiaceae) was capable of
quently may be modified, and, in some instances, quite com- producing a series of amines from ethylamine to undecylam-
plicated products arise. Among these easily derived com- ine in the presence of the corresponding aldehydes and ala-
pounds are simple amines, diamines, and simple aromatic nine (Fig. 28.2) (Smith, 1980).
and pyridine alkaloids. These short pathways, usually based Isobutylamine occurs as amides of various unsaturated
on common reactions, appear to have arisen many times in aliphatic and aromatic organic acids in the Asteraceae (along
the course of plant evolution. Simple aIkaIoids are found in with acetylenic amides), Piperaceae, and Rutaceae. Several

513
514 Simple Amines, Simple Aromatic and Pyridine Alkaloids

fH,CO,H
NH,
methylamine
glycine
CH,CH,NH, CH'fHCO,H histamine (11) histidine

(lr-r;H,
NH,
ethylamine
alanine tryptamine (1)
~N} ~

o
~CO'H

phenylethylamine (8) phenylalanine

~IH, tryptophan O:)(
: :,. . I I NH,
CO'H

NH

~
CO'H

~I Many N-methyl- and N,N-dimethyl-

derivatives of these amines also
HO :::,... NH, occur in nature.

tyramine (9)
tyrosine

Fig. 28.1. Simple amines.

of these derivatives are insecticidal (Jacobson, 1971; Smith, this compound is involved in causation of depression and,
1980). Other aliphatic amines are known to occur in plants, perhaps, of schizophrenia. Ingestion of large quantities of
especially in the above families, as well as in the Caryophyl- serotonin in plantains (Musa species) in West Africa may be
laceae, Rosaceae, and Staphyleaceae (Waterman and Gray, responsible for some human health problems (Smith, 1977a;
1987) (Fig. 28.3). Thin-layer chromatographic (TLC) and Smith 1981).
high-performance liquid chromatographic (HPLC) methods The corresponding N ,N-dimethyltryptamine derivative,
of analysis for alkamides are available (Bauer and Remiger, bufotenine (4), is hallucinogenic and is found in Anadenan-
1989). thera (Piptadenia) peregrina (Fabaceae, Mimosoideae)
Aromatic amines also are known to occur in plants. The (Schultes and Hofmann, 1973). Both serotonin and bufotenin
aromatic amine tryptamine probably arises from decarboxyl- have 5-hydroxy groups (Fig. 28.5).
ation of tryptophan, but, in addition, may arise from indole. N,N-dimethyltryptamine (5) occurs in the leaves, stems,
Tryptamine (1) appears to be a precursor of the plant and sap of Banisteriopsis argentea (Malpighiaceae), several
growth hormone fAA (indole acetic acid) (2) (Fig. 28.4). Virola species (Myristicaceae), and a number of mimosoid
This compound also is produced in the metathoracic glands legumes (Scbultes and Hofmann, 1973). This compound ap-
of the leafcutter ant, Atta sexdens, and is a factor in the ant's pears to serve as a hallucinogen only when used as a snuff
culture of fungi on fresh leaf material. Gall-forming wasps because it is metabolized by a monoamine oxidase when
and a number of other insects also synthesize fAA (Schild- taken orally (Smith, 1977a). Co-occurring l3-carboline alka-
knecht, 1976). loids may accentuate the activity, as they are monoamine
Another important compound derived from tryptamine, oxidase inhibitors.
5-hydroxytryptamine (3) or serotonin, is derived by hydrox- The grass Phalaris tuberosa accumulates mostly N,N-
ylation and is found in many plants (Fig. 28-5). Serotonin dimethyltryptamine (5), 5-methoxy-N,N-dimethyltrypta-
is widely distributed in nature and is a natural nerve transmit- mine, and bufotenin (4), whereas P. arundinacea accumu-
ter in the central nervous system, responsible, at least in lates mainly gramine (6) and hordenine (7). Although the
part, for sleep patterns. There also is some evidence that level of alkaloids is too low to cause acute poisoning in

CO,H CO,H

CH,CHNH,
I
CH,CH,CHO --+ CH,-c=O
I
alanine propanal pyruvic acid propylamine

Fig. 28.2. Biogenesis of propanal in Mercurialis.

 Simple Amines, Simple Aromatic and Pyridine Alkaloids SIS

methylamine herbivorous animals, there is a sufficient amount to de-

crease dry matter intake. However, sheep consume more
ethylamine Phalaris tuberosa dry matter with about 0.01 % gramine
than dry matter that lacks alkaloids (Waller and Nowacki,
CH3CH,CH,NH, n..propylamine 1978).
CH3CH,CH,CH,NH, n-butylamine Phenylalanine, tyrosine, and 34-dihydroxyphenylalanine
(DOPA) decarboxylases have been found in several plants.
CH3(CH,),CH,NH, n-amylamine Each causes the formation of the corresponding amine. Phen-
CH3(CHzl.CH,NH, n-hexylamine ylethylamine (8) and tyramine (9) are widespread among
plants and may occur at high levels. These amines are com-
CH3(CH')5CH,NH, n-heptylamine mon in the Cactaceae, Fabaceae, Rosaceae, and Rutaceae
(Wagner, 1988).
CH3(CH,),CH,NH, n..octylamine
~H3 Biological Activity of Amines in Plants
CH3CHNH, iso..propylamioe Several simple amines may be involved in protecting
fH 3
plants from various types of pathogens. A number of conju-
gates with cinnamic or fatty acids are known. Some of these
CH3CHCH,NH, iso-butylamine compounds, which appear to be widespread, have antifungal
fH3 activity.
CH3CHCH,CH,NH, iso-amylamine Aliphatic amines are especially common in algae and
CH3 particularly in the red-algae Rhodophyceae. These amines
I may protect the algae from microorganisms; several of
CH3CH,CHCH,NH, 2-methylbutylamine these amines are known to be toxic to a range of other
microbes.
HSCH,CH,NH, 2-thioethylamine
Several monoamines and diarnines are known to serve
CH3-S-CH,CH,CH,NH, 3-methyltbiopropylamioe as pollinating agents in plants. In general, these amines
are involved in the attraction of various types of fly. Plants
3-thiopropylamioe such as Helleborus joetidus, Heracleum sphondylium,
many members of the Asclepiadaceae, and a number of
ethanolamine plants in the Araceae are pollinated in this manner. The
pollination mechanisms used by Arum nigrum and A. mac-
2"propanolamine ulalum have been studied in some detail. The bright purple
spathe opens overnight to reveal the spadix in which
respiration is rapid and temperatures of up to 30°C have
Fig. 28.3. Aliphatic monoamines in plants.
been measured. The heat generated helps to volatilize the
amines that are released. Dung beetles and flies are at-
tracted, fall into the bottom of the flower, and are trapped.
The insects are essentially kept prisoners and, as such,

o:rr:
transfer pollen to the receptive styles (Harbome, 1982;

::::,..
I
NH
I NH
'
CO'H

----+-
CCJ(
I I °
::::,.. NH
COlH

tryptophan indole pyruvic acid

!
~Hl
~N} ~ - ~~
~N)
~CO'H
~N}
tryptamine (1) indole acetaldehyde indolyl 3-acetic acid (2)

Fig. 28.4. Biogenesis of IAA (modified from Smith, 1980; used with pennission of the copyright owner, Springer-Verlag, Berlin).
516 Simple Amines, Simple Aromatic and Pyridine Alkaloids

CtJ):,
I I
CO'H

~H'-
~N} ~
NH -
~ NH '

tryptophan
tryptamine (1)

H0'(t:0
~ I I H°'Cc)J1 --
"NH
NH
' -- ~ I NH
NHCH3

serotonin (3)

H0'Cc)J
~ I I ~(CH'),
NH
N(CH3l, ~N} ~
bufotenin (4) N,N·dimethyltryptamine (5)

~N(CH3l'
~N}
00 JYl
~
1
NH
I

HO
~ I N(CH,h

gramine (6) indole (to) hordenine (7)

Fig. 28.5. Formation of bufotenin in Anadenanthera (Piptadenia) peregrina and other simple amines (modified from Smith. 1980; llsed with
permission of the copyright owner, Springer-Verlag, Berlin).

Smith and Meeuse, 1966; Vogel, 1983), A similar system both the producing and the receiving organism) in the polli-
is found in Victoria (Nympheaceae) (Vogel, 1983). nation of a number of plants and is a fairly common compo-
As previously mentioned, tryptamine (1) serves as a pre- nent of flower essential oils. The biosynthesis of indole in
cursor for indoleacetic acid, a plant growth hormone. plants is uncertain. Indole apparently occurs as an enzyme-
Indole (10) is a synomone (i.e., the compounds benefits bound intermediate in the biosynthesis of tryptophan, but

R; (CH,hCHCH;CH(CH,k
(CH,l,CH(CH,l.-
(CH,hCH(CH,ls- '
C H -~O J Y NHCOR I
(CH,hCH(CH,l7-
HO ~
(CH,hCHCH;CH(CH,ls-
(CH,hCHCH,CH;CH(CH,k
CH,CH,(CH,)CH(CH,).-
CH,CH,(CH,)CHCH;CH(CH,l,-
CH,(CH,l.-
CH,(CH,h-
CH,(CH,l.-
CH,(CH,),-
,
C H -~ I
0 X ) " NHCO(CH')4CH;CHCH(CH,h

CH,(CH,l..-
HO ~
CH,CH,(CH,)CH(CH,l4-
capsaicin (12)
CH,(CH,lu-

Fig. 28.6. N-(4-Hydroxy-3-methoxybenzyl) amides or capsaicinoids from Capsicum species (Suzuki and Iwai, 1984: llsed with permission of the
copyright owner, Academic Press, Orlando, FL).
 Simple Amines. Simple Aromatic and Pyridine Alkaloids 517

appears to be derived from degradation of tryptophan in vates blood pressure. A drop in body temperature is pro-
other instances (Cordell, 1981). duced by intraperitoneal or subcutaneous administration that
Histamine (11) and serotonin (3) are found in the stinging may be associated with vasodilation (Suzuki and Iwai,
hairs of several species such as Cnidoscolus texanus (Eu- 1984).
phorbiaceae), Mucuna pruriens (Fabaceae), and Urtica di-
oica (Urticaceae), as well as vegetative materials of Cnidos- Diamines and Polyamines in Plants
colus urens, and fruits of the banana, Musa sapentium
(Lookadoo and Pollard, 1991; Seigler and Bloomfield, un- A number of diamines occur in plants, animals, and fungi
published data; Smith, 1980). (Smith, 1977a, 1977b, I 977c; 1984). Several are toxic to
The wings of bumet moths (Zygaena Jilipendulae) con- mammals, whereas others have interesting physiological
tain small amounts of histamine (11) and the cyanogenic properties. Putrescine (13) accumulates in plants with potas-
glycoside linamarin. Furthermore, the addition of small sium deficiency and under osmotic and other stress condi-
amounts of histamine to linamarin-containing solutions re- tions. This diamine is moderately toxic; it also serves as a
sulted in the rejection of solutions normally consumed by precursor for a number of alkaloid types in plants. Animals,
quail (Coturnix coturnix coturnix). It was suggested that yeasts, and most filamentous fungi synthesize putrescine by
histamine plays a role in defense of the moth from predatory decarboxylation of ornithine; plants and bacteria are able to
birds (Muhtasib and Evans, 1987). synthesize putrescine from either ornithine decarboxylase
or from arginine through a pathway initiated by arginine
Biological Activity of Amines in Animals decarboxylase (Fig. 28.7). Both systems can be specifically
inhibited (Slocum and Galston, 1987). Accumulation ofpu-
Several of these amines are found in animals and some trescine in barley, oat, wheat, and maize leaves in response
are involved in nerve transntission (see Chapter 27). When to osmotic shock is linked to a pathway from arginine (Fig.
plant amines are consumed by animals, they can be quite 28.8).
toxic. For example, phenylethylamine (8) in Acacia berlan- Putrescine is catabolized to "y-aminobutyric acid in both
dieri is poisonous to livestock (Smith, 1977b). The presence monocotyledonous and dicotyledonous plants, but by differ-
of amines in foods consumed by humans also has been noted. ent pathways (Flores and Filner, 1985).
Catecholamines, indoleamines, and histamine (11) fulfill im- Cadaverine (14), a five-carbon diantine, also is known to
portant metabolic functions, especially in the nervous system occur in plants and may be derived from either homoarginine
and in the control of blood pressure. The occasional presence or from lysine (Fig. 28.9). This compound serves as a precur-
of greater than usual amounts of tyramine in cheese can sor for several types of alkaloids. In the biosynthesis of most
cause severe episodes of hypertension, especially in the pres- alkaloids, cadaverine is derived from lysine.
ence of monoamine oxidase inhibitors, which often are used Several polyamines are widely distributed in plants. In
in the treatment of depression (Sntith, 1981). Amines can plants and in Escherichia coli, spermine (15) and spermidine
be formed from bacterial activity in foods (Smith, 1981). (16) are synthesized by aminopropyltransferases that use de-

Amines from Capsicum Species

NH, CO,H
The pungent principles of the fruits of pepper (Capsicum I I
species, Solanaceae) areN-(4-hydroxy-3-methoxybenzyl)al- NH,CNH(CH,),CHNH, arginine
kyl amides (Fig. 28.6). This plant should not be confused
with black pepper, Piper nigrum (Piperaceae) (see the dis-
cussion of pungent principles of black pepper in Chapter
29). The major and best known of these compounds is capsa-
icin (12) (Suzuki and Iwai, 1984). Capsaicin and its relatives f'
NH,CNH(CH,).(CH,).,NH, agmatine
comprise about 0.5-1.5% of some hot pepper cuItivars.
Other pepper cultivars have been selected for their very low
content of these compounds. Capsaicin is synthesized in the
fruit of Capsicum species (Suzuki and Iwai, 1984).
Disease resistance in peppers is correlated with the pres-
ence of capsaicin, although other compounds, such as the N..ca.rbamylputrescine
sesquiterpene capsidiol, may also be involved. In animals,
these compounds produce strong irritation and inflammation
of the skin, mucous membranes, and eyes, and can even be
fatal if large doses are given at one time. In smaller doses, putrescine (13)
capsaicin stimulates the flow of digestive juices and has a Fig. 28.7. Synthesis of putrescine from arginine (modified from Smith,
laxative effect on the colon. Consumption of capsaicinoids 1980; used with pennission of the copyright owner, Springer-Verlag,
causes increases in respiration and vasoconstriction and ele- Berlin).
518 Simple Amines, Simple Aromatic and Pyridine Alkaloids

HO,CCH,CHCO, H
HO,CCH,CHNHCNH(CH.J,CHCO,H
I I I I aspartate
NH.""';....-_ __
CO,HNH, NH,
arginosllccinate lyase
argininosuccinate OCHONDRIAL MATRIX

H1N»NH(CH.J'iHC01H
fumarate o NH,
o citrulline
CYTOSOL
II ornithine transearbamoylase
H, NCNH, urea ( 00
1111
H1NCNH(CH1)'IHC01~arginase IH1 H,NCOrOH carbamoyl phosphate

arginine Nil H
N H
1
H, N(CH.J,CHCO, H - - . - - - / OH
~ba::: pbosphate

\---
ornithine ~hase

~---
H NCNH(CH ) NH .....!..,. H1NCNH(CH1}4NHZ ~ HIN(CH~4NH2 ~ polyamine
1 II 14 1 II biosynthesis
NH 0 putrescine
agmatine N~carbamoylputresclne

... agmatine iminohydrolase ** N-carbamoylputresclne amJdohydrolase

Fig. 28.8. Biosynthesis of polyamines (Slocum et aI .• 1984; modified and used with pennission of the copyright owner, Academic Press, Orlando, FL).

carboxylated S-adenosylmethionine as the aminopropyl compass both basic simple amides and neutral diamides (Ca-
group donor (Slocum and Galston, 1987). Another system banne et aI., 1981; Martin-Tanguy et aI., 1978).
that catalyzes the formation of a Schiff base from putrescine Putrescine (13), cadaverine (14), spermine (15), and sper-
and aspartic j3-semialdehyde and subsequent reduction to midine (16) exhibit marked antisenescence activity when
spermidine has been isolated from Lathyrus sativa (Leete, applied to cereal grain leaves and isolated protoplasts (Slo-
I 983b). Spermine and spermidine are derived in animals cum et aI., 1984) (Fig. 28.9).
and yeast from putrescine and S-adenosylmethionine (Fig. At physiological pH in plants, polyamines exist as proton-
28.10). ated forms. These protonated forms bind to the negatively
Amides of polyamines with compounds such as feruIic, charged phospholipid head groups and other anionic sites in
p-coumaric, and caffeic acid are widespread among higher membranes and alter the stability and permeability character-
plants (e.g., caffeoylputrescine, feruloylputrescine, and p- istics of such membranes. Polyamines also stabilize chloro-
coumaroyltryptamine) and appear to be the main phenolic plast thylakoid membranes and retard chlorophyll loss in
constituents of the reproductive organs of many plants. Ad- senescing barley leaf tissue. Polyamines also influence mem-
ducts include those of most of the common diamines and brane fluidity, affect membrane localized proton pumps, and
polyamines, tyramine, and tryptamine. Known products en- alter plant hormone responses, perhaps by competing for

Fig. 28.9. Polyamines commonly found in plants.

 Simple Amines, Simple Aromatic and Pyridine Alkaloids 519

NH,

N~IHHNN"11 adenine
a-keto-y-methylthio- 5-methylthioribose- ~_,jLN
butyric acid ~ I-phosphate N
putrescine (13)

/ \ \ N:)NH' NH
H,CSCH,CHCO,H I I /)
kH' methionine
H,C(CHOH),CHCH,SCH,
Lo~ --
....N NCH(CHOH) CHCH SCH
Lo~"

1
S-adenosylmetbionine (SAM)
5'-methylthioribose

:)NH'
5'-methylthioadenosine

NH
NHl S-adenosylmethionine N...... I /) +
NH de<arboxylase...K ~N NCH(CHOH) CHCH SCH (CH,' NH
:) ,
N~ / L 'I ' , ", ,
~I.v ~ 0-' spermidine (16)
N NC0.::JHC, ~H'(C~HNH'CO'H de<arbosylated SAM

X
polyamine
biosynthesis
*ACC,ynthase ~ NH, NH

l-amlnocyclopropane- H,N
I-carboxylic acid (ACC)
CO,H t.~ )N NCH(CHOH),CHCH,SCH,
L-o-.:J H,N(CH'),NH(CH,)~(CH,),NH,
CH,=CH, +CN' + CO, spermine (14)
5'-methylthioadenosine
ethylene

Fig. 28.10. Spennidine and spennine biosynthesis (Slocum et a1., 1984; used with pennission of the copyright owner, Academic Press, Orlando. FL).

H02C(CH2hC0 2H succinate

* y-aminobutryaldehyde !
dehydrogenase HO,C(CH,),CHO
NH diamine oxidase
H,N~ '~
I
succinic semialdehyde

putrescine (13) ~o
'9' NH.
GADA-transaminase

- H,N(CH,),CO,H

'Y-aminobutyric acid
Al.pyrroline
H'N~NH~NH,
spermidine (16)

H,N(CH,),NH, - + - H,N(CH,),CO,H
l,3-diaminopropane p-alanine
/'0.. ./'.. /'-.. ./'.. ,NH /'-.. ,NH,
H,N' "-'" 'Nil "-'" "-'" "-'" "-'"

spermine (15) POLYAMINE

DEGRADATION
1-(3-aminopropyl)-pyrroline

Fig. 28.11. Oxidative degradation of polyamines by amine oxidases (Slocum et al., 1984; modified and used with pennission of the copyright owner,
Academic Press, Orlando, FL).
520 Simple Amines, Simple Aromatic and Pyridine Alkaloids

honnone-binding sites or by counteracting honnone-induced There is an absolute growth requirement for polyamines
changes in membrane penneability (Slocum et ai., 1984). in some microorganisms, mammalian cells, and higher
Polyamines also appear to regulate nucleic acid structure at plants. Active growth and cell division are correlated with
several levels of organization. Spennine-DNA complexes increased rates of macromolecular and polyamine synthesis.
have a regular confonnation that stabilizes the nucleic acid Increased polyamine biosynthesis during the G, phase of the
molecule against thennal denaturation in vitro. Polyamines cell cycle, just preceding the onset of DNA synthesis in
modify the confonnation and activity of bacterial and yeast dividing cells, appears to be a universal phenomenon in ani-
tRNAs. Spennidine is an intrinsic component of the RNA mals and plants (Slocum et ai., 1984).
core of a plant virus. Because the activities of many enzymes Spennine (15) and spennidine (16) stimulate growth of
appear to be regulated by reversible phosphorylation, polya- Helianthus tuberosus in explant cultures, but are effective
mine modulation of protein phosphorylation may indirectly spore inhibitors against a number of fungi, including Penicil-
result in the regulation of cellular enzymes. Spennidine or lium digitatum. Spennidine is used for stimulating lactic acid
spennine reversibly prevent the activation of the chloroplast bacteria in the dairy industry. Polyamines are converted into
enzyme, fructose-I,6-bisphosphatase, without affecting the a variety of products that are summarized in Fig. 28.11 (Slo-
catalytic activity of already activated enzyme. Polyamines cum et ai., 1984).
have long been known to increase protein synthesis when
added to cell-free systems. Onset of DNA and RNA synthe- POLYAMIl'IE ALKALOIDS
sis in plants is preceded by increased rates of polyamine
synthesis (Slocum et ai., 1984). Polyamines and ethylene Spennidine (16) has been reported to occur in a number of
appear to have antagonistic functions in plant cells, but the plants in small amounts. Among these are several cereals,
underlying mechanisms are presently unknown. spinach, cabbage, and artichokes. Sym.-homospennidine

12

lunarine (18) palustrine (19) palustridine

oncinotine (21)
inandenine A (22) inandenine B (23)

HN~N~NH&O
N" 0
H~\ ~

~&::~ ""
o "" T'I ?' OH
I I

codonocarpine (24) agrobactine (20)

(a)

Fig. 28.12. (continued)

 Simple Amines, Simple Aromatic and Pyridine Alkaloids 521

H 0
H 0

:~J g?
~~:J
HO
Sf
H
H
OH
HO

H
H
o

CSHll CsHlI

cannabisaliven. (28) anhydrocannablsativine (29)

NO
C_ if
H, r
~ h homalin. (27)

OHN~I
HN~Z:
CH,(CH,)~
o
~ IINH
N'~ ~NH

~NH~
0

""" I I ~
pitheroJlobine (26) "'" 0 Q
(main component)
OCH, OH

(b) isococionocarpioe (25)

Fig. 28.12 (a & b). Some representative polyamine aIka1oids.

(17) has been reported from the leaves of Santalum album loids is based on the biogenic amine spermidine (Guggisberg
(Santalaceae). The biosyntheses of these two componnds and Hesse, 1983). The other part of the molecule is derived
have been studied and they appear to be derived from putres- from a C l6 fatty acid. Isomeric componnds also are fonnd
cine (13), which is, in turn, derived from arginine or orni- in the same plant. The main alkaloids of Oncinous inan-
thine (see the discussion of simple amines above). densis are the 21 -membered ring lactones, inandenine A and
Alkaloids based on these structural units are widespread, B (22,23) (Fig. 28.12) (Guggisberg and Hesse, 1983).
but sporadically distributed, in nature; at least 125 alkaloids Another alkaloid related to lunarine, codonocarpine (24),
of this type are known (Verpoorte et aI., 1991). The first has been isolated recently from Codonocarpus australis. Al-
reported was lnnarine (18), from Lunaria biennis (Brassica- though this plant has been placed in the family Phytolacca-
ceae), which was reported in 1908, although the correct ceae, its true affinities are not known with certainty (Fig.
structure was established only in 1965 (Fig. 28.12). Palus- 28.12) (Guggisberg and Hesse, 1983). Isocodonocarpine
trine (19), from Equisetum palustre (Equisetaceae), also was (25) has been isolated from Capparis decidua (Capparida-
reported quite early (Badwani et al., 1973). ceae) (Ahmad et al., 1989).
Simple amine derivatives apparently occur in most, if not Pithecolobine (26) has been isolated from Pithecelobium
all, plants, and the alkaloids derived from them are wide- saman (syn. Samanea saman, Fabaceae: Mimosoideae). Pi-
spread. thecolobine consists of a substituted fatty acid and spermine.
Several polyamine alkaloids appear to be involved as sid- Recently, the structure has been clarified (Fig. 28.12) (Bad-
erophores (i.e., microbial iron-transport agents, in bacteria). wani et al., 1973). Similar alkaloids have been isolated from
For example, agrobactine (20) is thought to play this role in the related plantAlbizia amara (pezzuto et al., 1991). Certain
Agrobacterium tumefaciens (Fig. 28.12) (Guggisberg and of the isolates from this plant inhibit the catalytic activity
Hesse, 1983). of DNA polymerase, RNA polymerase, and HIV-l reverse
The main alkaloid of Oncinotis nitida (Apocynaceae) is transcriptase (Mar et al., 1991).
the spermidine alkaloid oncinotine (21). This group of alka- Other spermidine alkaloids occur in Homalium pronyense
 ux:
522 Simple Amines. Simple Aromatic and Pyridine Alkaloids

9' I
~
CO'H
CO'H
__
~I
::::,.. NH, HO """ NH,

- HO~ (Y:H,~ --
HO OJ
I::::,..

hordenine (7)
N(CH,>.

(32) N·methyltyramine (31)

88% retention of tritium

Fig. 28.13. Biogenesis of N~methyltyramine and hordenine (modified from Daly et al., 1972; used with pennission of the copyright owner, Birkhliuser
Verlag, Basel).

(Homaliaceae) (27) (Fig. 28.12). The Homaliaceae may be scription medicines. This compound stimulates the central
confamilial with the Flacourtiaceae. These alkaloids also are nervous system and elevates blood pressure. The effects are
known to occur in Aphelandra squarrosa (Acanthaceae), similar to epinepbrine. Most ephedrine·like compounds used
Cannabis sativa (Cannabaceae) (28, 29), Pleurostylia afri· today are prepared synthetically. The compound amphet·
cana (Celastraceae) (30), Chaenorrhinum spp. (Scrophulari· amine (34) is related structurally to ephedrine, but is not
aceae), and a number of other sources (Cordell, 1981; Gug· naturally occurring.
gisberg and Hesse, 1983). The biosynthetic pathway to ephedrine appears to be as
presented in (Fig. 28.14) (Cordell, 1981; Yamasaki et aI.,
1973).
SIMFLE AROJIIATIC ALKALOIDS Incorporation of ephedrine at 0.1 % into artificial diets
is totally lethal to the larvae of Callosobruchus maculatus
Hordenine (Janzen et al., 1977).

In barley (Hordeum vulgare, Poaceae), phenylalanine is

Kbat
converted to tyrosine and then to tyramine. Tyramine (1) is
subsequently methylated and converted to hordenine (7) The use of khat (Catha edulis, Celastraceae) is common
(Fig. 28.13). A related species, Hordeumdistichum, converts in several areas of northeastern Africa and in Yemen. AI·
labeled phenylalanine into N·methyltyramine (31), a reac· though the leaves and young stems are usually used as a
tion that is believed to procede via the intermediacy of the masticatory, a tea is sometimes prepared (Brenneisen and
so·called NIH shift (32) (Fig. 28.13). The half·life of ElSohly, 1992). The active compounds of khat are two
hordenine in barley is about 42 h (Waller and Nowacki, closely related compounds: d·pseudonorephedrine (35) and
1978). Hordenine has been suggested to playa role in alle· ( - )·{X·arninopropiophenone or cathinone (36) (Crombie et
lopathy in cultivated barley fields and is a feeding repellent al., 1990; Szendrei, 1980) (see also Chapter 36). These com·
for grasshoppers (Smith, 1977b). pounds have central nervous stimulant activity (Tyler et al.,
1981). (S)·Cathinone is thought to be responsible for the
Ephedrine cardiac stimulating activity of khat (Wagner, 1988).
Ephedrine (33) occurs in the Chinese drug "rna huang,"
Mescaline and Related Alkaloids
Ephedra sinica (Ephedraceae), a drug used in China for more
than 5000 years (Tyler et al., 1981). The active ingredient A series of simple alkaloids are found in a variety of
is ephedrine, a well·known adrenergic, bronchodilating members of the cactus family, the Cactaceae. One of these
agent that is a common ingredient of both patent and pre· compounds, mescaline (37), disturbs normal mental function
 Simple Amines, Simple Aromatic and Pyridine Alkaloids 523

o
o
~CO'H

~
CH'OH
Nu,-- ~I
"" NH,

-- d'LH' ~NH' ~NH'

OH

() ,"".H () , .••• H

ephedrine (33) ampbetamine (34) cathinone (36)

(+)-norpseudoephedrine (35)
• The two carbon unit may come from serine, glycln~ alanine, propionic
acid, aspartic acid, glucose or formate (Cordell, 1981),
Fig. 28.14. Biogenesis of ephedrine and related alkaloids (Cordell, 1981; modified and used with permission of the copyright owner Dr. Geoffrey
Cordell).

and causes hallucinations and euphoria. Unusual and bizarre reactions but which also occur in the Cactaceae will be dis-
color perception occurs (Schultes and Hofmann, 1973; Tyler cussed in Chapter 33).
et aI., 1981). Although most commonly isolated from the
peyote cactus, Laphaphara williamsii, mescaline occurs in Gramme
related species as well. Mescaline is hallucinogenic and is
used as an integral part of religious activities by many Amer- Gramine (6) is found in several Acer species (Aceraceae,
ican Indians. Mescaline is nonaddictive. maples), barley (Hardeum vulgare), and a number of other
Tyrosine and tyramine (9) are established firmly as the plants. The mechanism of the side-chain modification which
precursors of this group of aJkaloids. Dopamine (3,4-dihy- produces gramine from tryptophan is not well understood.
droxyphenethylamine) (38) is incorporated in peyote and 5- Tracer studies indicate that the indole unit is incorporated
hYdroxy-3,4-dimethoxyphenylethylamine has been detected intact, but the origin of the second nitrogen atom is in doubt.
in peyote (Fig. 28.14). The presence of 3-methoxy4-hy- The methylene carbon of gramine is the same as the methyl-
droxy- and 4-hydroxy-3,5-dimethoxyphenylethylamine (39) ene in the same position in tryptophan (Fig. 28.16).
in Trichacereus pachanai, another mescaline-producing spe- It is thought that hordenine (7) and gramine (6) are allelo-
cies, reveals another possible alternative pathway (Fig. pathic agents, as fields of barley are usually weed-free.
28.15). Gramine inhibits radicle growth of several plants (Wink,
Mescaline has an LDso p.o. (per as, or oral) of 100 mg/kg 1993). At 0.1 % in artificial diets, both gramine and nicotine
in Agelaius (red-winged blackbirds) (Wink, 1993). were totally lethal to the bruchid beetle Callasabruchus mac-
A number of other alkaloids which involve more complex ulatus (Janzen et aI., 1977). Gramine is a feeding deterrent

~
-
CO'H
HO~ _ _ CH"O~

HO
~ I
........
NH--+
, HO~ NH, HO~ NH,

dopamine (38)

CH'-OYJ CH"O~ CH'O~

HO:::'" I -
NH, HO:::'" , NH, -- CH,O :::".
, NH
,
OH OCH, OCH,
(39) mescaline (37)

Fig. 28.15. Proposed route of mescaline biogenesis (modified from Smith, 1980; modified and used with pennission of the copyright owner, Springer~
Verlag, Berlin),
524 Simple Amines, Simple Aromatic and Pyridine Alkaloids

~co,~
~NJ
+ CHO
Nu,
O:t:YtCO,H

"" NH
POCH
{1 - t. 0'

POCH~O' ~
f
1
u...~.( pyridoxal phosphate CH3
~+ CH3 H
H

~NH'
~N} -- -- ~N(CH3l'
~N}
gramlne(6)
0r
~NH~
isogramine (40)
N(CH3l,

Fig. 28.16. Biogenesis of gramme (modified from Cordell, 1981; modified and used with permission of the author).

Ct:J):
""'I
""
1
NH
NH
'
CO'H

-- ~
~N} N(CH3), -
OH HO, pH
l' . . o
~(CH3l'-
psilocin (41)
°
Ct:n(
psilocybin (42)
CH3l'

H0'(Jc:(J
"" 1 NH
1 NH,

serotonin (3) bufotenin (4)

,. (Y0H
(t:jN~
catJeoylputrescine p-coumaroyltryptamine

Fig. 28.17. Psilocybin and related compounds (modified from Geissman and Crout, 1969).
 Simple Amines, Simple Aromatic and Pyridine Alkaloids 525

and insecticide for certain aphids, has antibacterial and anti- Psilocybin has an LD5• i. v. of 285 mg/kg in mice (Wink,
fungal properties, and uncouples photophosphorylation 1993).
(Wink, 1993).
Central Asian camels refuse to eat a certain type of
"reed" due to the presence of gramine (6). When Erdtrnan PYRIDINE ALKALOIDS
and co-workers synthesized this a1kaloid and the isomeric
compound, isogramine (40), Erdtrnan tasted a small sample
Alkaloids that possess an aromatic pyridine ring often are
of isogramine. To his surprise, it tasted bitter sweet and
called pyridine alkaloids (Leete, 1980; Pinder, 1993; Strunz
numbed his tongue. Gramine, however, did not have local
and Findlay, 1985); approximately 250 alkaloids of this type
anesthetic properties (Sneader, 1985).
are known (Verpoorte et al., 1991). Although many are de-
PsDocybln rived from nicotinic acid (Fig. 28.18), others arise from aro-
matization of piperidine alkaloids and other pathways (Wal-
Species of the mushroom genus Psilocybe (as well as ler and Nowacki, 1978). This chapter includes several of
those of several other genera) produce metabolites such as the most important compounds of this type, although others
psilocin (41) and psilocybin (42) that have 4-hydroxyl (such as a number of monoterpene-derived a1kaloids, Nu-
groups (Fig. 28.17) (Antkowiak and Antkowiak, 1990). poor alkaloids, and sesquiterpene alkaloids of the Celastra-
These compounds have hallucinogenic properties similar to ceae) are discussed in Chapter 36.
those of LSD; psilocybin interacts with the serotonin recep-
tor (Wink, 1993). The properties of these mushrooms have
Nicotine
been recognized in Central and South America for at least
3000 years (Mann, 1987; Schultes and Hofmann, 1973; The most important a1kaloid of this group, nicotine (43),
Tyler et al., 1981). is common and found in many plant families (see Leete,
Tryptophan and tryptamine are incorporated into psilocy- 1979, 1980; 1983a, 1983b). (S)-Nicotine is best known from
bin. Psilocin is an apparent intennediate for the biosynthesis Nicotiana species (tobacco, Solanaceae). Nicotinic acid (44)
of psilocybin (Antkowiak and Antkowiak, 1990). and ornithine serve as efficient precursors for nicotine. Orni-

CHO /CO,H
1 CH,
bH------
CHOH
I ?, +
CH0r>' /\

r
H,N
CO,H
OH
glyceraldehyde aspartic acid quinolinic acid
-3-phosphate

aN
pbosphoribosyl pyrophosphate nicotinic acid
adenine

a - a
dinucleotide
CO,H NH,

L:X~
wO -P-O
CO,H a C O ' H / '" +1 ~~ CO,H

"N+
CHJ
I --
"'N
I
nicotinic acid H
0 N° o-p-o-p-°lcd
U . .
()H
"IOOHNN
+
0- OH HH
HHH
t
a
OHOH
nicotinic acid mononucleotide OR OH I HO OH

"'N+ 1
CONH, aCONH,

-- '" 1 _____ a 1
CONH
'
t NH,

N:XN~
ll}

1
. N '" 0 0 HN N

~CONH:iCOaCN ~CN JQr0-~_-o-t~o1QL NAD

O~NjJ ----... 0 ~ Il .,.lo

N nicotine, anabasine OH OR HO OR
eH] CH 3 eH 3 and ='fJ:idlne
N-methyl-S-carbamoyl nudiftorine ricinine
2-pyridone

Fig. 28.18. Biosynthesis of pyridine alkaloids (Waller 1966; used with pennission of the copyright owner, the American Society for Biochemistry and
Molecular Biology).
526 Simple Amines, Simple Aromatic and Pyridine Alkaloids

Cu'
:.:p/CO,H
In plants: H~~'CH CO,H aCO,H
I I
°
(
:9'} (
I,tH
--+

aH
HPOH --+ - - "N "-N
II CH CO,H
-O-P-o/ ~ CO,H
b-~ quinolinic acid nicotinic acid (44)
glyceraldehyde
-3-phosphate aspartic acid co~
H"'~' #
'\~
a
N+ N-methylpyrrolidine
I
H N b CH,

~.~Q.. )
H".J.() .. ~ NH H D 0;:D °

!
~ ~
~
N

0?IH
~N)
NH H''''? Q
!) N ! H

0 0
CH,

A 0- 0
(S)-(-)-anabasine(4S)

U
~
N
H
1
NHHH

"'"
N
IHHN "",IH~
N C~
anatabine (46) nornicotine (47) nicotine (43)

Fig. 28.19. Biogenesis of nicotinic acid and nicotine (modified from Leete, 1977; modified and used with pennission of the copyright owner,
Academic Press, London).

thine is incotporated via putrescine, a symmetrical interme- nicotine. Tobacco homworm larvae, and those of many other
diate. Nicotinic acid is not incorporated via a symmetrical insects that feed on tobacco, excrete nicotine rapidly. Several
intermediate (Fig. 28.19). Furthermore, nicotinic acid in other mechanisms that degrade residual amounts of nicotine
plants is not synthesized by the same biosynthetic pathway also exist (Brattsen, 1986). PA50 enzymes that convert nico-
as in animals. Many of the feeding studies involving Nicoti- tine to the much less toxic compound cotinine are important.
ana species have been summarized (Leete, 1983b). Nonetheless, plants of Nicotiana section Repandae are
An average 60-day-old tobacco plant contains about 250 highly toxic to this insect. Topical application of I mg of
mg of nicotine. Based on the net carbon dioxide fixation and the crude exudate of leaves of N. repanda to larvae caused
the rate of turnover of nicotine in the plant, maintenance of 100% mortality in 24 h. Plants of this species were shown
this amount of nicotine may represent as much as 10% of to contain N-acyl derivatives of nicotine such as (48) (Har-
the plant's total metabolism (Robinson, 1974). The half-life borne, 1989).
of nicotine in tobacco plants is about 22 b (Waller and No- Approximately 40-60 mg of nicotine (43) is a lethal dose
wacki, 1978). The alkaloid content of plants of Nicotiana for an adult human; the LD,o i.v. in mouse is 0.3 mg/kg
sylvestris undergoes a fourfold increase following damage (Wink, 1993). This base is the main pharmacologically ac-
to the leaves (Hartmann, 1991). The production of nicotine tive component of tobacco smoke and probably is responsi-
and related pyridine alkaloids such as anabasine (45), anatab- ble for the addictive nature of cigarettes. The pharmacology
ine (46), and nomicotine (47) in plant tissue, cell, and hairy of nicotine and other Nicotiana alkaloids has been reviewed
root cultures has been reviewed (Vetpoorte et al., 1991). (Fodor and Colasanti, 1985). Nicotine mimics acetylcholine
Nicotine (43) inhibits radicle growth in Lepidium and is and activates acetylcholine receptors (Wink, 1993).
toxic to Lemna (Wink, 1993). It is a useful insecticide and Plants of Nicotiana tabacum release nicotine through
probably serves as a defensive compound in the plant (Wink, their roots. Nicotine is stored in the roots and transported to
1993). Some coadapted insects, such as the tobacco hom- the upper parts of the plant. These alkaloids are accumulated
worm, Manduca sexta, are highly resistant to the effects of in all leaf parenchyma cells and in the vacuoles, extraplasmic
 Simple Amines. Simple Aromatic and Pyridine Alkaloids 527

CO'H
CONH
cr :::,. I __ c
:::,. r
I __' :::,. r
: :,.N+ r
1CN- c 1-CN

N c
N N I
CH,
nicotinic acid (SO)

a -t.)" --
I NON
CH3
CN ~CN
OH

CH3
0
~C~
N
OCH,

l~.A
I
0
CH,
~
~CN
t.AHN 0
+
I
.. senescent yellow leaves CH3
ricinine (49) •• fresh green leaves

o
D ~I
~
CN

0
bO~OH
OCH,

NH
CN
OH

OH
aCO'H uCO'CH'

N+
I
N
I
CH, CH, CH,
nudiftorine (50) acalyphin (51) trigonelline (54) arecoline (52)

~~ OH (48)

lNJ O~(CH~~
Fig. 28.20. Ricinine biogenesis and other pyridine alkaloids (modified from Waller et aI., 1966 and Skurskyet aI., 1969; used with pennission of the
copyright owner, the American Society of Biochemistry and Molecular Biology).

Q-- -
" ·6
CO,H

~
N

1# I ('~ {~
--~
NI

CO,H
o O-H H+
00.';
(53)
nicotinic acid

------.. ~CO'H
~7Y~ HN

~ ~ HO I
OHHO,C HO,C

dioscorlne

Fig. 28.21. Biogenesis of dioscorine (modified from Leete. 1980; modified and used with pennission of the copyright owner, Springer-Verlag, Berlin).
 cP:
528 Simple Amines, Simple Aromatic and Pyridine Alkaloids

space between the membranes of the plastid envelope, plas- OAc

tid vacuoles, and in the endoplasmic reticulum (Neumann,
H
1975).
Nicotine (43) and anabasine (45) are produced by hairy OAc '
root cultures of Nicotiana rustica (Fiores et aI., 1987). Dif- ~ 0 0
ferentiated cultures of Duboisia species produce nicotine
(0.2-0.9%) as well as tropane alkaloids (Ellis, 1988).
'"N I anibtne (55)
Nomicotine (47) (in which the N-methyl group is absent)

~
is the major pyridine alkaloid in many other Nicotiana spe-
cies. This alkaloid has an LDso i.p. 23.5 mg/kg in rat (Wink,
1993). Nornicotine is a feeding deterrent to Melanoplus. An-
abasine (45) (with two 6-membered rings) is the principal
alkaloid of Nicotiana glauca (Strunz and Findlay, 1985).
~NJHO~OCH'
duckein(Slt)

Ricinine and Related AIkaloids Fig. 28.22. Additional pyridine alkaloid>.

A series of compounds that appear to be derived from
nicotinic acid is found in a number of euphorbiaceous plants. to be a likely intermediate (Fig. 28.21) (Strunz and Findlay,
Ricinine (49), an alkaloid from Ricinus communis, is derived 1985; Specialist Periodic Reports 1978, 1979).
from nicotinic acid via nicotinamide (50). This alkaloid may
occur in quantities as great as 1% of the plant dry weight. other Pyridine AIkaloids
A crude enzyme preparation has been isolated that will con-
vert a series of pyridinium salts into ricinine and related Trigonelline (54), first isolated from fenugreek seed (Tri-
compounds (Fig. 28.20) (Fodor and Colasanti, 1985). This gonella foenum-graecum, Fabaceae), is now known to be
alkaloid disappears completely from senescent leaves. Ri- widespread in plants (Fodor and Colasanti, 1985). Trigonel-
cinine (49) has an LDso p.o. of 42 mglkg in Agelaius (Wink, line promotes cell arrest in G 2 of the cell cycle in plants
1993). (Wink, 1993).
A number of related alkaloids, such as nudiflorine (50), The Chinese plant Tripterygium wilfordii (Celastraceae)
are found in related plants such as those of the genus Trewia. contains compounds with insecticidal properties. On alkaline
Introduction of labeled nicotinic acid resulted in the intro- hydrolysis, these compounds give rise to wilfordic acid (57)
duction of label into several of these (Strunz and Findlay, and hydroxywilfordic acid (Strunz and Findlay, 1985).
1985). Anibine (55) and duckein (56), from Aniba duckei (Laur-
A cyanogenic compound, acalyphin (51), that appears to aceae), are components of commercial rosewood oil (Fig.
have a similar biosynthetic origin has been isolated from 28.22). These compounds appear to be derived by extension
Acalypha indica (Nahrstedt et al., 1982). of a nicotinoyl precursor by two or three acetate units, re-
spectively (Strunz and Findlay, 1985).
Areca Alkaloids Several pyridine alkaloids are known to occur in both
plants and insects (Nahrstedt, 1982).
Areca catechu (Arecaceae, betel nut) is the world's most
commonly used masticatory. This palm seed is used in con-
junction with Piper betle leaves, Acacia catechu bark, lime, REFERENCES
and other accompaniments (Gowda, 1951) by perhaps 1 bil-
lion of the world's inhabitants daily. Arecoline (52) is the AHMAD, V. U., N. ISMAlL, and A. AMBER, IsocodoDocarpine from
most commonly studied alkaloid of Areca catechu seeds. Cappari. decidua, Phytochemistry, 28, 2493-2495 (1989).
This alkaloid has muscarinic action (mimics acetylcholine ANTKoWIAK, R. and W. Z. ANTKOWIAK, Alkaloids from mush-
and binds to acetylcholine receptors) (Wink, 1993). Low rooms, in The Alkaloids, Vol. 40 (A. Brossi, ed.), 189-340,
doses produce vasodilation and a fall in both systolic and Academic Press, New York, 1990.
diastolic blood pressure. Arecoline exerts stimulatory effects BADWANI, M. M., K. BERNAUER, P. VAN DEN BROEK, D. GROOER,
on the gastrointestinal tract and enhances diaphoresis. Arec- A. GUGGISBERG, S. JOHNE, 1. KOMPIS, F. SCHNEIDER, H. VBITH,
oline has an LDso s.c. of 100 mglkg in mouse. This alkaloid M. HEsSE, and H. SCHMID, Macrocyclic spermidine and speIlD-
is a feeding deterrent to certain insects (Wink, 1993). iDe alkaloids, Pore Appl. Chern., 33. 81-108 (1973).
BAUER, R. and P. RsMlGER, 1LC and HPLC analysis of a1kamides
Dioscorine and Related Alkaloids in Echinacea drugs, Planta Medical, 55, 367-371 (1989).
BRAITSEN, L. B., Fate of ingested plant allelochemicals in herbivo-
A series of alkaloids have been isolated from members rous insects, in Molecular Aspects of Insect-Plant Associations
of the genus Dioscorea (Dioscoreaceae). These compounds (L. B. Brattsen and S. Ahmad, eds.), 211-255, Plenum Press,
are derived from nicotinic acid and compound 53 appears New York, 1986.
 Simple Amines. Simple Aromatic and Pyridine Alkaloids 529

BRENNEISEN, R. and M. A. ELSOHLY, Socia-economic poisons: acid, in Secondary Plant Products (E. A. Bell and B. V. Charl-
Khat, the natural amphetamine, in Phytochemical Resources for wood, eds.), 65-91, Springer-Verlag, Berlin, 1980.
Medicine and Agriculture (H. G. Nigg and D. S. Seigler, eds.), LEETE, E., Biosynthesis and metabolism of the tobacco alkaloids,
97-116, Plenum Press, New York, 1992. in Alkaloids. Chemical and Biological Perspectives, Vol. 1 (S.
CABANNB, F., M. A. DsLEBROUX, J. MARTIN-TANGUY, and C. MAR- W. Pelletier, ed.), 85-152, Wiley, New York, 1983a.
TIN, Hydroxycinnamic acid amides and ripening to flower of LEETE, E., The biosynthesis of alkaloids, Specialist Periodical Re-
Nicotiana tabacum var. xanthi n. c., Physiol. Plant, 53, 399-404 ports, Biosynthesis, 7, 102-223 (1983b).
(1981).
LoOKADOO, S. E. and A. J. POLLARD, Chemical contents of stinging
CORDELL, G. A., Introduction to Alkaloids, Wiley, New York, trichomes of Cnidoscolus texanus, 1. Chem. Beol., 17.
1981. 1909-1916 (1991).
CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual MANN, J., Secondary Metabolism, Oxford University Press, Ox-
Reports on NMR Spectroscopy, Vol. 13 (G. A. Webb, ed.), ford, 1978; 2nd edition, 1987.
60-120, Academic Press, London, 1982.
MAR, W., G. T. TAN, G. A. CORDELL, and J. M. PEzzuro, Biological
CROMBIE, L., W. M. L. CROMBIE, and D. A. WHITING, Alkaloids activity of novel macrocyclic alkaloids (budmunchiamines)
of khat (Catha edulis), in The Alkaloids, Vol. 39 (A. Brossi, from Albizia amara detected on the basis of interaction with
ed.), 139-164, Academic Press, New York, 1990. DNA, J. Nat. Prod., 54, 1531-1542 (1991).
DALY, J. W., D. M. JERlNA, and B. WITKOP, Arene oxides and MARTIN-TANGUY, J., F. CABANNE, E. i'ERDRlZET, and C. MARTIN,
the NIH shift: The metabolism, toxicity and carcinogenicity of The distribution of hydroxycinnamic acid amides in flowering
aromatic compounds, Experientia, 28, 1129-1149 (1972). plants, Phytochemistry,17, 1927-1928 (1978).
ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod. MUHTAsm, H. and D. L. EVANS, Linamarln and histamine in the
Rep., 5,581-612 (1988). defense of .adult Zygaena filipendulae, J. Chern. Bcol., 13,
FLoRES, H. E. and P. Fn.NER, Polyamine catabolism in higher 133-142 (1987).
plants: Characterization of pyrroline dehydrogenase, Plant NAHRSTBOT, A., StrokturelJe Beziehungen zwischen pflanzlichen
Growth Regul., 3, 277-291 (1985). und tierischen Sekundiirstoffen, Planta Medica, 44, 2-14
FLORES, H. E., M. W. Hoy, and J. J. PICKARD, Secondary metabo- (1982).
lites from root cultures, Tibtech, 5, 64-69 (1987). NAHRSTBOT, A., J. KANT, and V. WRAY, Acalyphin, a cyanogenic
FODOR, G. B. and B. COLASANTI, The pyridine and piperidine alka- glucoside from Acalypha indica, Phytochemistry, 21,101-105
loids: Chemistry and pharmacology, in Alkaloids: Chemical and (1982).
Biological Perspectives, Vol. 3 (S. W. Pelletier, ed.), 1-90, NEUMANN, D., Beitriige zur Physiologie der Alkaloide. VI. Ultra-
Wiley, New York, 1985. stroktur der Parenchymzellen und Speicherung von Alkaloid in
GElSSMAN, T. A. and D. H. G. CsOUT, Organic Chemistry of Sec- Blattstecklingen von Nicotiana rustica L., Biochem. Physiol.
ondary Plant Metabolism, Freeman Cooper, San Francisco, Pflanzen, 168, 511-518 (1975).
1969. PEzzUTO, J. M., W. MAR, L. LIN, G. A. CORDELL, A. NEZMELYI,
GoWDA, M., The story of pan chewing in India, Botanical Museum and H. WAGNER. DNA-based isolation and the structure elucida-
Leaflets, Harvard Univ., 14, (No.8), 181-214 (1951). tion of the budmunchiamines, novel rn.acrocyclic alkaloids from
GUGGISBERG, A. and M. HEsSE, Putrescine, spennidine, spennine, Albizia amara, Heterocycles, 32, 1961-1967 (1991).
and related polyamine alkaloids, in The Alkaloids, Vol. 22 (A. PINnER, A. R., pyrrole, pyrrolidine, pyridine, piperidine and related
Brossi, ed.), 85-188, Academic Press, New York, 1983. alkaloids, in Alkaloids and Sulphur Compounds (p. G. Water-
HARsoRNB, J. B., Introduction to Ecological Biochemistry, 2nd ed., man, ed.), Vol. 8 of Methods in Plant Biochemistry (P. M. Dey
Academic Press, New York, 1982. and J. B. Harbome, eds.), 241-269, Academic Press, London,
HARBORNB, J. B., Recent advances in chemical ecology, 7, 85-109 1993.
(1989). ROBINSON, T., Metabolism and function of alkaloids in plants, Sci-
HARTMANN, T., Alkaloids, in Herbivores: Their Interactions with ence, 184, 430-435 (1974).
Secondary Plant Metabolites, Vol. I, 2nd ed. (G. A. Rosenthal SCHn.DKNECJIT, H., Chemische Okologie-Ein Kapitel modemer
and M. R. Berenbaum, eds.), 79-121, Academic Press, San Naturstoffchemie, Angew. Chern., 88, 235-243 (1976).
Diego, CA, 1991. SCHULTES, R. E. and A. HOFMANN, The Botany and Chemistry of
HEoNAUER, R., Biochemistry, distribution, and taxonomic rele- Hallucinogens, C. C. Thomas, Springfield, IL, 1973.
vance of higher plant alkaloids, Phytochemistry, 27, 2423-2427 SKURSKY, L., D. BURLESON, and G. R. WALLER, Interconversion
(1988). of ricinine and N-demethylricinine in senescent and green castor
JACOBSON, M., The unsaturated isobutylamides, in Naturally Oc- plant leaves, J. BioI. Chern., 244, 3238-3242 (1969).
curring Insecticides (M. Jacobson and D. G. Crosby, eds.), SLOCUM, R. D., R. KAUR-SAWHNEY, and A. W. GALSTON, The phys-
137-176, Marcel Dekker, Inc., New York, 1971. iology and biochemistry of polyamines in plants, Arch. Bio-
JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary chern. Biophys., 235, 283-303 (1984).
compounds to the seed-eating larvae of the bruchid beetle Callo- SLOCUM, R. D. and A. W. GALSTON, Inhibition of polyamine bio-
sobruchus maculatus, Phytochemistry, 16, 223-227 (1977). synthesis in plants and plant pathogenic fungi, in Inhibition of
LEETE, E., Biosynthesis and metabolism of alkaloids, Planta Med- Polyamine Metabolism (p. P. McCann et al., eds.), 305-316,
ica, 36, 97-112 (1979). Academic Press, New York, 1987.
LEETE, E., Alkaloids derived from ornithine, lysine, and nicotinic SMITH, B. N. and B. J. D. MEEUSE, Production of volatile amines
530 Simple Amines, Simple Aromatic and Pyridine Alkaloids

and skatole at anthesis in some Arum lily species, Plant Physio!., istry, biochemistry, pharmacology, and food science of the pun-
41, 343-347 (1966). gent principle of Capsicum species, in The Alkaloids, Vo!. 23
SMITH, T. A., Tryptamine and related compounds in plants, Phyto- (A. Brossi, ed.), 228-299, Academic Press, New York, 1984.
chemistry, 16, 171-175 (1977a). SZSNDREI, K., The chemistry of khat, Bull. Narcot., 32, 5-35
SMITH, T. A., Phenethylamine and related compounds in plants, (1980).
Phytochemistry, 16, 9-18 (1977b). TYLER, V. E., L. R. BRADY, and J. E. ROBBERS, Pharmacognosy,
SMITH, T. A., Recent advances in the biochemistry of plant amines, 8th ed., Lea and Febiger, Philadelphia, 1981.
Prog. Phytochem., 4, 27-81 (1977c). VERPOORTE, R., R. VAN DER HEDDEN, W. M. VAN GUUK, and H.
SMITH, T. A., Plant Amines, in Secondary Plant Products (E. A. J. G. TEN HOOPEN, Plant biotechnology for the production of
Bell and B. V. Charlwood eds.), 433-456, Springer-Verlag, alkaloids: Present status and prospects, in The Alkaloids, Vo!.
Berlin, 1980. 40 (A. Brossi, ed.), 1-188, Academic Press, New York, 1991.
SMITH, T. A., Amines in food, Food Chern., 6, 169-200 (l98!). VOGEL, S., Ecophysiology of zoophilic pollination, in Physiological
Plant Ecology ill (0. L. Lange, P. S. Nobel, C. B. Osmond,
SMITH, T. A., Putrescin and inorganic ions, Rec. Adv. Phytochem.,
and H. Ziegler, eds.), 559-624, Springer-Verlag, Berlin, 1983.
18, 7-54 (1984).
WAGNER. H., Nonwsteroid, cardioactive plant constituents, in Eco-
SMITH, T. A., Amines, in Alkaloids and Sulphur Compounds, (P. nomic and Medicinal Plant Research, Vo!. 2 (H. Wagner, H.
G. Waterman, ed.), Vol. 8 of Methods in Plant Biochemistry Hilrino, N. R Farnsworth, eds.), 17-38, Academic Press, Lon-
(p. M. Dey and J. B. Harbome, eds.), 17-49, Academic Press,
don, 1988.
Loodon, 1993.
WALLER, G. R, K. S. YANG, R. K. GHOLSON, and L. A. HADWIGER,
SNEADER, W., Drug Discovery: The Evolution of Modem Medi- The pyridine nucleotide cycle and its role in the biosynthesis
cines, John Wiley & Sons, Ltd., Chichester, 1985. of ricinine by Ricinus communis L., 1. BioI. Chern., 241,
SPECIALIST PERIODIC REPoRTS, The Alkaloids, Vol. 8, Chemical 4411-4418 (1966).
Society, London, 1978. WALLER, G. R and E. K. NOWACKI, Alkaloid BiolOgy and Metabo-
SPECIAUST i'ERIODIC REPORTS, The Alkaloids, Vo!. 9, Chemical lism in Plants, Plenum Press, New York, 1978.
Society, London, 1979. WATERMAN, P. G. and A. I. GRAY, Chemical systematics, Nat.
SPENSER, I. D., Biosynthesis of alkaloids, in Chemistry of the Alka- Prod. Rep., 4, 175-203 (1987).
loids (S. W. Pelletier, ed.), 669-718, Van Nostrand Reinhold, WINK, M .• Allelochemical properties or the raison d'etre of alka-
New York, 1970. loids, in The Alkaloids, Vo!. 43 (G. A. Cordell, ed.), I-lOS
STRUNZ, G. M. and J. A. FlNDLAY, Pyridine and piperidine alka- Academic Press, New York, (1993).
loids, in The Alkaloids, Vo!. 26 (A. Brossi, ed.), 89-183, Aca- YAMASAKI, K., T. TAMAKl, S. UZAWA, U. SANKAWA, and S. SHI-
demic Press, New York, 1985. BATA, Participation of C(j-C 1 unit in the biosynthesis of ephed-
SUZUKI, T. and K. IwAI, Constituents of red pepper species: Chem- rine in Ephedra, Phytochemistry, 12, 2877-2882 (1973).
29
Pyrrolidine, Tropane, Piperidine,
and Polyketide Alkaloids

Introduction PYRROLIDIl'IE ALKALOIDS

Pyrrolidine Alkaloids
Biogenesis of Pyrrolidine Alkaloids The approximately 80 known pyrrolidine alkaloids possess a
Distribution of Pyrrolidine Alkaloids 5-membered nitrogen-containing ring (Massiot and Delaude,
Biological Activity of Pyrrolidine Alkaloids 1986; Pinder, 1993). Several subgroups of pyrrolidine alka-
Alkaloids of the Stemonaceae loids arise by condensation of these units with other mole-
Tropane alkaloids cules. Pyrrolidine bases usually are modified by additional
Biogenesis of Tropane Alkaloids Schiff-base formation, Mannich condensation, and aldol-
Cocaine type processes to yield other alkaloids of this general class.
Biological Activity of Tropane Alkaloids For example, condensation of pyrrolidine derivatives with
Distribution of Tropane Alkaloids nicotinic acid is involved in the formation of pyridine alka-
Piperidine Alkaloids loids such as nicotine (see Chapter 28). Pyrrolidine units
Alkaloids from Piper Species react with acetyl- or malonyl-CoA and condense via a Man-
Piperine Alkaloids Involving Condensation with nich condensation to form compounds such as hygrine and
Acetate/Malonate cuscohygrine and tropane alkaloids (see below).
Sedum Alkaloids Biogenesis of Pyrrolidine Alkaloids
Lobelia Alkaloids
Lycopodium Alkaloids Pyrrolidine alkaloids are derived via the intermediacy of
Piperidine Alkaloids in Insects putrescine (1). Putrescine is derived by decarboxylation of ar-
Polyketide-Derived Alkaloids ginine or ornithine; the enzyme ornithine decarboxylase (E.C.
Coniine 4.11.1.17) has been detected in tobacco roots (Leete, 1980)
Polyketide Alkaloid Phytotoxins (also see Chapter 30). Arginine decarboxylase (E.C. 4.1.1.19)
Polyhydroxy Piperidine Alkaloids first converts arginine to agmatine, and subsequently to car-
References bamoylputrescine, and then to putrescine (see Fig. 28.7 of
Chapter 28). In several studies, arginine, rather than ornithine,
appears to be the major precursor (Leete, 1990).
INTRODVcnOI'l
In tobacco, label is introduced into both the 2' and 5'
A number of alkaloids are derived from ornithine and lysine carbons of nicotine (2), indicating that the free diarnine is
by decarboxylation, conversion by amine oxidases to the cor- involved; that is putrescine occurs as a symmetrical interme-
responding aldehyde, and cyclization to a 5-membered ring diate at some stage of the biosynthetic process. Further, hy-
system (pyrrolidine alkaloids) and a 6-membered ring system grine and cuscohygrine formed from [5- 14C]ornithine are
(piperidine alkaloids). Several other alkaloids that coinciden- labeled equally at the C-2 and C-5 positions (Leete, 1990).
tally have similar structures have been discussed under the In instances in which an asymmetric intermediate is in-
groups of alkaloids to which they are most closely related. . volved, methylated and enzyme-bound forms of putrescine
Both lH_ and 13C_NMR spectra of the alkaloid types have been suggested. The N-methyl groups of N-methylpyr-
above and the nuclear magnetic resonance (NMR) and mass rolidine are derived from S-adenosyl-L-methionine (Fig.
spectroscopy (MS) analysis of tropane alkaloids have been 29.1) (Geissman and Crout, 1969) and their formation in-
reviewed (Crabb, 1982; Lounasmaa, 1988). volves putrescine-N-methyltransferase (Leete, 1980, 1990).

531
532 Pyrrolidine, Tropane, Piperidine, and Polyketide Alkaloids

,H,NH, ,H,NH, CH,NHCH, CH,NHCH, ~

I I
iH, iH, iH, iH,
CHz ---+- CHz CH, __ N OH
~ CHz ---+-
I
CHNH,
I I I I
CH,NH, CH,NH, CHO CH,
I
o
L
CO,H putrescine N.methylputrescine
(may be bound to H·OH

Q c;j (J"
ornithine
enzyme)

o
(or arginine)

N N+
N I I I
CH, CH, CH,
Al.pyrrolideine N·methylpyrrolldine N.methyl.Al.pyrroUnium cation (3)

Fig. 29.1. Biogenesis of N·methylpyrrolidine and modification of the N·methylpyrrolidine structure (modified from Leete, 1990; used with pennission
of the copyright owner, Georg Thieme Verlag, Stuttgart).

An enzyme that catalyzes the fonnation of 4-N-methyl- proposed to condense with acetoacetate (in torn, derived
aminobutanal from N-methylputrescine has been isolated from the condensation of two units of acetyl-CoA) (Herbert,
from several sources. The enzyme that fonns the N-methyl. 1986) to produce the pyrrolidine alkaloid (R)-hygrine (4)
b.1-pyrrolinium salt (3), N-methylputrescine oxidase, has that is known from several plant families. Acetic acid is
been isolated from tobacco roots (Leete, 1980, 1990). incolporated into this part of hygrine and tropane alkaloids,
Ornithine is incolporated into pyrrolidine rings unsym- but despite several efforts, an enzyme capable of catalyzing
metrically in several Datura species, but a symmetrical inter- this condensation has not been identified. The product is
mediate is involved in Nicotiana species, Erythroxylum fonned, in some instances, after denaturation of proteins.
coca, Duboisia leichardtii, Hyoscyamus albus, and Nicandra Labeling stodies for tropane alkaloids (see below) suggest,
physaloides (Leete, 1990). however, that acetate or malonate may be added in steps
The first product of the cyclization, N-methyl-b. l_pyrrol_ (Fig. 29.2) (Leete, 1990).
ideine (or the N-methyl-b. l-pyrrolinium cation) (3), has been Subsequent reaction of hygrine with another unit of N-

Q
o

!::NH, )-OH ~_~

n r:::. CH n
J
bH, COS·CoA
I -- N+# \- N S.CoA N
CHNH, I COSCoA I I
bo H CH, CH, -CH -CO,H CH,

[2.":1.
ornithine
(3) ~\OSCOA / (19)1
COS·CoA

l.,/ (fl ~
OJJ:) N N
~I+/
~
JH,
hygrine (4)

I I
CH, CH,
cuscobygrine (5)

Fig. 29.2. Biogenesis of hygrine and cuscohygrine (modified from Leete. 1990; modified and used with pennission of the copyright owner, Georg
Thieme Verlag. Stuttgart).
 Pyrrolidine, Tropane, Piperidine, and Polyketide Alkaloids 533

CH'.O~
I HO HO~
CH.O~HO
7 P,
~ , ~ h
CH,·O
o NH' 0 NH NH

ruspolinone (6) norruspolinone (7) norruspolin

OH

HO CH,O

OH 0
ftcin.(8) vocbysine (10)

~ .0

~v HN
CH,O
o
~oH
phyUospadin. (11) Isoneln. (9) H·odorlnol (11)

Fig. 19.3. Selected pyrrolidine alkaloids.

methyl-a l-pyrrolideine (3) yields cuscohygrine (5) (known In addition to a number of piperidine alkaloids, at least
only from the Convolvulaceae, Erythroxylaceae, and Sola- 19 pyrrolidine alkaloids have been reported from secretions
naceae) (Massiot and Delaude, 1986). Cuscohygrine fre- of thief ants and ftre ants of the genera Solenopsis and Mono-
quently is a by-product of the production of tropane alkaloids morium. In general, these compounds occur in minute
in tissue and cell cultures (Verpoorte et a1., 1991). amounts (50 fLglant) (Jones and Blum, 1983; Massiot and
Deprotonation of the I-methyl-a l_pyrrolinium salt (3) Delaude, 1986; Numata and Ibuka, 1987).
yields 1-methyl-a2-pyrroline, which is in equilibrium with
the salt (Leete, 1990). BiolOgical Activity of PyrroUdine Alkaloids

Distribution of PyrroUdine Alkaloids These alkaloids are minor products in many important
drug plants, especially those of the genus Erythroxylum
As these alkaloids are derived by a short biosynthetic (Erythroxylaceae) and of the Solanaceae. In general, the ac-
sequence involving reactions that appear to be widespread tivity of pyrrolidine alkaloids is overshadowed by the more
in plants, it is not surprising that pyrrolidine alkaloids are active tropane alkaloids with which they co-occur (Massiot
commonly encountered. Many of the families in which pyr- and Delaude, 1986).
rolidine alkaloids are found are not particularly closely re- (- )-Odorinol (12) (Fig. 29.3) from Aglaia odorata (Mel-
lated. Monocotyledenous and dicotyledenous angiosperms iaceae), a cinnamic acid and isovaleric acid diamide, has
and ferns are represented (Massiot and Delaude, 1986). In antileukemic properties (Nahrstedt, 1985).
most cases, these alkaloids are found only sporadically
among the various members of these families. Alkaloids of the Stemonaceae
Pyrrolidine alkaloids such as ruspolinone (6) and norrus-
Alkaloids of the roots of plants of the Stemonaceae have
polinone (7) have been found in RuspoJia hypercrateriformis
insecticidal activity. Several alkaloids have been isolated
(Acanthaceae) (Fig. 29.3) (Roessler et a1., 1978). These com-
from the root extracts that have been used to eradicate zoo-
pounds are similar to the proposed precursors of the alkaloids
parasites from man and agricultural animals. Among these
of Tylophora species (see Chapter 30).
are stemospironine (13), stemofoline (14), and stemonine
An unusual type of pyrrolidine alkaloids involving flavo-
(15) (Fig. 29.4) (Sakata et a1., 1978).
noid structures (8-11) has been reported from the Combreta-
ceae, Moraceae, Vochysiaceae, and Zosteraceae (Fig. 29.3)
(Houghton, 1987; Leete, 1982; Massiot and Delaude, 1986). TROPANE ALKALOIDS
Similar alkaloids are found in the Rubiaceae and Meliaceae
(Houghton, 1987); none of these families are particularly Alkaloids formed by "bridging" the 5-membered ring
related. The NMR spectra of these compounds have been with a three-carbon portion derived from acetate/malonate
reviewed (Houghton, 1987). are commonly known as "tropane" alkaloids (Fig. 29.5).
534 Pyrrolidine, Tropane, Piperidine, and Polyketide Alkaloids

...CH! /CH J

N'\r--(CO,CH, • :~' Ho?Q

~OCOC.,H,
H
7 4
3
H
• OCOCH(CH,OH)C.,H,
cocaine (16) hyoscyamine (18) (-)-tropic acid (22)

,CHJ ,CHJ

'~~oa;'~:J--
convoJamine
00 OCHJ tropine senecioic ester
0

valeroidine scopolamine (17) tropinone (20)

Fig. 29.5. Some representative tropane alkaloids.

About 300 compounds with the characteristic 8-azabi- and tropane-type alkaloids. The" production of tropane alka-
cycIo[3.2.11octane system are known (Leete, 1990; Ver- loids in plant tissue, cell, and hairy root cultures has been
poorte et al., 1991; Wooley, 1993). The methods forisolation reviewed (Verpoorte et al., 1991).
and purification of tropane alkaloids have been reviewed
(Wooley, 1993). Biogenesis of Tropane Alkaloids
Tropane alkaloids are produced in the roots of plants and
transported to the aereal portions. In grafting experiments, Tropane alkaloids result from addition of
when aereal portions of Datura ferox, D. stramonium, D. acetate/malonate to a precursor such as the 1-methyl-~ ,_
innoxia, and Atropa belladonna were grafted onto roots of pyrrolinium cation (3), followed by intramolecular cycIiza-
Cyphomandra betacea (normally not alkaloid containing), tion (Leete, 1990) (Figs. 29.2 and 29.6). (R)-Hygrine (4) in
the aereal portions lacked alkaloids. In the reverse situation, several plants, primarily those of the Solanaceae and Eryth-
aereal portions of C. betacea plants grafted onto roots of roxylaceae, may react intramolecularly to form tropane alka-
the other species contained tropane alkaloids (Waller and loids (Fig. 29.6). Pyrrolidine alkaloids, such as (R)-hygrine
Nowacki, 1978). (4) and cuscohygrine (5), frequently co-occur with tropane
Although the production of tropane alkaloids in tissue alkaloids.
culture of solanaceous plants usually is low, hairy root cul- Although all tropane alkaloids are derived from ornithine
tures of Duboisia leichhardtii produced 1.1 % (dry weight) or arginine, some differences in intermediates and labeling
of scopolamine (Ellis, 1988). Cultures of this genus have patterns have been observed. On introduction of [5- 14C]or-
been studied widely because they synthesize both nicotine- nithine into plants of Erythroxylum, the activity was divided

stemonine (15) stemofoline (14)

stemospironine (13)

Fig. 29.4. Alkaloids of the Stemonaceae.

 Pyrrolidine. Tropane. Piperidine, and Polyketide Alkaloids 535

between C-1 and C-5 of the bridgehead of cocaine (16). bert, 1988; Leete, 1984). The configuration of the f3-carbon
These studies indicate that a symmetrical intennediate (i.e., of phenylalanine is preserved in (S)-tropic acid (Herbert,
putrescine) is involved in the fonnation of tropane alkaloids 1988; Leete, 1984).
(Leete, 1983b, 1990). Incorporation of [1- 13C,methylamino- Large changes in the amounts of atropine [a mixture of
"N]-N-methylputrescine into scopolamine (17) in Duboisia (R)- and (S)-hyoscyamine] present in Atropa belladonna
provided evidence that free putrescine was not involved, at occur diurnally and seasonally (Waller and Nowacki, 1978).
least beginning with this intennediate, as only one 13C_15N The amount of atropine in leaves is greatest at about 6:00
coupling was observed in the 13C_NMR spectrum (Mann, P.M., whereas the atropine concentration of the froits peaks
1987). When [2-14C]ornithine is introduced into root cul- at about 4:00 P.M. The smallest amount of atropine in the
tures of Hyoscyamus albus, hyoscyamine (18) labeled at leaves occurs at 10:00 A.M. (Waller and Nowacki, 1978).
both bridgehead carbon atoms C-1 and C-5 is isolated. How- Although undifferentiated calli of this species do not produce
ever, in similar experiments, in which the same precursor is hyoscyamine (18), production resumes when roots fonn on
introduced into root cultures of Datura stramonium, hyoscy- the callus. Thus, the accumulation of this alkaloid seems to
amine (18) labeled at only bridgehead carbon atom C-l is be developmentally regulated. Root cultures of Hyocyamus
isolated (Leete, 1990). Clearly, plants differ in whether sym- niger also produce hyoscyamine (Flores et aI., 1987).
metrical or unsymmetrical intennediates are involved in the A series of polyhydroxylated tropane alkaloids have been
fonnation of tropane alkaloids. isolated from the leaves of Solanum tuberosum and from
Although it may appear that the double bond in the imi- both a sphingid moth and an ithomiine butterfly, the larvae
nium salt (3) can isomerize between the C-2 and the C-5 of which feed on Solanum species (Nash et al., 1993). These
positions (which could lead to resultant scrambling of iso- alkaloids are potent glycosidase inhibitors, in a manner simi-
topic label at these positions), this does not occur in either lar to certain hydroxypyrrolidine and indolizidine alkaloids
acidic or basic media (Leete, 1990). Furthennore, studies in (see Chapter 30).
which appropriately labeled intennediate (3) is introduced Hybrids of Datura ferox [virus susceptible and hyoscine
into plant cultures or intact plants indicate that incorporation (23) producing] and D. stramonium [virus resistant and hyo-
occurs in an unsymmetrical manner. For example, in biosyn- scyamine (18) producing] produced vigorous virus-resistant
thetic studies in which [2-14C]-I-methyl-,~.I-pyrrolinium plants that produced hyoscine, an alkaloid important in med-
chloride (3) was fed to plants of Erythroxylum coca, the icine. The F 1 plants inherited the high alkaloid level of D.
resultant cocaine was labeled at the C-5 and not the C-1 stramonium, but the oxidative ability of D.ferox. The segre-
position as suggested by previously proposed biosynthetic gation ratio in the F2 generation is 3 : 1 and hyoscine produc-
schemes. The presence of the carboxyl group of the methyl tion is dominant (Waller and Nowacki, 1978).
ester in cocaine indicates that the original attack is at C-2 of
the [2_14C]-I-methyl-A I-pyrrolinium chloride (3) precursor. Cocaine
Attack by a methyl group (or a singly activated methylene Probably the best known alkaloid of this group is cocaine
group) as previously proposed is without precedent, but the (16), derived from the coca plant (Erythroxylum coca, Eryth-
presence of label in C-5 (and not C-l) of cocaine precludes roxylaceae) (White, 1989). This polymorphic species is a
this mode of cyclization, in any case. In order to accommo- native of the Andes of nurthwestern South America. The
date these data, Leete (1990) proposed that the initial attack leaves of the plant, which contain 0.5-2% cocaine, are
involves one molecule of malonyl-CoA, followed by exten- chewed by many Andean Indians who are said to derive a
sion by a second unit, to yield the required intennediate (19) feeling of well-being and alleviation of hunger pangs
(Fig. 29.6). This saturated compound is then converted to thereby. There is no concomitant hallucinogenic effect. Most
the corresponding iminium compound (24). Intramolecular commercially cultivated coca for the pharmaceutical indus-
closure occurs between the methylene group of the side chain try comes from Peru and Bolivia (Boucher, 1991; Tyler et
(fonned by the sequential addition of two molecules of malo- al., 1981; White, 1989).
nate) and C-5 of the precursor. Cocaine binds and inhibits the dopamine uptake carrier
The results of a number of feeding studies leading to (Wink, 1993). This alkaloid currently is a drog of abuse in
tropane alkaloids have been tabnlated (Leete, 1983a, 1990). many countries. The widespread method for illicit
Tropinone (20) is a key intennediate in the synthesis of extraction/processing of cocaine is an interesting example
many tropane alkaloids, for example, hyoscyamine (18) [an of alkaloid isolation and properties. Leaves of the coca plant
ester of tropine (21) and (S)-tropic acid (22)] and scopolam- are dried and then soaked in a mixture of water and kerosene.
ine (17) (Leete, 1990). Scopolamine is fonned from 613- The resulting paste is then mixed, in tum, with dilute sulfuric
hydroxyhyoscyamine by a direct attack of the 6f3-hydroxyl acid, lime, potassium pennanganate, and more kerosene. Fi-
group at C-7, displacing the 7f3-hydrogen. The requisite en- nally the crude alkaloidal paste is extracted with ether and
zyme, 6f3-hydroxyhyoscyamine epoxidase, requires 2-keto- acetone to remove impurities (Anon., 1985; Boucher, 1991).
glutarate and oxygen as cofactors (Leete, 1990). Cocaine (16) has been used medicinally as a local anes-
Many tropane alkaloids occur as esters of (S)-tropic acid thetic. Although cocaine itself is not as widely used in medi-
(22), a moiety derived from phenylalanine (Fig. 29.7) (Her- cine as was fonnerly the case, the synthesis of a number of
 w- " -
536 Pyrrolidine, Tropane, Piperidine, and Polyketide Alkaloids

~ .(C"",-
CH, CH COSCOA(CO'H
COSCoA ''-..._

COSCoA HO,C
o

~.-~"
·
CoAS·CO

(24) 0 o OH

R.
tropinone (20) /
tropine (21)
I CH ,

-
N 2

,
H
,
OCOCH(CH,OH)C,H,

.p C,=~ .~
(. )-tropic acid (22) hyoscyamine (18)
<a)

.. ~ ~
~SCOA CH,
COSCoA

o o

~~.t;Jy~~
o o 0 0
(19)

/H, N
/CH,
N

~
CO'CH'
f r = - < C 0 2CH, _ _....
, 2

.~ ~OH • 4 OCOC,H,
H H'

(b)
methyl ecgonine cocaine (16)

Fig. 29.6 (a & b). Biogenesis of tropane alkaloids (modified from Leete, 1990; used with permission of the copyright owner, Georg Thieme Verlag,
Stuttgart).
 Pyrrolidine, Tropane, Piperidine, and Polyketide Alkaloids 537

H01C\ cholineesterase inhibitors such as physostigmine or organo-

~
T
phosphate insecticides. Atropine binds to muscarinergic ace-
I~ OH
tyl choline receptors (Wink, 1993). Scopolamine (17) has
similar properties. The LCso p.o. (per as, or oral) in rat is
"'" H H 750 mg/kg (Wink, 1993). Incorporation of atropine at 0.1%
into artificial diets is totally lethal to the larvae of Calloso-
(8)-(. )-tropic acid (22)
bruchus maculatus (Janzen et al., 1977). Atropine is a feed-
Fig. 29.7. Biogenesis of tropic acid (modified from Leete. 1990; used ing deterrent for several insects (Wink, 1993).
with pennission of the copyright owner, Georg Thieme Verlag, The corresponding 6,7-epoxide, scopolamine (17), is pro-
Stuttgart).
duced only in young leaves. This alkaloid inhibits germina-
tion of the seeds of several plants and inhibits growth of the
other local anesthetics was based on the structure of this radicle in Lepidium and Lactuca. Scopolamine has been used
alkaloid (Tyler et aI., 1981). Cocaine is generally considered as a premedication prior to surgery and as a powerful hyp-
too toxic for use by injection, but it is used as a 0.1 % solution notic in the treatment of the delirium tremens (DTs) (Cordell,
as a topical anesthetic and 10-20% solutions for other appli- 1981). Scopolamine also is widely used to lreat motion sick-
cations (Cordell, 1978). The LDso i.v. of cocaine is 17.5 ness (Tyler et al., 1981). This alkaloid is a feeding deterrent
mg/kg (Wink, 1993). for several insects (Wink, 1993).
Cocaine (16) inhibits germination of the seeds of certain
Biological Activity of Tropane Alkaloids plants and is a feeding deterrent for the insect Leptinotarsa
(Wink, 1993).
A number of Solanaceous plants have a long folkloric
history in many parts of the world. All are poisonous and
Distribution of Tropane Alkaloids
contain tropane alkaloids (Cutler, 1992). Among these are
Atropa belladonna, Mandragora officinarum, and species Tropane alkaloids are common in the Solanaceae, but are
of Hyoscyamus, Latua, Duboisia, Datura, Brugmansia, and found in several other plant families. Tropane alkaloids are
Brunjelsia. The juice of the fruits of belladonna was placed found in the tribes Ceslreae, Datureae, Nicandreae, Salpig-
in the eyes of women in former times in order to expand the lossideae, and Solaneae (Evans, 1979). These alkaloids also
pupils and make the eyes darker. This lead to the name are found in the Convolvulaceae, which most systematists
"bella donna" or beautiful lady (Cutler, 1992). Mandrake consider to be related closely to the Solanaceae (Waterman
(Mandragora) was considered by many as a miracle drug. and Gray, 1987). Other families that contain tropane alka-
The root of this plant was also considered an aphrodisiac. loids are the Brassicaceae (Cochlearia), Dioscoreaceae,
Several of these species contain alkaloids that imparted a Elaeocarpaceae, Erythroxylaceae, Euphorbiaceae (Phyllan-
feeling of flying and were involved in witchcraft in Europe thus), Orchidaceae, Proteaceae, and Rhizophoraceae (Dahl-
and in ritualistic uses in other parts of the world (Schultes grenetal., 1981; Romeike, 1978; Waterman and Gray, 1987;
and Hoffman, 1973). Datura stramonium has been reported Wooley, 1993). Because other types of alkaloids are found
to be used in Haiti in association with the catatonic state in several of these families, many reports should be con-
associated with zombies (Anon., 1987). A number of plants firmed (Romeike, 1978).
containing tropane alkaloids have been used in Chinese tra-
ditional medicine (Han et al., 1988).
These compounds possess potent physiological effects in FIPBRlDIl'IE ALKALOIDS
animals and have long been of interest in medicine. Many
have mydriatic activity (i.e., they cause dilation of the pupil Piperidine alkaloids arise from lysine in much the same way
of the eye). Others, such as cocaine, are anesthetics (Leete, that pyrrolidine alkaloids arise from ornithine. A total of
1990). about 700 alkaloids of these two groups are known (Ver-
Hyoscyamine (18) is produced in the roots of Datura poorte et al., 1991). The product of decarboxylation of ly-
(Solanaceae), but not in the leaves. This is the most common sine, cadaverine, generally seems to be incorporated as an
alkaloid in solanaceous plants (Cordell, 1978). This alkaloid unsymmetrical intermediate, but, in some instances, a sym-
inhibits germination of certain seeds, inhibits radicle growth metrical intermediate may be involved. This has been inves-
in Linum, and is toxic to Lemna (Wink, 1993). Hyoscyamine, tigated by feeding [6_I5NJ-Iysine and [2_1SNJ-Iysine (Fig.
usually obtained from Hyoscyamus species) is a powerful 29.8). A pathway involving a bound form of cadaverine (25)
anticholinergic drug. Hyoscyamine is a feeding deterrent for to pyridoxal that accounts for most observed chemical data
several insects (Wink, 1993). has been proposed (Spenser, 1970) (Fig. 29.8). Decarboxyl-
Atrnpine [a mixture of (R) and (S)-hyoscyamineJ, usually ation occurs with retention of configuration.
obtained from Atropa belladonna or from Hyoscyamus or The results of a number of feeding studies and the forma-
Duboisia species (all Solanaceae), is formed from hyoscya- tion of piperidine alkaloids have been tabulated (Leete,
mine in the process of extraction. Atrnpine is a potent anti- 1983a).
cholinergic drug that is sometimes used as an antidote for Piperidine alkaloids of the Apiaceae, such as coniine, and
S38 Pyrrolidine. Tropane, Piperidine, and Polyketide Alkaloids

~ L·lysine ~
H,N) H~~H~O'H H,N) H~~·CO'H

IXH
CHO
"
CH "
CH

~OH ~H
ll.,A
)\
N N / N

.~f:\N CH .J l
I Ie
~I OH H,N NH,
S-aminopentenal

o-
l1l>.ru~ cadaverine (25)

O
+
N:m'l face ('i H 0

00
N ~ ~NH~
l\,!.piperideine
~ from Fe face 2(R).a1ka1oids
(for example, (lR)-pelletierine)

H2N~( l .HA
O.NH

CO,H
2
NH ,::v
2(S)-aIkaloids
I (for example, (2S)·anabasine)
D·lysine ~

H'N~CO'H 6,1.piperideIne carboxylic acid (30) pipecolic acid (29)

Fig. 29.8. Biogenesis of piperidine alkaloids (modified from Leistner and Spenser, 1973; used with pennission of the copyright owner, the American
Chemical Society, Washington, DC).

those of coniferous gymnosperms are derived from polyke- size piperidine alkaloids. However, it is derived from D-
tide pathways (see below). lysine rather than L-Iysine and probably via 6,'-piperideine
In the tobacco aIkaloids anabasine (26) and anatabine carboxylic acid (30) (Fig. 29.8) (Leete, 1983a; Strunz and
(27), a lysine derivative is condensed with a derivative of Findlay, 1985).
nicotinic acid to yield a system reminiscent of nicotine (Fig.
29.9) (Leete, 1977, 1983a). Alkaloids of Piper Species
Anabasine (26) has antisinoking properties similar to lo-
beline (see below) (Cordell, 1978). Structurally similar com- Several representatives of the genus Piper (Piperaceae)
pounds are known from ants and other animals (Nahrstedt, are of economic importance. Some of these plants [e.g.,
1982). Anabasine is insecticidal and serves as a feeding de- Piper nigrum (black pepper)] include alkaloids that are par-
terrent for several insects and is teratogenic (Wink, 1993). tially responsible for the desirable pungent and preservative
A number of other compounds with similar structures properties of the plant. Other species are P. betle (widely
[e.g., ammodendrine (28) from Ammodendron conollyii, Fa- used as a masticatory), P. methysticum (widely used in the
baceae] are not derived from nicotinic acid, but wholly from South Pacific as a soporific drug), and P. guineense (ashanti
lysine derived units (Fig. 29.10) (Herbert, 1988). pepper). Most of these plants contain compounds that com-
L-Pipecolic acid (29) occurs in many species that synthe- bine phenylpropanoid compounds (see Chapter 8) and pyr-
 Pyrrolidine, Tropane, Piperidine. and Polyketide Alkaloids 539

D -
HaN .........
H02 T

L.[2.3Hllysine
NHCH3
HaN ""'~_
H
~
T
~ --0- TD
NHCH3 0
T NHCH3
7+
CH3

- +nicotinic

acid

anabasine (26) anatabine (27)

Fig. 29.9. Biogenesis of anatabine and anabasine (modified from Gupta and Spenser, 1970; modified and used with pennission of the copyright owner,
Elsevier Science Ltd., The Boulevard, Langford Lane, Kidlington OXS 1GB. UK),

rolidine alkaloid units. The pungent principle of black pep- produce a homologous series of alkaloids by intramolecular
per, piperine (31), has an all E (trans) structure (Fig. 29.11). cyclization. Internal cyclization produces alkaloids corre-
Chavicine (32), another component important in the taste sponding to the tropane alkaloids, but with a 6-membered
properties of pepper, has aZ (cis) structure. Piperlongumine ring system (Fig. 29.12). Many of these alkaloids have been
(33) and piperlonguminine (34) occur in Piper longum or isolated from Punica granatum L. (Punicaceae, pomegran-
long pepper. ate). Compounds analogous to hygrine and cuscohygrine
A number of Piper species, e.g., Piper longum, have been also are known. Examples are anaferine (35) and anahygrine
used in Ayurvedic medicine in India. This drug seems to (36) from Withania somnifera Dunal. (Solanaceae).
increase the bioavailability of other drugs. Piperine (31) is Pseudopelletierine (37) (similar to hygrine in structure)
similar in action to many central nervous system depressants is an active taeniacide and anthelminthic for tapeworm and
(Fodor and Colosanti, 1985). roundworm (Fodor and Colosanti, 1985).

Piperine Alkaloids Involving Condensation Sedum Alkaloids

with Acetate/malonate
A number of other piperidine alkaloids do not involve
The reaction of piperideine precursors, homologous to condensation with acetate or malonate (previously consid-
the N-methyl-.1 I-pyrrolinium cation (3) involved in tropane ered to occur as addition of acetoacetate). Several that occur
alkaloid formation, with acetyl-CoA or ma10nyl-CoA can in the genus Sedum (Crassulaceae) involve condensation of

o OI -- a,. I Q-
N NH NH aD N

- - cP )=0
N

ammodendrine (28)

Fig. 29.10. Biogenesis of ammodendrine (modified from Geissman and Crout, 1969).
S40 Pyrrolidine, Tropane. Piperidine. and Polyketide Alkaloids

o o
~~~OCH'
O ~O
~) ~O YOCH,
OCH,
piperine (31) piperlongumin (33)

piperlonguminine (34) chavicine (32)

Fig. 29.11. Piperine, piperlongumin, and piperolonguminine.

both lysine and phenylalanine moieties (Fig. 29.13). Label (Williams et al., 1987). Lobeline is a feeding deterrent for
is nonrandomly incorporated into sedamine (38), suggesting several insects. The alkaloid is toxic to Lemna and inhibits
the presence of an unsymmetrical intennediate. When cadav- radicle growth of Lepidium (Wink, 1993).
erine (25), chirally labeled with tritium at C-I, is used, the
I-pro-R hydrogen of the diamine is retained at C-2 of N- Lycopodium Alkaloids
methylpseudopelletierine and the I-pro-S hydrogen is lost Approximately toO alkaloids are known from the genus
(Strunz and Findlay, 1985). Lycopodium. These primitive seedless vascular plants, often
known as club mosses, are unusual among primitive plants,
LobeOa AJkaIoids as they contain alkaloids.
Two molecules of lysine and acetate are incorporated into
Another series of lysine derived alkaloids occurs in the lycopodine (40). It has been suggested that isopelletierine
genus Lobelia (Campanulaceae). One of these compounds, (41) plays a central role in the fonnation of these alkaloids
lobeline (39) (Fig. 29.13) (from Lobelia inflata, Indian to- (Fig. 29.14) (Blumenkopf and Heathcock, 1985; MacLean,
bacco), has been used in antismoking preparations. Both lo- 1986).
beline and nicotine are classed as ganglionic stimulants.
Therapeutically, lobeline is hardly used today (Fodor and Piperidine Alkaloids in Insects
Colosanti, 1985). Poisoning of cattle because of ingestion In addition to a number of pyrrolidine alkaloids, many
of Lobelia berlandieri has occurred in Texas and Mexico piperidine alkaloids have been reported from secretions of

Qyr
- CH·CO,H

~ -- ~y- \OSCOA
~d
r
COSCoA
) __ )'·.COH -+ CH·CO,H
H,N HNH,' I -----.. N
I
'-...- --+ --+-
CH, COSCoA CH, CO,H

~COSCOA

('i
~CH,

<:? ~
--COASOC~
c~ QClJvO
o
,
CH,

l..NH~HN,)
anaferine (35)0 pseudopelletierine (37) anagyrine (36)

Fig. 29.12. Biogenesis of pseudopelletierine.

 Pyrrolidine, Tropane, Piperidine, and Polyketide Alkaloids 541

lysine
0.
Vk.
OR o

u-
CO'R ~CO'R
~
CO'R

I ~
:::,... NR, I~-

o OR

N
I
CR,

(-)-sedamine (38) (-)-Iobeline (39)

Fig. 29.13. Biogenesis of sedamine (modified from Geissman and Crout, 1969),

0{)
~NR~
isopelletierine (41)

ro ~R

C)6; ~F ~~~H-
OR

~-~"~~"
j~ .<9-
~ N NR N 0
+- ~.OR OR

I N+ 0
CR, 0 - I
a-obscurine lycopodine (40)
• label from (2_14CJ-lysine

Fig. 29.14. Lycopodium alkaloids (modified from Geissman and Crout, 1969).
542 Pyrrolidine, Tropane, Piperidine, and Polyketide Alkaloids

H H

coccinelline (42) precoccinelline (43) convergine (44)

\f.
H

hippodemine (45) myrrhine (46) propyleine (47)

Fig. 29.15. Coccinellines.

thief ants and fire ants of the genera Solenopsis and Mono- ants and may be involved in deterring predation by other
morium. In general, these compounds occur in minute animals as well (Jones and Blum, 1983). Structurally similar
amounts (50 fLglant) (Jones and Blum, 1983; Massiot and compounds have been reported from the genus Poranthella
Delaude, 1986). The alkaloids often are components of the of the Euphorbiaceae (Nahrstedt, 1982; Numata and Ibuka,
venom, but they also serve as a repellent for other ant species 1987).
and possibly as trail pheromones (Jones and Blum, 1983;
Numata and Ibuka, 1987).
Other lysine-derived alkaloids known as coccinellines
(42-47) occur in many members of the family Coccinellidae POLYIillTIDE-DERIVBD ALKALOIDS
(ladybugs or ladybird beetles) (Fig. 29.15). Many of these
insects are also warningly colored. Cryptic members of the A number of polyketide-derived alkaloids are known to
same groups usually do not contain the alkaloids (Jones and occur. Probably best known among these are coniine (48)
Blum, 1983). These alkaloids also have been isolated from and related compounds, found primarily in the Apiaceae
a soldier beetle, Chauliognathus and from the boll weevil (mostly in the genus Conium) (Leete, 1971). Other alkaloids
Anthonomus grandis. Coccinellines are highly repellent to with similar origin are pinidine (49) from pines, the Galbuli-

HO,O
" ~, . . NH", (CH,JIOCOHCH,

cassine carnavaline prosopine

Cassia excelsa Cassia spec/obi/is Prosopis ajricaIUJ

H:Q NH (CH')12COCH,

isoprosopinine A julifloridine spectaline

Prosopis africana Prosopis juliflora Cassia spectabilis

Fig. 29.16. Piperidin-3-o1s from Cassia and Prosopis.

 Pyrrolidine. Tropane, Piperidine. and Po/yketide Alka/oids 543

[l.14C].acetate _ _ ('l __ ( ' l --+ H ('I --+

HO,t ~ HO,t o~ ;j o~

~~ N4
-----'>
~ • N •
H •

'Y~coniceine (51)
coniine (48)

r'l H . •OH

/~NH~
plnidine (49) pinidinol (50)

Fig. 29.17. Biogenesis of coniine and related alkaloids (modified from Cordell, 1981; used with consent of the author).

mina alkaloids from the family Himantandraceae, and alka- guinea pig is 150 mg/kg. Coniine and coniceine are terato-
loids from the genus Elaeocarpus of the Elaeocarpaceae. genic (Wink, 1993).
Alkaloids are uncommon in the Pinaceae. ( - )-Pinidine Both diurnal and seasonal variations in the content of
(49) from Pinus sabiniana and P. jeffreyi is an example of coniine and 'Y-coniceine in Conium maculalum have been
piperine alkaloids from this gymnospennous group. (-)- observed (Waller and Nowacki, 1978). The half-life of coni-
Pinidinol (50) occurs in Picea engelmanii, as well as other ine (48), 'Y·coniceine (51), andN-methylconiine is 1-4 days.
species of spruce (Stennitz et al., 1990). The biosynthesis of Coniine (48) is a feeding deterrent for several insects
these compounds from acetate has been demonstrated (Fodor (Wink, 1993).
and Colosanti, 1985; Schneider and Stermitz, 1990; Stermitz
et al., 1990; Strunz and Findlay, 1985). Polyketide Alkaloid Pbytotoxins
A series of substituted piperidine alkaloids have been iso-
lated from the legume genera Cassia (subfamily Caesalpini- Several compounds such as fnsaric acid (52) and a-picoli-
oideae) and Prosopis (subfamily Mimosoideae) (Fig. 29.16). nic acid (53) (Fig. 29.18) (from the pathogenic fungi Fu·
(Strunz and Findlay, 1985). Based on the structures of these sarium oxysporum and Pyricularia oryzae, respectively) are
compounds, it seems possible that they are derived from phytotoxins. Both are probably involved in chelation of fer-
acetate-malonate precursors. These compounds are quite ric and zinc ions (Stoessl, 1981).
similar in structure to the venoms of fIre ants (Nahrstedt,
1982). Indolizidine alkaloids also have been isolated from Polybydroxy Piperidine Alkaloids
Prosopis species (Howard and Michael, 1986). A series of piperidine alkaloids with hydroxyl substitu-
tions have been isolated from a number of higher plants
Coniine (54-56) (Fig. 29.19) (Herbert, 1986, 1988). Among these
Although the alkaloids coniine (48) and 'Y-coniceine (51) plant species are Lonchocarpus sericeus, L. coslaricensis
bear a structural resemblance to the piperidine alkaloids, and Derris elliplica (Fabaceae), Morus species (Moraceae),
these compounds are derived from a polyketide pathway F agopyrum esculenlum (Polygonaceae), and a fern of the
(Fig. 29.17). Lysine is a poor precursor, and early attempts
to show incorporation of this compound resulted in failure.

~CO'H
Acetate is a much better precursor. Coniine is a highly toxic
alkaloid and is one of the toxic components of poison hem-
lock (Conium maculalum, Apiaceae) (Cutler, 1992). Other-
wise, alkaloids are very uncommon in the Apiaceae. Coniine
does occur in several other plants, for example, Sarracenia
(Sarracenia). 'Y-Coniceine is found in several species of Aloe fusarie acid (52) a~picolinle acid (53)
(a~pycolic acid)
(LiJiaceae) (Dring et al., 1984). Coniine is toxic to the
aquatic plant Lemna (Wink, 1993). The LDlOo p.o. in the Fig. 29.18. Fusaric acid and a~picolinic acid.
544 Pyrrolidille, Tropane, Piperidine, and Polyketide Alkaloids

oo'(:c 00, Ceo"CH20~NH~CH20H

OH

NH
A~OH 00)::(
CH20H
OH

NH
OH

HOCH{ NH CH 20H

deoxymannojirimycin (54) deoxynojirimycin (55) fagomine (56) a fructose analog from Derris elliptica

Fig. 29.19. Polyhydroxy plant alkaloids.

Aspidiaceae (FeJlows et aI., 1986; Janzen et aI., 1990). These and Taxonomy of the Solanaceae (1. G. Hawkes, R. N. Lester,
compounds are similar in some regards to coniine (48), a and A. D. Skelding, eds.), Linnean Society of London Sympo-
nonhydroxylated piperidine alkaloid. In some legumes, sium, Series 7, 241-254, Academic Press, London, 1979.
polyhydroxy piperidine alkaloids make up as much as 2% FELLOWS, L. E., S. V. EVANS, R. J. NASH, and E. A. BELL, Polyby-
of the dry weight of the plant. These compounds are similar droxy plant alkaloids as glycosidase inhibitors and their possible
structuraJly to sugars and are potent inhibitors of glycosidase ecological role, in Natural Resistance of Plants to Pests (M. B.
Green and P. A. Hedin, eds.), ACS Symposium Series 296,
activity. Specificity varies and some compounds of this se-
72-78, American Chemical Society, Washington, DC, 1986.
ries inhibit mannosidases, others fucosidases, and so forth.
FLORES, H. E., M. W. Hoy, and 1. 1. PICKARD, Secondary metabo-
No glucosidase inhibitor has been reported to present (Fel-
lites from root cultures, Tibtech, 5, 64-69 (1987).
lows et aI., 1986) (also see Chapter 15).
FODOR, G. B. and B. COLOSANTI, The pyridine and piperidine alka-
loids: Chemistry and phannacology, in Alkaloids. Chemical and
Biological Perspectives, Vol. 3 (S. W. Pelletier, ed.), 1-90,
REFERENCES Wiley, New York, 1985.
GEISSMAN, T. A. and D. H. G. CROUT, Organic Chemistry of Sec-
ANON., Time, Feb. 25, 1985, p. 32.
ondary Plant Metabolism, Freeman Cooper, San Francisco,
ANON., American Scientist interviews Wade Davis, Am. Sci., 75. 1969.
412-417 (1987).
GUPTA, R. N. and I. D. SPENSER, Biosynthesis of the piperidine
BLUMENKOPF, T. A and C. H. HEATHCOCK, Synthesis of Lycopo- nucleus: The occurrence of two pathways from lysine, Phyto-
dium alkaloids, in Alkaloids: Chemical and Biological Perspec- chemistry, 9, 2329-2334 (1970).
tives, Vol. 3 (S. W. Pelletier, ed.), 185-240, Wiley, New York,
HAN, G., Y. CHEN, and X. LIANG, Distribution of alkaloids in tradi-
1985.
tional Chinese medicinal plants, in The Alkaloids, Vol. 32 (A.
BOUCHER, D. H., Cocaine and the coca plant, BioScience, 41, Brossi, ed.), 241-270, Academic Press, New York, 1988.
72-76 (1991).
HERBERT, R. B., The biosynthesis of plant alkaloids and nitrogenous
CORDELL, G. A., Alkaloids, in Kirk-Othmer Encyclopedia of
microbial metabolites, Nat. Prod. Rep., 3, 185-203 (1986).
Chemical Technology, Vol. I, 3rd ed., Wiley, New York, 1978.
HERBERT, R. B., The biosynthesis of plant alkaloids and nitrogenous
CORDELL, G. A., Introduction to alkaloids, Wiley, New York,
microbial metabolites, Nat. Prod. Rep., 5, 523-540 (1988).
1981.
HOUGHTON, P. 1., Chromone alkaloids, in The Alkaloids, Vol. 31
CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual
(A. Brossi, ed.), 67-100, Academic Press, New York, 1987.
Reports on NMR Spectroscopy, Vol. !3 (G. A. Webb, ed.),
60-210, Academic Press, London, 1982. HOWARD, A. S. and J. P. MICHAEL, Simple indolizidine and quino-
lizidine alkaloids, in The Alkaloids, Vol. 28 (A. Brossi, ed.),
CUTLER, H. G., An historical perspective of ancient poisons, in
183-308, Academic Press, New York, 1986.
Phytochemical Resources for Medicine and Agriculture (R. G.
Nigg and D. S. Seigler, eds.), 1-13, Plenum Press, New York, JANZEN, D. H., L. E. FELLOWS, and P. G. WATERMAN, What protects
1992. Lonchocarpus (Leguminosae) seeds in a Costa Rican dry forest?
DAHLGREN, R. M. T., S. ROSENDAHL-JENSEN, and B. J. NIELSEN, A
Biotropica, 22, 272-285 (1990).
revised classification of the angiosperms with comments on JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary
correlation between chemical and other characters, in Phyto- compounds to the seed-eating larvae of the bruchid beetle Callo-
chemistry and Angiosperm Phylogeny (D. A. Young and D. S. sobruchus maculatus, Phytochemistry, 16, 223-227 (1977).
Seigler, eds.), 149-204, Praeger, New York, 1981. JONES, T. H. and M. S. BLUM, Arthropod alkaloids: Distribution,
DRING, J. V., R. J. NASH, M. F. ROBERTS, and T. REYNOLDS, Hem- functions, and chemistry, in Alkaloids: Chemical and Biological
lock alkaloids in aloes. Occurrence and distribution of 'Y-conic- Perspectives, Vol. I (S. W. Pelletier, ed.), 33-84, Wiley, New
eine, Planta Medica, 50, 442-443 (1984). York, 1983.
ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod. LEETE, E., Biosynthesis of the hemlock and related piperidine alka-
Rep., 5, 581-612 (1988). loids, Ace. Chern. Res., 4, 100-107 (1971).
EVANS, W. c., Tropane alkaloids of the Solanaceae, in The Biology LEETE, E., The incorporation of [5,6- 13C 2 Jnicotinic acid into the
 PYl'rolidine. Tropane. Piperidine, and Polyketide Alkaloids 545

tobacco alkaloids examined by the use of 13C nuclear magnetic Ruspolia hypercrateriformis M. R.: Isolierung und Strukturauf-
resonance, Bioorg. Chern., 6, 273-286 (1977). kliirung von neuen Pyrrolidin-Alkaloiden, Helv. Chim. Acta,
LEETE, E., Alkaloids derived from ornithine, lysine, and nicotinic 61, 1200-1206 (1978).
acid, in Secondary Plant Products (E. A. Bell and B. V. Charl- ROMEIKE, A., Tropane alkaloids-occurence and systemati'c impor-
wood, eds.), 65-91, Springer-Verlag, Berlin, 1980. tance in angiosperms, Bot. Notiser, 131, 85-96 (1978).
LEETE, E., A one-step synthesis of ficine and isoficine, 1. Nat. Prod., SAKATA, K., K. AOIG, C. CHANG, A. SAKURAI, S. TAMURA, and
45, 605-607 (1982). S. MURAKOSHI, Stemospironine, a new insecticidal alkaloid of
LEETE, E., Biosynthesis and metabolism of the tobacco alkaloids, Stemona japonica Miq. Isolation, structural detennination and
in Alkaloids: Chemical and Biological Perspectives, Vol. I (S. activity, Agric. BioI. Chern., 42, 457-463 (1978).
W. Pelletier, ed.), 85-152, Wiley, New York, 1983a. SCHULTES, R. E. and A. HOPPMAN, The Botany and Chemistry of
LEETE, E., The biosynthesis of alkaloids, in Specialist Periodical Hallucinogens, CC Thomas, Springfield, IL, 1973.
Reports, Biosynthesis, Vol. 7 (R. B. Herbert and T. J. Simpson, SCHNEIDER, M. J. and F. R. STERMITZ, Uptake of host plant a1kaIoids
eds.), 102-223, Royal Society of Chemistry, London, 1983b. by root parasite Pedicularis species, Phytochemistry, 29,
LEETE, E., 1,2-Migration of hydrogen during the biosynthesis of 1811-1814 (1990).
tropic acid from phenylalanine, J. Am. Chern. Soc., 106, SPENSER, I. D., Biosynthesis of alkaloids, in Chemistry of the Alka-
7271-7272 (1984). loids (S. W. Pelletier, ed.), 669-718, Van Nostrand Reinhold,
LEETE, E., Recent developments in the biosynthesis of the tropane New York, 1970.
alkaloids, Planta Medica, 56, 339-352 (1990). STERMITZ, F. R., M. M. MILLER, and M. J. SCHNEIDER, The stereo-
LEISTNER, E. and I. D. SPENSER, Biosynthesis of the piperidine chemistry of ( - )-pinidol, a piperidine alkaloid from Picea en-
nudeus. Incorporation of chirally labelled [I-'H]cadaverine, J. ge/mannii, J. Nat. Prod., 53, 1019-1020 (1990).
Am. Chern. Soc., 95, 4715-4725 (1973). STOESSL, A., Structure and biogenetic relations: Fungal nonhost-
LoUNASMAA, M., The tropane a1kaIoids, in The Alkaloids, Vol. 33 specific, in Toxins in Plant Disease (R. D. Durbin, ed.),
(A. Brossi, ed.), 2-81, Academic Press, New York, 1988. 110-219, Academic Press, New York, 1981.
MAcLEAN, D. B., Lycopodium a1kaIoids, in The Alkaloids, Vol. STRUNZ, G. M. and J. A. FINDLAY, Pyridine and piperidine alka-
26 (A. Brossi, ed.), 241-298, Acadentic Press, New York, 1986. loids, in The Alkaloids, Vol. 26 (A. Brossi, ed.), 89-183, Aca-
MANN, J., Secondary Metabolism, Oxford University Press, Ox- demic Press, New York, 1985.
ford, 1978; 2nd edition, 1987. TYLER, V. E., L. R. BRADY, and J. E. ROBBERS, Pharmacognosy,
MASSIOT, G. and C. DELAUDE, Pyrrolidine alkaloids, in The Alka- 8th ed., Lea and Febiger, Philadelphia, 1981.
loids, Vol. 27 (A. Brossi, ed.), 270-322, Acadentic Press, New VERPooRTE, R., R. VAN DER HEuoEN, W. M. VAN GuUK, and H.
York,1986. J. G. TEN HooPEN, Plant biotechnology for the production of
NAHRSTEDT, A.. Strukurelle Beziehungen zwischen pflanzlichen alkaloids: Present status and prospects, in The Alkaloids, Vol.
und tierischen Sekundiirstoffe, Planta Medica, 44, 2-14 (1982). 40 (A. Brossi, ed.), 1-188, Academic Press, New York, 1991.
NAHRSTEDT, A., Neue phannakologisch interessante Naturstoffe WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and Metabo-
(1983-1984), Oesterreichische Apotheker-Z., 39, 733-741 lism in Plants, Plenum Press, New York, 1978.
(1985). WATERMAN, P. G. and A. l. GRAY, Chemical systematics, Nat.
NASH, R. 1., M. ROTHSCHILD, E. A. PORTER, A. A. WATSON, R. D. Prod. Rep., 4, 175-203 (1987).
WAlGH, and P. G. WATERMAN, Calystegines in Solanum and WHITE, P. T., An ancient Indian herb turns deadly. Coca, National
Datura species and the death's head hawk-moth (Acherontia Geographic, 8-47, (January 1989).
atropus), Phytochemistry, 34, 1281-1283 (1993). WtLUAMS, H. S., A. C. RAy, and H. L. KIM, a'-Piperideine alka-
NUMATA. A. and T. IBUKA, Alkaloids from ants and other insects, loids from the toxic plant Lobelia berlandieri, 1. Agric. Food
in The Alkaloids, Vol. 31 (A. Brossi, ed.), 194-315, Academic Chern., 35, 19-22 (1987).
Press, New York, 1987. WINK, M., Allelochemical properties or the raison d'etre of alka-
1'INDER, A. R., Pyrrole, pyrrolidine, pyridine, piperidine, and related loids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), 1-105,
alkaloids, in Alkaloids and Sulphur Compounds (P. G. Water- Academic Press, New York, 1993.
man, ed.), Vol. 8 of Methods in Plant Biochemistry (p. M. Dey WOOLEY, J. G., Tropane alkaloids, in Alkaloids and Sulphur Com-
and J. B. Harborne, eds.), 241-269, Academic Press, London, pounds (P. G. Waterman, ed.), Vol. 8 of Methods in Plant Bio-
1993. chemistry (p. M. Dey andJ. B. Harborne, eds.), 133-173, Aca-
ROESSLER, F., D. GANZINGER, S. JOHNE, E. SCHOPP, and M. HESSE, demic Press, London, 1993.
30
Pyrrolizidine, Quinolizidine, and
Indolizidine Alkaloids

Introduction loids defmitely are of distinct origins. Pyrrolizidine alkaloids

Pyrrolizidine Alkaloids are derived from arginine or ornithine and possess two fused
Biosynthesis 5-membered rings that share a nitrogen atom. Quinolizidine
Distribution of Pyrrolizidine Alkaloids alkaloids are derived from lysine and have two fused 6-
Pyrrolizidine Alkaloids in Animals membered rings that share a nitrogen atom. Indolizidine
Pyrrolizidine Alkaloids and Parasitic Plants rings have a 5-membered ring fused to 6-membered ring
The Role of Pyrrolizidine Alkaloids in Livestock and share a nitrogen atom. Pyrrolizidine, qninolizidine, and
Poisoning indolizidine alkaloids are involved in many plant-herbivore
Human Poisoning by Pyrrolizidine Alkaloids interactions, but are of anthropocentric importance chiefly
Medicinal Uses of Pyrrolizidine Alkaloids because of their involvement in livestock poisoning, al-
Quinolizidine Alkaloids though several have antitumor activity and others are used
Biosynthesis medicinally. The phenanthroindolizidine alkaloids are a
Complex Quinolizidine Alkaloids major subgroup of indolizidine alkaloids.
Systematic Usefulness of Quinolizidine Alkaloids Both the 'H- and "C-NMR (nuclear magnetic resonance)
Biological Activity of Quinolizidine Alkaloids spectra for several of the groups of alkaloids above have
Medicinal Uses of Quinolizidine Alkaloids been reviewed (Crabb, 1982).
Quinolizidine Alkaloids and Livestock Poisoning
Mescal-Hallucinogenic Properties of Quinolizidine
Alkaloids PYRKOLIZIDIl'IE ALKALOIDS
Indolizidine Alkaloids
Biosynthesis Pyrrolizidine alkaloids are derived from ornithine and char-
Biological Activity of Indolizidine Alkaloids acteristically have two fused 5-membered rings with a nitro-
Indolizidine Alkaloids in Insects gen at one of the common positions (Fig. 30.1). At least 560
Elaeocarpaceae Alkaloids alkaloids of this type are known (Verpoorte et al., 1991).
Phenanthroindolizidine and phenanthroquinolizidine Pyrrolizidine alkaloids rarely occur in the free form and are
Alkaloids found as complex esters and N-oxides. Although pyrroliz-
Biosynthesis idine alkaloids occur in all plant parts, the concentration
Biological Activity of Phenanthroindolizidine Alkaloids in at least one example, Senecio vulgaris, is highest in the
References inflorescence (Hartmann, 1991).
Cells of Senecio vulgaris in tissue culture lack the ability
to synthesize pyrrolizidine alkaloids, but retain the ability
INI'KODVCTlOI'l to take up and accumulate the corresponding N-oxides. The
translocation of exogenous N-oxides into the vacuole by car-
Pyrrolizidine, quinolizidine, and indolizidine alkaloids are rier-mediated transport has been demonstrated; cells of non-
chemically diverse and restricted in distribution. Some simi- alkaloid-producing plants do not take up pyrrolizidine N-
larities in structures and biosynthesis exist, but as the path- oxides (Hartmann, 1991).
ways become more clear, these three major groups of alka- Pyrrolizidine alkaloids are synthesized in the roots and

S46
 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids 547

~
H 14 \ HO HO ..fHO

:"'6)00
...",
~O
"

0~to160'-
, jO~
heliotridine H II H II
7 ; ~
i ~
N
N
senecionine (5) monocrotaline (8)
(retronecine + (retronecine +
senecic acid) monocrotalic acid)
retronecine (1)

CH30.
~(
/ ~O
-< H
~'''/
HO,fHO

ct5
heliotrine (15) heliosupine
(heliotridine + angelic
~~tt5,100
H

trichodesmine
(retronecine +
II;

and echimidinic acids) trichodesmic acid)

HO~
lr1'"
d5 toO to'
_ H
[0 HO ".•'HO ..,,,CH,OH o
'. 0 19
0
H

H '! II
i '\ o H II o H 0

N+
; ~ 6 7 ~ 1~ ,

I N
5 4
O'

indicine N-oxide (16) junceine gynuramine

Fig. 30.1. Some typical pyrrotizidine alkaloids.

transported to other parts of the plant. Because of the ready iodoplatinate or Ehrlich's reagent. Important modifications
uptake of these compounds by parasitic plants (see below), have been introduced by Mattocks and Jukes (1981),
transport probably occurs in the phloem (Hartmann, 1991),
Root cultures of Senecio vulgaris synthesize pyrrolizidine Biosynthesis
alkaloids and accumulate them as N-oxides (Herbert, 1988), Although most pyrrolizidine alkaloids have been re-
This synthesis parallels that of intact plant roots, Senecionine garded as coming from ornithine by the intermediacy of pu·
is synthesized and converted to the N·oxide and transported trescine, arginine probably is the actual precursor in most
throughout the plant. Plants of many species contain only instances (Hartmann, 1991), High incorporations of argi·
N·oxides of pyrrolizidine alkaloids (Hartmann et aI., 1989), nine, isoleucine, putrescine, and spermidine were obsetved
A sensitive method for the detection and quantitation of in many pyrrolizidine-alkaloid-containing plants (Herbert,
pyrrolizidine alkaloids based on stoichiometric reaction of 1988), In some Heliotropium species (Boraginaceae), pu·
protonated alkaloids with methyl orange may be useful, as trescine is derived from arginine through an alternate path·
these alkaloids sometimes give poor tests with Dragendorff's way (see Chapter 28), In contrast, no arginine decarboxylase
reagent (Birecka et aI., 1981), Most give good tests with activity could be shown in species of Senecio and Crotalaria
S48 Pyrrolizidine. Quinolizidine. and lndolizidine Alkaloids

that were examined (Birecka et aI., 1988). Most nonalkaloi- strated that homospermidine was incorpomted as a symmet-
dal plants have high arginine decarboxylase activity and al- rical intermediate (Khan and Robins, 1985a, 1985b; Spenser,
most no ornithine decarboxylase activity. 1985; Wrobel, 1985). [1,9-1 3CzlHomospermidine (2) is in-
Most pathways leading to the biosynthesis of pyrroliz- corporated intact into retronecine (1). Introduction of [1,4-
idine alkaloids involve a symmetrical intermediate (see zH.l-putrescine followed by zH_NMR revealed that the ma-
Chapter 29 for additional discussion of symmetrical versus jority of 2H-Iabel was located at C-3, C-5, C-8, and C-9
unsymmetrical intermediates) (Fig. 30.2). [1-Amino- 15N- (Fig. 30.3). The pattern was consistent with the proposed
!3Clputrescine was incorporated into retronecine (1) that was biosynthetic pathway including homospermidine (Khan and
isolated following hydrolysis of retrorsine (3) (Fig. 30.4). Robins, 1985a, 1985b). The formation of (9S)-retrorsine (3)
Homospermidine (2) was suggested as a symmetrical inter- with 9_zH indicates that a stereospecific addition (of lH from
mediate and also was shown to be incorporated into retronec- the re face to the carbonyl group of the aldehyde precursor)
ine (1). By means of l4C-Iabeled intermediates [such as has occurred. When putrescine (4) labeled at the (R)-[l-
HzN(CH3hNHI4CHz(CHzhI4CHzNHz)], it was demon- zHl- or (S)-[I-zH]-positions was introduced, equal label, at

arginine or
ornithine
~ putrescine
- bomospermidine (2) (1)

'.
I
Hs +
lossof

di N
HO ~HO~ CO yHsH
. :. . - rl----.-
~N:)
R' H

NH

* loss of HR or Hs
0

~
*'" attack from re-face

retronecine (1)
Fig. 30.2. Biogenesis of pyrrolizidine alkaloids (Spenser, 1985; used with peIll1ission of the copyright owner, the International Union of Pure and
Applied Chemistry, Oxford).
 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids 549

~'"' --
DH
(R)-[l.'HI·putrescine
(4)

--
retrorsine (3)

Fig. 30.3. Biogenesis of retrorsine from (R)-[1- 2H]putrescine (Wrobel, 1985; used with pennission of the copyright owner, Academic Press, Orlando,
FL).

positions 3/3, 50<, 80<, and 9-pro·S by the (R) precursor, other families. Of these plants, the genus Senecio, with about
whereas only the 30<- and 5/3 was found positions were la- 1200 species worldwide and about 50 species in the United
beled by the (S) precursor (Fig. 30.3). These results suggest States and Canada, is particularly important. The macrolide-
that the oxidation of putrescine to 4-aminobutanal requires type pyrrolizidine alkaloids, such as senecionine (5), are
the loss of the pro'S hydrogen, reduction of the intermediate found in the Asteraceae (Senecioneae) and the Fabaceae
imine to homospermidine (2) occurs at the si face, two oxida- (Crotalaria) (Waterman and Gray, 1987). Compounds with
tion steps leading to an aldehyde proceed with removal of pyrrolizidine ring systems also have been found in the Apo-
pro-S hydrogens, cycIization of the iminium ion to 8o<-pyr- cynaceae, Celastraceae, Convolvulaceae, Euphorbiaceae,
rolizidine proceeds at the re face of the ion, and reduction Linaceae, Orchidaceae, Poaceae, SantaIaceae, and Sapota-
of the aldehyde occurs via attack on the re face of the car- ceae (Culvenor, 1978; Jenell·Siems et aI.,. 1993; van Wyk
bonyl group (Wrobel, 1985) [see Leete (1983) for a review and Verdoom, 1988; Waterman and Gray, 1987). There also
of the biosynthesis and feeding studies of labeled precursors are reports from the genus Caltha (Ranunculaceae) and from
leading to pyrrolizidine a1kaloidsl. Castilleja (Scrophulariaceae) (see below). 1-Aminopyrroliz-
Pyrrolizidine alkaloids usually occur as complex esters idine derivatives are known from the Poaceae, the Fabaceae
of unique monobasic or dibasic acids called necic acids. (Papilionoideae: Genisteae), and Rhizophoraceae. Despite
Senecionine (5) is composed of both the necine base, retro- these occurrences, however, pyrrolizidine alkaloids are only
necine (1), and senecic acid (6) (Fig. 30.4). characteristics of the genus Parsonia (Apocynaceae), the
Senecic acid (6) is produced by condensation of two mol- Asteraceae, Boraginaceae, and Crotalaria (Fabaceae) (Cul-
ecules of isoleucine. The formation of the lO·carbon necic venor, 1978).
acid (7) is accompanied by loss of C-4-pro-R hydrogen from Study of the pyrrolizidine alkaloids of the Fabaceae, tribe
both isoleucine components (Wrobel, 1985). In retrorsine Crotalarieae, suggest that these data are useful for systematic
(3), two hydrogen atoms of (2S)-isoleucine at C-13 and one and phylogenetic study of this complex group of legumes
of the C-15 ethylidine group are derived from C-4 methylene (van Wyk and Verdoom, 1990).
hydrogen atoms of (2S)-leucine. These were later shown to
arise by loss of 4-pro-S hydrogen (or that the atoms above Pyrrolizidine Alkaloids in Animals
at C-13 and C-15 were derived from the 4-pro-R hydrogen
Although most of the toxic effects of pyrrolizidine a1ka·
of L-isoleucine) (Wrobel, 1985).
loids probably are produced by alkylation of DNA and pro-
teins, heliotrine inhibits cholinesterase, and monocrotaline
Distribution of PyrroIizidine Alkaloids
(8) modulates pulmonary Na+/K+ pumps. Many pyrroliz-
These alkaloids are found mostly in the Asteraceae (tribes idine alkaloids are mutagenic in Drosophila (Wink, 1993b).
Senecioneae and Eupatorieae), the Boraginaceae, Fabaceae Pyrrolizidine alkaloids of most plants occur primarily as
(Croralaria, Lotononis, and Buchenroedera), and a few the N-oxides. Conversion to the corresponding bases yields
550 Pyrrolizidine, Quinolizidine, and lndolizidine Alkaloids

~ HO ! HO (2S)-isoleucine H '.. HO

="~0ttS'o~/: ~TIN'
CLJ~:~ ~"'-"J
i '\ 0 OH t

8
N

rn~.lHrn~;,~,
C, HNH, CH,C,H _ 0 tt5°~H
+ OH
0
_ ~o
° Htt5~0
CO,H CHNH, ~ ; "
isoleucine I ' '\ N
+ CO,H N
retronecine (1)
Chap. 30,Fig. 4

Fig. 30.4. Biosynthesis of the necic acid of senecionine (Davies and Crout, 1974; Devlin and Robins, 1984; modified and used with permission of the
copyright owner. The Royal Society of Chemistry, London).

forms that can passively penetrate membranes (Hartmann, the nectar (Krasnoff and Dussourd, 1989). Some moths store
1991). Senecio alkaloids without modification. The larvae of the
Pyrrolizidine alkaloids are feeding deterrents and insecti- cinnabar moth, Tyria jacobaea and related species, accumu-
cida
for many species of insects (Schneider, 1987; Wink, late such alkaloids in their tissues by feeding on groundsel
1993b). Some insects, such as Melanchra persicariae and or ragwort, both rich in pyrrolizidine alkaloids and, thus,
Spodoptera littoralis feed on Senecio vulgaris, which con- become distasteful to predators (Fig. 30.5) (Harborne, 1982;
tains pyrrolizidine alkaloids, but do not sequester the alka- Mann, 1987). All stages of these moths are brightly colored
loids. The larvae are cryptically colored (Schneider, 1987). and are not eaten by most insectivores (Schneider, 1987).
Certain insects appear to be able to excrete these alkaloids Ithomiine butterflies, widely known as distasteful, show
without harm. For example, senecionine (5), its N-oxide, and warning coloration and are usually rejected by both verte-
hydrolytic products, including retronecine (1), were found brate and invertebrate predators. The Ithomiine larvae con-
in the honeydew of the aphid Myzus persicae when this in- sume plants of the Solanaceae, but do not accumulate the
sect fed on Senecio vulgaris (Molyneux et aI., 1990). alkaloids from those plants (see Chapter 36). However, adult
On the other hand, pyrrolizidine alkaloids are sequestered ithomiine butterflies feed on plants of the Asteraceae and
and have important ecological roles in a number of insects Boraginaceae that contain pyrrolizidine alkaloids. By a pro-
(Harborne, 1982; Nahrstedt, 1982; Schneider, 1987; Wink, cess of ejecting fluid through the proboscis onto the surface
1993b). The two main orders involved are the Danainae of of the plant and subsequently taking it up again, they acquire
Nortb America and the Ithomiinae of South America (Har- pyrrolizidine alkaloids from the plants (Schneider, 1987).
borne, 1989). Both larvae and adults of the Danainae may Adult males may contain as much as 10% dry weight pyrrol-
consume plant materials with pyrrolizidine alkaloids, but the izidine alkaloids, such as dehydropyrrolizidine alkaloid
Ithomiinae are unable to obtain these alkaloids at the larval monoesters and their N-oxides. These compounds also are
stage (Harborne, 1986). The nectar of some plants in the implicated in reproduction of the insect (Brown, 1984). The
Asteraceae (mostly of the tribe Eupatorieae) contains 2-4% males of some species of Ithomiidae secrete a lactone on
of the dry weight as alkaloids that can serve as a reward the hairs of hind wing fringes that serves as an aphrodisiac for
for pollination (Harborne, 1989) for ithomiine and danaine the female and/or as a male recognition signal. This lactone
butterflies, but inhibit feeding by general feeding insects probably is derived from the acid moiety of pyrrolizidine
(Masters, 1991). Adults of at least three genera of arctiid alkaloids (Boppre, 1978).
moths visit plants that produce dehydropyrrolizidine attrac- Despite the defenses that older adults have, freshly
tants, such as (- )-(R)- (9) and ( + )-(S)-hydroxydanaidal in emerged adults (that presumably lack these defenses) are
 Pyrro/izidine, Quinolizidine, and lndolizidine Alkaloids 551

eaten by various predators (e.g., the giant tropical orb spider, questers monocrotaline (8) obtained from its food plant Cro-
Nephila clavipes). talaria retusa (Bemays et aI., 1977).
Dead shoots of Heliotropium indicum (Boraginaceae) act In addition to the use of pyrrolizidine alkaloids as protec-
as a powerful attractant for male ithomiine and danaine but- tive chemicals, a number of species of three groups of Lepi-
terflies. The butterflies use alkaloids obtained from these doptera (Danainae, Ithomiinae, and Arctiidae) biosynthesize
plants for the production of chemicals with pheromonal ac- pheromones from these compounds thatthey acquire by phar-
tivity. Baiting with alkaloids and the esterifying necic acids macophagy or larval feeding (Schneider, 1987).
indicated that a volatile product derived from the necic acids Danaid butterflies can metabolically alter pyrrolizidine
attracts males to the plants, whereas the intact alkaloids then alkaloids without apparent harm. Individuals of the Ameri-
act as phagostimulants (Pliske et aI., 1976). can monarch butterfly, Danaus plexippus, captured in the
Moths of the families Ctenuchidae and Arctiidae also are field were found to contain pyrrolizidine alkaloids that were
attracted to plants containing pyrrolizidine alkaloids. In par- present in food plants at the capture sites. Adults of D. plex-
ticular, larvae of Nyctemera annulata (the magpie moth) ippus were able to store alkaloids for several days. Unlike
ingest pyrrolizidine alkaloids from a variety of Senecio spe- certain other species, however, these male butterflies do not
cies. Larvae that feed on Senecio spathulatus are able to secrete alkaloid-derived substances on their hairpencils, nor
store pyrrolizidine alkaloids. The alkaloids subsequently ap- do the hairpencils figure prominently in courtship. It has
peared in the adult moths and their eggs and in a parasite been suggested that the alkaloids may contribute to the lack
of the larvae of this species (Benn et aI., 1979; Goss, 1979; of palatibility of the butterflies to potential predators (Edgar
Schneider, 1987; Specialist Periodic Reports, Alkaloids, et aI., 1976; Pliske, 1975).
1978). Male butterflies of several other species of the genus Da-
The moth Utetheisa ornatrix consumes plants of the naus (17 out of 23 tested) ingest pyrrolizidine alkaloids and
genus Crotalaria that contain pyrrolizidine alkaloids. Al- convert them into pheromones such as danaidone (11), da-
though adults normally are not eaten by the spider Nephila naidal (10), and hydroxydanaidal (9) that are released from
c/avipes, moths raised in the laboratory were consumed. the abdominal hairpencil organs during courtship behavior
Males of this species also produce two pyrrolizidines that (Fig. 30.5) (Boppr", 1978; Schneider, 1987). Certain male
are closely related to danaidal (10) and three hydrocarbon danaid butterflies feed on plants in the Senecioneae (As-
pheromones. The pyrrolizidines are passed from the male to teraceae) and utilize pyrrolizidine alkaloids as a source for
the female and deposited in the eggs (Eisner and Meinwald, compounds such as (9-11). Compounds such as danaidone
1987). (11) are used by the male butterflies of the queen butterfly
The aposematic grasshopper Zonocerus variegatus se- (Danaus gilippus) (of Africa and Australia) as female flight

toxic form bound

to liver

pyrrole I

(R)-hydroxydanaidal (9)

HO CO,H

(14)
senecipbyJlin N-oxide (12)

Fig. 30.5. Compounds derived from pyrrolizidine alkaloids found in insects (modified from Harborne, 1982).
552 Pyrrolizidine, Quinolizidine, and lndolizidine Alkaloids

arrestants or as aphrodisiacs and thus play a vital part in necie acid than that of the host plant (L'Empereur et aI.,
the prelude to mating. The male deposits .. dust" of this 1989). The alkaloids of the plant occurred primarily as N-
compound and a viscous material on the antennae of the oxides.
female butterfly, and mating usually ensues (Eisner and Aphids that feed on Senecio species sequester pyrroliz-
Meinwald, 1987; Scbneider, 1987). Danaus chrysippus idine alkaloids. A ladybird beetle that feeds on the aphids,
males must dip their hairpencils into the wing pockets (scent in tum, sequesters these compounds (Hartmann, 1991).
organs) before pheromones (such as 14) are produced (Bop- Pyrrolizidine alkaloids also have been found in amphib-
pre et aI., 1978; Scbneider, 1987; Specialist Periodic Re- ians. Certain of these amphibian alkaloids also occur in in-
ports, The Alkaloids, Vol. 8,9, and to). When the butterflies sects. For example, the venom of the thief ant, Solenopsis
are raised in the laboratory away from the host plants, they xenovenenum, is an amphibian pyrrolizidine (Daly et al.,
lack the pyrrolizidine alkaloids and have limited reproduc- 1993).
tive success (Boppre, 1978; Schneider, 1987).
It has been hypothesized that the ancestral food plants of Pyrrollzidine Alkaloids and Parasitic Plants
these butterflies were plants that contained both pyrroliz-
idine alkaloids and cardiac glycosides (Edgar et al., 1974); Members of the genus Castilleja are hemiparasites that
attack many species of host plants, especially those in the
however, few plants today meet these criteria. A few species
Asteraceae and Fabaceae. Plants of Castilleja linariifolia
of apocynaceous plants have both types of compounds but
that utilize sagebrush (Artemisia species which lack alka-
have not been reported to serve as hosts for the insects.
loids) are devoid of alkaloids. Several species that parasitize
Clearly, we lack sufficient information to explain the origin
quinolizidine-containing legumes also contain similar alka-
of such complex pheromonal, reproductive, mimicry, and
loids, although not in identical concentrations. Other species
protective systems.
that attack Senecio species contain pyrrolizidine alkaloids.
In the case of species of the arctiid moth, Creatonos, not
The variability of alkaloid content in Castilleja species is
only are the pyrrolizidine alkaloid metabolites involved in
linked to that of the host plant (Stermitz and Harris, 1987).
courtship, but they are also involved in the growth and devel-
Plants from five species of Pedicularis (Scrophularia-
opment for the pheromone-producing organ in the pupa
ceae) take up alkaloids from their host plants. Those of Pe-
(Scbneider, 1987).
dicularis groenlandica and P. bracteosa take up senecionine
The stems and leaves of Castilleja rhexifolia (Scrophula-
(5) from the host Senecio triangularis. Those of Pedicularis
riaceae) contain senecionine (5) and its N-oxide (obtained
crenulata contain anagyrine (Fig. 30.1) (a quinolizidine al-
from Senecio triangularis) as their main alkaloids. The
kaloid) from the host Thermopsis divaricarpa. Plants of Pe-
major alkaloid of the blossoms and seeds is rhexifoline, an
dicularis racemosa contain quinolizidine alkaloids from a
iridoid-monoterpene-derived alkaloid, which is found in
Lupinus argenteus hybrid (Scbneider and Stermitz, 1990).
many Castilleja species. These plants serve as a host for the
plume moth, Platyptilia pica. The larvae primarily eat the
green seeds. Both the larvae and the adults of this insect lbe Role of Pyrrolizidine Alkaloids
contain rhexifoline (Roby and Sterrnitz, 1984). Plants of C. in Livestock Poisoning
hispida contain quinolizidine alkaloids, and occasional Many pyrrolizidine-alkaloid-containing plants are poi-
plants of C. sulphurea contain quinolizidine alkaloids, sonous to livestock (Hartmann, 1991). Over 300 plants from
whereas those of C. occidentalis do not contain alkaloids at least 30 genera and 6 families that contain this group of
other than rhexifoline. The iridoid monoterpene content of alkaloids are responsible for livestock poisoning. Probably
all these alkaloids is similar. The larvae were found to ex- the mOSt important of these are plants of the genera Crota-
crete the monoterpene iridoids (Stermitz et al., 1986). laria, Heliotropium, Senecio, and Symphytum (Soffness and
Larvae of the chrysomelid beetle Oreina cacaliae pro- Cordell, 1985). Pyrrolizidine alkaloids produce hepatotoxic
duce seniciphylline N-oxide (12) as a part of their defensive effects; the action tends to be cumulative. As little as 1-5%
secretions. This pyrrolizidine alkaloid probably is derived of the animal's weight in plant material is enough to be fatal.
from the alkaloids of the host plant Adenostyles leucophylla The LOsos of a variety of pyrrolizidine alkaloids range from
(Asteraceae: Senecioneae) (Pasteels et al., 1988). about 50 to 1100 mg/kg. Monocrotaline (8) has an LOso i.p.
Larvae of the arctiid moth Creatonotos transiens accumu- in rat of 175 mg/kg; retronecine (1) has an LOso i.p. of 634
late pyrrolizidine alkaloids when they feed on plants contain- mg/kg in mouse; senecionine (5) has an LOso i.p. of 50
ing these compounds. Other types of alkaloids tested were mg/kg in mouse (Wink, 1993b). The toxicity of the N-oxides
excreted in the frass (Wink and Scbneider, 1988). Pyrroliz- of these alkaloids may be about the same or less as the
idine alkaloids act as precursors for pheromones, but also alkaloids themselves (Suffness and Cordell, 1985). How-
as morphogens for the development of the male scent organ ever, N-oxides do not have the bitter taste associated with
(Wink and Schneider, 1988). the alkaloids (Wrobel, 1985).
Larvae of Gnophaela latipennis consume plant material The toxicity may be due to the formation of pyrrole esters
of Hackelia californica (Boraginaceae), butlarvae and emer- that have potent alkylating properties (Fig. 30.6). Hydroxy-
gent adults contain a pyrrolizidine alkaloid with a different danaidal (9) has been suggested as a major hepatotoxic com-
 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids 553

):.AoeOR ~eHO
~!J 80
E-4-bydroxy-1-enaI(4)

Fig. 30.6. Pyrrole esters (modified from Suffness and Cordell, 1985; used with pennission of the copyright owner, Academic Press, New York).

pound (Harborne, 1986). These esters are produced by me- cause of the cumulative toxic effects of these compounds
tabolism of pyrrolizidine alkaloids in the mammalian liver (Roder, 1992). Although the toxicity of specific compounds
and are able to bind to· certain sites in the liver as well as varies, the use of many herbal remedies involving plants of
with macromolecules such as DNA. Some studies suggest the Asteraceae and Boraginaceae that may contain pyrroliz-
that the toxicity of pyrrolizidine alkaloids lies not in their idine alkaloids should be avoided.
own structures but in the one major metabolite (13) to which A variety of pyrrolizidine alkaloids and their N-oxides
they are converted. This dehydrogenation product produced have antitumor activity (Wink, 1993b). Herbal material of
by mammals is much more toxic. Hydrolysis of the complex Senecio vulgaris (Asteraceae) has been used for hundreds
ester groups of these compounds yields retronecine, which is of years as a treatment for cancer. The active compounds
then dehydrogenated. More recently, however, a breakdown have been shown to be senecionine (5) and senecionine N-
product, E-4-hydroxyhex-2-enal (14), has been suggested to oxide. Indicine N-oxide (16) from Heliotropium indicum
be the actual reactive metabolite that binds to DNA in the (Boraginaceae) has pronounced antitumor activity. Mono-
liver (Harborne, 1986). crotaline (8), from several Cratalaria species (Fabaceae),
has similar activity (Blasko and Cordell, 1988; Suffuess and
Human Poisoning by Pyrrolizidine Cordell, 1985). Alkylation seems to be involved in the antito-
Alkaloids mor activity. As the N-oxides cannot serve as enamines and
Human health problems have been attributed to the acci- hence cannot be directly involved in alkylation, these com-
dental consumption of plant materials that contain pyrroliz- pounds should only be active to the extent they are converted
idine alkaloids (Beier and Nigg, 1992). The consumption of to the free bases. At least in the case of indicine N-oxide
wheat containing seeds of H eliotropium popovii has caused (16), this does not appear to be the case (Suffness and Cor-
a large outbreak of veno-occlusive disease in Mghanistan. dell,1985).
The main alkaloid present was heliotrine (15). In India, simi- Senecionine (5) and senecionine N-oxide have been iden-
lar problems have occurred with Crotalaria species. tified as the antifertility agents from Senecio vulgaris (Tu
Pyrrolizidine alkaloids also occur in honey made from et a1., 1988).
Senecio jacobaea (tansy ragwort) and in milk of cattle that
feed on this plant.
Herbal teas are responsible for poisoning problems as QUII'IOLJZlDII'IE ALKALOIDS
well (Beier and Nigg, 1992). In Arizona, a number of cases
of veno-occlusive disease have been attributed to the con- Quinolizidine alkaloids are derived from lysine and have
sumption of tea from Senecio longilobus, whereas a similar two fused 6-membered rings that share a nitrogen atom. In
poisoning in West Germany is thought to have arisen from contrast to the pyrrolizidine alkaloids, quinolizidine alka-
senecionine in a Petasites species (Robins, 1993). Tea from loids usually occur free. At least 570 quinolizidine alkaloids
comfrey leaves (Symphytum spp., Boraginaceae) has long are known (Verpoorte et a1., 1991).
been used (Schneider, 1987); however, comfrey leaves and This group of alkaloids primarily is found in the subfam-
roots cause tomors when fed in the diet to rats. Pyrrolizidine ily Papilionoideae of the family Fabaceae; quinolizidine al-
alkaloids have been identified as one of the causes of primary kaloids are the most commonly encountered alkaloids in this
liver cancer in countries of central Africa, where the inci- family (Howard and Michael, 1986; Kinghorn and Ba-
dence of this cancer is the highest in the world. Many pyrrol- landrin, 1984). Quinolizidine alkaloids have been reported
izidine alkaloids are carcinogenic and have been demon- from all plant parts, but in most plants, they are synthesized
strated to induce tumors in a number of different animal in young photosynthetic tissues (Wink, 1987).
systems. Lupine alkaloids are among the simplest representatives
of this group; they are common in the genus Lupinus (Fig.
Medicinal Uses of Pyrrollzidine Alkaloids 30.7) (Aslanov et aI., 1987; Wink, 1984b).
The medicinal uses of pyrrolizidine alkaloids and pyrrol- Studies with cell suspension cultures have established
izidine-alkaloid-containing plants bave been questioned, be- that alkaloid biosynthesis undergoes a diurnal rhythm (Kin-
554 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids

oY
(-)-sparteioe (17) angustifoline (21) rhombifoline (28) matrine (26)

/OH

CO'
'<::: NH H i

I N
° N

(-)-anagyrine (29) (+)-17-oxosparleine (25) lamprobine

Fig. 30.7. Some representative quinolizidine aIkaloids.

ghorn and Baiandrin, 1984; Wink and Witte, 1984). Maxi- Alkaloids in Chapter 29) (Fig. 30.7) (Howard and Michael,
mum alkaloid synthesis occurs during the day and dimin- 1986).
ishes at night. This confirms suggestions that lupine alkaloid Most of the legume species that have been introduced into
biosynthesis undergoes light-dependent regulation (Kingh- tissue culture produce only small amounts of quinolizidine
orn and Balandrin, 1984; Wink and Witte, 1984). The alka- alkaloids (Ellis, 1988).
loids are synthesized in the leaves and transported to the Techniques for analysis (Kinghorn and Balandrin, 1984;
seeds, pods, and roots. Ready uptake of quinolizidine alka- Wink, 1987, 1993a) and NMR and mass spectra (Aslanov
loids by parasitic plants suggests that this transport occurs et al., 1987; Wink, 1987) of quinolizidine alkaloids have
via the phloem (Hartmann, 1991). During senescence of been reviewed.
leaves, the concentration of alkaloids drops by as much as
90% (Kinghorn and Balandrin, 1984). Biosynthesis
The level of quinolizidine alkaloids in seeds decreases Quinolizidine alkaloids are derived from lysine. Studies
greatly during germination, suggesting utilization of the al- with labeled precursors indicate that a symmetrical interme-
kaloids as a uitrogen source. Yoimg plants with fully devel- diate, cadaverine (20), is involved in their formation (Her-
oped leaves are able to synthesize these compounds. Lupin bert, 1988; Kinghorn and Balandrin, 1984; Leete, 1983;
cell cultures can survive and grow on media that contain Spenser, 1985), although no intermediate comparable to the
sparteine (17) as the only nitrogen source (Wink and Witte, dimeric form plays a role in the formation of pyrrolizidine
1985). alkaloids is involved (Spenser, 1985). Much recent informa-
Cutting up leaflets of Lupinus polyphyllus induces a rapid tion is based on cell suspension cultures of Lupinus polyphyl-
increase in quinolizidine alkaloids of up to 400% within Ius, Baptisia australis, and Sarothamnus scoparius (all Faba-
2.5-4.0 h. This response occurs under a variety of conditions ceae). Lysine decarboxylase is localized in leaf chloroplasts
and is neither a constitutive defense nor the usual type of (Wink 1987; Wink and Hartmann, 1982, 1984); the presence
induced response (Wink, 1983a). of a diamine oxidase does not appear to be involved. Lysine
Certain other groups of alkaloids that coincidentally have decarboxylase is found in all parts of Lupinus plants.
a quinolizidine ring system are discussed in other chapters. [3,3-2H21 Cadaverine is incorporated into ( + )-Iupanine
For example, the alkaloids of the Lythraceae are discussed (21) and (+ )-13a-hydroxylupanine (23) in Lupinus pol-
in Chapter 37 and those of the genus Nuphar are discussed yphyllus andL. angustifolius, respectively, into (+ )-angusti-
in Chapter 36. Yet other quinolizidine alkaloids (18 and 19) foline (22) in L. angustifolius and into (- )-Iupinine (24),
are known from the Ericaceae (Vaccinium myrtillus) and and (- )-sparteine (17) in L. luteus (Fig. 30.8). The labeling
the Euphorbiaceae (Poranthera corymbosa) (see Piperidine patterns were determined by 2H_NMR spectroscopy. Car
 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids 555

bon-3 of cadaverine remains unaffected by events during (IR)-[1-2HIlCadaverine afforded sparteine (17) that was
condensation. Retention of deuterium at position 13 of 13a- labeled with deuterium at C-2a, C-6J3, C-lla, C-15a, and
hydroxylupanine indicates that the introduction of oxygen C-17a, whereas (IS)-[1-2HIlcadaverine gave sparteine that
at this site cannot involve keto or enol intermediates. Further, was labeled with deuterium at C-2J3, C-lOa, and C-17a
the labeling of C-13 of angustifoline (22) by deuterium indi- (Fraser & Robins, 1987). The appropriate results were also
cates that the allylic moiety comes from cadaverine (Herbert, obtained for lupanine and angustifoline. These results were
1988; Spenser, 1985). compatible with the hypothesis that these alkaloids are modi-

C - D5P:
(a)
lysine cadaverine (20)

D NH, D N

D 3 D D. D
NH, N A D

(-)-sparleine (17)

~ ~
D'N DN
D D D
D D,,, D
N ~ D N Ii OH

°(+)-lupanine (21) ° (+)-13a-hydroxylupanine (23)

:ro'
CH,OH
D~D'NH
D n
N ~ D

(-)-lupinlne (24)
° anguslifoline (22)
D H

('-'NH'

yNH'
D H
(R)-[I-'Hlcadaverine

~
('-'NH'
'xNH,
H D
(S)-[l-'H]cadaverine
- D H D
(b)

Fig. 30.8 (a & b). Labeling studies for quinolizidine alkaloid biosynthesis (modified from Herbert. 1988 and Spenser. 1985; used with pennission of
the copyright owners, The Royal Society of Chemistry, Cambridge and the International Union of Pure and Applied Chemistry, Oxford. respectively).
556 Pyrrolizidine, Quinolizidine. and Indolizidine Alkaloids

£:)
C:B'~ Q~. -
o

si attack at C-2 from the face tbat is si

cadaverine (20)

~
.HR atC-3

Hs
~ ~N~ NH
N~ -

loss of lO(pro-1OR)-proton

l~~~ -
~~--~-
l~HV ·.··k!!' il
--........" ~
re-attack at C-ll from the face
that is si at C-9

H
re attack ofH- at ColO and C-17 sparteine (17)
Fig. 30.9. Proposed biosynthesis of quinolizidine alkaloids (Spenser, 1985; modified and used with pennission of the copyright owner, the International
Union of Pure and Applied Chemistry, Oxford).

fied trimers of a I-piperideine. [2-2HJ-a l-Piperideine gave In whole plants, leaf chloroplasts convert cadaverine di-
labeling at C-613, C-ll"', and C-17", (Herbert, 1986). Be- rectly into lupanine (21) without releasing free intermediates
cause lupanine and sparteine retain deuterium from (lR)-[I- in a channeled process involving membrane-associated en-
2Hdcadaverine and from [2-2H]-a 1-piperideine at C-17", at zymes (Kinghorn and Balandrin, 1984). It is possible that
a similar level to that at positions 11", and 613, both 17- matrine (26) and other related types of quinolizidine alka-
oxosparteine (25) and the 1,2-didehydrospartenium ion are loids are synthesized by a similar channeled process (King-
excluded as intermediates (Fraser and Robins, 1987 Herbert, horn and Balandrin, 1984; Wink, 1987).
1986). Similar evidence excludes lO-oxosparteine or 1,2- Cytisine (27), the second most widely distributed quino-
didehydrosparteniurn ion as intermediates and excludes the lizidine alkaloid (after sparteine), is derived from sparteine
possibility that lupanine is a direct precursor of sparteine (17) in Lupinus luteus by loss of ring D (Fig. 30.9). Interme-
(Herbert, 1986). The results of feeding studies leading to diate compounds in this process such as angustifoline (21)
the formation of quinolizidine alkaloids have been tabulated and rhombifoline (28), which are intermediate in this pro-
(Leete, 1983). cess, are known also.
 Pyrrolizidine, Quinolizidine. and Indolizidine Alkaloids 557

Quinolizidine alkaloids are accumulated in the epidermal Systematic Usefulness of QuinoIizidine

cells of leaves, petioles, and stems. These plant parts often Alkaloids
contain as much as 5-6 mg/g fresh weight. Although the
Quinolizidine alkaloids occur in 10 of the most primitive
alkaloids are accumulated in the epidermis, they are synthe-
tribes of the Papilionoideae (Fabaceae) (Kinghorn and Ba-
sized in chloroplasts of mesophyll cells. Specific mecha-
landrin, 1984). They are especially common in the Sopho-
nisms for movement of the alkaloids into the vacuoles of
reae, Podalyrieae sensu lato, and the Genisteae (Wink,
the epidermal cells appear to involve carrier proteins (Mende
1987). Isolated occurrences have been reported from Anaba-
and Wink, 1987; Wink, 1985b).
sis aphylla (Chenopodiaceae), Caulophyllum thalictroide,
Leontice spp. (Berberidaceae), Chelidonium majus (Papav-
Complex QulnoIizidine Alkaloids eraceae), Castilleja miniata (Scrophulariaceae), Orobanche
rapum-genistae (Orobanchaceae), Osyris alba (Santala-
Complex quinolizidine alkaloids are found in other leg- ceae), Solanum lycocarpum (Solanaceae), and in species of
ume genera, such as those in the genus Omwsia (tribe So- the Ranunculaceae (Wink, 1987).
phoreae) (Fig. 30.10). In view of recent biosynthetic findings Cell suspension cultures of Lupinus polyphyllus produce
on the biosynthesis of other quinolizidine alkaloids, it seems quinolizidine alkaloids when foreign alkaloids are added to
possible that these alkaloids may be synthesized directly the cultures (Brooks and Watson, 1985), or with a number of
from four cadaverine units (Kinghom and Balandrin, 1984). other "disturbances." Quinolizidine alkaloid accumulation

-
OH
OH /OH /

CO-D
1( ~~!0 ~ii0
fJ
N .......,,'"
N ~ N+ _.""",,, HN
~) HN_ ~ UN -- ~-.. -
NH+ N~- H~_
H~.". -

H~ HOO H
H ;~/"-,

~ j !j.""'~.J
H,~ ...
NJiN - H ......~
NH

ormosanine
H H

penamine
Fig. 30.10. Biogenesis of Ormosia alkaloids (Geissman and Crout, 1969).
SS8 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids

sparteine (17) lupaolne (21) thermopsine (30) N·methylcytislne (31)

~~l ~ll ~j
~k~OH ~N~'OH ~N~
o o
18-hydroxysparteine baptifolene cytisine (27)

Fig. 30.11. Quinolizidine alkaloids from Baptisia.

also was demonstrated in cell cultures of Atropa belladonna, sine (31) or a combination was dominant. The presence of
Chenopodium rubrum, Conium maculatum, Daucus carota, almost the same alkaloids in the genus Thermopsis suggests
Spinacia oleracea, and Symphytum officinale. Nonnally, a close relation of these two genera.
these plants produce other types of alkaloids or none at all. The distribution of many of the same alkaIoids in the
This suggests that the genes for the production of quinolizi- genus Lupinus correlated closely with taxonomic groupings.
dine alkaloids are not restricted to the Fabaceae, but are The patterns of inheritance of many quinolizidine alkaloids
more widely distributed among higher plants. Nonetheless, have been examined (Waller and Nowacki, 1978). In gen-
quinolizidine alkaIoids are accumulated to measurable levels eral, these patterns are simple and easily explainable by
only in legumes under most conditions (Wink and Witte, means of genetics.
1983). The presence of quinolizidine alkaIoids in the African
Alkaloids of the genus Baptisia (Fabaceae) have been genus Camoensia suggests that this genus belongs to the
studied by Mabry and co-workers (Mears and Mabry, 1971; subfamily Papilionoideae, rather than to the Caesalpini-
Waller and Nowacki, 1978). These compounds are highly oideae (Watennan and Gray, 1987).
variable within this genus, which contains about 16 species
and occurs widely in the eastern half of North America.
Hundreds of populations of these species have been studied. Biological Activity of Quinolizidine
Several quinolizidine alkaloids are found in Baptisia species Alkaloids
(Fig. 30.11).
A variety of quinolizidine alkaloids inhibit phenylalanine
Ten plants of Baptisia leucophaea from two populations
tRNA binding to ribosomes or aspects of translation (Wink,
approximately 120 miles apart proved to be similar in alka-
1993b). These effects probably explain most of the pro-
loid chemistry at a given developmental stage, but differ-
ences did occur during development of the plant. As the nounced toxicity of this group of alkaloids. Anagyrine (29)
plants matured, the amount of cytisine (27) decreased and and cytisine (27) are teratogenic. Angustifoline (22), 13-
that of anagyrine (29) (Fig. 30.1) increased. The amount of hydroxylupanine (23), lupanine (21), sparteine (17), and 13-
total alkaIoids decreased from 1.02 to 0.65 to 0.08% dry tigloyloxylupanine have activity against gram-positive bac-
weight of the plant. Differences in individual plants also teria. Lupanine, sparteine, and 13-tigloyloxylupanine have
were observed. The inheritance of alkaloids in hybrid plants antifungal activity against the fungus Erysiphe graminis
was complex but did clearly indicate that hybridization was (Wink, 1993b).
taking place within the populations studied. Under the same Quinolizidine alkaloids strongly inhibit gennination of
situations, the inheritance of flavonoids in hybrids tended lettuce seeds. Alkaloid esters, for example, 13-tigloyloxylu-
to be additive. In many of the hybrids, anagyrine (29) or panine, inhibited gennination completely at a concentration
thennopsine (30) was dominant, whereas the parental types of 6 tuM (Wink, 1983b). Sparteine (17) and cytisine (27)
contained a tricyclic-type alkaloid such as cytisine or N· inhibit radicle growth in Lepidium (Wink, 1993b).
methylcytisine (31) (Mabry and Mears, 1970). Variations The importance of quinolizidine alkaIoids for protection
were observed in different parts of plants as well. The most of plants against pathogens and herbivores has been re-
striking changes were observed in reproductive parts (i.e., viewed (Wink, 1988). Incorporation of sparteine (17) sulfate
buds, flowers, fruits, and seeds). at 0.1 % into artificial diets is totally lethal to the larvae of
The widespread occurrence of the same alkaloids within Callosobruchus maculatus (Janzen et aI., 1977).
the genus precluded separation of the species into phyletic A number of quinolizidine alkaloids have been demon-
groups. In all but one species, cytisine (27) or N-methylcyti- strated to serve as feeding deterrents (or to be lethal to var-
 Pyrrolizidine, Quinolizidine, and lndolizidine Alkaloids 559

ious insects). Others are nematicidal and molIuscicidaI dine alkaloid) from the host Thermopsis divaricarpa. Plants
(Wink,1993b). of the parasite Pedicularis racemosa contained quinolizidine
The quinolizidine alkaloids lupanine (21), sparteine (17), alkaIoids from a Lupinus argenteus hybrid (Schneider and
lupinine (24), 13-hydroxylupanine (23), and 17-hydroxylu- Stermitz, 1990). Plants of Pedicularis semibarbata from
panine are toxic to and inhibit the growth of the hemipterous California were shown to contain quinolizidine alkaloids that
aphid Acyrthosiphon pisum that normally feeds on quinolizi- they obtained via root pamsitism from Lupinus fulcratus.
dine-free peas and beans (Harborne, 1986; Kinghorn and Other plants were alkaloid free (Stermitz et al., 1989).
Balandrin, 1984). Lupinine (24) has been shown to be a Plants of the parasite Cuscuta reflexa (Cuscutaceae or
feeding deterrent and growth inhibitor for the two-striped Convolvulaceae) acquire quinolizidine alkaloids from the
grasshopper Me/anop/us bivittatus (Orthoptem). In general, host Spartiumjunceum (Hartmann, 1991). The species Cus-
aphids avoid feeding on plants that contain quinolizidine cuta reflexa and C. platyloba take up alkaloids from a variety
alkaloids, but some phloem-feeding aphids were also found of legume host plants, including Cytisus praecox, Chamae-
to contain these alkaIoids (Kinghorn and Balandrin, 1984). cytisus hirsutus, Petteria ramentacea, andSpartiumjunceum
Sparteine (18) in Sarothamnus scoparius acts as a feeding (Baumel et aI., 1994).
stimulant for the specialized aphid Acyrthosiphon spartii. Polyphagous molluscs such as Helix pomatia and Arion
Larvae of the blue Iycaenid butterfly, Plebejus icarioides, rufus generally do not feed on plants containing alkaloids.
are obligate feeders on several alkaIoid-rich Lupinus species. These species avoided the alkaloids cytisine (27) and N-
The butterfly readily tolemtes the alkaIoids, but these com- methylcytisine (31) more than lupanine (21). The molluscs
pounds do not appear to be essential for the diet of the adult clearly preferred alkaIoid-free artificial diets (Wink, 1984a).
insect (Kinghorn and Balandrin, 1984). The larvae, however, The presence of these alkaloids also has been shown to in-
will not accept other types of plants and succumb without hibit feeding of mammals and insects and to inhibit growth
lupines in their diet (Waller and Nowacki, 1978). of bacteria and fungi (Wink, 1985a). The ranges of alkaIoids
In the aphid genus Aeyrthyosiphon, two closely related in the plants tested were higher than those shown to be active
species feed on legumes. Individuals of one species, A. pisii, in the labomtory.
develop well only when they feed on plants lacking quinoliz-
idine alkaloids. Those of the second species, A. spartii, only lIIedicinal Uses of Quinolizidine Alkaloids
develop when they feed on plants containing these alkaloids. Both matrine (26) and cytisine (27) have established ame-
This aphid actually appears to be a!!mcted by sparteine (17) bicidal activity (Phillipson and O'Neill, 1987). Matrine has
(Waller and Nowacki, 1978). antitumor activity against Ehrlich ascites tumor (Wink,
The aphid Macrosiphon albifrons is attracted to Lupinus 1993b).
species with a chamcteristic alkaloid pattern. Individuals that
had low alkaloid content were not attacked, nor were plants Quinolizidine Alkaloids and Livestock
with different quinolizidine alkaloid composition. This aphid Poisouing
excretes a portion of the alkaloids in the honeydew and re-
tains a portion in its body (Wink and Romer, 1986). Another Alkaloids of this class have long been known for their
aphid, Aphis cytisorum, which infests broom plants (Cytisus toxicity to range animals (Hartmann, 1991; Kinghorn and
scoparius), accumulates up to 0.5 mg alkaloid per gram fresh Balandrin, 1984). Sheep seem to be especially susceptible.
weight. The alkaloids are similar in composition to those of Poisoning with quinolizidine alkaloids does not appear to
the host plant, 17-oxosparteine (25), sparteine (17), 12,13- be cumulative. Most problems occur in the fall when seeds
dehydrosparteine, and iupanine (21) (Wink et al., 1982). (which often accumulate the alkaloids in large quantities)
Some plants of the genus Castilleja are hemiparasitites are produced (Hartmann, 1991). Cytisine (27) and anagyrine
that attack many species of host plants, especially those in (29) are particularly poisonous and have temtogenic activity
the Asteraceae and Fabaceae (McCoy and Stemitz, 1983). in higher animals (Kinghorn and Balandrin, 1984). LDso
Plants of Castilleja linariifolia that ntilized sagebrush (Arte- values (i.p.) range from about 10 to 600 mg/kg. The LDso
misia species which lack alkaIoids) were devoid of alkaIoids. i.p. for cytisine (27) in mouse is 9.3 mgikg, that for lupanine
(21) 80 mg/kg, and that for sparteine (17) 55 mg/kg (Wink,
Several species that parasitized quinolizidine-containing leg-
1993b).
umes contained similar alkaloids, although not in identical
concentrations. Other Castilleja species that attacked Sene-
cio species contained pyrrolizidine alkaIoids. The variability lIIescal-Hallucinogeuic Properties of
of alkaloid content in Castilleja species is linked to that of
Quinolizidine Alkaloids
the host plant (Stermitz and Hams, 1987). The seeds of Sophora secundiflora, the mescal bean or
Plants from five species of Pedicularis (Scrophularia- colorin, have long been considered hallucinogenic by Indi-
ceae) were found to take up alkaloids from their host plants. ans of Mexico and the southwestern United States (Schultes
Those of Pedicularis groenlandica and P. bracteosa took and Hofmann, 1973). The main alkaIoid in these seeds is
up senecionine from the host Senecio triangularis. Those of cytisine (27). Genista canariensis also has been considered
Pedicu/aris crenulata contained anagyrine (29) (a quinolizi- to be mildly hallucinogenic (Kinghorn and Balandrin, 1984).
560 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids

Il'IDOLIZIDIl'IE ALKALOIDS ter 32), Amaryllidaceae (alkaloids derived from both phenyl-
alanine and tyrosine, Chapter 33), and certain steroidal alka-
This group of bicyclic alkaloids has a fused 5- and 6-memb- loids of the Solanaceae (alkaloids derived from terpenes,
ered ring system and is intermediate in that regard to the Chapter 36).
pyrrolizidine and quinolizidine alkaloids (Fig. 30.12). At Amphibians (probably best known are frogs of the genus
least 170 indolizidine alkaloids are known (Verpoorte et al., Dendrobates and toads of the genus Melanophryniscus) are
1991). In contrast to those groups, however, these alkaloids known to produce a number of indolizidine alkaloids. Some
usually have not been identified clearly as a distinct group representative alkaloids of this origin are illustrated in Fig.
of alkaloids (Blbein and Molyneux, 1987; Molyneux, 1993). 30.13 (Daly, 1982; Daly and Spande, 1986; Daly et al., 1993;
Indolizidine alkaloids often do not react with Dragendorff Howard and Michael, 1986; Witkop and Gossinger, 1983).
reagent or with ninhydrin and have often been missed in Subcutaneous doses of pumiliotoxin B as low as 20 ,...g cause
screening programs. death in mice. Pumiliotoxin B (32) interacts with voltage-
The techniques for isolation, purification, and characteri- dependent sodium channels. Although pumiliotoxin A (33)
zation of indolizidine alkaloids have been reviewed (Moly- type class alkaloids originally were thought to be unique to
neux, 1993). dendrobatid frogs, they are now known to occur in a variety
Other groups of alkaloids that coincidentally have this of amphibians (Dalyet al., 1993).
ring structure will be discussed under better known group- Several indolizidine alkaloids are known from Dendrob-
ings [e.g., the alkaloids of Aspidosperma (indole alkaloids, ium (Orchidaceae). Dendroprimine (34) is representative of
Chapter 34), Erythrina (benzylisoquinoline alkaloids, Chap- these compounds (Howard and Micheal, 1986).

;lU
~-,)J-OO ep
HO H

N+

O·
I
yH _.•, OH

swainsonine (35) swainsonine N-oxide (37) castanospermine (36)

s1aframine (38)
'Cb '.c:±> ,
i
dendroprimine (34)
5
J

~'

julisporine (39)

Fig. 30.12. Some representative indolizidine alkaloids.

 Pyrrolizidine. Quino/izidine, and Indolizidine Alkaloids 561

\\ .S?
The alkaloid slaframine (38) is produced by cultures of

~
Rhizoctonia leguminocola grown on nontoxic red clover ex-
tract. This alkaloid is responsible for a series of characteris-
tics such as slobbering (excessive salivation) and lacrima-
tion. This .. disease" has been detennined to result from
consumption of the fungal mycelia (Elbein and Molyneux,
pPllllliDtoxin23'1A

Lt;
gephyrotcWnWAB
1987; Fellows et aI., 1989; Howard and Michael, 1986).
Several hydroxypiperidine alkaloids share properties with
hydroxyindolizidine alkaloids. These compounds are dis-
cussed in Chapter 29 in the section Piperidine Alkaloids.

Biosynthesis.
OR
OH
The discovery of the indolizidine alkaloid swainsonine
pumillotoxlnU7C allopumlllotoxln3J9A (35) and its N-oxide (37) in the leguminous genera Astraga-
lus, Oxytropis, and Swainsona suggests a biosynthetic route
somewhat resembling those of pyrrolizidine and quinolizi-
dine alkaloid biosynthesis (Harris et aI., 1987; Kinghorn and
~~
H

OR OK Balandrin, 1984). The similarity of certain indolizidine alka-

OU N H OR N H
loids to coniine-related compounds, such as 8-coniceine (not
naturally occurring) has been observed.
pumlllotonlnB('u)
The biosynthesis of slaframine (38) and swainsonine (35)
pumilioloxinA (J3)
in Rhizoctonia leguminicola involves lysine and proceeds
Fig. 30.13. Simple indolizidine alkaloids from amphibians (modified via the intennediacy of pipecolic acid. Carbon atoms 2 and
from Howard and Michael, 1986; used with pennission of the copyright 3 have been demonstrated to come from acetate via malonate
owner, Academic Press. Orlando. FL).
(Fig. 30.14). The biosynthesis of swainsonine and related
compounds in plants does not appear to have been investi-
gated (Elbein and Molyneux, 1987; Harris et aI., 1987; How-
The two most studied hydroxy indolizidine alkaloids are
ard and Michael, 1986).
swainsonine (35) and castanospennine (36). Swainsonine
Differences in configuration of swainsonine (35) and sla-
occurs in Swainsona canescens (of Australia) and Astragalus
framine (38) appear to arise by involvement of an iminium
and Oxytropis (western United States) at levels of about
ion similar to the situation involved in conversion of ( + )-
0.001-0.003%. These plants have long been known to pro- reticuline to (- )-reticuline in the benzylisoquinoline alka-
duce characteristic livestock poisoning, both in the United loid series (see Chapter 32) (Harris et aI., 1987).
States and Australia (Elbein and Molyneux, 1987; Harris
et aI., 1987; Hartmann, 1991; Howard and Michael, 1986; Biological Activity of IndoIizidine Alkaloids
Molyneux and James, 1982). The similarity in chemistry of G1ycosidases are important in life processes in all organ-
Castanospermum and a South American genus Alexa sug- isms. These compounds catalyze the hydrolysis of the sugar
gest that the two are closely related (Fellows et aI., 1989). moieties of numerous types of glycosides (see Chapter 1
The symptoms of this intoxication are similar to those of and 15). The glycosidases of animals, fungi, and bacteria
mannosidosis. Swainsonine (35), along with. another alka- degrade large molecules to prepare them for uptake by the
loid of this type, is found in the fungus Rhizoctonia legumini- organism. Glycosides also are involved in cells as compo-
cola which is pathogenic on red clover. Swainsonine N- nents of catabolic systems. In many organisms, these en-
oxide (37) occurs in Astragalus lentiginosus in addition to zymes are thought to be involved in the production of new
swainsonine. Castanospennine (36) occurs in the Moreton cell wall materials and expansion of existing cell walls. Fur-
Bay chestnut or black bean (Castanospermum australe, Fa- thennore, glycosidases are important in the fonnation and
baceae). Castanospermine occurs at about 0.3% in mature processing of glycoproteins (Elbein and Molyneux, 1987;
seeds and has also been detected in the leaves and bark of Howard and Michael, 1986).
the tree. Consumption of these seeds causes severe gastroin- Castanospennine (36) is a potent inhibitor of almond em-
testinal pain. As is true for locoweed poisoning, some ani- ulsin l3-glucosidase. This alkaloid gives almost complete in-
mals develop a preference for them, despite the often fatal hibition at 50 fLg/ml. At five times that level, castanosper-
gastroenteritis that develops. Both swainsonine and casta- mine did not inhibit a-glucosidase, a- or l3-galactosidase, a-
nospennine are soluble in water, but are not highly soluble mannosidase, l3-glucuronidase, I3-N-acetylglucosaminidase,
in nonpolar solvents (Elbein and Molyneux, 1987; Fellows or a-fucosidase. Castanospennine proved to be a competi-
et aI., 1989). tive inhibitor of amyloglucosidase and almond emulsin 13-
Castanospennine (36) is a component of the honeydew glucosidase. This alkaloid also inhibited glycoprotein pro-
of mealybugs (Pseudococcus longispinus) living on Casta- cessing. Castanospennine inhibits plant root elongation, cy-
nospermum australe (Molyneux et aI., 1990). tomegaloviruses, and retroviruses (Wink, 1993b).
562 Pyrrolizidine. Quinolizidine. and lndolizidine Alkaloids

O
OH

--
H
i CO,H

NH ......:m;;:.;;:I.;;:D;;:.t;;:e_ ( ) - \ ..•"'OH
~~J
!

65-
OCOCH,

~d) 00

slaframine (38)
swainsonine (35)

Fig. 30.14. Biosynthesis of slaframine and swainsonine in Rhizoctonia leguminocola (modified from Howard and Michael, 1986; used with pennission
of the copyright owner, Academic Press, Orlando, FL),

Castanospennine (36) (1,6,7 ,8-tetrahydroxyoctahydroin- tasis, intracellular transport, and viral infectivity. Both casta-
dolizine) is a potent phytotoxin for dicotyledonous plants nospennine (36) and swainsonine (35) have been of interest
at 300 ppb, and for monocotyledonous plants at 200 ppb. for study of mV-l (AIDS) (Fellows, 1988).
Swainsonine is ineffective as an inhibitor of root elongation. Castanospennine (36) is a feeding deterrent for aphids
(Stevens and Molyneux, 1988). and greenbugs (Wink, 1993b). Castanospennine is lethal tu
Animal systems contain three distinct types of a-mannos- the larvae of the bruchid beetle Callosobruchus maculatus
idases. Swainsonine (35) inhibits one of these completely at and the flour beetle Tribolium confusum at 0.03% and 0.1 %
concentrations as low as 20 11m. At much higher levels (more in the diet (Fellows et al., 1986).
than 50 times), this compound did not exhibit inhibitory The unusual indolizidine alkaloid julisporine (39) from
activity toward \3-mannosidase, a- or \3-glucosidase, a- or \3- Prosopis juliflora (Fabaceae) possessed in vitro activity
galactosidase, \3-N-acetylhexosaminidase, or a-L-fucosidase against several dennatophytic fungi (Clark and Hufford,
(Elbein and Molyneux, 1987). The alkaloid also alters nor- 1992).
mal processing of N-linked oligosaccharides and biosyn-
thesis of glycoproteins. Indollzldine Alkaloids In Insects
These compounds promise to be of great value for the
examination of the structure activity relationship of glyco- Indolizidine alkaloids are produced by the Pharaoh ant,

9&
proteins and their role in the immune response, cancer metas- Monomorium pharaonis. This tropical ant is now a common

HO=OHOH (bH ,:Y

(b: ,:Y

N N N

elaeocarplne (42) (+)-Isoelaeocarplne (43) (+)-elaeokanine (-)-e1aeokanlne B

(-)-elaookanine C isoelaeocarpine elaeokanine E peduncularine (44)

Fig. 30.15. Alkaloids of the Elaeocarpaceae,

 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids 563

pest in North America and Europe where it has been intro- Zealand). Although little biosynthetic work has been done,
duced. The alkaloids monomorine VI (40) and monomorine the precursors have been suggested to be tryptamine and a
I (41) are found in the poison gland and in the Dufour's monoterpene such as nerol. The alkaloids of a third genus,
gland of the insect (Fig. 30.12). These alkaloids act as attrac- Aceralium, have not been studied.
tants for the Pharaoh ant (Howard and Michael, 1986).
PbenantbroindoUzidine Alkaloids
Elaeocarpaceae Alkaloids
This group of about 20 alkaloids, best known of which
The primarily tropical family Elaeocarpaceae contains is tylophorine (45), are primarily found in the genus Tylo-
about 350 species in 7 genera. The genus Elaeocarpus con- phora, but also occur in the genera Cynanchum (Vinceloxi-
tains indolizidine alkaloids almost exclusively, for example, cum), Anlitoxicum, and Pergularia of the family Asclepi-
elaeocarpine (42) and isoelaeocarpine (43) (Fig. 30.15). adaceae. Some representatives of these alkaloids possess
Another genus, Aristotelia, contains indole alkaloids simple indolizidine structures. Both these simple types and
[e.g., peduncularine (44)], but these appear to be, at most, phenanthroindolizidine alkaloids are also known to occur
only distantly related to those of Elaeocarpus (Bick and Hai, in the Moraceae (Ficus), Urticaceae (Boehmeria), and the
1985). This genus is found in the temperate regions of the Lauraceae (Cryptocarya) (Bick and Sinchai, 1981; Herbert,
southern hemisphere (Argentina, Australia, Chile, and New 1985; Howard and Michael, 1986).

~
I
""
-
.I ~
~"----£-.l
&'
CO,R
n 0 ,R

°
OCR,
!
.
CR,O
--

JQ7 " (46)

~ I
0) RN
,(CO,R

•
CR,O "" °
OR t
~NR'

RO
~ CO,R

CR,O
~ OR OCR,
tylophorine (45)
OCR,
CR
C R ' °' Z m °
I""
,6

1 N

CR,O
OR OCR,
CR,O :'
tylophorinlne (47) cryptopleurine (48)

Fig. 30.16. Proposed biogenesis of tylopborine (Bick and Sinchai. 1981; used with pennission of the copyright owner, Academic Press, Orlando, FL);
cryptopleurine.
564 Pyrrolizidine, Quinolizidine, and Indolizidine Alkaloids

Biosynthesis BIRECKA, H., M. BIRECKI, and M. W. FROHLICH, Arginine and orni-

thine decarboxylases and polyamines in pyrrolizidine a1kaloid-
In terms of biosynthesis, these alkaloids are derived front containing plants, Abstracts, Phytochem. Soc. North America,
tyrosine, phenylalanine, and ornithine (Fig. 30.16). Phenyl- Annual Meeting, 1988.
alanine appears to be converted to cinnamic acid before in- BIR.ECKA, H., J. L. CATALFAMO. and R. N. EISEN, A sensitive method
corporation. Phenacylpyrrolidines (such as 46) are defined for detection and quantitative determination of pyrroIizidine al-
as key intermediates. Doubly-labeled compounds of this type kaloids, Phytochemistry, 20, 343-344 (1981).
were incorporated intact into tylophorinine (47). Oxidative BLASKO, G. and G. A. CORDELL, Recent developments in the chem-
coupling steps yield tylophorine (45) and tylophorinine (47) istry of plant-derived anticancer drugs, in Economic and Medic-
(Bick and Sinchai, 1981; Herbert, 1985; Specialist Periodic inal Plant Research, Vol. 2 (H. Wagner, H. Hikino, and N. R.
Reports, Alkaloids, 1978, 1979). Farnsworth, eds.), 119-191, Academic Press, London, 1988.
BoPPRE, M., Chemical communication. plant relationships, and
Biological Activity of mimicry in the evolution of danaid butterliies, Ent. Exp. Appl.,
PhenanthroindoUzidlne Alkaloids 24, 264-277 (1978).
BOPPl<l', M., R. L. PETIT, D. SCENEIDER, andJ. MElNWALD, Behav-
Ty10phorine (45) has anti-inflammatory properties. Leaf iourally mediated contacts between scent organs: Another pre-
extracts of Ty/ophora species have been used to treat asthma requisite for pheromone production in Danaus chrysippus males
and rheumatism (Wagner and Proksch, 1985). Several alka- (Lepidoptera), J. Compo Physio!., 126, 97-103 (1978).
loids of this group have pronounced antitumor activity BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from
(Blasko and Cordell, 1988; Gellert, 1982). The activities of plants, in Phytochemical Resources for Medicine and Agricul-
tylocrebrine, tylophorine (45), and cryptopleurine (48) (a ture (H. G. Nigg and D. S. Seigler, eds.), 117-158, Plenum
phenanthroquinolizidine aIkaloid) (Fig. 30.16) are all less Press, New York, 1992.
than those of emetine, which they resemble in observed ac- BROOKS, C. J. W. and D. G. WATSON, Phytoalexins, Nat. Prod.
tivity (Blasko and Cordell, 1988). Tylophorine is toxic to Rep, 2, 427-459 (1985).
Paramecium caudatum at 1: 50,000 dilution, but has low BROWN, K. S., JR., Adult-obtained pyrrolizidine alkaloids defend
toxicity to mice and gninea pigs. Tylophorine is more than ithomiine butterflies against a spider predator, Nature, 309,
twice as potent against cultured Entamoeba histo/ytica as is 707-709 (1984).
emetine, a common drug for treating amebic dysentery CLARK, A. M. and C. D. HUFFORD, Antifungal alkaloids, in The
(Borris and Schaeffer, 1992). Alkaloids, Vol. 42 (G. A. Cordell, ed.), 117-150. Academic
Press, New York, 1992.
CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual

REFERENCES Reports on NMR Specttoscopy, Vo!' 13 (G. A. Webb, ed.),

60-210, Academic Press, London, 1982.
ASLANOV, KH. A., Yu. K. KUSHMURADOV, and A. S. SADYKOV, CULVENOR, C. C. J., Pyrrolizidine alkaloids-Occurrence and sys-
Lupine alkaloids, in The Alkaloids, Vol. 31 (A. Brossi, ed.), tematic importance in Angiosperms, Bot. Notiser, 131, 473-486
118-192, Academic Press, New York, 1987. (1978).
BAUMEL, P., L. WrITE, F. CzYGAN, and P. PROKSCH, Transfer of DALY, J. W., Alkaloids of neotropical poison frogs (Dendrobati-
quinolizidine alkaloids from various host plants of the Fabaceae dae), Fortschr. Chern. Org. Naturst., 41, 205-340 (1982).
to parasitizing Cuscuta species, Biochem. Syst. Ecol., 22, DALY, J. W. and T. F. SPANDE, Amphibian alkaloids: Chemistry,
647-656 (1964). pharmacology, and biology, in Alkaloids. Chemical and Biolog-
BEIER, R. C. and H. N. NIGG, Natural toxicants in foods, in Phyto- ical Perspectives, Vo!' 4 (S. W. Pelletier, ed.), 1-274, Wiley,
chemical Resources for Medicine and Agriculture (H. G. Nigg New York, 1986.
and D. S. Seigler, eds.), 247-376, Plenum Press, New York, DALY, J. W., H. M. GARRAFFO, and T. F. SPANDR, Amphibian
1992. alkaloids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.),
BENN, M., J. DE GRAVE, C. GNANASUNDERAM, and R. HurcHINs, 186-288, Academic Press, New York, 1993.
Host-plant pyrrolizidine alkaloids in Nyctemera annulata Bois- DAVlES, N. M. and D. H. G. CROUT, Pyrrolizidine alkaloid biosyn-
duval: Their persistance through the life cycle and transfer to thesis. Relative rates of incorporation of the isomers of isoleu-
a parasite, Experientia, 35,731-732 (1979). cine into the necic acid component of senecionine, J. Chern.
BERNAYS, E., J. A. EDGAR, and M. ROTHSCHILD, Pyrrolizidine alka- Soc., Perkin Trans. I, 2079-2086 (1974).
loids sequestered by aposematic grasshopper, Zonocerus varie- DEVLIN, J. A. and D. J. ROBINS, Pyrrolizidine alkaloids. biosyn-
gatus, J. Zool. (Lond.), 182, 85-87 (1977). thesis of trichodesmic acid. 1. Chern. Soc., Perkin Trans. I,
BICK, I. R. C. and M. A. 1iAI, Aristotelia alkaloids, in The Alkaloids, 1229-1332 (1984).
Vol. 24 (A. Brossi, ed.), 113-151, Academic Press, New York, EDGAR, J. A., P. A. COCKRUM, and J. L. i'RAEN, Pyrrolizidine alka-
1985. loids in Danaus plexippus L. and Danaus chrysippus L., Expe-
BICK, I. R. C. and W. SINCHAI, Phenanthroindolizidine and phe- rientia, 32, 1535-1537 (1976).
nanthroquinolizidine alkaloids, in The Alkaloids, Vol. 19 (R. EDGAR, J. A., C. C. J. CULVENOR, and T. E. PUSKE, Coevolution
G. A. Rodrigo, ed.), 193-220, Academic Press, New York, of danaid butterliies and their host plants, Nature, 250, 646-648
1981. (1974).
 Pyrrolizidine, Quinolizidine. and Indolizidine Alkaloids 565

EISNER, T. and J. MEINWALD, Alkaloid-derived pheromones and HERBERT, R. B., The biosynthesis of plant alkaloids and nitrogenous
sexual selection in lepidoptera, in Pheromone Biochemistry (G. microbial metabolites, Nat. Prod. Rep., 3, 185-203 (1986).
D. Prestwich and G. J. Blomquist, eds.), 251-269, Academic HERBERT, R. B., The biosynthesis of plant alkaloids and nitrogenous
Press, Orlando, FL, 1987. nticrobial metabolites, Nat. Prod. Rep., 5, 523-540 (1988).
ELBElN, A. D. and R. J. MOLYNEUX, The chemistry and biochemis- HOWARD, A. S. and J. P. MICHAEL, Simple indolizidine and quino-
try of simple indolizidine and related polyhydroxy alkaloids, in lizidine alkaloids, in The Alkaloids, Vol. 28 (A. Brossi, ed.),
Alkaloids. Chemical and Biological Perspectives, Vol. 5 (S. W. 183-308, Academic Press, New York, 1986.
Pelletier, ed.), 2-54, Wiley, New York, 1987.
JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary
ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod. compounds to the seed-eating larvae of the bruchid beetle Callo-
Rep., 5, 581-612 (1988). sobruchus maculatus, Phytochentistry, 16, 223-227 (1977).
FmLows, L. E., Castanospermine, swainsonine, and related poly- JENETI-SlEMS, K. M. KAwGA, and E. EICH, Ipangulines, the first
hydroxy alkaloids from plants, Phytochem. Soc. North America, pyrrolizidine alkaloids from the Convolvulaceae, Phytochemis-
Annual Meeting, 1988. try, 34, 437-440 (1993).
FELLOWS, L. E., S. V. EVANS, R. J. NASH, and E. A. BELL, Polyhy- KHAN, H. A. and D. J. ROBINS, Pyrrolizidine alkaloid biosynthesis.
droxy plant alkaloids as glycosidase inhibitors and their possible Synthesis of 13C-Iabelled putrescines and their incorporation
ecological role, in Natural Resistance of Plants to Pests (M. B. into retronecine, J. Chem. Soc., Perkin Trans. I, 101-105
Green and P. A. Hedin, eds.), ACS Symposium Series 296, (l985a).
72-78, American Chentical Society, Washington, DC, 1986.
KHAN, H. A. and D. J. ROBINS, Pyrrolizidine alkaloid biosynthesis.
FELLOWS, L. E., G. C. KITE, R. J. NASH, M. S. J. SIMMONDS, and Synthesis of 13C-Iabelled putrescines and their incorporation
A. M. SCOFIELD, Castanospennine, swainsonine and related into retronecine, J. Chem. Soc., Perkin Trans. I, 819-824
polyhydroxy alkaloids: Structure, distribution, and biological (1985b).
activity, in Plant Nitrogen Metabolism (J. E. Poulton, J. T.
KINGHORN, A. D. and M. F. BALANDRIN, Quinolizidine alkaloids
Romeo, and E. E. Conn, eds.), Recent Advances in Phytochem-
of the Leguminosae: Structural types, analyses, chemotaxon-
istry Vol. 23, 395-427, 1989.
omy, and biological properties, in Alkaloids. Chentical and Bio-
FRASER, A. M. and D. J. ROBINS, Application of2H NMR spectros-
logical Perspectives, Vol. 2 (S. W. Pelletier, ed.), 105-204,
copy to study the incorporation of enantiomeric 2H-Iabelled ca-
Wiley, New York, 1984.
daverines into quinolizidine alkaloids, J. Chern. Soc., Perkin
KRASNOFF, S. B. and D. E. DUSSOURD, Dihydropyrrolizine attrac-
Trans. I, 105-109 (1987).
tants for arctiid moths that visit plants containing pyrrolizidine
GElSSMAN, T. A. and D. H. G. CaOUT, Organic Chentistry of Sec-
alkaloids, J. Chern. Ecol., 15, 47-60 (1989).
ondary Plant Metabolism, Freeman Cooper, San Francisco,
1969. LEETE. E., The biosynthesis of alkaloids. in Specialist Periodical
Reports, Biosyntheis, Vol. 7 (R. B. Herbert and T. J. Simpson,
GELLBRT, E .. Iudolizidine alkaloids, J. Nat. Prod., 45, 50-72
eds.), 102-223, The Royal Society of Chemistry, Oxford, 1983.
(1982).
L'EMPEREUR, K. M., Y. LI, F. R. STERMrrZ, and L. CRABTREE,
Goss, G. 1., The interaction between moths and plants containing
Pyrrolizidine alkaloids from Hackelia californica and Gno-
pyrrolizidine alkaloids, Environ. Eutomol., 8, 487-493 (1979).
phaela latipennis, an H. californica-hosted arctiid moth, 1. Nat.
HARBORNE, J. B., Iutroduction to Biochentical Ecology, 2nd ed., Prod., 52, 360-366 (1989).
Academic Press, New York, 1982.
MABRY, T. J. and J. A. MEARs, Alkaloids and plant systematics,
HARBORNE, J. B., Recent advances in chentical ecology, Nat. Prod. in Chemistry of the Alkaloids (S. W. Pelletier, ed.), 719-746,
Rep., 3, 323-344 (1986). Van Nostrand Reinhold, New York, 1970.
HARBORNE, J. B., Recent advances in chentical ecology, Nat. Prod. MANN, J., Secondary Metabolism, Oxford University Press, Ox-
Rep., 7, 85-109 (1989).
ford, 1978; 2nd edition, 1987.
lIARRrs, C. M., M. J. SCHNEIDER, F. S. UNGEMACH, J. E. lIrLL, and
MASTERS, A. R., Dual role of pyrrolizidine alkaloids in nectar, J.
T. M. HARRIs, Biosynthesis of the toxic indolizidine alkaloids
Chem. Ecol., 17, 195-205 (1991).
slaframine and swainsonine in Rhizoctonia leguminicola: Me-
tabolism of I-hydroxyindolizidines, J. Am. Chern. Soc., lID, MATTOCKS, A. R. and R. JUKES, Improved field tests for toxic
940-949 (1987). pyrrolizidine alkaloids, J. Nat. Prod., 50, 161-166 (1987).
HARTMANN, T., ALKALOIDS, in Herbivores: Their Interactions with McCoy, J. W. and F. R. STERMrrZ, Alkaloids from Castilleja mini-
Secondary Plant Metabolites, Vol. I, 2nd ed. (G. A. Rosenthal ata and Penstemon whipp/eanus, two host species for the plume
and M. R. Berenbaum, eds.), 79-121, Academic Press, San moth, Ambryptilia (Platyptilia) pica, 1. Nat. Prod., 46, 902-907
Diego, CA, 1991. (1983).
HARTMANN. T., A. EHMKE, U. EILERT, K. VON BORSTEL, and C. MEARS, J. A. and T. J. MABRY, Alkaloids in the Leguminosae, in
THBURING. Sites of synthesis, translocation, and accumulation Chemotaxonomy of the Leguminosae (J. B. Harbome, D. Boul-
ofpyrrolizidine alkaloidN-oxides in Senecio vulgaris L., Planta, ter, and B. L. Turner, eds.), 73-178, Acadentic Press, London,
177, 98-107 (1989). 1971.
HERBERT, R. B., The synthesis of indolizidine and quinolizidine MENDE, P. and M. WINK, Uptake of the quinolizidine alkaloid lu-
alkaloids of Tylophora, Cryptocarya, Ipomoea, Elaeocarpus panine by protoplasts and isolated vacuoles of suspension-cul-
and related species, in Alkaloids. Chentical and Biological Per- tured Lupinus po/yphyllus cells. Diffusion or canier mediated
spectives, Vol. 3 (S. W. Pelletier, ed.), 241-273, Wiley, New transport, J. Plant Physiol., 129, 229-242 (1987).
York, 1985. MOLYNEUX, R. J., Polyhydroxy indolizidine and related alkaloids,
566 Pyrrolizidine, Quinolizidjne, and lndolizidine Alkaloids

in Alkaloids and Sulphur Compounds (P. G. Watennan, ed.), STERMITZ, F. R., G. N. BBLOFSKY, D. NG, and M. C. SINGBR,
Vol. 8 of Methods in Plant Biochemistry (P. M. Dey and J. B. Quinolizidine alkaloid, obtained by Pedicularis semibarbata
Harborne, eds.), 511-530, Academic Press, London, 1993. (Scrophulariaceae) from Lupinus fulcratus (Legumino,ae) fail
MOLYNEUX, R. I. and L. F. lAMES, Loco intoxication: Indolidizine to influence the specialist herbivore Euphydryas editha (Lepi-
alkaloids of spotted locoweed (Astragalus lentiginosus), Sci- doptera) oviposition. J. Chern. Bcol., IS, 2521-2530 (1989).
ence, 216, 190-191 (1982). STBRMITZ, F. R., G. H. HARRIS, and W. JING, lridoids and
MOLYNEUX, R. I., B. C. CAMPBELL, and D. L. DREYER. Honeydew alkaloids from Castilleja (Scrophulariaceae) host plants for
analysis for detecting phloem transport of plant natural prod- Platyptilia pica (Lepidoptera: Prerophoridae). Rexifoline con-
ucts, J. Chern. Beol., 16, 1899-1909 (1990). tent of P. pica, Biochem. Syst. Ecol., 14, 499-506 (1986).
NAHRSTEDT, A., Strukturelle Beziehungen zwischen pflanzlichen STEVENS, K. L. and R. I. MOLYNEUX, Castanospennine-A plant
und tieri,chen Sekundiirstoffen, Planta Medica, 44, 2-14 growth regulator, J. Chern. Beol., 14, 1467-1473 (1988).
(1982). SUFFNESS, M. and G. A. CORDEU., Antitumor alkaloids, in The
PASTEELS, J. M., M. ROWELL-RAHIER, T. RANooux, J. C. BRABK- Alkaloids, Vol. 25 (A. Brossi, ed.), 3-355, Academic Press,
MAN, and D. DALOZE, Pyrrolizidine alkaloid, of probable New York, 1985.
host-plant origin in the pronatal and elytral secretion of Tu, Z., C. KONNO, D. D. SOEIARTO, D. P. WALLER, A. S. BINGEL,
the leaf beetle Oreina cacaliae, Ent. Exp. Appl., 49, 55-58 R. J. MOLYNEUX, J. A. EOOAR, G. A. CORDELL, and H. H.
(1988). S. FaNG. Identification of senecionine and senecionine N-
PHn.!.IPsON, J. D. and M. J. O'NBILL, Antimalarial and arnoebici- oxide as antifertility constituents in Senecio vulgaris, I. Pharm.
dal natural products, in Biologically Active Natural Products Sci., 77, 461-463 (1988).
(K. Hostettmann and P. J. Lea, eds.), 49-64, Clarendon Press, VAN WYK, B. and G. H. VBRDOORN. The chemotaxonomic signifi-
Oxford, 1987. cance of integerrimine in Buchenroedera and Lotonis section
PLISKE, T. E., Courtship behavior and use of chemical communi- Krebsia, Biochem. Sys!. Beol., 16, 287-289 (1988).
cation by males of certain species of ithomiine butterflies, VAN WYK, B. and G. H. VERDOORN, Alkaloids as taxonomic
Ann. Entornol. Soc. Am., 68, 935-942 (1975). characters in the tribe Crotalarieae (Fabaceae), Biochem. Syst.
PLISKE, T. E., J. A. EOOAR, and C. C. J. CuLVENOR, The chemical Ecol., 18, 503-515 (1990).
basis of attraction of ithomiine butterflies to plants containing VERPOORTE, R., R. VAN DER HaDDEN, W. M. VAN GULIK, and
pyrrolizidine alkaloids, J. Chern. Beol., 2, 255-262 (1976). H. J. G. TEN HooPBN, Plant biotechnology for the production
ROBINS, D. J., Pyrrolizidine alkaloids, in Alkaloids and Sulphur of alkaloids: Present status and prospects, in The Alkaloids,
Compound, (P. G. Waterman, ed.), Vol. 8 of Methods in Vol. 40 (A. Brossi, ed.), 1-188, Academic Press, New York,
Plant Biochemi,try (P. M. Dey and J. B. Harborne, eds.), 1991.
175-195, Academic Press, London, 1993.
W AGNBR, H. and A. PROKSCH, Immunostimulatory drugs of fungi
ROBY, M. R. and F. R. STERMITZ, Pyrrolizidine and pyridine and higher plants, in Economic and Medicinal Plant Research,
monoterpene alkaloid, from two Castilleja plant hosts of the Vol. 1 (H. Wagner, H. Hikino, and N. R. Farnsworth, eds.),
plume moth, Platyptilia pica, J. Nat. Prod., 47, 846-853 113-153, Academic Pre,s, London, 1985.
(1984).
WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and
RODBR, E., Pyrrolizidinalkaloidhaltige Arzneipflanzen, Deutsche Metabolism in Plants, Plenum Press, New York, 1978.
Apoth. Zeit., 132, 2427-2435 (1992).
WATBRMAN, P. G. and A. I. GRAY, Chemical systematics, Nat.
SCHNEIDBR, D., The 'trange fate of pyrrolizidine alkaloids, in Prod. Rep., 4, 175-203 (1987).
Perspective, in Chemoreception and Behavior (R. F. Chap-
WINK, M., Wounding-induced increase of quinolizidine alkaloid
man, E. A. Bemays, and J. Stoffalono Jr., ed,.), 123-142,
accumulation in lupin leaves, Z. Naturforsch., 38c, 905-909
Springer-Verlag, New York, 1987.
(1983a).
SCHNEIDER, M. J. and F. R. STERMITZ, Uptake of host plant
alkaloids by root parasitic Pedicularis species, Phytochemis- WINK, M., Inhibition of seed germination by quinolizidine alka-
try, 29, 1811-1814 (1990). loids, Planta, 158, 365-368 (1983b).
SCHULTES, R. B. aod A. HOFMANN, The Botany and Chemi,try WINK, M., Chemical defense of lupins. Mollusc-repellent proper-
of Hallucinogens, CC Thomas, Springfield, IL, 1973. ties of quinolizidine alkaloids, Z. Naturforsch., 39c, 553-558
(1984a).
SPECIAUST PERIODIC RaPORTS, The Alkaloid" Vol. 8, Royal
Chemistry Society, London, 1978. WINK, M., The biochemistry and chemical ecology of lupin
SPECIALIST PERIODIC REPORTS, The Alkaloids, Vol. 9, Royal alkaloids, in Proceedings 3rd International Lupine Congress,
I.L.A., La Rochelle, France 1984b.
Chemistry Society, London, 1979.
SPEClAUST PERIODIC REPoRTS, The Alkaloids, Vol. 10, Royal WINK, M., Chemische Verteidigung der Lupinen: Zur biolog-
Chemistry Society, London, 1978. ischen Bedeutung der Chinolizidinalkaloide, Plant Syst. Evol.,
ISO, 65-81 (1985a).
SPENSER, I. D., Stereochemical aspects of the biosynthetic routes
leading to the pyrrolizidine and the quinolizidine alkaloids, WINK, M., Storage of quinolizidine alkaloids in epidennal tissues,
Pure Appl. Chern., 57, 453-470 (1985). Z. Naturforsch., 41c, 375-380 (1985b).
STBRMITZ, F. R. and G. H. HARRIS, Transfer of pyrrolizidine WINK, M., Quinolizidine alkaloids: Biochemistry, metabolism,
and quinolizidine alkaloids to Castilleja (Scrophulariaceae) and function in plants and cell suspension cultures, Planta
hemiparasites from composite and legume host plants, I. Medica, 53, 509-514 (1987).
Chern. Beol., 13, 1917-1925 (1987). WINK, M., Plant breeding: Importance of plant secondary metabo-
 Pyrrolizidine. Quinolizidine, and Indolizidine Alkaloids 567

lites for protection against pathogens and herbivores, Theoret. exploiting moth Creatonotos, Naturwissenschaften, 75,
Appl. Genet., 75, 225-253 (1988). 524-525 (1988).
WINK, M., Allelochemical properties or the raison d'etre of WINK, M. and L. WrITE, Evidence for a wide·spread occurrence
alkaloids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), of the genes of quinolizidine alkaloid biosynthesis, FEBS
I-lOS, Academic Press, New York, 1993a. Lett., 159, 196-200 (1983).
WINK, M., Quinolizidine alkaloids, in Alkaloids and Sulphur WINK, M. and L. WrrrE, Turnover and transport of quinolizidine
Compounds (P. G. Waterman, ed.), Vol. 8 of Methods in alkaloids. Diurnal fluctuations of lupanine in the phloem sap,
Plant Biochemistry (P. M. ney and J. B. Harborne, eds.), leaves and fruits of Lupinus albus, Planta, 161, 519-524
197-239, Academic Press, London, 1993b. (1984).
WINK, M. and L. WlTI'E, Quinolizidine alkaloids as nitrogen
WINK, M. and T. HARTMANN, Localization of the enzymes of
source for lupin seedlings and cell cultures, Z. Naturforsch.,
quinolizidine alkaloid biosynthesis in leaf chloroplasts of Lupi-
40c, 767-775 (1985).
nus polyphyllus, Plant Physio!., 70, 74-77 (1982).
WINK, M., T. HARTMANN, L. WlTI'E, and J. RHEINHElMER, interre-
WINK, M. and T. HARTMANN, Enzymology of quinolizidine alka- lationship between quinolizidine alkaloid producing legumes
loid biosynthesis, in Natural Products Chemistry 1984 (14th and infesting insects: Exploitation of the alkaloid-containing
IUPAC Symposium on the Chemistry of Natural Products) phloem sap of Cytissus scoparius by the broom aphid Aphis
(R. I. Zalewski and J. J. Skolik, eds.), Elsevier, Amsterdam, cytisorum, Z. Naturforsch., 37c, 1081-1086 (1982).
1984.
WITKOP, B. and E. GasSINGER, Amphibian alkaloids, in The
WINK, M. and P. ROMER, Acquired toxicity-the advantages of Alkaloids, Vo!. 21 (A. Brossi, ed.), 139-253, Academic Press,
specializing on alkaloid-rich lupins to Macrosiphon albifrons New York, 1983.
(Aphidae), Naturwissenschaften, 73, 210-212 (1986). WROBEL, J. T., Pyrrolizidine alkaloids, in The Alkaloids, Vo!.
WINK, M. and n. SCHNEIDER, Carrier-mediated uptake of pyrroliz- 26 (A. Brossi, ed.), 327-384, Academic Press, New York,
idine alkaloids in larvae of the aposematic and alkaloid- 1985.
31
Alkaloids Derived from Anthranilic Acid

Introduction completely blocks transcription from DNA, and as cancer

Quinazoline and Pyrroloquinazoline Alkaloids cells usually divide faster than normal body cells, they are
Biosynthesis preferentially killed by this antibiotic. Actinomycin D is one
Distribution of Pyrroloquinazoline Alkaloids of the most potent antitumor agents known.
Biological Activity An additional group of alkaloids that arises from anthra-
Quinazolinocarboline Alkaloids nilic acid, the harman alkaloids, is discussed in Chapter 35.
Quinoline, Furoquinoline, Acridone, and 4-Quinoline Avenalumins, 2,4-dihydroxy-7-methoxy-I ,4-benzoxazin-3-
Alkaloids one (DIMBOA), and related compounds are treated in Chap-
Chemosystematics and Phylogeny ter 7.
References Both the I H- and 13C_NMR (nuclear magnetic resonance)
spectra and techniques for the isolation and characterization
of several groups of alkaloids derived from anthrauilic acid
Il'ITRODVCTlOI'l have been reviewed (Crabb, 1982; Gray, 1993). The results
of feeding studies leading to several alkaloids derived from
Anthrauilic acid (1) is an intermediate in the biosynthesis anthrauilic acid have been tabulated (Leete, 1983).
of tryptophan (see Chapter 7) (Fig. 31.1). A number of alka- In addition to the relatively simple anthranilic-derived
loids, best known from the family Rutaceae, are biosynthes- metabolites mentioned above, several distinctive types of
ized from this important primary compound. alkaloids are derived from this amino acid. Most of these
Small amounts of N-methylanthranilate occur in a num- are associated with the Rutaceae and related families of the
ber of essential oils (Grager, 1980). Another simple repre- Rutales. Among these groups of alkaloids are the quinazo-
sentative, darnasceuine (2), is found in the seeds of Nigella line, pyrroloquinazoline, quinazolinocarboline, quinoline,
damascena L. (Ranunculaceae) (Fig. 31.2). This alkaloid has furoquinoline, and acridone types.
been shown to come from shikimic acid via anthranilic acid
(Cordell, 1981). [I4CH3J-Methiouine is an efficient precur-
sor of all three bound methyl groups (Grager, 1980; Geiss- QlJ1l'IAZOLIl'IE Al'ID FYRROLOQlJ1l'IAZOLIl'IE
man and Crout, 1969). ALKALOIDS
Dianthalexin (4) and related N-benzoylanthranilic acid
derivatives are formed in response to fungal attack in Dian- About 60 simple quinazoline alkaloids have been isolated
thus (Caryophylaceae) (Ponchet et aI., 1984). from several plant families, microorganisms, and auimals
Other compounds apparently are derived from 3-hydrox- (Grager, 1980). Arborine (5), from leaves of Glycosmis spe-
yanthranilic acid (3), for example, several compounds from cies (Rutaceae), is an example of this group of alkaloids
Tecoma stans (Bignoniaceae), are catabolic products oftryp- (Fig. 31.3) (Johne, 1986).
tophan (Grager, 1980; Geissman and Crout, 1969). 3-Hy- The biosynthesis of quinazoline alkaloids is not resolved,
droxyanthranilic acid is involved in the biosynthesis of sev- but anthrauilic acid, probably as N-acetylanthranilic acid, is
eral fungal pigments and in actinomycins from strains of a precursor (Cordell, 1981, Grager, 1980). Both anthranilic
Streptomyces. Actinomycin A was isolated soon after peni- acid and phenylalanine serve as precursors for arborine (5)
cillin, but it was too toxic for general use. Actinomycin D in Glycosmis arborea (Cordell, 1981).

568
 Alkaloids Derived from Anthranilic Acid 569

HO,C)) HO,C))
, 1 ~
~CO,H .0 h

-0 -
~ HO~ 0 HN HO,PO N Amadori

"" NH, - ~~H rearrange;ent

OH OH HoN0 H
cbormlsmic acid anthranilic acid (1)

HO,C~ Q NH, NH,

HO'POWHN
)l) ~HO,Pol-ACO'H
<" OP~
!dH
CO,H

OPO,H <"
(Jc(t:
1 1
H

NH
CO,H

H - 1 "",I OooH - ~ NH '

HO ~ NH
H OH 0
indole-3-g1ycerol phosphate L-tryptophan

Fig. 31.1. Biosynthesis of anthranilic acid and tryptophan (Atta-or-Rahman and Basha, 1983; used with pennission of the copyright owner, Oxford
University Press, Oxford),

(XI
::,....
CO,H

NH,
- Y Y
::,....
OH
1
CO'H

NH,
- ::,....
OCH,
1
CO'H

-
NH,

anthranilic acid (3) damascenine (2) dianthalexin (4)

Fig. 31.2. Biogenesis of damascenine (Geissman and Crout, 1969); dianthalexin.

Quinazoline alkaloids are known from the Acanthaceae Febrifugine (6) (from Dichroa and Hydrangea, Hydran-
(Adhatoda, Anisotes), Araliaceae (Mackinlaya), Arecaceae geaceae) is a febrifuge and has antimalarial activity (Cordell,
(Daemonorops), Fabacea (Galega), Hydrangeaceae (Hy- 1981; Johne, 1986). Although about 100 times more active
drangea, Dichroa), Malvaceae (Sida), Rutaceae [Euxylo- than quinine against some types of malaria, this compound
phora, Glycosmis, Hortia, Tetradium, Zanthoxylum (Fa- is too toxic for clinical use. Glomerin (7) has been isolated
gara)], Scrophulariaceae (Linaria), and Zygophyllaceae from the millipede Glomeris marginata.
(Peganum) (Griiger, 1980). The pyrroloquinazoline-type alkaloids are represented by

o
~~ r?)~N ~N
UNAvV~N~ UN)
~NH
V~A,
I I I ~ 0
CH, CH, CH, CH,

arborine (5) g10merin (7) glyeorine glyco.smtcine glycosminine

0( 1~NH 0 NH ,r,
(J(:,OCW:~
: ,. . Y U~N-;:~
.J." N" OH "" N '" ""
OH
febrifuglne (6) chrysogine pegamine glycophymoline

Fig. 31.3. Some representative quinazoline alkaloids (Jaime, 1986; used with pennission of the copyright owner, Academic Press, Orlando, FL.).
570 Alkaloids Derived from Anthranilic Acid

(rI CO'H

-+
~HN-r-CO'H
~NHCOCH,Lco,H
::::,.. NH,

anthranilic acid aspartic acid

!
cc~ HO
vasicinone (9) deoxyvasicinone (10) vasicine (8)
(peganine)

Fig. 31.4. Selected pyrroloquinazoline alkaloids and the biogenesis of vasicine (modified from Johne, 1986; used with pennission of the copyright
own6l", Academic Press, Orlando, Fl.).

vasicine (8), vasicinone (9), and desoxyvasicinone (10) species of the genera Araliopsis, Euxylophora, Hortia, Tet-
which occur together in Peganum harmala L. (Zygophylla- radium, Vepris, and Zanthoxylum (Fagara).
ceae) (Fig. 31.9). Pyrroloquinazoline alkaloids co-occur with Feeding experiments with [3- 14C]tryptophan, [14C]for_
alkaloids of the harmine type in this genus. mate, [ 14CH3 jmethionine, and ['Hjanthranilic acid in the
plant Tetradium (Euodia) rutaecarpa, indicated that all la-
Biosynthesis bels were incorporated into rutaecarpine (11) and evodia-
Although several proposals and studies of the biosyn- mine (12). The C 1 unit of methionine or formate was incor-
thesis of vasicine (8) have been made, the complete bio- porated into the 13b position of both alkaloids and into the
synthetic route is not known with certainty. [carboxy N-methyl group of evodiarnine (Bergman, 1983; Cordell,
14C]Anthranilic acid is incorporated into vascicine in Adha- 1981).
toda vasica with incorporation of the carboxyl label (Johne, The dried fruits of Tetradium (Euodia) rutaecarpa have
1986). It is established that anthranilic acid, probably as the long been used in Chinese medicine for the treatment of
N-acetyl derivative, also is incorporated. The acetyl portion headaches, abdominal pain, dysentery, postpartum hemor-
accounts for the origin of C-3 and C-lO. Aspartic acid may rhage, and amenorrhea (Bergman, 1983). The alkaloids from
explain the origin of the remaining portion of the molecule these fruits have marked uterotonic effects (King et aI.,
(Johne, 1986) (Fig. 31.4). 1980).

Distribution of Pyrroloquinazoline
Alkaloids
QVIl'IOLOl'IE. FUKOQVIl'IOLIl'IE. ACRlDOl'lE. Al'ID
Pyrroloquinazoline alkaloids have been reported from the 4-QVIl'IOLOl'IE ALKALOIDS
Acanthaceae (Adhatoda vasica, Anisotes), Araliaceae
(Mackinlaya), Fabaceae (Galega), Malvaceae (Sida), Scro-
phulariaceae (Linaria), and Zygophyllaceae (Peganum) Alkaloids derived from anthranilic acid are represented com-
(Johne, 1986): monly in the Rutaceae. The largest groups of these com-
pounds are the quinolone, furoquinoline, pyranoquinoline,
Biological Activity acridone, and 4-quinolone alkaloids. Quinolone alkaloids
arise by extension of the carboxyl group of anthranilic acid
Adhatoda vasica has long been used in medicine in India
with one acetate (malonate) group and subsequent cycliza-
(Johne, 1986). Vasicine (8), vasicinol (9), and desoxyvasi-
tion. These compounds have a 2,4-dioxygenated quinoline
cine (10) are antifeedant to several insects (Wink, 1993).
structure. Furoquinolines and pyranoquinolines are based on
prenylated quinolone nuclei. Both angular and linear pyrano-
QUINAZOLINOCARBOLIl'IE ALKALOIDS quinoline alkaloids are known. Acridone alkaloids involve
extension of the carboxyl group of anthranilic acid by three
More than 20 representatives of this group of alkaloids units of acetate (malonate) and cyclization. Anthranilic acid
(found only in the Rutaceae) are known (Fig. 31.5) (Berg- also may be extended with acetate-derived chains or by
man, 1983). These alkaloids primarily are found in a few phenylpropanoids to yield 4-quinolone-type alkaloids.
 Alkaloids Derived from Anthranilic Acid 571

Quinolone Alkaloids additional quinoline alkaloid has been reported from Limon-
ium (Plumbaginaceae).
A number of alkaloids that possess the quinoline ring
system as part of their structure have been reported, primar-
Furoquinoline and PyranoquinoHne
ily from plants in the Rutaceae (Fig. 31.6) (Grundon, 1979;
Alkaloids
1988).
Evidence from a number of incorporation studies indi- Many quinolone alkaloids are subject to prenylation and
cates that quinolone alkaloids are derived from anthranilic give rise to the furoquinoline or pyranoquinoline alkaloids.
acid and acetate or malonate. The structure of casimiroine The furan rings of furoquinoline alkaloids arise by a similar
(13) requires only the usual processes of hydroxylation, process to thosli of furanocoumarins that have been previ-
methylation, and closure of the methylenedioxy ring at ap- ously discussed (Fig. 31.8). (see also Chapter 9) (Geissman
propriate stages of the pathway. and Crout, 1969; Grundon, 1979, 1983).
Quinolone alkaloids are found most commonly in the Ru- Anthranilic acid is incorporated intact into the quinoline
taceae, but occasionally are encountered in other families. portion of furoquinoline alkaloids, such as dictamnine (14),
Alkaloids with a quinoline ring structure occur in four of in the same manner as for quinolone alkaloids. The biosyn-
the seven subfamilies of the Rutaceae (Aurantioideae, Flin- thesis of furoquinoline alkaloids involves formation of the
dersioideae, Rutoideae, and Toddalioideae) (in addition, a quinoline ring system before introduction of the side chains.
pyranoquinolone, N-methylflindersine, occurs in the 4-Hydroxy-2-quinoline is a specific precursor of skimmi-
Spathelioideae). Quinolone alkaloids are found in the anine (15) in Ruta graveolens. The quinolone base is preny-
subfamilies Rutoideae, Toddalioideae, and Aurantioideae, lated, the prenyl residue cyclized, and the isopropyl group
but also have been reported from Xyloearpus (Meliaceae). removed. The existence of furoquinoline alkaloids contain-
In addition to quinine and related alkaloids (see Chapter ing C, prenyl groups elsewhere in the molecule and the co-
34), other quinoline alkaloids occurin Tylophora asthmatiea occurrence of furoquinoline alkaloids and isopropyldihydro-
(Asclepiadaceae) (see Chapter 30). Two quinoline alkaloids furoquinolines supports this proposed route. Alkaloids such
have been reported from Meloehia (Sterculiaceae) (Special- as lunasine (18) and lunacrine (16) co-occur with hydroxy
ist Periodic Reports 1978, 1979). An alkaloid isolated from derivatives such as (17) in Lunasia amara and Pte/ea trifoli-
Broussonetia (Moraceae), 4-formyl-8-hydroxyquinoline, ata (Figs. 31.7 and 31.8). Epoxides may be intermediates in
may not be closely related to other quinoline alkaloids. An the formation of isopropyldihydrofuroquinolines. The C-4

(J( CO,H
"'I
,... NH, (')--(l
(t:():
I I
<;;"
CO,H
S-adenosylmethionine~ ~ ____ ~NH~~'N0
'" NH NH, ~NH~~ COR N ~
~
P'p 2 ruta«arpiDe (11) "'" 1
~NHCH3

~ OQQ
' ";~ ~::c;
euxylophoriDe
YOCH,
OCH,
YOCH,
OCH, ~
O::Q/N~N~~/ CH, ""
0

~NH~N 0 evodiamiDe (12)

CHfl

I" N~:: CH'O~~N

I;ON'

14N
,0

'9' .N V
N:
0 NH lOCH,
t9

I 0 O::1)r.oN0
... I' CH, I CH ,;7
• "" "" ""I
hortiacine hortlamine paraensin rhetsinine

Fig. 31.5. Typical quinazolinocarboline alkaloids (modified from Bergman, 1983; used with permission of the copyright owner, Academic Press,
Orlando, FL).
572 Alkaloids Derived from Anthranilic Acid

((
V
CoO _

~ wO~H'
".. I '-:: CHCOSCoA
NH, to,H
U",~-"..I
NOHO NO
'-0 I
I DMAPP CH,
.. casimiroine (13)

~~OH~0c21
- -V~)lOJ<:: U"'~~.. ~
U",.:l~ ~ I
!
+- V

1
oc2
NHO OCH' NOH"..
'CR, NH 0
alanine (+ HR)-isobalfouridine (21) flindenine (22)

~
:H'h
"..
I
N O ' "
I 0
~O
I I
V
VI NO
,,,..
'"
CR, '- 0 OCH,CH, NH 0
N-methylatanine oxirine
iunacrine (16) ~ihYdroninderSine

~'OH
! OCR, """ OCH,

UNJlok ~~
U",.J.)
CH,OYN0. 0 "
CH, OCH, N 0

(-)-(S)-ribaiinine (20) lunacridine skimmianine (15) diciamnine (14)

Fig. 31.6. Quinoline, furoquinoline. and pyranoquinoline alkaloids (modified from Geissman and Crout, 1969).

o o o

~-,"~OH
y~A.!\
OCR,
-- ~R,
~-"'<
y~~j\
OCH, bR,
~"'-'"<
y~~j'H
OCR, bH,

(-)-(R)-iunacrine (16)

i
~ __ 9¢c( --
OCH'
0CR' OR

~
,;:r """
~ I """
.&
~I 'H
N 0 ::::". N 0
OCH, CH,
I I
OCH, CH, OCH, bR,

(-)-<R)-IUl188ine (18)

Fig. 31.7. Biogenesis of lunasine and lunacrine (modified from Grundon, 1979; used with pennission of the copyright owner, Academic Press,
Orlando, FL).
 Alkaloids Derived from Anthranilic Acid 573

(X
""I
COlH CHCOSCoA
1- ~- ~
OH

9'
CH'

'" .&
NH, CO,H
UNH~O I
::,.. NH 0

cxn
;?'

""
1
OCH
'<::

NH
'

0
H

(+)-(R)-platydesmine (17)
OCH,

~ ~
CH,o¥N-:::-lO)J
yN-:::-lO)J
OCH,
OCH,
dictamnine (14) 'Y~fagarine (19) skimmianine (15)

evoxine choisylne

Fig. 31.S. Biogenesis of furoquinoline alkaloids (modified from Grundon. 1979; used with permission of the copyright owner, Academic Press,
Orlando, FL).

and C-5 of mevalonic acid is incorporated specifically into ucts require that alternate mechanisms be involved (Grun-
to C-2 and C-3 of skimmianine (15) in Zanthoxylum (Fa- don, 1983).
gara) coco (Fig. 31.6) (Grundon, 1979; 1983). Furthermore, The antifeedant activity of extracts of Zanthoxylum (Fa-
3-prenylquinolones labeled at C-l' with 14C were good pre- gara) chalybaea, Z. holstii, andXylocarpus granatum (Meli-
cursors of dictamnine (14) and skimmianine (15) aceae) has been shown by Kubo and Nakanishi (1977) to be
(3.6-4.8%). (±)-[14C]Platydesmine (17) was an excellent due to the presence of N-methylflindersine [flindersine (22)].
specific precursor for dictamnine (14) (18.8% incorporation) Both of these alkaloids are widespread in the Rutaceae.
in Skimmia japonica. DictantOine is a specific precursor for Skimmianine intercalates with DNA and, as is true for
skimmianine in Choisya ternata (incorporation 1.3-6.6%). several furoquinoline alkaloids, undergoes photoaddition re-
The order of introduction of hydroxylation and methylation actions with DNA as well (Wink, 1993). DictantOine is pho-
is more complicated (Griiger, 1980; Grundon, 1979, 1983). totoxic to both bacteria and fungi (Gray, 1993).
Furoquinoline and pyranoquinoline alkaloids primarily Dimers produced by Diels-Alder reactions between py-
are known from the Rutaceae and occur in four of the five ranoquinolone and dihydrofuroquinolone alkaloids occur in
alkaloid-containing subfamilies (Aurantioideae, Flindersi- a number of rutaceous species. These compounds are known
oideae, Rutoideae, and ToddaIioideae). The most common from Araliopsis, Euxylophora, Geijera, Oricia, Ptelea, and
is skimmianine (15) (Grundon, 1979). Vepris (Waterman, 1986).
Although unusual, these alkaloids also are known from
unrelated plant families. For example, Tylophora asthmatica Acridone Alkaloids
(AscJepiadaceae) contains -y-fagarine (19) and skimmianine Acridone alkaloids are formed by the addition of three
(15) (Etherington et al., 1977). Skimmianine also has been acetate (or malonate) units, followed by cyclization (Figs.
reported from Datura stramonium. 31.9 and 31.10) (Geissman and Crout, 1969; Griiger, 1980).
pyranoquinoline alkaloids, such as ribalinine (20) and More than 100 alkaloids of this type are known (Gray, 1993).
isobalfourodine (21), appear to arise from the same epoxide Representatives of this group of alkaloids are widespread
intermediate involved in the formation of furoquinoline alka- in the Rutaceae, especially in the genera Acronycia, Atalan-
loids. However, differences in stereochemistry of the prod- tia, Balfourodendron, Citrus, Euodia (which should be used
574 Alkaloids Derived from Anthranilic Acid

o OCR, o OR

~OCR'
~N~OCR' I
CR,

l,3-dimethoxy-N-methylacridone acronyclne (23) arborinine

o OR
o OCR, 0 OCR,

o(6)~O)
~fYO NR 0
CR, OCR,

rutacrldone epoxide (24)

o
evoxantbidine me1icopidine

Fig. 31.9. Some acridone alkaloids of the Rutaceae.

in preference to Evodia), Glycosmis, Melicope, Ruta, Teclea, been stated that' 'acronycine interfered primarily with struc-
and Zanthoxylum (Fagara), as well as in other species of ture, function andlor turnover of cell membrane compo-
the subfamily Toddalioideae (Groger, 1980) (Fig. 31.9). nents." The effect involves nucleoside transport across
Several acridone alkaloids have antitumor activity in a plasma membranes (Suffuess and Cordell, 1985). Acronyc-
number of cell lines; some are inhibitory to growth of Plas- ine (23) also has been reported to be carcinogenic (Gerzon
modium yoelii (Borris and Schaeffer, 1992; Wink, 1993). and Svoboda, 1983).
Acronycine (23) (from Acronycia, an Australian and Asian Rutacridone epoxide (24) from Ruta graveolens was
genus of the Rutaceae) has been shown to possess antitumor shown to possess antimicrobial activity against several or-
activity (Gerzon and Svoboda, 1983; Suffness and Cordell, ganisms (Nahrstedt et aI., 1980; Wink, 1993). Cultures of
1985). Acronycine (23) proved to possess the broadest ex- Ruta graveolens produced 50-fold more rutacridone epoxide
perimental antitumor spectrum of any alkaloid isolated from and hydroxyrutacridone epoxide when treated with fungi not
higher plants (Blasko and Cordell, 1988; Gerzon and Svo- specific to the host (Brooks and Watson, 1985; Ellis, 1988).
boda, 1983). The compound was active both interperitone-
ally and orally against 12 of 17 experimental tumor models 4-Quinolone Alkaloids
and appears to be active against long established tumors.
The LDso in mice as a single dose (orally) is from 350 to 4-Quinolones are rather rare alkaloids, known only from
680 mg/kg. The mechanism of antitumor activity appears to Tetradium, Ruta, Vepris, and Ptelea (Ng et aI., 1987) (Fig.
be different than that of most other antitumor drugs. It has 31.11).

(X1 - - (XICO,R COSCOA

+ 3
CO,R
I ---.
"", NR, "", NR, CR,COSCoA

1,3-dihydroxyacridone

Fig. 31.10. Biogenesis of acridone alkaloids (Geissman and Crout, 1969).

 Alkaloids Derived from Anthranilic Acid 575

from Galipea oJficinalis (Rutaceae) from Ruta graveo1ens

o>
OCR, o

o
)
o
cusparine eduleine

Fig. 31.11. Representative 4-quinolone alkaloids.

CHEIIIOSYSTBMATICS AI'ID PHYLOGBNY subfamilies from which alkaloids have been reported (Aura-
ntioideae, Flindersioideae, Rutoideae, and Toddalioideae).
The Rutaceae has been subdivided into several subfamilies Acridone alkaloids are not known from sources outside this
by various workers (Da Silva et al., 1988; Watennan, 1983). family. Most reports of quinolone and furanoquinoline alka-
The subfamilies Aurantioideae, Flindersioideae, Rutoideae, loids also are from this family. In contrast, benzylisoquino-
and Toddalioideae possess many common features and are line alkaloids are found in many plants of the Magnoliidae
considered by most workers to be closely related. Three and in the Rhamnaceae, Euphorbiaceae, Celastraceae, Sym-
other proposed subfamilies are more doubtful in relation- plocaceae, and other families (Seigler, 1977; Watennan and
ship. The Spathelioideae has been placed in the Simarou- Grundon, 1983).
baceae but contains N-methylflindersine, an alkaloid charac- Alkaloids of the I-benzyltetrahydroisoquinoline type are
teristic of the Rutaceae. The Dictylomatoideae also has been assumed to be primitive in the Rutaceae and the plants pro-
transferred to the Simaroubaceae by some. Another, the ducing them belong to the most printitive extant genera of
Rhabdodendroideae, probably is a close relative of the Phy- the family. As anthranilate-derived alkaloids are found in
tolaccaceae and should be transferred from the Rutaceae some of the same genera, it appears that the evolutionary
(Airy-Shaw, 1973; Prance, 1968). trend was for direct replacement of one type with another
However, several workers have felt that the present (Watennan and Khalid, 1981). The taxa that do not have 1-
subfamilies are not in accord with all lines of evidence (e.g., benzyltetrahydroquinoline alkaloids [e.g., tribes Diosmeae
floral anatomy and phytochemistry). A revised scheme of and Boronieae (Rutoideae) and the subfamily Auranti-
classification has been proposed (Watennan, 1975, 1983). oideaej, appear to be relatively advanced.
The Rutaceae is one of the most interesting families in Alkaloids of the benzylisoquinoline type occur mostly in
regard to alkaloid chemistry as well as the fonnation of fla- the genera Fagaropsis, Phellodendron, Tetradium, Toddalia
vonoids, mono-, sesqui-, and triterpenes, furocoumarins, and (Toddalioideae), and Zanthoxylum (Fagara) (Rutoideae); a
other secondary compounds (Watennan and Orondon, group of genera considered to represent the "proto-Ruta-
1983). Many chentical features support the view that the ceae" (Ng et ai., 1987; Watennan and Khalid, 1981).
Rutaceae is a distinct and homogenous group. Essential oils Plants of subfamily Aurantioideae contain other types of
and coumarins are found in at least four subfantilies. Furo- alkaloids. Members of the genera Merrillia and Micromelum
quinoline alkaloids are essentially ubiquitous in the family, and several Murraya species contain indole alkaloids such
and acridones are common. as yuechukene (see Chapter 35) (Kong et ai., 1986, 1988a,
Members of the family contain alkaloids based on several 1988b). Other Murraya species produce carbazole alkaloids
major pathways such as benzylisoquinoline (tyrosine de- (Kong et al., 1986, 1988b). Carbazole alkaloids have been
rived), quinolone, furoquinoline, quinazoline (all from an- reviewed (Bhattacharyya and Chakraborty, 1987; Chakra-
thranilic acid), imidazole (histidine), indoloquinazoline borty and Roy, 1991) (see also chapter 35).
(tryptophan), and both simple aliphatic and aromatic amines. Plants of the Rutoideae, tribe Diosmeae also contain quin-
In the tribe Ruteae (Rutoideae), no fewer than five types of olone and furoquinoline alkaloids (Campbell et ai., 1987).
alkaloids are common to the three major genera. Quinolone, The family Rutaceae has been regarded by most system-
furoquinoline, and acridone alkaloids are especially wide- atists as closely related to the Cneoraceae, Meliaceae, Ptaer-
spread within the family, being found in four of the five oxylaceae, and Simaroubaceae. Although alkaloids largely
576 Alkaloids Derived from Anthranilic Acid

are lacking in the last three families, the biosynthetic path- CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual
ways leading to limonoids, quassinoids, and related Reports on NMR Spectroscopy, Vol. 13 (G. A. Webb. ed.),
triterpenes link the Rutaceae to the Meliaceae, Simarou- 60-210, Academic Press, London, 1982.
baceae, and Cneoraceae. The presence of chromones and DA SILVA, M. F. D. G. F .. O. R. GoTTLIEB, and F. EHRENDORFER,
coumarins link this group with the Ptaeroxylaceae (Water- Chemosystematics of the Rutaceae: Suggestions for a more nat-
man and Khalid, 1981). It has been suggested that this last ural taxonomy and evolutionary interpretation of the family,
family has links to Rutaceae, subfamily F1indersioideae. Af- Plant Syst. Evol.,161. 97-134 (1988).
finities to the Burseraceae and Zygophyllaceae also have ELLIS, B. E., Natural products from plant tissue culture. Nat. Prod.
been suggested. The latter relationship has been suggested Rep., 5. 581-612 (1988).
largely because of the genus Peganum. This genus has cer- ENGLER, A., RUTACEAE, Nat. Pflanzenfam. III, 4. 195-201 (1896).
tain similarities to the Rutaceae and has been placed in that ETHERINGTON, T., R. B. HERaBRT, and F. B. JACKSON, Furoquino-
family by some (Engler, 1896). Species of Peganum contain line alkaloids from Tylophora asthmatica, Phytochemistry. 16.
alkaloids of the harmine and quinazoline types that resemble 1125-1126 (1977).
those of some rutaceous plants. GBISSMAN, T. A. and D. H. G. CROUT, Orgartic Chemistry of Sec-
As many of the primitive Rutaceae contain benzylisoqui- ondary Plant Metabolism, Freeman Cooper, San Francisco,
noline alkaloids, it is probable that the ancestors of this fam- 1969.
ily possessed them. The presence of these alkaloids and a GBRZCN, K. and G. H. SVOBODA, Acridone a1ka1oids: Experimental
variety of morphological features suggests that the Rutaceae antitumor activity of aeronycine, in The Alkaloids, Vol. 21 (A.
Brossi, ed.), 1-28, Academic Press, New York, 1983.
is derived from ancestors in the Magnoliidae, near the Ra-
nunculales or the PapaveraIes (Seigler, 1977; Waterman and GRAY. A. I., Quinoline a1ka1oids related to anthranilic acid, in Alka-
Khalid, 1981). The alkaloids produced by Fagaropsis ango- loids and Sulphur Compounds (p. G. Waterman, ed.). Vol. 8
of Methods in Plant Biochemistry (P. M. Dey and 1. B. Har-
lensis are completely typical of those produced by the Ra-
borne, eds.), 271-308, Academic Press, London, 1993.
nales, in particular by the Papaveraceae (Waterman and
GROOER, D., AlkaIoids derived from tryptophan and anthranilic
Khalid, 1981). Benzylisoquinoline alkaloids are uncommon
acid, in Secondary Plant Products (E. A. Bell and B. V. Charl-
in the Rosidae and this group would not seem tu be probable wood, eds.), Encyclopedia of Plant Physiology, N. S., Vol. 8,
as ancestral to the Rutaceae. 128-159, Springer-Verlag, Berlin, 1980.
GRUNDON, M. F., Quinoline alkaloids related to anthranilic acid,
in The Alkaloids, Vol. 17 (R. H. F. Manske and R. G. A. Ro-
REFERENCES drigo, eds.), 105-198, Academic Press, New York, 1979.
GRUNDON, M. P., Aspects of the biosynthesis of coumarins and
AIRy-SHAW, H. K., J. C. Wn.us' Dictionary of Flowering Plants quinoline alkaloids in the Rutaceae, in Chemistry and Chemical
and Ferns, 8th ed., Cambridge University Press, Cambridge, Taxonomy of the Rutales (P. G. Waterman and M. F. Grundon,
1973. eds.), 9-30, Academic Press, London, 1983.
AITA-uR-RAlIMAN and A. BASHA, Biosynthesis of Indole Alka- GRUNDON, M. F., Quinoline a1ka1oids related to anthranilic acid,
loids, Oxford University Press, Oxford, 1983. in The Alkaloids, Vol. 32 (A. Brossi, ed.), 341-439, Academic
BERGMAN. J., The quinazolinocarboline alkaloids, in The Alkaloids. Press, New York, 1988.
Vol. 21 (A. Brossi, ed.), 29-54, Academic Press, New York, JOHNE, S., Quinazoline alkaloids, in The Alkaloids, Vol. 29 (A.
1983. Brossi, ed.), 99-140, Academic Press, New York, 1986.
BHATrACHARYYA, P. andD. P. CHAKRABORTY, Carbazole alkaloids, KING, C. L., Y. C. KONG, N. S. WONG, H. W. YEUNG, H. H. S.
Fortscbr. Chem. Org. Naturst., 52, 159-210 (1987). FONG, and U. SANKAWA, Uterotonic effect of Evodia rutaecarpa
BLASKO, G. and G. A. CORDELL, Recent developments in the chem- a1ka1oids, J. Nat. Prod., 43. 577-582 (1980).
istry of plant-derived anticancer agents, in Economic and Me- KONG, Y., K. CHENG, K. NG, P. P. BUT, Q. LI, S. Yu, H. CHANG,
dicinal Plant Research, Vol. 2 (H. Wagner, H. Hikino, and N. R. C. CAMBm, T. KINOSHITA, W. KAN, and P. G. WATERMAN,
R. Farnsworth, eds.), 119-191, Academic Press, London, 1988. A chemotaxonomic division of Murraya based on the distribu-
BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from tion of the a1ka1oids yuehchukene and girinimbine. Biochem.
plants, in Phytochemical Resources for Medicine and Agricul- Syst. Ecol., 14. 491-497 (1986).
ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum, KONG, Y., P. BUT, K. NG, K. CHENG, K. CHANG, K. M. WONG, A.
New York, 1992. I. GRAY, and P. G. WATBRMAN, The biochemical systematics
BROOKS, C. W. J. and D. G. WATSON, Phytoalexins, Nat. Prod. of Merrillia; in relation to Murraya, the Clauseneae and the
Rep .. 2. 427-459 (1985). Aurantioideae, Biochem. Syst. Ecol., 16. 47-50 (1988a).
CAMPBELL, W. E., K. P. FINCH, P. A. BEAN, and N. l'rNKBLsTElN, KONG, Y., P. P. BUT, K. NG, Q. L~ K. CHENG, and P. G. WATBRMAN,
Alkaloids of the Rutoideae: Tribe Diosmeae, Phytochemistry. Micromelum: A key genus in the chemosystematics of the
26. 433-434 (1987). Clauseneae, Biochem. Syst. Ecol., 16. 485-489 (l988b).
CHAKRABORTY, D. P. and S. RoY, Carbazole a1ka1oids. III. Fortscbr. KUBO, I. and K. NAKANISHI, Insect antifeedants and repellants from
Chem. Org. Naturst., 57, 71-152 (1991). African plants, in Host Plant Resistance to Insects (P. A. Hedin,
CORDELL, G. A., Introduction to Alkaloids, Wiley-Interscience, ed.), ACS Symposium Series 62,165-178, American Chemical
New York, 1981. Society. Washington. DC. 1977.
 Alkaloids Derived from Anthranilic Acid 577

LEETE, E., The biosynthesis of alkaloids, Specialist Periodical Re- SUFFNESS, M. and G. A. CORDELL, Antitumor alkaloids, in The
ports, Biosynthesis, 102-223, The Royal Society Chemistry, Alkaloids, Vol. 25 (A. Brossi, ed.), 3-355, Academic Press,
Cambridge, 1983. New York, 1985.
NAHRSTEDT, A., U. Ell.ERT, B. WOLTERS, and V. WRAY, RutRCri- WATERMAN, P. G., Alkaloids of the Rutace..: Their distribution
done-epoxide, a new acridone alkaloid from Ruta graveo/ens, and systematic significance, Biochem. Syst. Bcol., 3,149-180
Z. Naturforsch., 36c, 200-203 (1980). (1975).
NG, K. M., P. P. BUT, A. I. GRAY, T. G. HARTLEY, Y. KONG, and WATERMAN, P. G., Phylogenetic implications of the distribution
P. O. WATERMAN, The biochemical systematics of Tetradium, of secondary metabolites within the Rutales, in Chemistry and
Euodia, and Me/icope and their significance in the Rutaceae, Chemical Taxonomy of the Rutales (P. G. Waterman and M.
Biochem. Syst. Bcol., 15, 587-593 (1987). F. Grundon, eds.), 377-400, Academic Press, London, 1983.
PaNCHET, M., J. MARTIN-TANGUY, A. MARAls, and A. POUPET, WATERMAN, P. G., The dimeric alkaloids of the Rutace.. derived
Dianthramides A and B, two N-benzoylanthranilic acid deriva- by Diels-Alder addition, in Alkaloids: Chemical and Biological
tives from elicited tissues of Dianthus caryophyllus, Phyto- Perspectives, Vol. 4 (S. W. Pelletier,ed.), 331-387, Wiley,New
chemistry, 23, 1901-1903 (1984). York,1986.
PRANCE, G. T., The systematic position of Rhabdodendron Gilg. WATERMAN, P. G. andM. F. GRUNDON, eds., Chemistry and Chemi-
and Pilg., Bull. Jard. Bot. Nat. Belg., 38,127-146 (1968). cal Taxonomy of the Rutales, Academic Press, London, 1983.
SEIGLER, D. S., Plant systematics and alkaloids, The Alkaloids, WATERMAN, P. G. and S. A. KHALID, The biochemical systematics
Vol. 16 (R. H. F. Manske, ed.), 1-82, Academic Press, New of Fagaropsis angoiensis and its significance in the Rutales,
York,1977. Biochem. Syst. Bcol., 9, 45-51 (1981).
SPECIALIST 1'ERlODlC REPoRTS, Alkaloids, Vol. 8, Chemical Society, WINK, M., Allelochemical properties or the raison d'elre of alka-
London, 1978. Alkaloids, Vol. 9, Chemical Society, London, loids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), 1-105,
1979. Academic Press, New York, 1993.
32
Isoquinoline and Benzylisoquinoline
Alkaloids

Introduction Curare
Isoquinoline Alkaloids Erythrina Alkaloids
Biosynthesis Biosynthesis
Dimers and Oligomers of Isoquinoline Alkaloids Biological Activity
Cryptostyline Alkaloids Distribution of Benzylisoquinoline Alkaloids
Distribution of Isoquinoline Alkaloids Ipecacuanha Alkaloids
Biological Activity Biosynthesis
Interactions of Cactus Species, Yeasts, and Drosophila Distribution of Ipecac Alkaloids
Benzylisoquinoline Alkaloids Medical Uses of Ipecacuanha Alkaloids
Tetrabydrobenzylisoquinoline Alkaloids Other Biological Activity
Biosynthesis References
Biological Activity
Azafluoranthene and Tropoloisoquinoline Alkaloids
Aporphine Alkaloids
Biogenesis INTRODUCTION
Biological Activity
Aristolactams and Aristolochic Acid Alkaloids that possess an isoquinoline moiety in their struc-
Eupomatia Alkaloids ture are among the most common of all alkaloids. More
Proaporphine Alkaloids than 4000 compounds of many structural types but with this
Morphinandienone Alkaloids (Promorphinanes) structural feature are known (Verpoorte et al., 1991). Those
Hasubanan Alkaloids treated in this chapter are all derived from a 3,4-dihydroxyty-
Morphine Alkaloids ramine (dopamine) precursor that is linked to an aldehyde or
Biosynthesis ketone of several different origins. One series of isoquinoline
Pharmacological Activity alkaloids involves glyoxylic acid, pyruvic acid, and an a-
Other Biological Activity keto acid derived from leucine as the carbonyl portion. These
Protoberberidine Alkaloids compounds are largely, but not exclusively, found in the
Biosynthesis Cactaceae.
Biological Activity A second major structural type, commonly known as ben-
Protopine Alkaloids zylisoquinoline (BIQ) or tetrabydrobenzylisoquinoline
Benzophenanthridine Alkaloids (TBIQ), alkaloids constitute one of the two most numerous
Phthalideisoquinoline Alkaloids types of alkaloids. As is true for the simple isoquinoline
Rhoeadane Alkaloids alkaloids mentioned above, all are derived from a 3,4-dihy-
Dibenzopyrrocoline Alkaloids droxytyramine (dopamine) precursor condensed with a car-
Pavine and Isopavine (Argemone) Alkaloids honyl compound. In this case, the precursor of most, if not
Cularlne Alkaloids all, benzylisoquinoline alkaloids is 4-hydroxyphenylacetai-
Bisbenzylisoquinoline Alkaloids dehyde. Benzylisoquinoline alkaloids are found in a limited
Biological Activity group of plant families; some have great medical impor-

578
 lsoquinoline and Benzylisoquinoline Alkaloids 579

tance. The origins of several major subgroups occur are de- are glyoxylic acid, pyruvic acid, and an ",-ketoacid derived
scribed below. from leucine (Fig. 32.1).
A third major group, including emetine and a series of The application of both 'H- and 13C-NMR (nuclear mag-
structurally related alkaloids, involves condensation of the netic resonance) spectroscopy and other spectroscopic meth-
same amine precursor (3,4-dihydroxyphenylethylamine or ods to the study of isoquinoline alkaloids has been reviewed
dopamine) with a carbonyl group from secologanin-an iri- (Crabb, 1982; Guinaudeau and Bruneton, 1993; Hughes and
doid monoterpene precursor. Most of these compounds have MacLean, 1981; Mata et aI., 1983).
been isolated from two families, the Rubiaceae and the Alan-
giaceae. Biosynthesis
Isoquinoline alkaloids are derived from tyrosine through
the intermediacy of 3,4-dihydroxyphenylethylamine (dopa-
ISOQUIl'IOLIl'IB ALKALOIDS mine) and a carbonyl unit of various origins. The genesis of
tyramine, and dopamine from tyrosine has been described
About 130 alkaloids based on a 3,4-dihydroxytyramine (do- previously (see Chapter 28). In contrast to benzylisoquino-
pamine) precursor linked to an aldehyde or ketone of several line alkaloids (see below), the formation of isoquinoline al-
different origins are known (Guinaudeau and Bruneton, kaloids usually involves the amine and an ",-ketoacid instead
1993). Among the usual origins of this carbonyl compound of an amine and aldehyde. However, several compounds can

CH"0:roJ _
1
CHJO~ ~ CH'OW

~ NH, CHj'O~ /'f~ CH,O "'- ' NH

CH,O COCO,H HO / 'CO,H HO CO,H
HO ,
CH,
peyoruvic acid (3)

~
CH'O~ CH'OW' CHJoy?

CH3 0 ::::....
, NH
CH,O
"'- N,
CH,
~ I "",N
2 CHJO
OCH, HO HO

mescaline (4) pellotine (2) 3,4·dihydro-6,7-dimefhoxy

-1-methyl-8-hydroxyisoquinoline

CH'O~,
~O "'- NH,-
CH,'O

HO
,y

~ I
+HCOCO H
2
CHJO
W HO COzH
g -+
CH"OW

CH,O
~l
HO
NH

SAM also is incorporated peyoxylic acid

anhalamine (1)

~
CH"0y) CHJOW
,yl CH','0:Q)
, N
~ N ..... ",-' NH
CH,'O CH, CH,O CH,-O ~ h
HO HO HO

anhalidine {IO} anhalonidine (11)

3,4-dihydro-6,7-dimethoxy
-8-hydroxyisoquinoJine

Fig. 32.1. Origin of some representative isoquinoline alkaloids (modified from Lundstrom. 1983; used with permission of the copyright owner,
Academic Press, Orlando, FL).
580 Isoquinoline and Benzylisoquinoline Alkaloids

serve as precursors in specific instances. In one case, closure ing studies reveal that both leucine and mevalonic acid can
of the nitrogen-containing ring to produce anhalamine (1) act as precursors. Previous studies indicate that leucine can
involves methionine (Fig. 32.1). A related alkaloid, pellotine be converted into l'I-hydroxY-I'I-methylglutaric acid (7), a
(2), is probably derived from pyruvate. A carboxylic acid probable precursor of mevalonic acid. Mevalonic acid can
of similar structure, peyoruvic acid (3), suffers oxidative be converted to the required aldehyde and condensed with
decarboxylation and serves as a precursor for other isoquino- dopamine to afford lophocereine (Fig. 32.2).
line alkaloids (LundstrOm, 1983; Leete, 1983). The results of a number of feeding studies leading to
The order of methoxylation and oxidation steps varies. isoquinoline alkaloids have been tabulated (Leete, 1983).
In some instances, the 3,4-dihydroxyphenylethylamine (do-
pamine) precursor may be modified before condensation oc- Dlmers and OUgomers of Isoquinoline
curs. Despite their structural similarity, pellotine (2) and an- AJkaIoids
halarnine (1) are probably synthesized along distinct In addition to simple monomeric units with isoquinoline
pathways from mescaline (4). All three alkaloids co-occur structures, a number of dimeric and oligomeric alkaloids of
in Lophophora williamsii. this series are known. An example of this type, pilocereine
Lophocereine (5), a related compound with a five-carbon (8) (Fig. 32.3), known from Pachycereus, Pilocereus, and
unit inserted into the nitrogen-containing ring, is found in Lophocereus (all Cactaceae), is derived by coupling of iso-
another cactus species, Lophocereus schottii. The usual car- quinoline alkaloid units (probably by a free-radical mecha-
bonyl precursor probably is 4-methyl-2-oxopentanoic acid nism, although proof for this is lacking), much in the same
(6), an entity that probably arises by transamination of leu- manner as the dimerization process leading to bisbenzyliso-
cine (Geissman and Crout, 1969) (Fig. 32.2). However, feed- quinoline alkaloids, a topic discussed below.

:::(rlNH'~O~I :~,y
(22)
I NH~O
CHO HO HO HO

~ Y
n
lophocereine(S)

H CH30 '" CHy~H3

HO
--
CO,H HO
~I_NH+- CH,
I
_fHNH,
CO,H
mevalonic acid leucine

! \ decarboxylation

1 ?O,H 0
~NH- I I( -+
'~CO,H~
1 COSCoA - I""", COSCoA
~
leucine (6)

-
(
~
CO,H
~
COSCoA - +
X
CO,H
H
__
COSCoA

p.hydroxy·~.methylgl.laryl CoA
X
bO,H l
mevalonic acid
OH

(7)

Fig. 32.2. Biogenesis of lopbocereine (modified from Geissman and Crout, 1969).
 isoquinoline and Benzylisoquinoline Alkaloids 581

CHro~1 CH'-oX:Y CH'-O~

HO ~ NH 1 N 1
.CH,-O ~ . . . CH, CH,-O ~ NH

HO
lophocereine (17) carnegeine (18) anhalanine (ll)

~
OCH'CH'·°7Yr1
HN I""" HO ~ NH

& HN 1
"'"
""" OCH,
o

pllocereine (8l

CH'-O:yo CH'-OXPI CHrow

o ~I N . . . CH 0 ~ NH
0
~I N '- ./
'--0 ' Lo '--0 .......
lopbopborine (14) anhalonine (13) peyophorine (15)

Fig. 32.3. Biologically active isoquinoline alkaloids.

CryptostyUne Alkaloids Other families known to possess isoqninoline alkaloids

include the Alangiaceae, Chenopodiaceae (a family related
A small group of isoquinoline alkaloids, in which substi-
to the Cactaceae), Fabaceae, Musaceae, Nymphaeaceae, and
tuted benzaldehyde derivatives such as 9 appear to be in-
Sterculiaceae (LundstrOm, 1983). At least four species of
volved in the biosynthetic process, occurs in the genus Cryp-
the Chenopodiaceae contain isoquinoline alkaloids, whereas
tostylis (Orchidaceae) (Fig. 32.4). Dopamine serves as a
other species contain (3-phenylethylamines. Many of the
precursor for both aromatic rings of the system (LundstrOm,
plants that contain benzylisoquinoline alkaloids also contain
1983).
smaller amounts of isoquinoline alkaloids. Among these are
plants belonging to the Annonaceae, Euphorbiaceae, Puma-
riaceae~ Hernandiaceae, Menispermaceae, Monimiaceae,
DlSTRlBUI101'1 OF ISOQUIl'IOLIl'IE ALKALOIDS Papaveraceae, Ranunculaceae, and Rharnnaceae (Lunds-
trOm, 1983). The distribution of benzylisoquinoline alka-
Isoquinoline alkaloids are found primarily in the Cactaceae, loids is discussed in more detail below.
Chenopodiaceae, and Fabaceae (Guinaudeau and Bruneton,
1993; Mata and McLaughlin, 1982). The Cactaceae also
elaborate (3-phenylethylamines (see chapter 28) that co-
occur with tetrahydroisoquinoline alkaloids. The latter ap- BIOLOGICAL ACDVITY
pear to have evolved by cyclization of variously substituted
phenylethylamines with suitable ring-closing units. A com-
prehensive listing of isoquinoline alkaloids in cacti has been In contrast to (3-phenylethylamines such as mescaline (4),
published (LundstrOm, 1983). isoquinoline alkaloids do not have hallucinogenic activity.
Peyote, Lophophora williamsii, appears to be the most These alkaloids are toxic to many animals and exhibit a
prolific cactus, in terms of alkaloid production. In addition number of other physiological effects. Both peyophorine
to mescaline and other (3-phenylethylamines, this cactus con- (15) and camegeine (18) have positive inotropic effects and
tains anhalamine (1), anhaladine (10), anhalonidine (11), modify heart action in mammals (Wagner, 1988). Other iso-
pellotine (2), anhalanine (12), anhalonine (13), lophophorine quinoline alkaloids have been reported to possess antitumor
(14), and O-methylanhalonidine (Lundstrom, 1983). activity (Lundstrom, 1983).
582 Isoquinoline and Benzylisoquinoline Alkaloids

HO~

CH,-O
~ NH,
dopamine (22)

~
4.0·methyldopamine

CH,-O CH'_O~
I H~O
~NH,-tt-;H-O
po
A)I ' ~ NCH
, ~ I ""

00
I "" ~
~
OH
3-0-melhyldopamlne / 0 OCH,
0-1 isovanillin

CH'_O~
~ I ",N (+)-(S)-cryploslyline I
CH,O
(9)

""
l,b
o
0--.1
Fig. 32.4. Proposed biosynthesis of cryptostyline alkaloids (modified from Lundstrom, 1983; used with pennission of the copyright owner, Academic
Press. Orlando. FL).

Interactions of cactus Species. Yeasts. host specificity. For example, D. nigrospiracula is found
and DrosopblJa only in decaying tissues of saguaro (and one other closely
related cactus), whereas D. pachea occurs solely in the senita
The interactions of certain Drosophila species and cacti
cactus. D. mojavensis breeds in organ pipe cactus in Arizona
in the southwestern United States have been discussed in
cbapter 23. These insect species feed on rotting limbs of and Sonora, where the agria cactus is absent, but is found
cacti and have a requirement for scbottenol, a steroidal mate- preferentially in the agria cactus, in areas where both plants
rial that occurs in the species involved (Kircher, 1982). The exist. This insect species also occurs on other cactus species
saguaro cactus (Carnegiea giganfea) contains primarily two on occasion (Kircher, 1982).
isoquinoline alkaloids: dopamine and the phytosterols camp- The restriction of D. pachea to the senita cactus is a result
esterol and sitosterol. The senita cactus (Lophocereus schot- of the strict 11.7 -sterol requirement of the fly and its tolerance
tii) contains a number of unusual alkaloids and sterols. Lo- of the senita alkaloids. Although schottenol (7-stigmastenol)
phenol and schottenol are among the sterols, and and lophenol (4a-methyl-7-cholestenol) were the main ste-
lophocereine (17) and pilocereine (8) are among the alka- rols present, the insect utilizes and requires 7-cholestenol,
loids (Fig. 32.3). The organ pipe cactus, Sfenocereus thur- which is present in smaller quantities (Kircher, 1982). Lo-
beri, previously known as Lemaireocereus thurberi, con- phocereine (17) and pilocereine (8) occur in the senita cactus
tains no alkaloids, but does possess large quantities of lipids and act as feeding deterrents to other Drosophila species.
and triterpene glycosides. Cholesterol, campesterol, and si- Only Drosophila pachea and D. mettleri are impervious to
tosterol are among the sterols. The agria cactus, Stenocereus their effects (Harborne, 1982; Meyer and Fogelman, 1987).
gummasus (formerly known as Machaerocereus gummosus) The last species typically breeds in soil beneath saguaro
has not been as thoroughly analyzed, but contains no alka- cacti.
loids. The cina cactus. Sfenocereus alamosensis, formerly The close association of D. nigrospiracula with saguaro
known as Rathbunia alamosensis, is rich in triterpene glyco- (and another cactus, cardon) can be explained only in that
sides. A common species of prickly pear, Opuntia ficus- this fly is excluded frum other potential hosts. Compounds
indica, produces mucilage, but no alkaloids or saponic mate- (alkaloids and medium-chain fatty acids) frum other poten-
rials. The organ pipe, agria, and cina cacti all contain large tial hosts are lethal to this insect (Kircher, 1982). The sagu-
quantities (25-45%) of water-soluble triterpene glycosides aro cactus contains camegeine (18), which acts as a repellent
and lipids. The saguaro and senita cacti contain alkaloids to D. pachea but not to D. nigrospiracula (a species that
(1-10%) (Kircher, 1982). feeds on saguaro) (Harborne, 1982)_
Many species of Drosophila (fruit flies) show significant Individuals of D. mojavensis cannot breed in the senita
 /soquinoline and Benzylisoquinoline Alkaloids 583

cactus because of the alkaloids present. In the saguaro, this have been observed. The three Drosophila species examined
species does not appear to be able 10 coexist with D. ni· showed consistent differences in tolerance toward given
grospiracula (Kircher, 1982). plant species. It was concluded that the yeasts hydrolyzed
The above observations were based on the identification the glycosides to produce aglycones that are selectively toxic
of a relatively small number of alkaloids and terpenoids. to D. mojavensis, but that the alkaloids themselves are not
Recently, the diversification of plant compounds has been generally toxic to these Drosophila species at the levels en-
discovered to be much greater than previously thought (Gib- countered in the plants. Furthermore, is was noted that nona-
son et aI., 1986; Spencer, 1987). Early studies considered dapted insects do not exhibit selective tolerance toward any
the activity of the aglycones of the steroidal compounds, but given compound arrays and may suffer toxication by alka-
not that of the glycosides themselves. Cactophilic yeasts are loids or aglycones or both. Triterpenoid glycosides may spe-
able to hydrolyze the glycosides and produce the correspond- cifically affect Drosophila, whereas the alkaloids may be
ing aglycones under certain conditions. Specific differences more general toxins (Spencer, 1987).
in toxicity to mixtures (consisting of alkaloids, glycosides, Despite the fact that chemical factors are important in
sugars, and aglycones) that had suffered hydrolysis by yeasts defining the interactions of Drosophila cactus species, not

, ,
oo~
HO ",' :H
rn,o~
HO '"
~o~HO '"
• N , ,H N'CH,
HO "" , ,H 'CH,
CH,O CH,O
HO '" '
0 0

rn'02f,
(. HS)·norlaudanosoline (26) salutaridine (52) sinoaeutine (53)

HO~
",' rn,o~
HO "" 1 N'CH CH,'O '" "N
0 H '
CH,'O ""
"'. H N'CH, CH,'O cY
HO" h ""I CH,O '"
(+)-isolbebalne (49) papaverine (31)
0

<
0

<0 0

0 OCH,
) OCH,
0
0 prolopine (60) berberine (71)

<
CH,O
0
0 <0 HO
OCH,

OCH, OCH,
0 0 0
OCH, (+)·rhoeadine (59) -.../ eryptaustoline (92)

CH"O~
1CIb l "" OCH,
CH'01Jg:foH
-- c:?
CH,'O '" 'N '" I HO ~ "N'CH,:::,... \
OCH, 0 : D OCH,
(.).argemonine (93) (:;,.. 1 N
o thalielidine

CH,O a
erythraline (106)
Fig. 32.S. Some major types of benzylisoquinoiine alkaloids.
584 lsoquinoline and Benzylisoquinoline Alkaloids

all aspects of the story can be explained by chemical factors Both the IH_ and 13C_NMR spectra for benzylisoquino-
(Kircher, 1982). line alkaloids have been reviewed (Crabb, 1982; Janssen et
al., 1989). The results of a number of feeding studies leading
BEl'IZYLlSOQUIl'IOLIl'IE ALKALOIDS to benzylisoquinoline alkaloids have been tabulated (Leete,
1983).
Benzylisoquinoline alkaloids differ from isoquinoline alka-
loids in that a different carbonyl donor unit, 3,4-dihydroxy- TetrabydrobenzyUsoquinoline Alkaloids
phenylacetaldehyde (19), is involved in their biosynthesis The simplest alkaloids of this series are those in which
(Herbert, 1988; Schumacher et al., 1983). The plants that the nitrogen-containing ring of the isoqniuoline system is
contain benzylisoquinoline alkaloids often are poisonous completely saturated (Fig. 32.7). Quaternary bases also are
(see references in Chapter I and Kingsbury, 1964). At least found. Tetrahydrobenzylisoquinoline alkaloids are wide-
4000 benzylisoquinoline alkaloids belonging to many spread and are found in almost all the families that possess
subgroups are known (Figs. 32.5 and 32.6) (Verpoorte et benzylisoquinoline alkaloids. About 100 alkaloids of this
al., 1991; Waller and Nowacki, 1978). type are known (Guinaudeau and Bruneton, 1993).

C~7¥'0 <:~,..
o
,...
'"
OCH3
: I N.CH
-
3
HO,...

CH30 ' "

I

I
H
N. CH

\
3
HO

O.
-
Ho~··rn,CH'O
r

:,..
I
CH30~
1
HO '"

1 1H
0
N· CH3

proaporphine. ~hines morphinane./ protomorphinanes

CH30h~oCH,
CH,O~ __
CH~:2?'" 1 N. CH
3
CH30
~7OH
OCH, HO".. -HO~/-CH \
pavines ,.,. 1 '-.J ,... OCH3
CH3 0 isopavines
(+)- and (-)-rellcu6ne (20 and 21)

CH,O OCH3
CH 0
3

~ '"

r
0 '
,
NTH
H '

/
CH,O ,.,.
OCH3 OCH3
benzophenanthridines protoberberines cularines
t
CH,<)

o HO
OCH3
o> OCH3
OCH3
OCH3
protopines pbtbalideisoquinolines dibenzopyrrocolines
t o ,...
<0 " I H
CH,O I
HN
"" OCH,
o OCH3

secophthaiideisoquinolines

Fig. 32.6. Biosynthetic relationships between alkaloids derived from reticuline (modified from Preininger, 1986; used with pennission of the copyright
owner, Academic Press, Orlando. FL).
 lsoquinoline and Benzylisoquinoline Alkaloids 585

CH,'O

C H ' . 012 7 CH'-027

HO:::'" NH :::,.. 1 N
2' CHr O 'CH
HO ,:? I'
H H 3

" 1 ,:71 ,:71

CH"O.,:::'" "
5' HO :::,.. HO :::,..

(+)-reticuline (2) (+)-(S)·coc:laurine (12) (-)-armepavloe (13)

.O~ rnro~ ~,o~

:::,.. 1
HO
N+-CH,
H .... CH,
HO:::'" HNH HO:::'" NH

HO ,:7 HO ,:7
CH,-O ,:7 1 H
1 :::,..1
HO :::,.. CH,-O
HO :::,..
(-)-norlaudanosoline (26) (+)-nororientallne tembetarine

CH~OW
HOW CHro:cQ
HO:::'" NH
1

d
HO :::,.. NH 1 N
! H HO:::'" , .... CH,
CH"O~ I H
,:71 H
HO :::,..
HO~
HO :::,..
HO :::,..

(R)-norcoclaurine (-)-nororientaline (27) 6-0-methylnorlaudanosoline (28)

<a> [(+)-(R)-demelhylcoclaurlne )

H
HO 0 2 1
NH ~ I HO~
~
HO
I N
H.... CH'

HO ~
~I I
HO ~ HO ~

demethylcoclaurine (l2) {+)-Iaudanosoline (33)

(b) (higenamine)

Fig. 32.7 (a & b). Some representative tetrahydrobenzylisoquinoiine alkaloids.

From the biogenetic point of view, the most important Phylica rogersii (Rbamnaceae). In the opium poppy, Pa-
tetrahydrobenzylisoquinoline alkaloids is undoubtedly ( + )- paver somniferum, both enantiomers of reticuline occur with
reticuline (20). This alkaloid is the precursor of a number the ( + )-enantiomer predominating. Morphine alkaloids are
of more elaborate compounds (Fig. 32.6) and is widely dis- derived from ( - )-reticuline (21).
tributed. (+ )-Reticuline is ftrst synthesized and then con-
verted to ( - )-reticuline (21) in the plant (Fig. 32.8) (Bat- Biosyntbesis
tersby and Foulkes, 1965; Beecher and Kellelior, 1988).
Reticuline occurs in the ( + )-form in Annona reticulata (An- Benzylisoquinoline (BIQ) or tetrahydrobenzylisoquino-
nonaceae), Cinnamomum camphora (Lauraceae), and in line (THBIQ) alkaloids are formed by condensation of dopa-
586 lsoquinoline and Benzylisoquinoline Alkaloids

H HOm HOm
I I H-O~
'-.. '-AI
~
i COzH NH ,;;-'
I
-CC(22)CH~Od-
HO 0 z::::.... N ::::.... ..... NH+

HO ~ NHz H O i llJl
'<::: ,;;-' ,;;-' I
L-tyrosine I ::.. I ::..
HO ~ (19) HO HO

H.:J~~
CH3-027'
HO;:::"" NH HO
~ NH
H

~, ~I
HO ::,... HO ::::....
(S)-norcocJaurine (23)
(S)-coclaurine (124)

CH3::27'"
*I >:-rn:~",
I <,,,,,--':~;:::,...' :'CH3

,
3'~ HO ~ _ HO ~ _
, ;:::,... ;:::,... ,
HO ;:::,... HO CH3-O
(S)-N-methyl-(S)-coclaurine 3'-hydroxy-N-methyl c..laurine (25) (+)-(S)-reticuline (20)

CH 0 CH3-0~

3:0~::::"" , hN"!:"' CH3 HO~:'CH3

HO ~ ---" HOXY

CH3- 0 ;:::,... I 1,2-dehydroreticuline CH3- 0 (-)-(R)-reticuUne (21)

Fig. 32.8. Biosynthesis of (+ )-(S) and (- )-(R)-reticuline (modified from Franzel and Zenk, 1990b and used with pennission of the copyright owners,
Elsevier Science Ltd., The Boulevard, Langford Lane, Kidlington OXS 1GB, UK).

mine (22) (Fig. 32.9) and 4-hydroxyphenylacetaldehyde (19) Several types of data based on feeding experiments, isola-
(Fig. 32.8) (Frenzel and Zenk, 1990b). tion of intermediates and in vivo NMR experiments, indicate
The lack of strict specificity for the enzymes involved that norcoclaurine (23) serves as a precursor to both coclau-
in the initial conversion to (S)-reticuline (20) suggests that rine (24) and (S)-reticuline (20) (Stadler et al., 1987, 1989),
stereochemical control does not occur at this stage ofbiosyn- and that (S)-coclaurine (24) serves as a specific precursor
thesis (Zenk, 1985). The use of plant cell cultures (both cal- to other classes of benzylisoquinoline alkaloids, such as the
lus and suspension types) from different species of plants protoberberines, benzophenanthridines, and morphinandie-
that produce benzylisoquinoline alkaloids has proven to be nones, as well as for pavine and benzophenanthridine alka-
a powerful approach for the isolation of enzymes and inter- loids in intact plants (Stadler et al., 1987, 1989). The five
mediates in the pathways leading to these compounds (Zenk enzymes involved in the conversion of dopamine and 4-
et al., 1985). For example, the use of cell-free extracts of hydroxyphenylacetaldehyde (19) to (S)-reticuline (20) have
Berberis cultures permitted Zenk and co-workers to isolate been elucidated (Frenzel and Zenk, 1990b).
and characterize all eight enzymes involved in the conver- The first step of this biosynthetic sequence involves con-
sion of dopamine (22) and 4-hydroxyphenylacetaldehyde version of the amine and aldehyde to (S)-norcoclaurine (23)
(19) to (S)-reticuline and, subsequently, to berberine. with (S)-norcoclaurine synthase. This condensation is fol-
 Isoquinoline and Benzylisoquinoline Alkaloids 587

£)tC~DJ~
H
i reticuline derived

HO
1 ~
b
NH
, HO
NH, alkaloids

L-tyrosine tyramine 4-hydroxyphenylacetaldehyde (10)

H o : c r t ; CO,H H o m
~
H

--I
__ H0)CJ
I"'" CHO

1# NH, # NH, HO #
HO HO

L-DOPA dopamine (22) 3,4·dihydroxyphenylacetaldehyde

HO
D! '
I
//
'" 0
CO H H o x Y Y C 0 2 H these two compounds ~ve no~

HO
1 ~

b
been shown not to be mtermed18tes
for the synthesis of benzylisoquinoline
0
alkaloids

4·hydroxyphenylpyruvate 3,4-dihydroxyphenylpyruvate

Fig. 32.9. Fonnation of the precursors of benzylisoquinoline alkaloids (modified from Zenk et aI., 1985; used with pennission of the copyright owner,
the American Society of Phannacognosy Downers Grove, lllinois).

lowed by methylation at the 6 position in the presence of S- zyme, (S)-noriaudanosoline synthase, that catalyzes the for-
adenosyl methionine, N-methylation, and hydroxylation at mation of (S)-norlaudanosoline (26) from dopamine (22)
C-3' to yield 3'-hydroxy-N-methylcoclaurine (25) (Fig. and 3,4-dihydroxyphenylacetaceldehyde (19) was isolated
32.8). The enzyme controlling the N-methylation has been from the soluble protein fraction of Eschscholtzia tenuifolia
isolated and studied (Frenzel and Zenk, 1990a). The enzyme cell suspension cultures (Schumacher et aI., 1983), but no
controlling the 3'-hydroxylation was a phenolase also capa- reaction was observed with cell-free extracts and 3,4-dihy-
ble of hydroxy1ating tyrosine to dopa, and tyramine to dopa- droxyphenylpyruvic acid. No apparent cofactor require-
mine, in the presence of ascorbate and oxygen. It is proposed ments were demonstrated (Schumacher et aI., 1983). The
that the phenolase plays a multiple function in Berberis sto- enzyme consisted of four isoenzymes upon further elec-
lonifera cultures (Loeffler and Zenk, 1990). The fmal meth- trophoretic separation. A series of relatively specific methyl-
ylation reaction involves methylation of the C-4' hydroxyl transferases have been isolated from cultures of Argemone
group of 3'-hydroxy-N-methylcoclaurine with S-adenosyl- platyceras (Papaveraceae) and Berberis. In the presence of
L-methionine (SAM): 3'-hydroxy-N-methyl-(S)-coc!aurine- SAM, the first enzyme was reported to convert norlaudano-
4'-O-methyltransferase and SAM to produce (S)-reticuline soline (26) to 6-0-methylnoriaudanosoline (28) (Zenk,
(20) (Frenzel and Zenk, 1990b). 1985). A second enzyme was reported to convert 6-0-meth-
It has long been known that differential incorporation of ylnoriaudanosoline to the corresponding 4,6-0-methyl de-
tyramine into the two portions of the molecule occurs (Bee- rivative, and another enzyme to methylate the 5'-position to
cher and Kelleher, 1988). When DOPA (3,4-dihydroxyphe- yield nororientaline (27) (Fig. 32.7). N-Methyltransferases
nylalanine) was fed to callus cultures of Berberis cana- were found in cultures of several plants and in latex of Pa-
densis, 91 % was incorporated into the isoquinoline moiety, paver somniferum that produced compounds such as reticu-
whereas tyrosine was almost equally incorporated into both line and orientaline (29) (Fig. 32.17) (Zenk, 1985).
portions of the molecule (Riiffer and Zenk, 1986), supporting Apparently, the reason that noriaudanosoline (which had
the pathway outlined above. never been isolated from any plant source) is easily incorpo-
Although norlaudanosoline (26) (Fig. 32.7) was previ- rated is because of the relative nonspecificity of the enzymes
ously assumed to be the precursor of (S)-reticuline (20) of the reticuline pathway (Hartmann, 1991). This nonspecif-
(based on the condensation of dopamine and 3,4-dihydroxy- icity contrasts with the high specificity found among the
phenylacetaldehyde), this route of biosynthesis is no longer enzymes involved in the synthesis of more complex struc-
considered to account for the formation of benzylisoquino- tures (Hartmann, 1991).
line alkaloids. In previous reports, it was noted that an en- Tetrahydrobenzylisoquinoline alkaloids such as lauda-
S88 Isoquinoline and Benzylisoquinoline Alkaloids

HO~
HOI ::::,.. :H
CH3"O~1
~HO NH
-- H
HO 9' HO ,;?
I ~I
HO ~ CH,-O

(-)·(S)·norlaudanosoline (26) (-)-norreticullne

+ +
CH'_O~I CH'_O~I CH'_O~I
~
HO::::'" NH CH,.O::::'" -",N
CH,.O::::'" NH
H --+
CH,-O 9'
CH,-O 9' I CH,-O 9' I H
I
~ CH,'O ~
HO CH,'O ~

(0 )onororientaline (-)-tetrahydropapaverine papaverine (31)

Fig. 32.10. Biogenesis of papaverine (Cordell, 1981; used with pennission of the author).

nosine (30) often are dehydrogenated to the corresponding dients of arrow poisons by the Juris of the Rio Japuras basin,
aromatic compounds, in this instance, papaverine (31) (Fig. Amazonas, Brazil (Fig. 32.11) (Buck, 1984). Alkaloids of
32.10). this structural type from an African tree, Cleistopholis patens
(Annonaceae), had inhibitory effects on Candida.albicans
Biological Activity
in concentrations as low as 1.56 ,...glml (Clark and Hufford,
Tetrahydrobenzylisoquinoline and derived alkaloids have 1992).
physiological effects in most animals (Schiff, 1987).
In general, quaternary benzylisoquinoline alkaloids can·
not pass through membranes (Zenk, 1988). (S)-Reticuline
APORPllll'lE ALKALOIDS
(2) and (S)-scoulerine (17) are transported into vacuoles of
Fumaria capreolata. This transport involves an active A TP-
requiring, stereospecific carrier system that transports only The simplest derivatives of the tetrahydrobenzylisoquinoline
the (S)-enantiomers and not the (R)-enantiomers (Zenk et alkaloids are the aporphine alkaloids (Fig. 32.12). Aporphine
al., 1985). alkaloids (about 650) are widespread and occur in almost the
An aromatized alkaloid of this derivation, papaverine (31) same families as benzylisoquinoline alkaloids (Guinaudeau
is the most important alkaloid of the BIQ group from the and Bruneton, 1993; Kametani and Honda, 1985). Several
pharmacological point of view. Papaverine inhibits aldol re- subgroups of aporphine alkaloids are known [noraporphines,
ductase, GABA and glucose response in chemosensory cells, dehydroaporphines (34), oxoaporphines (35), dimeric apor-
and phosphodiesterase (Wink, 1993). In vivo, this alkaloid phines, and phenanthrenes (36)] (Fig. 32.13) (Cave et al.,
decreases the tonus of the smooth muscle, increases coronary 1987). The aporphine alkaloids present in members of the An-
artery flow, and causes dilation. Papaverine has a beneficial nonaceae and in the genus Thalictrum (Ranunculaceae) have
effect on angina pectoris. This aromatic isoquinoline alka- been reviewed (Cave et al., 1987; Schiff, 1987).
loid is neither narcotic nor addictive (Cordell, 1981). Papav-
erine has an LDso Lv. in mouse of 27.5 mglkg. Demethylpa- Biogenesis
paverine inhibits aldose reductase (Wink, 1993).
Papaverine (31) inhibits root growth and induces poly- Aporphine alkaloids may arise by an oxidative coupling
ploidy in Allium. This alkaloid is a feeding deterrent to sev- reaction that can occur at positions ortho or para to phenolic
eral insects (Wink, 1993). groups (Fig. 32.14) (Battersby, 1967). Rotation about the
single bond by which the benzyl group is attached before
Azafluorantbene and Tropoloisoquinoline cyc1ization gives rise to two series of aporphine alkaloids,
Alkaloids as represented by bulbocapnine (32) on the one hand, and
This group of alkaloids occurs in the genus Abuta (Men- glaucine (28) on the other. Methylation can reduce the num-
ispermaceae). Crude extracts of the plants are used as ingre- ber of possible coupling sites, as coupling only occurs ortho
 Isoquinoline and Benzyiisoquinoline Alkaloids 589

OCH,
OCH,
CH,-O

CH,-O

OCH, OCH,

imerubrine imeluteine

tropoloisoquinoline alkaloids azatluoranthene alkaloids

Fig. 32.11. Azafluoranthene and tropoloisoquinoline alkaloids.

(+)-bulbocapnine (37) (+)-glaucine (38) (+ )-ocotelne (40)

CH,-O
HO

CH,-O
CH,-O

HO

HO
HO
OCH,
OCH,

{+)-lirioferine (43)
(+)-liriotulipiferine (44)

H-stephanlne (47) laureline (48) (+).isothebaine (49)

Fig. 32.12. Some representative aporphine alkaloids.

590 /soquinoline and Benzylisoquinoline Alkaloids

""w CH,-O
CH'°Z?I
CH3~O ~ h ..... CH3 o

:?'I 00
CH,-O "'" OCH,
OCH,
oxoglaucine (35) taspine (41)
1,2-dihydroxy-7-oxoaporphine
(Iiriodendronine)
OH

HO~ 0
CH,-O "'" I .,..:'CH'<O

:?'I
CH,-O "'"
OH

boldine (42) dehydrothalicmine (34) N-methylthaliglucine (36) liriodenine (45)

Fig. 32.13. Subgroups of aporphine alkaloids.

or para to free hydroxyl (phenolic) groups. The methylation high levels. Similar experiments with reticuline labeled in
pattern is of great importance in aporphine alkaloid biosyn- several positions demonstrated that direct coupling of reticu-
thesis (Fodor, 1980). line occurs to yield the aporphine skeleton. Furthermore,
The label from N-CH3 labeled ( + )-(S)-reticuline is incor- reticuline is incorporated into isoboldine (39) (Kametani and
porated into bulbocapnine (37) in Papaver somniferum at Honda, 1985; Santavy, 1979).

CH"O~I
o N'CH
CH'O~IN'CH HO "" CH'-OifN
. ~H ' 'H 'CH3-0 "" ' '''H 'CH3
-------"-
---+- "" ~
/ I I I"" :P I
CH,·o CH,O CH,-O ""
CH,O o OH
OCH,
HO isoboldine (39) (+ )-glaucine (38)

CH,H: : I 6'
(+).reticuline (20)
\CHJO~I 0
CH'Ogl
N . . . CH HO ~ N "CH
: 'H 3 "H 3

o ---+-HO ~ -+- ---...

I I
CH,- 0 "" CH,- 0 '"
(+ )-bulbocapnine (37)

Fig. 32.14. Biogenesis of aporphine alkaloids such as bulbocapnine and glaucine (modified from Geissman and Crout, 1969).
 Isoquinoline and Benzylisoquinoline Alkaloids 591

Biological Activity Aristolochic acids (46) and the related aristolactams are
Aporphine alkaloids exhibit several types of biological thought to be derived from aporphine alkaloids, although
activity in other organisms. For example, these compounds there is little experimental evidence to support this proposal.
display a variety of effects in mammals (Corde\1, 1981; Tyrosine. DOPA, and noradrenaline are precursors. A bio-
Schiff, 1987). Most are toxic; they often exhibit antagonistic synthetic pathway has been proposed (Fig. 32.15) (Cordell.
effects to dopamine (Castro, 1993). The anticonvulsant ac- 1981).
tion of several aporphines acting at dopamine receptors has Aristoloehia clematitis has been used to treat snakebites
been documented. Glaucine (38) induces narcosis and con- and wound infections. One of the major alkaloids of this
vulsions in mammals (Schiff, 1987). Ocoteine (40) (thalic- plant is aristolochic acid (46). The plant extract is not directly
mine) has antitussive, hypotensive, spasmolytic, and adreno- antimicrobial. but it produces an enbancement of phagocyto-
lytic activity in cats and has been noted to produce sis of leucocytes and peritoneal microphages. Aristolochic
hyperglycemia in rats (Schiff, 1987). Bulbocapnine (37) in- acid is suspected to be carcinogenic (Pezzuto et al.. 1988;
hibits peripheral dopamine receptors and has an LDso p.o. Wagner and Proksch, 1985) and has been reported to have
in mouse of 413 mg/kg and glaucine (Lv.) of 98 mglkg. antitumor activity. The LDso i.v. of aristolochic acid (46) in
Isoboldine (39) inhibits aldose reductase (Wink, 1993). mouse is 38-70 mg/kg (Wink. 1993).
Other members of this group of alkaloids exhibit a variety Larvae of the butterfly, Paehlioptera aristoloehiae, which
of activities. Taspine (41) is the active component of the sap feed exclusively on plants of the family Aristolochiaceae,
of Croton species used by the Jivaro Indians for its wound- accumulate aristolochic acid (46) (Levinson, 1976). Aris-
healing properties (Lewis, 1992). Demethylcoclaurine (hi- tolochic acids have also been isolated from severallepidop-
genamine, norcoclaurine) (23) has positive inotropic activity teran caterpillars Md are transferred to the pupae. Imagos
at a concentration of 10-7 g/ml (Wagner, 1988). Laudanoso- of the swallowtail butterflies, Battus polydamas and B. phi-
loline (23) is antifeedant and exhibits growth inhibition to lenor. contain aristolochic acid (Nahrstedt, 1982; Urzua et
larvae of Hyphantria. Spodoptera, and Lymantria. Lauda- al., 1987), but do not appear able to adjust the levels of
nosine (30) modulates glycine neurochemical activity toxins, as do monarch butterflies (Harborne, I 986).
(Wink, 1993). A mixture of aristolochic acids and inositol was found to
Several of these alkaloids are involved in plant-insect in- be an oviposition attractaot for an Aristolochiaceae-feeding
teractions. Isoboldine (39) is known to be an inbibitor of insect swallowtail, Atrophaneura alcinous. and induced a response
feeding (Cordell, 1981). Several aporphine alkaloids are re- identical to that of the host plant (Nishida and Fukami.
pellent or toxic to insects. Among these are boldine (42), a 1989).
feeding deterrent to polyphagous Syntomis larvae. glaucine Aristolochic acid is a feeding deterrent and produces
(38), which is a feeding deterrent and growth inhibitor for lar- growth inhibition in the larvae of Hyphantria. Spodoptera.
vae of Hyphantria. Spodoptera, and Lymantria, and isobol- and Lymantria (Wink, 1993).
dine (39), which is a feeding deterrent for Prodenia and Orae-
sia (Wink, 1993). The pharmacological effects of many Eupomatia Alkaloids
aporphine alkaloids have been reviewed (Castro, 1993). This small group of alkaloids is only known from the
Two previously undescribed aporphine alkaloids, liriofer- genus Eupomatia (Eupomatiaceae). These compounds ap-
ine (43) and liriotulipiferine (44) as well as several other simi- pear to be derived from oxoaporphine alkaloids, although
lar alkaloids were produced in the sapwood of Liriodendron little experimental work has been done to establish this pro-
tulipifera (tulip tree, Magnoliaceae) upon injury to the tree. posal (Fig. 32.16) (Taylor, 1985).
Under normal conditions, only glaucine could be detected
(Chen et al., 1976). Liriodenine (45), from the same source,
Proaporpbine Alkaloids
has broad-spectrum antimicrobial activity, including activity
against Candida albieans. Aspergillus niger, and several Although the formation of most aporphine alkaloids is
other fungi. Because of its relatively low toxicity and its good straightforward, the formation of others, such as stephanine
in vivo efficacy, liriodenine may have potential as an antifun- (47), laureline (48), and isothebaine (49) (Fig. 32.17) is more
gal agent (Clark and Hufford. 1992). This compound also is complicated (Mann, 1987). In this case. the ortho or para
active against nasopharyngial tumors and is cytotoxic (Wink. oxygen function reqnired for direct oxidative coupling is
1993). Boldine is toxic toLemna (Wink, 1993). absent in the probable precursor. Formation of this type of
alkaloids involves the intermediacy of compounds called
Aristolactams and Aristolocbic Acid proaporphine alkaloids, such as orientalinone (50). The for-
This group of alkaloids is known primarily from the genus mation of aporphine alkaloids with apparently anomalous
Aristoloehia (Aristolochiaceae). Several of the compounds structures has been postulated to occur by the biological
contain nitro groups. Despite their unusual structures, aris- equivalent of the dienone-phenol and the dienol-benzene
tolochic acids are derived from benzylisoquinoline precur- rearrangement (Fig. 32.17) (Geissman and Crout, 1969).
sors. Alkaloids from the genus Aristoloehia (Aristolo- Following coupling of the position para to the hydroxyl
chiaceae) have been reviewed (Chen and Zhu, 1987). of the benzyl group of N-methylcoclaurine (51) to produce a
592 Isoquinoline and Benzylisoquinoline Alkaloids

aristolocbic acid·U (46) &ristolocbic acid·I

t
CHHi¥2~?":"" "Hi§9:
~ 9' : I N'CH3CH30R:9'
HO I N, CH3
0.&
OCH3 HO.&OCH -- :9':",. I OCH
orientalinone (50)
3 3
Fig. 32.15. Biogenesis of aristolochic acids and aristolactams (modified from Cordell, 1981; used by permission of the author).

5-membered ring, ring expansion occurs by dienone-phenol Thalictrum (Ranunculaceae) have been reviewed (Santavy,
rearrangement to yield laureline (48) (Fig. 32.17) (Geissman 1979; Schiff, 1987).
and Crout, 1969). The in vivo conversion of (R)-reticuIine to morphinandie-
In a similar manner, coupling and ring closure occur with nones such as salutaridine (52) (promorphinanes) has been
( - )-orientaline (29) to yield isothebaine (49). 10 this in- demonstrated (Santavy, 1979) (Fig. 32.18). An enzyme from
stance, however, ring expansion occurs via a dienol-ben- a cell-free extract of Papaver somniferum utilized H20 2 to
zene rearrangement (Fig. 32.17) (Geissman and Crout, produce salutaridine in high yield. Morphine alkaloids (see
1969). Migration of the alkyl rather than the aryl group in below) are derived from the morphinandienone alkaloid sa-
an intennediate leads to an aporphine from which stephanine lutaridine.
is readily derived (Fig. 32.17). Both enantiomers of the morphinan skeleton occur in na-
Although the mechanisms above were postulated before ture.
compounds resembling the proposed intermediates were
known, compounds such as orientalinone (50) were later Hasubanan Alkaloids
isolated. Both (- )-orientaline (29) and (+ )-orientalinone
Hasubanan alkaloids (about 40 in total), in which the ni-
(50) (a proaporphine alkaloid) serve as precursors for iso-
trugen is attached at position C-14 (rather than C-9), are
thebaine (49) in Papaver orientale (Fig. 32.12) (Geissman
derived from sinoacutine (Guinaudeau and Bruneton, 1993).
and Crout, 1969).
Proaporphine alkaloids are known from the Euphor-
( + )-Reticuline (20) gives rise to sinoacutine (53). 10 Sino-
menium acutum, sinoacutine is the precursor of sinomenine
biaceae, Lauraceae, Menispermaceae, Monimiaceae, Ne-
(54), a compound enantiomeric to morphine.
lumbonaceae, and Papaveraceae (Santavy, 1979).
Hasubanonine-like alkaloids are known from the genera
Sinomenium and Stephania (Menispennaceae). A biogenetic
l'fORPIIINAI'IDmI'lOl'lE ALKALOIDS pathway leading to the fonnation of hasubanonine (55) and
(PROl'fOKPHIl"lAl'lES ) protostephanine (56) has been proposed (Battersby et al.,
1974, 1977, 1981a, 1981b, 1981c) (Fig. 32.19).10 experi-
Morphanandienone alkaloids are widely distributed in the ments in which labeled precursors were fed into whole
families Papaveraceae and Meuispennaceae. Alkaloids of plants, Battersby and co-workers found that simple tyramine
this structural type in the Papaveraceae and in the genus derivatives such as 2-(3,4-dihydroxy-5-methoxyphenyl)eth-
 Hte
lsoquinoline and Benzylisoquinoline Alkaloids 593

OOg2.
If. -
HO ::,... 1 "N oo 1h N
o 1 N
--+ --+ "
~ 1 0 ~ 1 0 ~ 1

~t2 -- 021:
0
::,... ::,... ::,...

o~ -
1'<::
HN I h
N N~ ~N
HN ""N
--+ ~
~ 1 0 ~ 1 0
~ 1 0
::,... ::,...
::,... 0
0

--
0

&2.
~
::,...
.0
1
""N

0
--
g1~
~.o N ~ 0

~ 1 0

~
OCH,
::,...

ci2J
eupolauramine
o ::,... N

l..,p ::,.. 1 1.0

"" 0
N
eupolauritline

Fig. 32.16. Eupomatia alkaloids (modified from Taylor, 1985; used with pennission of the copyright owner, Academic Press, Orlando. Fl.),

ylamine (57) were incorporated into hasubanonine and pro- morphine (58) itself is only known to occur in the opium
tostephanine, as were a series of l-benzyltetrahydroisoqui- poppy, Papaver somniferum (Bisset, 1985). Another species,
nolines that possessed two phenolic oxygens in one of the P. bracteatum, synthesizes quantities of thebaine (59) that
rings undergoing oxidative coupling during the biosynthesis can be converted chemically to morphine (Verpoorte et aI.,
(Battersby et aI., 1981a, 1981b, 1981c). 1991).
The role of alkaloids from the genus Stephania in Chinese Opium is produced by lacerating the immature capsules
traditional medicine has been discussed (Han et aI., 1988). of the plant. Care must be taken not to cut through the peri-
The NMR spectra of hasubanan alkaloids have been re- carp. Mter the latex has exuded and dried, the dark, resinous
viewed (Matsui, 1988). material is collected. This crude opium, which contains as
much as 12-14% morphine, is then used directly or further
purified (Tyler et aI., 1981). The estimated legal world pro-
JIIOKPHll'lE ALKALOIDS duction of opium is more than 1000 metric tons; from this,
about 660 metric tons of codeine and 200 metric tons of
Poppies have long been used by man as a medicinal plant morphine are prepared (Verpoorte et aI., 1991).
and as a drug of abuse. The use of opium is mentioned by Because of the economic importance of morphine alka-
several ancient writers; among them are Theopbrastus and loids, many attempts to produce them in plant tissue cultures,
Dioscorides (Tyler et aI., 1981). Morphine-type aIkaIoids usually Papaver somniferum or P. bracteatum have been
are found in several members of the genus Papaver, but made (Ellis, 1988; Verpoorte et aI., 1991). Cultures of both
S94 /soquinoline and Benzylisoquinoline Alkaloids

CH30~
I CH3 0 ' r3
HO '-' N' CH3 ',-,
H HO 1Z
CH3·O r CH3·O
1- 11 r
HO '-' 0 h

(-j.orientaline (29) (+ )-orienlalinone (50) dienol-benzene (+)-lsotbebalne (49)

rearrangement

N-methylcoclaurine (51)

CH30~
HO '-' N' 1 CH
H ....:.....
r l
HO '-'

dienone-phenol laureline (48)

rearrangement

N CH3-0
'CH3 HO '-'
1 » :N'
CH
<O~I
° '-' N.
H CH3
(+ )-orienlaline (29) H H 3_ r
- r I
'-' 1 '-' OCH3
OCH3 OCH3
stephanine (47)

Fig. 32.17. Fonnation of aporphine alkaloids via the intennediacy of proaporphine alkaloids (modified from Geissman and Crout, 1969).

species are excellent producers of sangninarine. In summary, among plants, this alkaloid has been isolated in trace
none of these attempts has successfully produced commer- amounts from the skin of a bufonid toad, Bufo marinus (Daly
cially interesting amounts of morphine alkaloids. Radio- et al., 1993). Morphine and thebaine have been reported from
tracer studies, however, suggest that the failure of these cul- mammals (Brossi, 1993). The biosynthetic pathway seems
tures to accumulate morphine alkaloids may be due to high to be similar to that of plants, although some differences
turnover rather than to an inability to synthesize the com- have been noted.
pounds of interest (Ellis, 1988). Other alkaloids from earlier
steps in the pathway are present. These include isothebaine Biosynthesis
(49), orientaIidine (50), protopine (60), cryptopine, and, usu-
ally as the main product, sanguinarine (61). Treatment of Robinson and Gulland (1925) were the first to observe
suspension cultures of Papaver somniferum with a homoge- that the foonation of aporpine alkaloids was by oxidative
nale of the fungns Botrytis increased the amount of sanguina- coupling of benzylisoquinoline precursors. They also
rine 150-fold. Tissue cultures have been extremely useful pointed out that by a different mode of coupling, the basic
for the isolation of key enzymes of this pathway (Hutchin- carbon skeleton of the morphine alkaloids thebaine (59),
son, 1986). codeine (62) and morphine (58) could arise (Fig. 32.20).
Although morphine has an extremely limited distribution (- )-(R)-Reticuline serves well as a postulated precursor be-
 Isoquinoline and Benzylisoquinoline Alkaloids 595

(-).(R)·reticuline (21)

salutaridine (52) morphinan system

CH30~
I
HO
?"
~ N 'CH
CH30~
HO
~ I CH30
0.
I 2t
__
"H ~ ......' \. ~ .......' ....
= ?" N N
HO P I ~ I H ' CH3 I I H ' CH3
CH3 0 ~ CH30 CH30
OH 0
(+ HS)·reticuline (20)

CH30~,
CHP~
HO ~
, 3

4
HO ~
.."
., ~"

, ......, H \'-;3- 5 16
H N'CH3
o h
CH3 0 OCH3
o
sinoacutine (53) hasubanan system sinomenine (54)

Fig. 32.18. Fonnation of motphinandienones from (+)- and (- )-reticuline (modified from Battersby, 1967; modified and used with pennission of the
copyright owner, Marcel Dekker, Inc., New York).

cause the methylation pattern precludes formation of unde- Quadruply-labeled (- )-reticuline (21) was specifically in-
sirable coupling modes and favors the desired pathway (Cor- corporated into thebaine. Tritium-labeled salutaridine (52)
dell, 1981). also was specifically incorporated. Salutaradine first was dis-
The results of numerous labeling and feeding studies and covered in Croton salutaris (Euphorbiaceae) from Brazil,
other experimental results have established that the steps of but later was demonstrated to occur in Papaver somniferum
the pathway are as indicated (Fig. 32.20). 2-14C-Tyrosine in low concentrations (Geissman and Crout, 1969). This
produces morphine labeled at C-9 and C-16. 1.1 4 C-Dopa- compound gives salutaradinol I (64) by reduction (Fodor,
mine gives morphine labeled at C-16. 3,4-Dihydroxyphenyl· 1980). In the above study, it was demonstrated that the oxy-
alanine is readily incorporated into morphine (58), thebaine gen bridge is formed by an SN2' displacement of the oxygen
(59), narcotine (63), and papaverine (31) (Santavy, 1979). on salutaradinol rather than by addition to the dienone sys-
(R,S)-Norcoclaurine (23) and (R,S)·coclaurine (24) (both lao tem of salutaridine (Fodor, 1980).
beled at C-I) were incorporated into thebaine and morphine N-Demethylation is the most frequently observed step in
(labeled at position C-9) when fed to Papaver samniferum alkaloid catabolism (Waller and Nowacki, 1978). Not only
plants (Loeffler et aI., 1987). Noriaudanosoline (26) (Fig. is morphine formed by this process, but morphine is con-
32.5) is an efficient precursor of morphine. Studies of the verted into normorphine quite readily.
rates of incorporation of radioactivity demonstrated the se- Over 40 alkaloids are known from opium. Several of
quence thebaine (59) to codeine (62) to morphine (58). these are of importance. Among them are morphine (58),
S96 Isoquinoline and Benzylisoquinoline Alkaloids

CH,-O
CH'-OW
CH'-0)QJ
I

--
N HO
HO
I "'" NH, HO ~ ,'CH,

""it
HO~H --

CH'-O~ CH,-O
o
HO

CH,-O CH,-O
CH'.O~
- --
I
HO

CH,-O OCH,
--
··"".::::::::'N+....... CH
3
HO

CH,- 0
""'-

I ······,·""."N ..... CH3

OCH,
CH,-O

o
o o OCH,

(0) hasubanonine (55)

CH'.O~OCH'
~I
17 N-CH, I
HO~~ NH,

~ I HO
CH,· 0 OCH, OCH,

(b) protostephanine (56) 2--(3,4-dihydroxy-5-methoxyphenyl)ethylamine (57)

Fig. 31.19. Proposed biosynthesis of basubanonine (Battersby et aI .• 1981c; used with pennission of the copyright owner, The Royal Society of
Chemistry, Cambridge).

codeine (67), thebaine (59), a-narcotine (noscapine) (Fig. are most rapid during the first days of capsule development,
32.27) (63), and papaverine (11). at which time the pericarp volume is increasing rapidly.
Two of the principal alkaloids of opium are morphine When morphine was fed to the stem below the developing
(4-21 %) and a a-narcotine (noscapine) (4-8%) (63). These capsules, 8% of the morphine was metabolized in a few
alkaloids occur partially bound to meconic acid, which is days. Diurnal changes in the content of alkaloids in Papaver
diagnostic for opium. Morphine (58) and its salts are classi- somniferum occur (Waller and Nowacki, 1978). The half-
fied as narcotic analgesics. Codeine (62) also occurs in life of morphine in several species of poppies is 7.5 h.
opium, in the concentration range 0.7-2.5%. Most codeine
(62) is prepared by methylation of morphine. Pharmacological Activity
Morphine does not continuously accumulate in the latex
of P. somniferum. The isolated latex of the opium poppy The pharmacological actions of morphine (58) are com-
sustains alkaloid synthesis and further metabolizes morphine plex. Some specific central nervous system functions are
in vivo to as yet unidentified products. The alkaloids are depressed; others are stimulated. Typically, morphine pro-
synthesized in the vesicles and are translocated to the devel- duces analgesia, drowsiness, changes in mood, and mental
oping capsule by the upward movement of the latex. The clouding. Analgesia occurs without loss of consciousness.
metabolites pass out of the latex into the pericarp and into In humans, death from morphine poisoning is almost always
the ovules (Santavy, 1979). Translocation from the stem to due to respiratory arrest. Morphine (58) and its analogs are
the developing capsule is well established. These transfers powerful depressants of the cough center (antitussives).
 Isoquinoline and Benzylisoquinoline Alkaloids 597

(-)-(R)-reticuline (21) salutaridine (52)

salutaridinol I (64) tbebalne (59) neopinone

codeinone codeine (62) morphine (58)

Fig. 32.20. Biogenesis of morphine (modified from Geissman and Crout, 1969).

Morphine has an LDso i.v. in mouse of 226-318 mglkg (Cordell, 1978a). This substance has an LDso Lp. in mouse
(Wink, 1993). Codeine (62) is also an important analgetic of 20 mglkg (Wink, 1993).
and antitussive drug. Codeine is the most widely used mor-
phine type aIkaloid; it has an LDso Lp. in mouse of 300 otber Biological Activity
mg/kg (Wink, 1993). This aIkaloid is derived from opium Although the biological properties of these alkaloids in
(0.2-0.7%) and synthesized by methylation of morphine humans are well known, their role in other biological interac-
(Tyler et al., 1981). Heroin (65) (Fig. 32.21), a synthetic tions has been little studied. Morphine inhibits root growth
diacelate of morphine, crosses the blood-brain barrier more and induces polyploidy in Allium. Narcotine (63) inhibits
rapidly than morphine and is highly euphoric and analgetic. seed germination (Wink, 1993). Several morphine alkaloids
The compound methadone (66) appears to have the struc- have activity in plant-insect interactions. Morphine (58) is
tural features needed to bind to opiate receptors in the brain. a phagorepellent for Pieris and Leptinotarsa. Codeine (62)
This compound is active orally. Naturally occurring peptides is a feeding deterrent for Phormia; narcotine (noscapine)
called enkephalins (67 and 68) have been isolated from brain (63) is a feeding deterrent for polyphagous Syntomis larvae
tissues of certain animals, including man (Fig. 32.21). These (Wink, 1993).
compounds are derived from larger peptides. Both have anal-
gesic effects and apparently act at the same receptors as
opiates. Morphine and related aIkaloids may also occur in PROTOBERBERINE ALKALOIDS
animal tissues (Kosterlitz, 1987).
Thebaine (59) is of special interest, as this alkaloid can The protuberberine aIkaloids are one of the most widely
be converted into morphine. Thebaine is almost the sole distributed groups of the benzylisoquinoline alkaloids (a
alkaloid (up to 26%) of the latex of Papaver bracteatum subgroup of these is called "berberine aIkaloids"). Approxi-
 %
598 lsoquinoline and Benzy/isoquinoline Alkaloids

o
"""
1
H
N(CH,h
J-O~
"""I
o
• N
~I
)-0....... Q Ii 'CH,

""" o
methadone (66) heroin (65)

H,N·tyr·gly·gIy·pbe·met·CO,H (67)
enk:ephalins
H,N.tyr.gly.gly.pbe·leu·CO,H (68)

Fig. 32.21. Methadone and enkephalins.

mately 100 alkaloids of this series are known (Beecher and 1969). (S)-Scoulerine (72) is the first tetrahydroberberine
Kelleher, 1988; Guinaudeau and Bruneton, 1993) from at formed. This compound is subsequently methylated at the
least 10 plant families: Alangiaceae, Annonaceae, Berberi· hydroxyl group in position 9 (S-adenosyl methionine) to
daceae, Fabaceae (Erythrina), Fumariaceae, Lauraceae, yield (S)-tetrahydrocolumbarnine (73), which is then aroma-
Menispermaceae, Papaveraceae, Ranunculaceae, and Ruta· tized to form columbarnine (70) (Hartmann, 1991). The
ceae (Bhaknni and Jain, 1986). Berberine also has been reo methyltransferase that reacts with (S)-scoulerine (72), S-ad-
ported from the Juglandaceae and Rubiaceae (Suffness and enosyl-L-methionine: (S)-scoulerine 0-9-methyltransferase,
Cordell, 1985). is highly specific (Herbert, 1986; Zenk et aI., 1985). Colum-
Berberine (71) is produced by several plant cell cultures barnine (70) gives rise to berberine (71) by formation of a
(Ellis, 1988). Suspension cultures of Coptis japonica pro· methylenedioxy ring at ring A (Amann et al., 1988). A highly
duce large quantities (8% by dry weight) of berberine. Some specific Fe2+-containing enzyme with 32,000 MW converts
cultures excrete berberine directly into the culture medium columbamine into the corresponding methylenedioxy deriv-
whereupon it crystallizes (Ellis, 1988). Cell cultures of some ative, berberine (49)(Herbert, 1986; Riiffer and Zenk, 1985).
Berberis species synthesize jatrorrhizine (69) and colum· The C-8 bridge of berberine is derived from the N-methyl
bamine (70) as the principal alkaloids, especially during the of the tetrahydrobenzylisoquinoline precursor. The results
exponential growth phase and stationary phase (Verpoorte of double-labeling experiments suggest that the process is
et al., 1991). Many of these same cultures produce berberine similar to the formation of methylenedioxy systems.
as the major alkaloid during lag·phase growth. All of these enzymes are associated with specific vesicles,
most likely derived from smooth endoplasmic reticulum. Be-
Biosyntbesis cause of the charge of the quaternary protoberberine alka-
loids, the alkaloids do not leave the vesicles, but ultimately
Based on work with cell·free extracts, the enzymology end up in the vacuole when the vesicle membrane fuses with
of the biosynthesis of protoberberine alkaloids has been reo the tonoplast (Hartmann, 1991).
solved to a great extent (Herbert, 1986). (S)·Reticuline (20) Berberine biosynthesis in Coptis cultures involves forma-
is converted intu berberine (71). (S)·Coclaurine (24) is an tion of a methylenedioxy bridge into tetrahydrocolum-
early precursor for protoberberine alkaloids (Stadler et al., barnine (73) to yield canadine (74) (Fig. 32.23). The pathway
1987). This compound gives rise to a later precursor of pro· differs in Berberis cultures. This should be a warning not
toberberines, (S)- or (+ )-reticuline (10), a major branch to generalize pathways of secondary metabolism unless the
point in the synthesis of many types of benzylisoquinoline enzymatic steps have been elucidated for each species (Hart-
alkaloids. The additional methylene group of protoberberine mann, 1991). (- )-Canadine (SO) was efficiently incorpo-
alkaloids has been demonstrated to come from methionine rated into berberine in Hydrastis canadensis (Bhaknni and
(Fig. 32.22) and is called "the berberine bridge" in many Jain, 1986).
publications. The "berberine-bridge" enzyme converts (S)- An enzyme, (S)-tetrahydroprotoberberine oxidase, from
reticuline (20) into (S)-scoulerine (72) (Hartmann, 1991). suspension-cultured cells of Berberis wilsoniae catalyzes the
The N-methyl group is transformed into an iminium function removal of four hydrogen atoms from a number of tetrahy-
by oxidation and followed by ring closure (Beecher and Kel- droprotoberberine alkaloids in the presence of oxygen
leher, 1988; Bhaknni and Jain, 1986; Geissman and Crout, (Amann et al., 1988).
 Isoquinoline and Benzylisoquinoline Alkaloids 599

-
CH3·O

HO

OCH3

(+)-(S)-reticuline (20)

CH30~
I CH30~
I CH3 0

~ ~ ~OCR'
'"
HO'" N HO'" N ~ ",I N 3
H __ H HO-
o ",OH H OCH
I I '" 3
OCR, OCR,

(S)-scoulerine (72) (S)-tetrahydrocolumbamine (73)

<
CH3 0 CH3-O o
4
HO -0
OCH3

OCH3

columbamlne (70) berberine (71)

1: berberine bridge enzyme, 50,000 daltons, pH optimum 8.9, Oz

2: (S)-scoulerine-9-Q-metbyltransferase, 60,000 daltons, pH optimum 8.8, SAM
3: (S)-tetrahydroproloberberine oxidase, 100,000 daftoos, pH optimum 8.9, 0,
4: berberine synthase, 52,000 daltons, pH optimum 8.9, O2

Fig. 32.22. Proposed biosynthesis of berberine (modified from Amann et aI., 1988; used with pennission of the copyright owner, Springer-Verlag.
Berlin).

It has been demonstrated that berberine is incorporated ine has general antimicrobial, trypanocidal, antiamebic, anti-
into jatrorrhizine (69) (Beecher and Kelleher, 1983). Cul- fungal, anthelminthic, leishmanicidal, and tuberculostatic
tures of Berberis aggregata also yielded a specific methyl- properties. This alkaloid inhibits the growth of Trypanosoma
transferase that converts tetrahydrocolumbamine (73) to tet- cruzi (Wink, 1993). It inhibits reverse transcriptase, interca-
rahydropalmatine (75) and an oxidase that converts lates with DNA, and inhibits aldose reductase, acetylcholin-
tetrahydroberberine (76), tetrahydrocolumbamine (73), and esterase, alcohol dehydrogenase, aldehyde reductase, di-
tetrahydropalmatine (75) to their quaternary counterparts amine oxidase, tyrosine decarboxylase, and RNA synthesis
(Beecher and Kelleher, 1984). (Wink, 1993). This alkaloid is widely used for the treatment
of malaria, amoebiasis, and leishmaniasis. The ICs• against
Biological Activity Plasmodiumfalciparum is from 0.14 to 0.36 ILg/mi. How-
Berberine (71) inhibits growth of radicle in Lepidium and ever, berberine lacks in vivo activity in mice (Borris and
Lactuca (Wink, 1993). Schaeffer, 1992; Phillipson et al., 1993). The LDso of berber-
The phannacological activity of berberine and protober- ine in mice (intraabdominal) is 27.5 mgikg. Further, berber-
berine alkaloids has been reviewed (Schiff, 1987). Berberine ine has antitumor properties (Schiff, 1987; Suffness and Cor-
has a wide range of interesting biological activities. Berber- dell, 1985). Berberine is sold in India as an antidiarrheal and
600 lsoquinoline and Benzylisoquinoline Alkaloids

RO

CR,-O :::~:~71 N

OCR "'" OCR,

, I
OCR, h OCR,

jatrorrhizine (69) tetrahydroberberine (76) tetrahydropalmatine (75)

canadine (74) palmatine (78) tetrahydrojatrorrhizine (77)

Fig. 32.23. Biologically active protobererine alkaloids.

has been shown to be effective for this pwpose (Beecher creased the amplitude of uterine contractions in rabbits and
and Kelleher, 1988). 1n Japan, berberine and the rhizomes guinea pigs and appears to be useful as an oxytocic. Proto-
of the plant Coptis japonica are widely consumed as a stom- pine (60) has smooth-muscle relaxant, hypotensive, and anti-
ach tonic (Ranunculaceae). Berberine contracts the uterine microbial activity against Gram-positive bacilli as well as
muscle and is used to stop uterine bleeding (Verpoorte et oxytocic properties (Schiff, 1987). Protopine has an LDso
al., 1991). Berberine (71) is a feeding deterrent and is toxic i.p. in mouse of 36-102 mg/kg (Wink, 1993).
to a number of insects (Wink, 1993).
Jatrorrhizine (69) and palmatine (78) are active against
Plasmodium Jalciparum. Jatrorrhizine inhibits butyrylchol-
BENZOPHEI'IANTHRIDIl'IE ALKALOIDS
inesterase (Wink, 1993). Palmatine inhibits reverse tran-
scriptase, aldose reductase, and acetylcholinesterase (Wink,
1993). The benzophenanthridine skeleton is encountered in approx-
Columbamine iodide and certain other protoberberine al- imately 30 alkaloids, principally of the family Papaveraceae
kaloids are active inhibitors of the mv-1 reverse tran- (Cordell, 1978a).1n contrastto the biosynthesis of protopine
scriptase system (Tan et al., 1991). Columbamine (70) also alkaloids, phenanthridine alkaloids are synthesized in the
inhibits butyrylcholinesterase (Wink, 1993). cytoplasm (Hartmann, 1991). This type of system arises
Canadine inhibits aldose reductase (Wink, 1993). from a protoberberine precursor by fission of the C-6-N
bond and recyclization. The biogenetic sequence leading to
chelidonine (SO) biosynthesis in Chelidonium majus has
been supported by feeding experiments with multiply-la-
PROTOPIl'IE ALKALOIDS beled (+ )-reticuline [(S)-reticulinej (20) and with labeled
stylopine (79) (Fig. 32.25) (Hutchinson, 1986; Simanek,
Berberine or protoberberine alkaloids [such as stylopine 1985; Tanabashi and Zenk, 1988). (S)-Z-N-Methylstylopine
(79) j may be converted into alkaloids of the protopine type and protopine (60) have been shown to be metabolites in
(60) (Fig. 32.24) (Geissman and Crout, 1969; Hartmann, this pathway. Reticuline is oxidatively cyclized to (-)-
1991). Highly specific microsomal cytochrome P-450-de- scoulerine (72). Formation of two methylenedioxy groups
pendent enzymes from cells of Eschscholtzia californica results in the formation of stylopine (79) (Hartmann, 1991).
(and other species of the Papaveraceae and Fumariaceae) Oxidation of the methylene group at C-6 to the aldehyde
introduce two methylenedioxy bridges into (S)-scoulerine (81) involves stereospecific removal of the pro-S hydrogen.
(72) to form (S)-stylopine (79), which is, in tum, an impor- A microsomal cytochrome P-450 NADPH-dependent en-
tant precursor of the protopine, phthalideisoquinoline, and zyme which hydroxylates C-6 of protopine (60) and leads
benzophenanthridine groups (Hartmann, 1991). to 6-hydroxyprotopine (S2) has been discovered. This en-
Allocryptotine (80) produces antiarrhythmic effects in zyme is dependent on NADPH and molecular oxygen and
humans in doses of 60-120 mglpatient. This alkaloid in- probably is a P-450-linked monooxygenase (Tanabashi and
 lsoquinoline and BenzyJisoquinoline Alkaloids 601

stylopine (79) protopine (60)

OCR,

OCR,
11

allocryptopine (80)

Fig. 32.24. Biogenesis of protopine alkaloids (modified from Geissman and Crout. 1969).

Zenk, 1990). 6-Hydroxyprotopine (82) spontaneously rear- Some benzophenanthridine alkaloids possess a wide
ranges to dihydrosanguinarine (83) (Tanahashi and Zenk, range of biological activity, including antifungal, antibacte-
1988, 1990). Another enzyme (dihydrobenzophenanthridine rial, anti-inflammatory, and antitumor effects. Chelerythrine
oxidase) has been isolated from elicited cell suspension cul- and sanguinarine are used in human medicine (Simanek,
tures of Eschscholtzia californica that catalyzed the oxida- 1985). Chelythrine (84) has an LDso s.c. in mouse of 95
tion of dihydrobenzophenanthridines to the quaternary alka- mgikg, chelidonine (SO) (i. v.) of 35 mgikg, and sanguinarine
loids in the presence of oxygen (Schumacher and Zenk, (61) (i.v.) of 16 mglkg. Chelythrine has antitumor activity,
1988; Tanahashi and Zenk, 1988, 1990). The pro-R hydro- intercalates with DNA, and inhibits reverse transcriptase and
gen at C-13 is retained in chelidonine (80) (Simanek, 1985). alanine and aspartate aminotransferases; chelidonine (SO) in-
Although protopine (60), sanguinarine (61) and a number hibits reverse transcriptase and microsomal monooxygen-
of related alkaloids were found in callus cultures of certain ases and is cytotoxic (Wink, 1993). Both dehydrocheler-
papaveraceous plants, chelerythrine (84) and protoberberene ythrine and dehydrosanguinarine inhibit reverse
alkaloids (that occur in intact plants) were not produced (Si- transcriptase. Nitidine (86) intercalates with DNA and inhib-
manek, 1985). Treatment of suspension cultures of Papaver its reverse transcriptase, DNA polymerase, !RNA methyl-
somniferum with a homogenate of the fungus Botrytis in- transferase, and (Na+,K+)-ATPase (Wink, 1993). Nitidine
creased the amount of sanguinarine 150-fold (Ellis, 1988; (S6), sanguinarine (61), and chelerythrine (S4) have been
Flores et al., 1987), and polypeptide antibiotics induced for- shown to possess antitumor properties (Suffness and Cor-
mation of sanguinarine (61), chelerythrine (84), chelirubrine, dell, 1985). Chelidonium majus (papaveraceae) has folkloric
chelilutine, and macarpine in cell cultures of Eschscholtzia use for the treatment of cancer. The alkaloid chelidonine
california (Schumacher et al., 1987). Sanguinarine, cheler- sulfate, isolated from this plant, has been used for gastric
ythrine, and macarpine (85) (Fig. 32.26) also were formed cancer (Cordell, 1978b). Both fagaronine (S7) and nitidine
after treatment of suspension cultures of this species with a (S6) (Fig. 32.26) are active in the P-388 and L-1210 tumor
yeast elicitor. The relative proportions of the three alkaloids cell lines (Blasko and Cordell, 1988; Cordell, 1978b). Fagar-
in this last study depeuded on the growth stage of the cells onine is a potent inhibitor of RNA-directed DNA polymerase
when elicited (Collinge and Brodelius, 1989). in several viral systems and binds to DNA (Pezzuto et al.,
Benzophenanthridine alkaloids occur in five plant fami- 1983). Both of these alkaloids inhibit reverse transcriptase
lies: Fumariaceae, Papaveraceae, Rutaceae, Caprifoliaceae and have potent activity in the mv-1 reverse transcriptase
(Symphoricarpos), and Meliaceae (Kyiocarpus) (the last system (Tan et al., 1991).
two reports should be confirmed). Most alkaloids of this Bloodroot (Sanguinaria canadensis, Papaveraceae) has
series occur in the first three of these families (Simanek, been used as a plant drug. The active ingredients are sanguin-
1985). arine (61) (about 1%), chelerythrine (84), protopine, and
Chelidonine (S4) and sanguinarine (61) inhibit growth of allocryptopine (SO). The salts of these compounds are col-
the radicle in Lepidium (Wink, 1993). ored, hence, the name bloodroot. The roots of the plant serve
602 /soquinoline and Benzylisoquinoline Alkaloids

CH30

HO

(+ HS)-reticullne (20)
protopine (60)

o o

<o < o

o
)
-- 0

)
dihydrosangulnarine (83)

;;
o OCH3

OCH3
HO 11

chelidonine (80) sanguinarine (61) chelerythrine (84)

Fig. 32.25. Biogenesis of benzophenantbridine alkaloids (modified from Tanahashi and Zenk, 1988; modified and used with pennission of the
copyright owner, Elsevier Science Ltd., The Boulevard, Langford Lane, IGdlington, OXS 1GB, UK).

OCH3
o
CH3-O ) CH3-O
o
CH;-O

nitidine (86) fagaronine (87) macarpine (85)

Fig. 32026. Biologically active benzophenanthridine alkaloids.

 lsoquinoline and Benzylisoquinoline Alkaloids 603

CH3 0 CH3 0 CH30

HO HO HO

OCH3

(+)·(S)·reticuline (20) (-)-(S)..coulerine (72)

-- OCH3
OCH3 OCH3
OH

Darcotine (63) (-)-~.hydrasdne (88)

Fig. 32.27. Biogenesis of pbthalideisoquinoline alkaloids (modified from Geissman and Crout, 1969).

as a stimulating expectorant and have emetic properties narcotine (63) biosynthesis in Papaver somniferum involves
(Tyler et al., 1981). This alkaloid uncouples respiration and (+ )-(S)-reticuline (20) and (- )-(S)-scoulerine (72) (Fig.
oxidative phosphorylation in mitochondria and photosyn- 32.27) (Geissman and Crout, 1969; MacLean, 1985). Scoul-
thetic photophosphorylation, inhihits reverse transcriptase, erine is a precursor of a-narcotine in Papaver somniferum.
and intercalates with DNA (Wink, 1993). Sanguinarine has Only 14-S-scoulerine is incorporated and the H -14(C) retains
tuberculostatic, antifungal, antiyeast, and antimicrobial ac- its stereochemical integrity. The 13-pro-S hydrogen of scoul-
tivity against gram-positive and gram-negative bacteria and erine is removed in the conversion to narcotine (MacLean,
has been reported to have antitumor activity (Verpoorte et 1985).
al., 1991; Wink, 1993). Because of its antiplaque properties, The major alkaloid of this series is narcotine (63). This
sanguinarine has been included in some toothpastes (Water- alkaloid possesses antitussive activity, depresses smooth
man, 1993). Sanguinarine (61) exhibits deterrency to several muscles, and, in spite of its name, is not a narcotic (Cordell,
insects and growth inhibition to larvae of Hyphantria, 1978a).
Spodoptera, and Lymantria (Wink, 1993). The active principle of goldenseal (Hydrastis canadensis,
Species of the genus Argemone have been introduced Ranunculaceae) is (- )-(3-hydrastine (88), a phthalideisoqui-
from the New World into the Old World. Seeds of some noline alkaloid. Hydrastis has long been used as a medicinal
of these species (in particular, A. mexicana) often contami- plant and is used to treat inflammation of the mucous mem-
nate oil seeds and produce outbreaks of poisoning in India. branes (Tyler et al., 1981).
This poisoning has various symptoms, but a primary high- A second series of phthalideisoquinoline alkaloids, the
tension glaucoma that leads to permanent blindness often secophthalideisoquinoline alkaloids, involves additional
has been observed (Bisset, 1985). The alkaloids are soluble cleavage (Fig. 32.28). These compounds are found only in
in the oil expressed from the seeds. Sanguinarine (61) the Papaveraceae and Fumariaceae (MacLean, 1985).
seems to be the principal alkaloid involved (Bisset, 1985).
Sanguinarine also exhibits a positive inotropic action by
inhibition of (Na+,K+)-ATPase (Steinegger and Hiinsel, RHOEADAl'IE ALKALOIDS
1992; Waguer, 1988).
Chelidonine (80) is a feeding deterrent to polyphagous This group of about 40 alkaloids is known only from the
Syntomis larvae (Wink, 1993). Papaveraceae (Guinaudeau and Bruneton, 1993; Ronsch,
1986). These compounds probably are derived from proto-
berberine precursors (Santavy, 1979). (13R)- and (13S)-
mTIfALIDEISOQUINOLIl'IE ALKALOIDS scoulerine (72) are converted in plants of Papaver rhoeas
into rhoeadine (89) (Fig. 32.29). This alkaloid had lost 79%
Phthalideisoquinoline alkaloids are frequently observed of the tritium label from the (13S) precursor, whereas the
in the Papaveraeeae and occasionally in the Berberidaceae alkaloid retained 74% of the label from the (13R) precursor.
and the Ranunculaceae. The pathway established for ( - )-a- Although the starting materials were not completely confor-
604 Isoquinoline and Benzylisoquinoline Alkaloids

CH,O r N(CH,J,

CH,O ~

5,OCH3
"o OCH, OCH,
OCH,

narceineimide N·methylhydrastine fumaramidine

Fig. 32.28. seco-PhthalideisoquinoIine alkaloids.

(S)-reticuline (20) -+- CH'O~ I

<o
HO ~ _., N -+ --+- 0----"
H'

o>
'" 0"
I
h OCH,

(S)-scoulerine (72) protopine (60)

o o o
~<
<o
---
~O ___ 0

o
>
o

o
<o
o o
OH 0-1 OCH,O---l

.
{+)·rhoeadine (89)

• CH,-O
•
~CO,H

r --
HO~ NH,

.
CH,SCH,CH,CHCO,H
H2

alpigenine (90)

Fig. 32.29. A biogenetic sequence for rhoeadine alkaloids (modified from Ronsch, 1986; used with permission of the copyright owner, Academic
Press, Orlando, FL).
 /soquinoline and Benzylisoquinoline Alkaloids 605

HO HO HO

HO - - -....
·'HO
HO
OH OH

OH OH

CH,'O
CH,'O

HO
HO
OCH,

cryptaustoline (91) cryptowolline (92)

Fig. 32.30. Biogenesis of dibenzopyrrocoline alkaloids (modified from Geissman and Crout, 1969).

mationally pure, these values suggest that a stereospecific to the genera Papover and Argemone have been investigated
loss of the pro-S hydrogen occurs from C-13 of scoulerioe (Stermitz, 1968). The phannacological effects of certain
(Santavy, 1979). By introduction of I'C-Iabeled muramine, pavine and isopavine alkaloids have been reviewed (Gozler,
it was shown that a protopine (60) (rather than a phthalidei- 1987; Schiff, 1987).
soquinoline) alkaloid is the precursor of alpinigenine (60)
in Papaver bracteatum (Ronsch, 1986). A biogenetic
scheme that accounts for most of the available labeling data CULARII'IE ALKALOIDS
has been proposed (Ronsch, 1986).
So far only C-C bond formation has been considered. A
number of alkaloids also are known in which C":'O bond
OmEI'IZOFYRKOCOLIl"IB ALKALOIDS formation occurs. Among the compounds of this type are
the cularine alkaloids [such as cularine (96), which occur in
Early attempts to simulate the synthesis of the aporphine species of Corydalis, Dicentra, and other genera of the fam-
and morphine alkaloids often met with failure. Efforts to ily Fumariaceae (Fig. 32.32). This faruily is sometimes
oxidize laudanosoline (30), for example, did not give the merged with the Papaveraceae (Castedo and Suau, 1986;
desired type of coupling, but resulted in the formation of Preininger, 1986; Santavy, 1979).
dibenzopyrrocoline structures (Fig. 32.30) (Elliott, 1987;
Geissman and Crout, 1969). Alkaloids of this general struc-
ture were discovered in the Australian plant Cryptocarya BISBEI'IZYLISOQVIl'IOLIl"IB ALKALOIDS
bowiei (Lauraceae). Among these were cryptaustoline (91)
and cryptowolline (92). Intermolecular coupling of benzylisoquinoline units is
known to occur, as well as intramolecular coupling. An ex-
tensive series (about 400) of alkaloids formed by intermolec-
PAYII'IE AI'ID ISOPAYII'IE (ARGEMOIm) ular coupling of different monomeric units, of the benzyliso-
ALKALOIDS quinoline, tetrahydroisoquinoline, aporphine, and pavine
series, are known (Battersby, 1967; Guinaudeau and Bru-
Various species of Argemone and Eschscholtzia (papavera- neton, 1993; Schiff, 1987). One major group involves ben-
ceae) and Thalictrum (Ranunculaceae) contain pavine alka- zylisoquinoline and tetrahydroisoquinoline subunits. These
loids (about 20) and isopavine alkaloids (about 10) with a dimers are called bisbenzylisoquinoline alkaloids (Fig.
tetracyclic nucleus derived from benzylisoquinoline alka- 32.33) (Geissman and Crout, 1969). The parts of these alka-
loids (Gozler, 1987; Guinaudeau and Bruneton, 1993; loids often are linked by one or two diphenylether bridges.
Schiff, 1987). The formation of argemouine (93) and related The majority of bisbenzylisoquinoline alkaloids appear to
alkaloids is rationalized as follows (Fig. 32.31) (Geissman be derived from norcoclaurine (23) or a suitably methylated
and Crout, 1969). An alternate mode of cyclization also has derivative. Tubocurarine (97), a constituent of South Ameri-
been observed in the formation of ( - )-eschscholtzine (94) can tube curare, exhibits a type of "head-to-tail" coupling.
and ( - )-muuitagine (95). Only a few bases arise by C-C coupling. Ocotine (98) from
Systematic applications of pavine and isopavine alkaloids Ocotea rodiaei (Lauraceae) is one of these. Bisbenzylisoqui-
606 lsoquinoline and Benzylisoquinoline Alkaloids

CH,O CH,O
'"
CH,O
'" H"-
N· CH, _CH,O C H ' ° nH
CH" Ittc
"" OCH,
V CH,O::"" N::,...
I o
'" OH
OCH,

--+-
CH'O~
CH,O::""
I CH,,'
._", N
'"
'" I
OCH, <
0"
r
2 ::,...
OCH, 0
11

(. )-argemonine (93) (-)-eschscholizine (94) (-)-munatagine (95)

a pavine alkaloid

12 11

thalictidine
an isopavine alkaloid

Fig. 32.31. Biogenesis of pavine and isopavine alkaloids (modified from Geissman and Crout, 1969),

noline alkaloids occur in many of the same families as ben- (+ )-Tetrandrine (99) is a component of the Chinese drug
zylisoquinoline alkaloids. plants Stephania tetrandra and Menispermum dauricum, as
well as other plants. This alkaloid interacts with the plasma
Biological Activity membrane and is highly cytotoxic to KB cells (EDso 0.1
The pharmacological activity of certain bisbenzylisoqui- mg/kg). In clinical trials, the drug proved to be quite toxic
noline alkaloids has been reviewed (Buck, 1987; Schiff, and to lack significant antitumor properties (Suffness and
1987). Many of these compounds have weak to moderate Cordell, 1985). This alkaloid also had blood pressure lower-
antimicrobial activity. Two types of bisisoquinoline alka- ing and anti-inflammatory effects. Thalicarpine (100) also
loids possess antitumor activity. These are bisbenzylisoqui- has hypotensive and antitumor properties (Suffness and Cor-
noline and aporphine-benzylisoquinoline alkaloids [e.g., dell, 1985). Thalidasine (101) also has pronounced antimi-
tetrandrine (99) and thalicarpine (100), respectively]. Both crobial and antitumor activity. The crude extract of Thalic-
have pronounced side effects (Cordell, 1978b). trum faberi (Ranunculaceae) has been used to treat stomach

CH,-O CH,-O

--
CH,-O

CH,-O CH,-O
CH,-O
OR OCH,
OH

cularine (96)

Fig. 32.32. Cularine alkaloid biogenesis (modified from Geissman and Crout, 1969),
 /soquinofine and BenzyJisoquinoline Alkaloids 607

CH::27:::,..
I N'CH,
17 --
I
HO :::,..
(+)- and (-)-N-methylcoclaurine
OCH, HO
magnoline

oondrodendrine (102)

isochondrodendrine (103)

(a)

Fig. 32.33 (8 & b). Bisbenzylisoquinoline alkaloids.

cancer in China (Schiff, 1987). Chondrodendrine (curine, misinin (see Chapter 21), tetrandrine is said to provide long-
bebeerine) (102) inhibits growth of Leishmania, whereas acting and synergistic activity (Phillipson et aI., 1993).
isochondrodendrine (103) is active against nasopharyngial
carcinoma (Wink, 1993). Five bisbenzylisoquinoline alka- Curare
loids from Triclisia patens (Menispermaceae) bad ICso val- Curare is a mixtore of plant products that is used to poison
ues against Plasmodium Jalciparum at levels of 0.15-1.43 the tips of arrows for hunting and sometimes for warfare.
IJ.g1ml (chloroquine phosphate has an ICso of 0.14 IJ.g/ml The peoples that made these combinations of ingredients
under the same conditions) (Borris and Schaeffer, 1992; used many types of plants, but several of these plants contain
Phillipson and Wright, 19911; Phillipson et aI., 1993). BIQ and bisbenzylisoquinoline alkaloids (Bisset, 1985; Cor-
Phaenthine, also from a Triclisia species, was twice as potent dell,1981).
against a cbloroquine-resistant strain as against a chlo- So-called tobe curares (because they are stored in bamboo
roquine-sensitive strain with 50% inhibitory concentrations tubes) often are prepared from the menispermaceous plant
of 360 nM (Ekong et al., 1991). The bisbenzylisoquinoline Chondrodendron tomentosum. These extracts are prepared
alkaloid tetrandrine (99) and structurally related alkaloids and used in localized areas of the upper Amazon and Colom-
are more active against chloroquine-resistant strains that bia. Most of the active alkaloids are bisbenzylisoquinoline
against chloroqnine-sensitive strains. These alkaloids are ef- alkaloids. The major alkaloid is tubocurarine (97), a quater-
fective in reversing resistance in chloroquine-resistant Plas- nary species.
modium Jalciparum. When used in combination with arte- Injection of tubocurarine rapidly blocks neuromuscular
608 /soquinoline and Benzylisoquinoline Alkaloids

CH,O

CH,O

CH,O

thalicarpine (100)

thalidasine (101)

tubocurarine (97)
(b) (+ )-tetrandrine (99)

Fig. 32.33. (continued)

action, causing respiratory failure and death. A paralyzing Erythrina (Fabaceae) and in Cocculus (Menispermaceae)
dose for man is in the range 3-7 mg; the LDso p.o. (per as. or (Fig. 32.34). Alkaloids of this type have been reported from
oral) in mouse is 33.2 mg/kg (Wink, 1993). The compound a Dysoxylum species (Meliaceae), but some question about
has been used in small doses to enhance the action of anes- the identify of the plant material exists (Waterman, 1993).
thetics because it causes paralysis of the abdominal muscles Seeds of Erythrina species typically contain about 0.1 % of
without stopping the natural movement of the intestines. Tu- the alkaloids, but these compounds are also known from the
bocurarine is also used to control convulsions of strychnine leaves, stalks, stems, bark, roots, pods, and flowers (Dyke
poisoning and of tetanus (Tyler et a1.. 1981). The drug is inac- and Quessy, 1981).
tive orally. The di -O-methy1 ether of tubocurarine is interest-
ing as it has three times the skeletal-muscle relaxant activity Biosynthesis
but does not cause respiratory depression (Cordell, 1981).
Erythrina alkaloids are the products of a lengthy pathway
involving a dienone-phenol rearrangement and several rear-
rangement steps (Fig. 32.34) (Geissman and Crout, 1969).
ERYTJIRIlVA ALKALOIDS Labeled erysodiene (104), erysopine (105), erythraline
(106), and erysodienone (107) are converted to a- and 13-
A series of at least 60 complex alkaloids derived from ben- erythroidine (Fodor, 1980). Label from [2-14C]tyrosine was
zylisoquinoline alkaloid precursors is found in the genus found to be incorporated equally into C-8 and C-lO of 13-
 Isoquinoline and Benzy/isoquinoline Alkaloids 609

H02Y
-
CH•. O "" 1 NH

CH.·O
'"""I
OH
N~normethylprotosinomenine (108)

_CH'O~;~~~C::
OH

CH'O~
""I

CH.O
"'I
""
NH

CH.O
1 1
CH.O
o
-
OH
o /
(110) erysodienone (107)

- o

::X(),
OH
erythratine

fX),
'CH.O'v CH.OrJLll"'"U
CH.O"'"
erisodiene (104) erysoplne (lOS) cocculidlne (109)

Fig. 32.34. Biogenesis of Erythrina alkaloids (modified from Dyke and Quessy, 1981; used with pennission of the copyright owner, Academic Press,
Orlando, FL).

erythroidine (Dyke and Quessy, 1981). (+ )-(S)-Norprotosi- cursor of erythraline and both Cl- and /3-erythroidine (105,
nomenine (108) was incorporated 100 times more effectively 106).
than the enantiomeric compound, suggesting that this com-
pound is a specific precursor of erythraline. TIte intennedi- Biological Activity
acy of compound 110 was also established. Further, eryso-
diene (104) has heen isolated from some of the species that Many Erythrina alkaloids possess curare-like action and
synthesize these alkaloids. Introduction of [14C]4'-methoxy- interact with acetylcholine receptors. Extracts of many
norprotosinomenine in Erythrina christa-galli produced Erythrina species are used in indigenous medicine, particu-
erythraline (106) in which the methoxyl and methylenedioxy larly in India. Cocculine and cocculidine nitrates have been
group carbon atoms were equally labeled, confrnning the reported to show hypotensive action in dogs (Dyke and
presence of a symmetrical intennediate such as (111) (Dyke Quessy, 1981). The insecticidal alkaloid cocculolidine (109)
and Quessy, 1981). (- )-(5S)-Erysodienone (107) is a pre- has been isolated from Cocculus trilobus and C. carolinus
610 !soquinoline and Benzylisoquinoline Alkaloids

(Dyke and Quessy, 1981; Wink, 1993). IncOlporation of There also has been considerable discussion concerning
Erythrina flabelli/ormis seed alkaloids at 0.1 % into artificial whether the Fumariaceae should be included in the Papaver-
diets is totally lethal to the larvae of Callosobruchus macu- aceae. The two families share many types of alkaloids (e.g.,
latus (Janzen et al., 1977). the protoberberine, benzophenanthridine, and protopine
groups). Characteristic for the Fumariaceae are the spiroben-
zylisoquinoline, indenobenzazene, secophthalideisoqnino-
DlSTRIBVTIOl'l OF BEl'IZYLlSOQUIl'IOLIl'IE line, phthalideisoquinoline, and cularine alkaloids (Preinin-
ALKALOIDS ger, 1986).
The Rutaceae is one of the more interesting and complex
The distribution of benzylisoquinoline alkaloids is restricted
families with regard to alkaloid chemistry (see Chapter 31).
among higher plants (Guinaudeau and Bruneton, 1993;
The family contains alkaloids derived from several different
Seigler, 1977; Waterman, 1993; Waterman and Gray, 1987).
pathways, such as acridine, benzylisoqninoline, furoquino-
Most of these alkaloids occur in certain families of the Mag-
line, imidazole, indoloqninazoline, quinazoline, quinoline,
noliales and Ranunculales, although sporadic occurrences
and both simple aliphatic and aromatic amines. Furoquino-
are known among related plant groups and from others that
line and quinoline alkaloids are especially widespread within
are only distantly related. Among the families that have been
the family and are found in four of the five subfamilies from
reported to have these alkaloids are Annonaceae, Aristolo-
which alkaloids have been reported. The Rutaceae appears
chiaceae, Berberidaceae, Eupomatiaceae, Hemandiaceae,
to be a relatively homogeneous group by both morphological
Fabaceae (Erythrina, Andrachne), Fumariaceae, Lauraceae,
and chemical criteria. Alkaloids of the benzylisoquinoline
Magooliaceae, Menispermaceae, Monimiaceae, Nelum-
type are found in the genera Fagaropsis, Phellodendron,
bonaceae, Papaveraceae, Ranunculaceae, Winteraceae, Eu-
Tetradium, Toddalia, and Zanthoxylum (including Fagara),
phorbiaceae (Croton), Rhamnaceae (Rhamnus, Phylica,
three closely related genera (Waterman, 1993). Alkaloids of
Colletia, Colubrina, Discaria, Retanilla, Talguenea, andZi-
the I-benzyltetrahydroisoqninoline type are assumed to be
zyphus), Phellinaceae (Phelline), Symplocaceae (Sym-
primitive within the family, and the genera that produce them
plocos), Rutaceae, Combretaceae, Araliaceae (Hedera),
appear to be primitive within the family as well. These alka-
Apiaceae (Heracleum), Caprifoliaceae (Symphoricarpos),
loids appear to have been lost in relatively advanced taxa
Rubiaceae (Cephaelis), and Araceae (Lysichiton) (Bisset,
and to have been replaced by other types of alkaloids
1985; Dahlgren et al., 1981).
(Seigler, 1977; Waterman and Gray, 1987). If this assump-
Most botanists agree that the orders and families of the
tion is true, the family is likely derived from benzylisoquino-
Magooliidae (sensu Cronquist) are related and derived from
line-alkaloid-containing ancestors. The alkaloids and many
common ancestors. This conclusion is based on both mor-
morphological features of the Rutaceae are similar to such
phological and chemical data. The families Annonaceae,
families as the Berberidaceae, Menispermaceae, and Papav-
Aristolochiaceae, Berberidaceae, Eupomatiaceae, Hernandi-
eraceae (Seigler, 1977; Waterman and Gray, 1987). Benzyl-
aceae, Fabaceae (Erythrina, Angrachne), Fumariaceae,
tetrahydroisoquinoline-derived alkaloids have been studied
Lauraceae, Magnoliaceae, Menispermaceae, Monimiaceae,
as systematic markers (Rezende et al., 1975).
Nelumbonaceae, Papaveraceae, Ranunculaceae, and Win-
teraceae contain benzylisoqninoline alkaloids and their de-
rivatives. The families Himantandraceae, Myristicaceae, Ca- IPECACVAl'IHA ALKALOIDS
lycanthaceae contain other types of alkaloids. Some other
families such as the Degenariaceae, Austrobaileyaceae, Tri- Various groups of South American Indians used ipecac root
meniaceae, Lardizabalaceae, Corynocarpaceae, and Coria- long before their contact with Europeans. Ipecacuanha was
ceae have been tested and shown (at least usually) not to brought to Europe late in the 17th century. This preparation
contain alkaloids. Several other smaller families have appar- is derived from the root of Cephaelis ipecacuanha (Rubia-
ently not been tested. The statos of the non-alkaloid-contain- ceae). Ipecac (from Brazil) consists of about 2% alkaloids,
ing families within the group is unclear. Probably many of 60-70% of which is emetine (111) and about 25% cephae-
these families have lost the ability to synthesize benzyliso- line (112) (Fig. 32.35).
quinoline alkaloids and, in some instances, gained the ability Despite the importance of this material, work on cell and
to synthesize other types of alkaloids (Seigler, 1977). plant tissue culture of this plant is limited. Callus, root, and
The placement of the families Fumariaceae and Papavera- root suspension cultores contain emetine (111) and cephae-
ceae near the Magooliales, Piperales, Ranuncnlales, and so line (112) at low levels. Root suspension cultures had ap-
forth has become generally accepted in recent years (Seigler, proximately the same amount on a dry weight basis as the
1977; Waterman and Gray, 1987). Alkaloids of these two parent plant (Verpoorte et al., 1991).
families are rather similar and are similar to those of other A series of related compounds are found in several other
families of the order Ranunculales. These two families are plants of the Rubiaceae. The compounds differ from emetine
especially close to the families Berberidaceae, Menisperma- in that one or both of the dopamine-derived portions are
ceae, and Ranuncnlaceae, in terms of the types of alkaloids replaced with tryptamine. These compounds are discussed
they possess. in Chapter 34.
 Isoquinoline and BenzyJisoquinoline Alkaloids 611

tO~H OH ~ I strictosidine

~1"'~~oH~ NHH ", P

",HH OH
CH,02C HO 0N~ NH2
"""O~OH
o HO 0
seCOllogani~~~~ • CH,02C o

Ho0 ~H2 ~ indole alkaloids

dopamine (22) HO
deacetylisoipecoside (116)
HO NHp
HO

NHp ""HH OH
HO
""O~OH
"",HH OH o HO HO
"""O~OH
H"
'" 0 HO HO \
desacetylipecoside
CH'O~ '"

jH
CH'O~ ~ I"
\
protoemetine CH,O N

HO "" IH N '" 0 HO~ H" ,,~/

H HO~ NH2 CHO
o
I H ""H OH

alangiside(llS) ~~OH CH'O~

CH,o~HO CH,O '" IH" N H

CH . 0 : I N H'" ""./
HO , H" H \H
H" ''''./ HN~OCH'
HO
• H ~OH
"",HH OH C o ' OCH,
H'" '''''O~OH HN I '"
cephaeline (ll2)
CH,O,C '" 0 HO HO '"" OCH,
ipecoside (ll4) emetine (lll)

Fig. 32.35. Proposed biogenesis of emetine (modified from Geissman and Crout, 1969).

Biosynthesis graphic methods and by correlation with known compounds

(Kapil and Brown, 1979),
Ipecacuanha alkaloids arise by pathways similar to those Feeding experiments with [2-14C]geraniol and [0-
previously discussed for the origin of isoquinoline and ben- methyl-'H,6-'Hzlloganin showed that the C IO unit of ipeco-
zylisoquinoline alkaloids with the exception that a different side and the C9 unit of cephaeline were of iridoid monoter-
aldehyde moiety is involved. In this case, the condensation pene origin, Furthermore, [0-methyl-'H,6-'H2 1secologanin
involves dopamine (22) and secologanin (113). The origin and [3'-14C]deacetylipecoside served as specific precursors
of secologanin was discussed previously in Chapter 20, Em- for ipecoside (114) (Kapil and Brown, 1979) and alangiside
etine (114) and an alkaloidal glycoside, ipecoside (114), co- (115) (Fujii and Ohba, 1983). Contrary to some earlier re-
occur in Cephaelis ipecacuanha (Kapil and Brown, 1979), ports, however, deacetylisoipecoside (116) (Fig, 32.37)
The presence of ipecoside, an acetylated derivative of the (with a la-H configuration) is the precursor of cephaeline
first proposed intermediate, lends credence to the proposed (112) and emetine (111) (Fujii and Ohba, 1983),
biogenetic pathway (Fig, 32.36). The absolute stereochemis- Alkaloids with both types of stereochemistry at C-I are
try of ipecoside has been determined by x-ray crystallo- known to occur. For example, alangiside (115) (from Alan-
612 Isoquinoline and Benzylisoquinoline Alkaloids

CH,O CH,O

CH,O CH,O

OCH, OCH,
CH,O
OCH, OH
CH,O

OCH,

OH

CH,O

HO

OH
HO

HO CH'O~I
CH,O::"" N
H H

ipecoside (114) protoemetine CHO

Fig. 32.36. Ipecacuanha and related alkaloids (modified from Fujii and Ohba, 1983; used with permission of the copyright owner, Academic Press,
Orlando. FL).

gium lamarckii) has a C-II3-proton (Fig. 32.36). This glyco- ranged secologanin moiety as well as quinoline and isoqui-
side was found in the unripe fruit, roots, and leaves of the noline alkaloids. Isoquinoline alkaloids [such as emetine
plant (Kapil and Brown, 1979). (114)] are also found in Pogonopus (Rondeletieae), Toco-
vena (Gardenieae), some members of the Rubioideae, for
Distribution of Ipecac Alkaloids example, Psycho tria and Cephaelis of the Psychotrieae,
Most ipecac alkaloids are found in the family Rubiaceae, Bothriospora (Hamelieae), Borreria, Spermacoce, Richard-
often in the genus Cephaelis but they are known to occur sonia (Spermacoceae), and Manettia (Hedyotideae). Within
in other genera as well. These alkaloids have been found in the Hillioideae, Hillia species contain only emetine-type al-
the genera Borreria. Bothriospora, Capirona, Ferdi- kaloids.
nandusa, Hillia, Psychotria and Uragoga (synonyms of These alkaloids also have been found in the Alangiaceae
Cephaelis), Remija, and Tocoyena (Rubiaceae). (Alangium lamarckii) and Araliaceae (Hedera helix) (Fujii
According to the classification of Leeuwenberg (1980), and Ohba, 1983). The last report should be confirmed, as it
the Rubiaceae may be divided into four subfamilies: the Rub- is from a most unusual source.
ioideae, Cinchonoideae, Guettardoideae, and Hillioideae.ln-
dole alkaloids of the corynanthean type are the most common Medical Uses of Ipecacuanha Alkaloids
in the family. The tribe Cinchoneae of the subfamily Cincho- Emetine (114) was first demonstrated to be toxic to the
noideae is the only group to contain alkaloids with a noorear- microorganism Entamoeba histolytica in 1912; today, eme-
 lsoquinoline and Benzylisoquinoline Alkaloids 613

HO

-
HO

HO
,:r
H - HO
H OH
""O~O~OH
OHO~
HO

deacetylisoipecoside (116)

HO

HO

CHO CHO

protoemetine

Fig. 32.37. Emetine and related biologicany active alkaloids.

tine (114) and tubulosine (118) are used widely to treat ame- REFERENCES
bic dysentery caused by Entamoeba histolytica (Borris and
Schaeffer, 1992; Phillipson and O'Neill, 1987; Phillipson et AMANN, M, N. NAOAKURA, and M. H. ZENK, Purification and prop-
aI., 1993). World consumption of plant material of Cephaelis erties of (S)-tetrahydroprotoberberine oxidase from suspension-
cultured cells of Berberis wilsoniae. Eor. J. Biochem., 175,
ipecacuanha plant material is 10-100 metric tons (Ver-
17-25 (1988).
poorte et aI., 1991). Emetine has an LDso i.v. in mouse of
BAITERSBY, A. R, Phenol oxidations in the alkaloid field. in Oxida-
135 mg/kg, but 12.1 mg/kg in rat, and is an active inhibitor
tive Coupling of Phenols (W. I. Taylor and A. R. Battersby,
of protein biosynthesis (Wink, 1993). This compound also eds.), 119-165, Mercer Dekker, Inc., New York, 1967.
has been used to treat schistosomiasis, leishmaniasis, and BATI'ERSBY, A. R. and D. M. FOULKES, Alkaloid biosynthesis. Part
malaria (Borris and Schaeffer, 1992). Emetine has many VIII: Use of optically active precursors for investigations on
toxic side effects: among these are cardiotoxicity, muscle biosynthesis of morphine alkaloids, J. Cbem. Soc., 3323-3332
weakness, and gastrointestinal problems as diarrhea, nausea, (1965).
and vomiting. Toxic effects often are observed at therapeutic BA1TERSBY, A. R., R. C. F. JoNES, R. KASLAUSKAS, C. POUPET,
doses. This alkaloid has a profound reversible effect on DNA C. W. ThORNBER, S. S. RUCHlRAWAT, and J. STAUNTON, The
synthesis and produces essentially total inhibition at concen- biosynthesis of hasubanonine and protostephanine, 1. Chern.
trations of 1 fLM. The aIkaloid does not, however, penetrate Soc., Cbem. Commun., 773-775 (1974).
the cell wall of yeast or bacteria. Both emetine (114) and BA1TERSBY, A. R., R. C. F. JONES, R. KAzLAUSKAS, C. W.
THORNBER, S. RUCHIR.AWAT, and J. STAUNTON, Biosynthesis.
tubulosine (118) inhibit protein biosynthesis (Borris and
Part 24. Speculative incorporation experiments with 1-benzyli-
Schaeffer, 1992; Wink, 1993). soquinolines and a logical approacb via C.-Cz and C.-C, pre-
Emetine also has some antitumor activity but is quite cursors to the biosynthesis of hasubanonine and protostepha-
toxic and does not appear highly promising (Suffness and nine, J. Chern. Soc. Perkin Trans. I, 2016-2029 (1981a).
Cordell, 1985). Psychotrine (117) and O-methylpsychotrine BA1TERSBY, A. R., R. C. F. JONES, R. KAzLAUSKAS, A. P. OrnuooE,
are selective inhibitors of mv-1 reverse transcriptase (Tan C. POUPAT, andJ. STAUNTON, Biosynthesis. Part 23. Degradative
et aI., 1991a, 1991b). Alangium lamarkii has long been used studies on the alkaloids hasubanonine and protostephanine from
in Indian medicine (Fujii and Ohba, 1983). Stephaniajaponica, J. Cbem. Soc. Perkin Trans. I, 2010-2015
(l981b).
BATI'ERSBY, A. R., R. C. F. JONES, A. MINTA, A. P. OrnuooE, and
Other Biological Activity 1. STAUNTON, Biosynthesis. Part 25. Proof that hasubanonine
and protostepbanine are biosynthesized from the 1-benzyliso-
Emetine is toxic to Lemna and is a feeding deterrent to quinoline system, J. Cbem. Soc. Perkin Trans. I, 2030-2039
polyphagous Synromis larvae (Wink, 1993). (1981c).
614 Isoquinoline and Benzylisoquinoline Alkaloids

BAlTERSBY, A R., A MINTA, A. P. OrmIDGB, and J. STAUNTON, COLLINGE, M. A and P. E. BRODBLlUS, Dynamics of benzophe-
Biosynthetic derivation of hasubanonine and protostephanine nanthridine alkaloid production in suspension cultures of
from the 1-benzylisoquinoline system, Tetrahedron Lett., 18, Eschscholzia cali/ornico after treatment with a yeast elicitor,
1321-1324 (1977). Phytochemistry,28, 1101-1104 (1989).
BEECHER, c. W. W. and W. J. KEu.mmR, The incorporation of CORDELL, G. A, Alkaloids, in Kirk-Othmer's Encyclopedia of
berberine into jatrorrbizine, Tetrahedron Lett., 24, 469-472 Chemical Technology, Vol. 1, 3rd ed., 883-943, Wiley, New
(1983). York, 1978a.
BEECHER, C. W. W. and W. J. KELLEHER, Enzymatic study of the CORDBLL, G. A, Anticancer compounds from plants, in Progress
late stages of protoberberine alkaloid biosynthesis, Tetrahedron in Phytochemistry, Vol. 5 (L. Reinhold, J. B. Harborne, and T.
Lett., 25, 4595-4598 (1984). Swain, eds.), 273-316, Pergamon, London, 1978b.
BBECHER, c. W. W. and W. J. KELLEHER, The biosynthesis of the CoRDELL, G. A, Introduction to Alkaloids. A Biogenetic Approach,
protoberberine alkaloids, in AJkaioids: Chemical and Biological Wiley-Interscience, New York, 1981.
Perspectives, Vol. 6 (S. W. Pelletier, ed.), 297-337, Wiley, New CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual
York,1988. Reports on NMR Spectroscopy, Vol. 13 (G. A Webb, ed.),
BHAKUNI, D. S. and S. JAIN, Protoberberine alkaloids, in The Alka- 60-210, Academic Press, London, 1982.
loids, Vol. 28 (A Brossi, ed.), 95-181, Academic Press, New DAHLGREN, R. M. T., S. ROSBNDAL-JBNSEN, and B. J. NIELSEN, A
York,1986. revised classification of the Angiospenns with comments on
BISSET, N. G.. Plants as a source of isoquinoline alkaloids, in The correlation between chemical and other characters, in Phyto-
Chemistry and Biology of Isoquinoline Alkaloids (J. D. Phil- chemistry and Angiospenn Phylogeny (D. A Young and D. S.
lipson, M. F. Roberts, and M. H. Zenk eds.), 1-22, Springer- Seigler, eds.), 149-204, Praeger, New York, 1981.
Veriag, Berlin, 1985. DALY, J. W., M. GARRAFFO, and T. F. SPANDB, Amphibian alka-
BLASKO, G. and G. A. CoRDELL, Recent developments in the chem- loids, in The AJkaioids, Vol. 41 (G. A Cordell, ed.), 185-288,
istry of plant-derived anticancer agents, in Economic and Me- Academic Press, New York, 1993.
dicinal Plant Research, Vol. 2 (H. Wagner, H. Hikino, and N. DYKE, S. F. and S. N. QuEsSY, Erylhrina and related alkaloids, in
R Farnsworth, eds.), 119-153, Academic Press, London, 1988. The AJkaioids, Vol. 18 (R H. F. Manske and R. G. A Rodrigo,
BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from eds.), 1-98, Academic Press, New York, 1981.
plants, in Phytochemical Resources for Medicine and Agricul- EKONG, R, S. J. PARTRIDOB, M. M. ANDERSON. G. C. KIRBy, D.
ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum C. WARHURST, P. F. RUSSBLL, and J. D. PHU-LJPSON, Plasmodium
Press, New York, 1992. falciparum: Effects of phaeanthine, a naturally-occurring bisbe-
BROSSI, A, Manunalian alkaloids II, in The AJkaioids, Vol. 43 (G. nzylisoquinoline alkaloid. on choroquine-resistant and -sensi-
A Cordell, ed.), 119-183, Academic Press, New York, 1993. tive parasites in vitro. and its influence on chloroquine activity,
BUCK, K. T., Azafluoranthene and tropoloisoquinoline alkaloids, Ann. Trop. Med. Parasitol., 85, 205-213 (1991).
in The AJkaioids, Vol. 23 (A Brossi, ed.), 301-325, Academic ELLIS, B. E., Natural products through tissue CUlture, Nat. Prod.
Press, New York, 1984. Ref., 5, 581-612 (1988).
BUCK, K. T., The bisbenzylisoquinoline alkaloids, in The Alka- ELLIOTT, I. W., Dibenzopyrrocoline alkaloids, in The AJkaioids,
loids, Vol. 30 (A Brossi, ed.), 1-222, Academic Press, New Vol. 31 (A Brossi, ed.), 103-116, Academic Press, New York,
York,1987. 1987.
CASTEDO, L. and R. SUAU, The cularine alkaloids, in The Alkaloids. FLORES, H. E., M. W. Hoy. and J. J. PICKARD, Secondary metabo-
Vol. 29 (A Brossi, ed.), 287-324, Academic Press, New York, lites from root cultures, Tibtech, 5, 64-69 (1987).
1986. FODOR, G. B., Alkaloids derived from phenylalanine and tyrosine,
CASTRO, O. C., Chemical and biological extractives of Lauraceae in Secondary Plant Products (E. A. Bell and B. V. Charlwood,
species in Costa Rican tropical forests, in Phytochemical Poten- eds.), 92-127, Springer-Verlag, Beriin, 1980.
tial of Tropical Trees (K. R. Downum, J. T. Romeo, and H. A FRENZEL, T. and M. H. ZBNK, Purification and characterization of
Stafford, eds.), Recent Advances in Phytochemistry Vol. 27), three isoforms of S-adenosyl-L-methiouine: (R,S)- Tetrahy-
65-87, Plenum Press, New York, 1993. drobenzylisoquinoline-N-methyl transferase from Berberis koe-
CAvE, A, M. LEBoEUF, and P. G. WATERMAN, The aporphinoid lineana cell cultures, Phytochemistry, 29, 3491-3497 (1990a).
alkaloids of the Annonaceae, in Alkaloids: Chemical and Bio- FRENZEL, T. and M. H. ZBNK, S-Adenosyl-L-methionine:3'-hy-
logical Perspectives, Vol. 5 (S. W. Pelletier, ed.), 133-270, droxy-N-rnethyl-(S)-coclaurine-4'-O-methyl transferase, a re-
Wiley, New York, 1987. gio- and stereoselective enzyme of the (S)-reticuline pathway,
CHEN, Z. and D. ZHU, Arislolochia alkaloids, in The AJkaioids, Phytochemistry,29, 3505-3511 (l990b).
Vol. 31 (A. Brossi, ed.), 133-270. Academic Press, New York, Fum, T. and M. OHBA, Ipecac alkaloids and f3-carboline congeners,
1987. in The Alkaloids, Vol. 22 (A Brossi, ed.), 1-50, Academic
CHEN, C., H. CHANG, E. B. COWLING, C. Hsu, and R P. GATES, Press, New York, 1983.
Aporphine alkaloids and lignans formed in response to injury GmssMAN, T. A. and D. H. G. CROUT, Organic Chemistry of Sec-
of sapwood in Liriodendron lulipi/era, Phytochemistry, 15, ondary Plant Metabolism, Freeman Cooper, San Francisco,
1161-1167 (1976). 1969.
CLARKE, A. M. and C. D. HUFFORD, Antifungal alkaloids, in The GIBSON, A C., K. C. SPENCER, R. BAJAJ. and J. L. McLAUGHLIN.
AJkaioids, Vol. 42 (G. A Cordell, ed.), 117-150, Academic The everchanging landscape of cactus systematics. Ann. Mo.
Press, New York, 1992. Bot. Gard., 73, 532-555 (1986).
 [soquinoline and Benzylisoquinoline Alkaloids 615

GOZLBR, B., Pavine and isopavine alkaloids, in The Alkaloids, Vol. Phytochemical Resources for Medicine and Agriculture (H. N.
31 (A. Brossi, ed.), 317-389, Academic Press, New York, 1987. Nigg and D. S. Seigler, eds.), 33-74, Plenum Press, New York,
GUlNAUDEAU, H. and J. BRUNETON, Isoquinoline alkaloids, in Alka- 1992.
loids and Sulphur Compounds (P. G. Waterman, ed.), Vol. 8 LoEFFLER, S. and M. H. ZBNK, The hydroxylation step in the biosyn-
of Methods in Plant Biochemistry (P. M. Dey and J. B. Har- thetic pathway leading from norcoclaurine to reticuline, Phyto-
borne, eds.), 373-419, Academic Press, London, 1993. chemistry, 29, 3499-3503 (1990).
HAN, G., Y. CHEN, and X. LlANG, Distribution of alkaloids in tradi- LoEFFLER, S., R. STADLBR, N. NAGAKURA, and M. H. ZBNK, Nor-
tional Cbinese medicinal plants, in The Alkaloids, Vol. 32 (A. coclaurine as biosynthetic precursor of thebaine and morphine,
Brossi, ed.), 241-270, Academic Press, New York, 1988. J. Chern. Soc., Chern. Commun., 1160-1162 (1987).
HARBoRNE, J. B., Introduction to Ecological Biochemistry, 2nd ed., LUNDslROM, J., Simple isoquinoline alkaloids, in The Alkaloids,
Academic Press, New York, 1982. Vol. 21 (A. Brossi, ed.), 255-327, Academic Press, New York,
HARBORNE, J. B., Recent advances in chemical ecology, Nat. Prod. 1983.
Rep., 3, 323-344 (1986). . MAcLBAN, D. B., Pbthalideisoquinoline a1ka1oids and related com-
HARTMANN, T., Alkaloids, in The Herbivores, Their Interactions pounds, in The Alkaloids, Vol. 24 (A. Brossi, ed.), 253-286,
with Metabolites, Vol. I (G. A. Rosenthal and M. R. Beren- Academic Press, New York, 1985.
baum, eds.), 79-121, Academic Press, San Diego, CA, 1991. MANN, J., Secondary Metabolism, Oxford University Press, Ox-
HERBERT, R. B., The biosynthesis of plant a1ka1oids and nitrogenous ford, 1978; 2nd edition, 1987.
microbial metabolites, Nat. Prod. Rep., 3, 185-203 (1986). MATA, R. and J. L. McLAUGHLIN, Cactus a1ka1oids. 50. A compre-
HERBERT, R. B., The biosynthesis of plant alkaloids and nitrogenous hensive tabular survey, Rev. Latinoamer. Quim., 12, 95-117
microbial metabolites, Nat. Prod. Rep., 5, 523-540 (1988). (1982).
HUGHES, D. W. and D. B. MAcLBAN, The 13C_NMR spectra of MATA, R., C. CHANG, and J. L. McLAUGHLIN, 13C NMR analysis
isoquinoline a1ka1oids, in The Alkaloids, Vol 18 (R. H. F. Man- of some simple tetrahydroisoquinolines, Phytochemistry, 22,
ske and R. G. A. Rodrigo, eds.), 217-262, Academic Press, 1263-1270 (1983).
New York, 1981. MATSUI, M., Hasubanan a1ka1oids, in The Alkaloids, Vol. 33 (A.
Brossi, ed.), 307-347, Academic Press, New York, 1988.
HUTCHINSON, C. R., Biological methods for studying the biosyn-
thesis of natural products, Nat. Prod. Rep., 3,133-152 (1986). MEYER, J. M. and J. C. FOGLEMAN, Significance of saguaro cactus
alkaloids in ecology of Drosophila mettleri, a soil breeding,
JANSSBN, R. H. A. M., R. J. J. C. LoUSBERG, P. WUKBNS, C. !{aUK,
cactophilic drosophilid, J. Chern. Bcol., 13, 2069-2081 (1987).
and H. O. THEUNs, Assignment of IH and 13C NMR resonances
of some isoquinoline alkaloids, Phytochemistry, 28,2833-2839 NAHRSTEDT, A., Strukturelle Beziehungen zwischen pflanzlichen
(1989). und tierischen Sekundiirstoffen, Planta Medica, 44, 2-14
(1982). .
JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary
compounds to the seed-eating larvae of the bruchid beetle Ca/lo- NISHIDA, R. and H. Ft1KAMI, oviposition stimulants of an aristolo-
sobruch... I7UJculatus, Pbytochemistry, 16, 223-227 (1977). chiaceae~feeding swallowtail butterfly, Atrophaneura alcinous,
1. Chern. Bcol., 15, 2565-2575 (1989).
KAMETANI, T. and T. HONDA, Aporphine alkaloids, in The Alka-
loids, Vol. 24 (A. Brossi, ed.), 153-251, Academic Press, New PazzuTO, J. M., S. K. ANroSlAK, W. M. MasSMER, M. B. SLAITOR,
York,1985. and G. R. HONIG, Interaction of the antileukemic a1ka1oid 2-
hydroxy-3,8,9-trimethoxy-5-methylbenzo[ c Jphenanthridine
!CAPn., R. S. and R. T. BROWN, Monoterpene a1ka1oid glycosides,
(fagaronine), with nucleic acids, Chern. BioI. Interact., 43,
in The Alkaloids, Vol. 17 (R. H. F. Manake and R. G. A. Rodrigo
323-339 (1983).
eds.), 546-588, Academic Press, New York, 1979.
PazzuTO, J. M., S. M. SWANSON, W. MAR, C. CHE, G. A. CORDELL,
KINGSBURY, J. M., Poisonous Plants of the U.S. and Canada, Pren-
and H. H. S. FONG, Evaluation of the mutagenic and cytostatic
tice-Hall, Englewood Cliffs, NJ, 1964.
potential of aristolochic acid (3,4-methylenedioxy-8-methoxy-
KIRCHER, H. W., Chemical composition of cacti and its relationship I O-nitrophenanthrene-I-carboxylic acid) and several of its rela-
to Sonoran desert Drosophila, in Ecological Genetics and Evo- tives, Mutat. Res., 206, 447-454 (1988).
lution (J. S. F. Barker and W. T. Starmer, eds.), 143-158, Aca-
I'Hn.iJpsON, J. D. and M. J. O'NED-L, Antimalarial and amoebicidal
demic Press, Sydney, 1982.
natural products, in Biologically Active Natural Products (K.
KOSTERLITZ, H. W., Biosynthesis of morphine in the aninIa1 king- Hostettmann and P. J. Lea, eds.), 51-64, Clarendon, Oxford,
dom, Nature, 330, 606 (1987). 1987.
LEETE, E., The biosynthesis of alkaloids, in Specialist Periodic Re- I'Hn.iJpSON, J. D. and C. W. WRIGIIT, Can ethnopharmacology con-
ports, Biosynthesis, Vol. 7 (R. B. Herbert and T. J. Simpson, tribute to the development of antimalarial agents? J. Ethnophar-
eds.), 102-223, The Royal Society of Chemistry, 1983. macol.,32, 155-165 (1991).
LEEUWENBBRG, A. 1. W., The taxonomic position of some genera in Pmu.IPSON, 1. D., C. W. WRIGHT, G. C. KIRBy, and D. C. W ARH-
the Loganiaceae, Apocynaceae, and Rubiaceae, related families URST, Tropical plants as sources of antiprotozoal, in Phytochem-
which contain indole a1ka1oids, in Indole and Biogenetically ical Potential of Tropical Trees (K. R. Downum, J. T. Romeo,
Related Alkaloids (J. D. Pbillipson and M. H. Zenk, eds.), 1-10, and H. A. Stafford, eds.), Recent Advances in Pbytochemistry
Academic Press, New York, 1980. Vol. 27, 1-40, Plenum Press, New York, 1993.
LEVINSON, H. Z., The defensive role of alkaloids in insects and PRmNINGER, V., Chemotaxonomy of Papaveraceae and Fumaria-
plants, Experientia, 32, 408-411 (1976). ceae, in The Alkaloids, Vol. 29 (A. Brossi, ed.), 1-98, Academic
LEWIS, W. H., Plants used medically by indigenous peoples, in Press, New York, 1986.
616 lsoquinoline and Benzy/isoquinoline Alkaloids

REZENDE, C. M. A. D. M., O. R. GOTILIEB, and M. C. MARX, SUFFNESS, M. and G, A. CORDELL, Antitumor alkaloids, in 'The
Benzyltetrahydroisoquinoline derived alkaloids as systematic Alkaloids, Vol. 25 (A. Brossi, ed.), 3-355, Academic Press,
markers, Biochem. Syst. Beol., 3, 63-70 (1975). New York, 1985.
ROBINSON, R. and]. M. GULLAND, The constitution of codeine and TAN, G. T., A. D. KINGHORN, S. H. HUGHES, and J. M, PEZZUTO,
thebaine, Mem. Proc. Manchester Lit. Phil. Soc., 69, 79-86 Psychotrine and its Owmethyl ether are selective inhibitors of
(1925). human immunodeficiency virus-l reverse transcriptase, J, BioL
RONSCH, H., Rhoeadine alkaloids, in The Alkaloids, Vol. 28 (A. Chern., 266, 23529-23536 (1991a).
Brossi, ed,), 1-93, Academic Press, New York, 1986. TAN, G. T" 1. M, PEzzuTO, and A. D. KINGHORN, Evaluation of
RUFFER, M. and M, H. ZENK, Berberine synthase, the methylenedi- natural products as inhibitors of human immunodeficiency virus
oxy group forming enzyme in berberine synthesis, Tetrahedron type 1 (HIV -1) reverse transcriptase, J. Nat. Prod., 54, 143-154
Lett., 26,201-202 (1985). (1991b).

RUFFER, M, and M. H. ZENK, Distant precursors ofbenzylisoquinow TANAHASHI, T. and M. H. ZENK, One step enzymatic synthesis
line alkaloids and their enzymatic fonnation, Z. Naturforsch" of dihydrosanguinarine from protopine, Tetrahedron Lett, 29,
5625-5628 (1988).
42c, 319-332 (1986).
TANAHASHI, T, and M. H. ZENK, Elicitor induction and characteriza-
SANTAVY, F., Papaveraceae alkaloids, in The Alkaloids, Vol 17
tion of microsomal protopinew6 whydroxylase, the central en-
(R. H. F. Manske and R. G. A. Rodrigo eds.), 385-544, Aca-
zyme in benzophenanthridine alkaloid biosynthesis, Phyto-
demic Press, New York, 1979,
chemistry,29, 1113-1122 (1990).
SCHIFF, P. L., JR., The Thalictrum alkaloids: Chemistry and phar-
TAYLOR, W. c., Eupomatia alkaloids, in The Alkaloids, Vol. 24
macology, in Alkaloids: Chemical and Biological Perspectives,
(A. Brossi, ed.), 1-23, Academic Press, New York, 1985.
Vol. 5 (S. W. Pelletier, ed.), 271-637, Wiley, New York, 1987.
(Numerous references to other review articles are cited in this TYLER, V. E., L. R. BRADY, and J. E. ROBBERS, Pharmacognosy,
reference.) 8th ed., Lea and Febiger, Philadelphia, 1981.

SCHUMACHER, H. M, and M, H. ZENK, Partial purification and char- URZUA, A., R, RODRiGUEZ, and B. CASSELS, Fate of ingested aris w
acterization of dihydrobenzophenanthridine oxidase from tolochic acids in Battus archidamas, Biochem, Syst. EcoL, 15,
687-689 (1987).
Eschscholtzia californica cell suspension cultures, Plant Cell
Rep., 7,43-46 (1988). VERPOORTE, R, R. VAN DER HEDDEN, W. M. VAN GULIK, and H.
1. G. TEN HOOPEN, Plant biotechnology for the production of
SCHUMACHER, H. M., H. GUNDLACH, F. FIEDLER, and M. R. ZENK,
alkaloids: Present status and prospects, in The Alkaloids, Vol.
Elicitation of benzophenanthridine alkaloid synthesis in
40 (A. Brossi, ed.), 1-188, Academic Press, New York, 1991.
Eschscholtzia cell cultures, Plant Cell Rept., 6, 410-413 (1987).
WAGNER, H., Non-steroid, cardioactive plant constituents, in Eco-
SCHUMACHER, H. M., M, RUFFER, N. NAGAKURA, and M, H. ZENK,
nomic and Medicinal Plant Research, Vol. 2 (R. Wagner, H.
Partial purification and properties of (S)~norlaudanosoline syn-
Hikino, and N. R. Farnsworth, eds,), 17-38, Academic Press,
thase from Eschscholtzia tenuifolia cell cultures, Planta Medica,
London, 1988.
48, 212-220 (1983).
WAGNER, H. and A. PROKSCH, Immunostimulatory drugs of fungi
SEIGLER, D. S., Alkaloids and plant systematics, in The Alkaloids, and higher plants, in Economic and Medicinal Plant Research
Vol. 16 (R. H. F. Manske, ed.), 1-82, Academic Press, New
(H. Wagner, H. Hikino, and N. R. Farnsworth, eds.), 113-153,
York, 1977.
Academic Press, London, 1985,
SIMANEK, V., Benzophenanthridine alkaloids, in The Alkaloids, WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and Metabo-
Vol. 26 (A. Brossi, ed.), 185-240, Academic Press, New York, lism in Plants, Plenum Press, New York, 1978,
1985.
WATERMAN, P. G" Phytochemical diversity in the order Rutales,
SPENCER, K C, Specificity of action of allelochemicals: Diversifi- in Phytochemical Potential of Tropical Plants (K. R. Downum,
cation of glycosides, in Allelochemicals: Role in Agriculture 1. T. Romeo, and H. A. Stafford, eds,), Recent Advances in
and Forestry (G. R. Waller, ed.), ACS Symposium Series 330, Phytochemistry Vol. 27), 203-233, Plenum Press, New York,
275-288, American Chemical Society, Washington, DC, 1987. 1993.
STADLER, R, T, M. KUTCHAN, and M, H. ZENK, (S)-Norcoclaurine WATERMAN, p, G, and A. I. GRAY, Chemical systematics, Nat.
is the central intermediate in benzylisoquinoline alkaloid biow Prod. Rep., 4, 175-203 (1987).
synthesis, Phytochemistry, 28, 1083-1086 (1989).
WINK, M., Allelochemical properties or the raison d'etre of alka-
STADLER, R., T. M, KUTCHAN, S, LoEFFLER, N, NAGAKURA, B. loids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), 1-105,
CASSELS, and M. H. ZENK, Revision of the early steps of reticu w Academic Press, New York, 1993.
line biosynthesis, Tetrahedron LeU., 28, 1251-1254 (1987). ZENK, M, H., Enzymology of benzylisoquinoline alkaloid forma-
STEINEGGER, E. and HANSEL, Phannakognosie, 5th ed., Springer- tion, in The Chemistry and Biology ofIsoquinoline Alkaloids (1.
Verlag, Berlin, 1992. D. Phillipson, M. F. Roberts, and M. H. Zenk, eds.), 241-256,
STERMITZ, p, R, Alkaloid chemistry and the systematics of Papaver Springer-Verlag, Berlin, 1985.
and Argemone, in Recent Advances in Phytochemistry VoL 1 ZENK, M. R" M. RUFFER, M, AMANN, and B. DEUS-NEUMANN,
(T. J. Mabry, R. E. Alston, and V. C. Runeckles, eds.), 161-183, Benzylisoquinoline biosynthesis by cultivated plant cells and
Appleton-Century-Crofts, New York, 1968. isolated enzymes, J. Nat. Prod., 48, 725-738 (1985).
33
Alkaloids Derived from Both Tyrosine
and Phenylalanine

Introduction found in the leaves and mature seed. At least 19 species of

Colchicine and Other Tropolonic Alkaloids Colchicum and 10 other genera (including Androcymbium,
Biosynthesis Bulbocodium, Gloriosa, Iphigenia, Kreysigia, and Merend-
Biological Activity of Colchicine era) contain colchicine and related alkaloids. These genera
Medicinal Uses of Colchicine belong to the subfamily Wurmbaeiodeae, with the exception
Use of Colchicine in Horticulture of the Australian genus Kreysigia (Capraro and Brossi,
Amaryllidaceae Alkaloids 1984).
Biosynthesis The amide nitrogen of colchicine is not basic, and some
Biological Activity have argued that colchicine is not a true alkaloid. However,
Medicinal Uses of Amaryllidaceae Alkaloids as the precursors and the pathway leading to this compound
Mesembryanthemum and Sceletium Alkaloids are typical for those of alkaloids, colchicine is considered
Cephalotaxus and Homoerythrina Alkaloids an alkaloid for present purposes. Although this compound
Biosynthesis of Cephalotaxus Alkaloids is the best known of the group, more than 30 other tropolonic
Biosynthesis of Homoerythrina Alkaloids alkaloids occur in Colchicum autumnale and related plants
Systematic Value of Cephalotaxus and Homoerythrina (Capraro and Brossi, 1984). The physical constants and spec-
Alkaloids tral data of colchicine and related alkaloids have been tabu-
Antitumor Activity lated (Tojo, 1989).
References Natural (7S)-( - )-colchicine (1) displays molecular sym-
metry derived from a noncoplanar arrangement of the tropo-
INTRODUCTION Ionic ring C and the benzenoid ring A (Fig. 33.1). These
rings are twisted out of the plane with a torsion angle of
The biosynthesis of several types of alkaloids involves both about 53°. This feature proves to be critical for the binding
phenylalanine and tyrosine. These alkaloids occur in the of colchicine to tubulin (Boye and Brossi, 1992). Natural
closely related families Amaryllidaceae and Liliaceae and colchicine has the two aromatic moieties arranged in a coun-
in the Cephalotaxaceae and Mesembryanthemaceae. The last terclockwise helicity, which implies an (as) absolute con-
two families are not closely related to each other, nor to the ftguration (Boye and Brossi, 1992; Geissman and Crout,
ftrst two families. 1969).
Both the IH_ and 13C_NMR (nuclear magnetic resonance)
spectra for several of these alkaloids have been reviewed Biosynthesis
(Crabb, 1982). The results of feeding experiments leading
to colchicine, Amaryllidaceae, and Cephalotaxus alkaloids Colchicine (1) is derived from both tyrosine and phenylal-
have been tabulated (Leete, 1983). anine (Fig. 33.1). One of the major steps in unraveling the
biosynthetic sequence leading to colchicine was the isolation
COLCHICJl'IE AND Ol'HEK TKOPOLOl'IIC and recognition that androcymbine (2) is related to an inter-
ALKALOIDS mediate in the pathway (Fig. 33.2). [1- 13C]Autumnaline (3)
is incorporated into colchicine. Androcymbine is the de-
Colchicum is one of the most ancient plants known to man methylated form of the intermediate O-methylandrocymbine
both as a drug and as a poisonous plant. Colchicine (1) is (4) in the pathway. Attack on the bridged system occurs,

617
618 Alkaloids Derived from Tyrosine and Phenylalanine

OCH,

'Ok",
CH,O

I '
'CO,H
+-- HO'C~O
.~. NHl ~ 1~1H2 ~
o OH

phenylalanine tyrosine

o
colchicine (1)
Fig. 33.1. Colchicine (modified from Boye and Brossi, 1992; used with pennission of the copyright owner, Academic Press, New York).

followed by ring expansion of the aromatic tyrosine-derived of microtubules) has long been known (Capraro and Brossi,
nucleus. Inclusion of the first carbon atom of the side chain 1984). Colchicine binds to tubulin <limers of 100,000 molec-
(C-12 of colchicine) generates the tropolone ring. This ular weight in a 1: 1 stoichiometry at the same site as podo-
scheme was established by introduction of [3- 14C]tyrosine phyllotoxin (Boy" and Brossi, 1992). Treated cells are ar-
and degradation studies of the colchicine obtained (Capraro rested at metaphase because microtubules are essential for
and Brossi, 1984; Geissman and Crout, 1969). moving chromosomes during mitosis. The transport of vesi-
cles also is affected. Colchicine inhibits the incorporation
Biological Activity of Colchicine of uridine into RNA. This alkaloid has many other types of
The ability of colchicine to interfere with microtubule- activity in animal systems (Capraro and Brossi, 1984).
dependent cell functions and to bind to tubulin (the subunit Colchicine is a feeding deterrent for Locusta, polypha-

OCB, OH
(S)-autumnaUne (3) O-mefhylandroeymblne (4)

CH.lO~HO) CH30~~
CHiO HO
NHCH I ') NHCH, I -<.cno
n ~ 'l! H--+ CR~O ~ 'I ~ H ---+
,---+
CH3 0 'CH,O
OCR) OCH ~ 0
o '~r 0 OCR;
~3 cn, CH,O

CH;JO

o
c:oIdddne(1) o democolclne OCR,

Fig. 33.2. Biogenesis of colchicine (modified from Capraro and Brossi, 1984; used with pennission of the copyright owner, Academic Press, New
York).
 Alkaloids Derived from Tyrosine and Phenylalanine 619

gous Syntomis larvae, and Agelaius (red-winged blackbird) plants. This effect has been useful in many studies of plant
(Wink, 1993). Incorporation of colchicine at 0.1 % into artifi- genetics. A number of plant varieties of economic impor-
cial diets is totally lethal to the larvae of Collosobruchus tance have been produced by use of this alkaloid.
maculatus (Janzen et aI., 1977). This alkaloid is insecticidal
for Leptinotarsa (Wink, 1993).
AlllARYLLIDACEAE ALKALOIDS
Medical Uses of Colchicine
Colchicum autumnale has long been used to treat gout, The approximately 100 alkaloids of this series occur in the
for which it is efficacious, although the drug exhibits toxic family Amaryllidaceae. This family is considered closely
manifestations. The colchicine content of Colchicum autum- related to the Liliaceae and some taxonomists (e.g., Cron-
nale (Liliaceae) is about 0.25-0.6%. At a dose of 1 mg every quist, 1981) have merged the two into one large family. The
2 h for 8 hours and a maintenance dose of 500 ILg twice a alkaloids of the Amaryllidaceae are similar to colchicine and
day, colchicine is still an effective treatment for gout. Colchi- other related alkaloids of the Liliaceae in that both phenylala-
cine has an LDso i.v. in mouse of 4.11 mglkg (Wink, 1993). nine and tyrosine are involved in their synthesis. AIl Amaryl-
This alkaloid also may be useful for the treatment of cirrhosis Iidaceae alkaloids, however, are derived from a single inter-
of the liver, amyloidosis, and familial Mediterranean fever mediate, norbelladine (4). The three major structural Iypes
(Brossi, 1992). Colchicine is an anticancer agent (Blasko are found at times in individuals of the same species.
and Cordell, 1988; Brossi, 1992), but without practical im-
portance because of its toxicity. The inhibition of cell divi- Biosynthesis
sion is not specific for malignant cells.
Amaryllidaceae aJkaJoids are derived from both iyrosine
Use of Colchicine in Horticulture and phenylalanine. The adduct of these two amino acids,
norbelladine (5), is an intermediate in the three major Iypes
Colchicine is used to introduce polyploidy in plants and of Amaryllidaceae aJkaJoids (Fig. 33.3). This adduct arises

·.'
causes doubling or increase in the chromosome number of by prior conversion of phenylalanine to 3,4-dihydroxybenz-

•m
• ::,...
I •
NH,
CO,H

-+
H{:N0"
HO'-'HN~
~ 1· . ' U OH + -
HO,C
~•
NH,
•

'~OH
•

phenylalanine norbelladine (5) tyrosine

VTu
,
;,.. I NH,-
CO,H
~:;,.. CO,H
~
I -+
HO~:;'"
HO ~
1 __
COH
, HO:oCHO

HO
~I
,...

phenylalanine
/ (6)

norbelladine (5)

Fig. 33.3. Biogenesis of norbelladine and the origin of the three major types of AmarylIidaceae alkaloids (modified from Geissman and Crout. 1969).
620 Alkaloids Derived from Tyrosine and Phenylalanine

.;=;
OR H+

HO ~
~ NH~ ;,
OH
CH,-O
~~
Q
'" C
fi

norbelladine (5) ~ HO, 4 1.& HN '-....

11 7 1"HO CH,-O H
CH,-OjCG'"" .. / I"'"
1 "''' I
HO • .& HN HO
• 7

O-methylnorbelladine (13) OH

norpluviine (9) Iycorine (8)

CH,-O.
!~;W _ ~HO'"
HO
H I! H 1 CH,-O
I"'"
H
N
1
CH,-O

CH,-O
QT.) CH,-O'&
Cor -
CH,-O
H~ \
CH,s-
pluviine (17)

~
o OCH,

CH,-O
CH,-O '" I
-+-
CH,-O 1.& N
CH,-O
o
Iycorenine (10) homolycorine (11) galanthine (12)

Fig. 33.4. Biogenesis of lycorine and structurally related alkaloids (modified from Geissman and Crout, 1969).

aldehyde (6) and conversion of tyrosine to tyramine (7), Tyrosine with 'H label ortho to the hydroxyl group and
followed by Schiff-base fonnation and reduction. with 14C in position 2 in the side chain (which should pro-
Synthesis of alkaloids of the first structural type [such as duce norbelladine labeled in positions 2 and 4) was incorpo-
lycorine (8), norpluviine (9), lycorenine (10), and homoly- rated into norpluviine and into lycorine with loss of one-
corine (11)1 in various daffodil varieties may occur as indi- half of the 'H label. The 'H atom retained in lycorine was
cated (Fig. 33.4). Radioactively labeled norbellidine (5) is located at C-2. Thus, it appears that all of the 3H was retained
incorporated into norpluviine (9). in the derived lycorine (8). This suggests a stereospecific
[3H1Norpluviine (9) is incorporated into lycorine (8) in course in both protonation to fonn the allylic methylene of
daffodils (Fuganti, 1975; Martin, 1987). Feeding studies norpluviine (9) and hydroxylation to form lycorine (8). The
with [8-3H1norpluviine to Narcissus psewionarcissus (King 'H atom in norpluviine at C-2 has a ~-configuration. Proton-
Alfred daffodils) afforded mostly lycorenine (10) and homo- ation of the aromatic precursor of norpluviine (9) at C-2
lycorine (11), although there was some incorporation into takes place from the ( l side of the molecule and hydroxyla-
pluviine (17), galanthine (12), and methylpseudolycorine. In tion of C-2 of norpluviine occurs with inversion to fonn
other experiments with Narcissus poeticus, norpluviine was lycorine (Fuganti, 1975).
incorporated into lycorine (8), galanthine (12), methylpseu- Incorporation of [4-'H;~-I4C]cinnamic acid into norpluv-
dolycorine, and narcissidine, but no label was incorporated iine resulted in 50% loss of the 'H label. The same loss was
into lycorenine (10) (Martin, 1987). observed with the [3,5-2H2;4-'H;~-I4C]-isomer. It appears
 Alkaloids Derived from Tyrosine and Phenylalanine 621

that para-hydroxylation involves complete migration and re- (16), respectively, have been examined. [6_3H~-!4C]Hae­
tention of tritium and a second hydroxylation at one of the manthamine (14) was converted into haemanthidine (16)
two equivalent positions causing loss (without migration) of (Fig. 33.5) without 3H loss in Spreckeliaformosissima. [7-
half the remaining tritium. The 3H atom retained is at posi- 3H-5-!4C]Norpluviine (9) was incorporated into lycorenine
tion 11 of norpluviine (9). Enzymatic control appears to be (10) (Fig. 33.4) without loss of 3H. O-Methylnorbelladine
involved in the elimination (Fuganti, 1975). (13) randomly labeled in the benzylic positions, was incorpo-
The genesis of the second structural type is illustrated by rated into pluviine (17) (Fig. 33.4) without loss of tritium
the pathway leading to haemanthamine (14) (Fig. 33.5). 1n and into lycorenine (10) with loss of half the tritium. 1n
this case a para-para instead of an ortho-para coupling is similar experiments, haemanthamine (14) and haemanthid-
observed. The framework is completed by nitrogen addition ine (16) lost approximately half the activity (Fig. 33.5).
to the dienone system, as before. Haemanthamine (14) may These experiments indicate that incorporation of protoca-
then be converted into pre-tazettine (15). techualdehyde (6) (3,4-dihydroxybenzaldehyde) into the
The stereochemistry of protonation involved in the incor- C.-C! unit of the alkaloids and both the protonation to form
poration of protocatechualdehyde (3,4-dihydroxybenzalde- the benzylic methylene of norbelladine (5) [and hence of
hyde) into haemanthamine (14) and the stereochemical norpluviine (9) and haemanthamine (14)] and the subsequent
course of the hydrogen removal during the hydroxylation Ol hydroxylation of norpluviine (9) and haemanthamine (14)
to the nitrogen atom in the conversion of norpluviine (9) are stereospecific processes. The hydrogen introduced in the
and haemanthamine into lycorenine (10) and haemanthidine process or formation of norpluviine (9) and haemanthamine

HO~
- NH

norbeUadine (5)
v6 ~
"-...
h
OH

OH

elf ____
OH CH,- 0
HO
-03
,r
"'" I C
"(

NH
~H+
1.0

--
CH,-O~ ~
HO~NH
O-methylnorbelladine (I3)

, OCH,

CH'-O~
9" H 9"
O @ OOH 11 ---l_'-I--OH

<
CH,-O 9" ___ 0 109"

HO ""
1 N
-- HO ""
1 NO,"'"
• 7
haemanthamine (14)

~
OCH' ~OCH'
<~ <::
OCH'
H 9" OH H 9" 1 CH,
o

o '::,T[: (if ~
9"
""I
o
~
N

H
--+

CH,S-
I
oY
I
0'-
H
haemanthidine (16) pretazzettine (15)

Fig. 33.S. Biogenesis of haemanthamine, pre-tazettine. and tazettine (modified from Geissman and Crout, 1969).
622 Alkaloids Derived from Tyrosine and Phenylalanine

(14) is the same as that removed in the oxidation step gen atom in haemanthamine takes place during the hydroxyl-
(Fuganti, 1975). In the oxidation process a pro-R hydrogen ation and, because the absolute stereochemistry of haema-
at C-7 is removed (Martin, 1987). These results establish nthamine (14) is known, must proceed with retention of
that in the incorporation of protocatechualdehyde (6) into configuration (Fuganti, 1975).
the aromatic C6 -C I unit of the alkaloids, protonation takes Biosynthesis of the third type of Amaryllidaceae alka-
place from the re face of the molecule and that oxidation loids is represented by the formation of galanthamine (18)
of the amine proceeds with the stereospecific removal of (Fig. 33.6).
an a-hydrogen from the benzylic position (Fuganti, In daffodil plants, galanthamine is biosynthesized from
1975). compounds 19-21, but not from norbelladine (5) [which is
Hydroxylation of the methylene at position 6 (13 to the a good precursor of haemanthamine (14) and Iycorine (8) in
nitrogen atom) to form haemanthamine (14) occurs late in the same plants] (Fig. 33.6). These experiments suggest a
the sequence, once the ethanophenanthridine skeleton has definite order of methylation of norbelladine (5) during the
been constructed from O-norbelladine (5) [O-methyInorbel- biosynthesis of galathamine. In feeding experiments with
ladine (13)]. O-Norbelladine stereospecifically labeled at po- the plant Leucojum aestivum, [2,4-3H2 ; '4C_O_methyl]O_
sition 6 (mixed with identical 14C-labeled samples) was fed methylnorbelladine (13) is a precursor of galanthamine (18)
to daffodil plants. High tritium retention was observed in that retains the same 3H114C ratio as in the precursor. There
haemanthamine (14) when the (6S)-isomer of O-norbellidine is a 50% loss of tritium in the biosynthesis of Iycorine (8).
was administered, whereas loss of tritium was observed with The tritium labels were found in positions 2 and 4 of galan-
the (6R)-isomer. Thus, 3H loss from position 13 to the nitro- thamine (18) (Fuganti, 1975).

OH CH+

HO~NH~OH OH CH30mH'~ 0

...... OHg 7 --+

norbelladine (5) CH3o'(C' ___ ::,... I N,

::,...' N CH3

m
\
CH,
N,O-metbylnorbelladine (20)

o 0
CHJ 0 -:9' CHjO 10

::;,.. I ~
N
, CH,
17
.::;,..

narwedine galanthamine (18) crinine (27)

HO

CH'-0:cG IJ CH~O~ ~.
HO

10 . ; ; ) HO~'
7 H0
72, ;

HO CH,
(19) (20)

"",
HO,()

HO~.~)
narcicJasine (22) H02;
HO~ :.

HO~I HO~
CH,
(21) norbelladine (5)

Fig. 33.6. Biogenesis of galanthamine (modified from Geissman and Crout, 1969),
 Alkaloids Derived from Tyrosine and Phenylalanine 623

pseudolycorine (26)
Iycoricidine (23) narclclasine (24)
Oycoricidinol)

Fig. 33.7. Biologically active Amaryllidaceae alkaloids.

Biological Activity blastosis virus, a typical C-type virus, by binding with the
polymerase itself (Martin, 1987). Pretazettine inhibits
Lycoricidine (23) and Iycoricidinol (24) are antifeedant Rauscher virus NIHl3T3 and herpes simplex virus (Wink,
and galanthamine (18) is insecticidal to the yellow butterfly, 1993).
Eurema hecabe mandarina (Lepidoptera) (Fig. 33.7) (Mar- The LDso Lv. of tazettine (25) in mouse is 100 mg/kg
tin, 1987). An extract of the bulbs of Lycoris radiata was (Wink, 1993). Lycorine (8), haemanthamine (14) and pseu-
found to exhibit antifeeding activity when tested against the dolycorine (26) are toxic to Rauscher virus NIHl3T3 cells
yellow butterfly, Eurema hecabe mandarina (Martin, 1987). and inhibit protein biosynthesis (Wink, 1993).

Medicinal Uses of Amaryllidaceae Alkaloids

/fIBSBMBBYAJYTIIEMlJM Al'ID SCELETIlJIII
Galanthamine (18) has been of interest, as it has powerful ALKALOIDS
cholinergic activity and is reputed to have analgesic activity
comparable to morphine. This alkaloid was used in the for- A novel group of alkaloids is found in a number of species
mer Soviet Union for the treatment of myasthenia gravis, of the genus Mesembryanthemum (Mesembryanthaceae or
myopathy, and diseases of the nervous system (Cordell, Aizoaceae). Many taxonomists have now subdivided this
1981). Galantharnine inhibits cholinesterase, causes brady- genus into a number of segregates. Alkaloids are found in
cardia and atrioventricular conduction distorbances, and has many of these species, but especially in those plants now
an LDso Lv. in mouse of 8 mg/kg (Wink, 1993). considered to comprise the genus Sceletium. This group of
A number of AmaryUidaceae alkaloids are of interest as plants is characteristic of a number of regions of South Af-
potential antitumor agents. The most important are lycorine rica, especially the South-West Cape Province. A number
(8), narcic1asine (22), and pretazettine (15). Lycorine is of species were used by the Hottentots and Bushmen of Na-
widely distributed in the family and is known from at least maqualand for preparation of a drug called' 'channa" (Jeffs,
30 genera. Pretazettine is not so widely distributed but is 1981; Schleiffer, 1979).
found in at least 18 genera. Narciclasine (22) is known from The approximately 25 compounds of this group possess
only a few species of the family (Martin, 1987; Suffness and several types of ring systems; the mesembrine alkaloids are
Cordell, 1985). major among these.
Lycorine (8) inhibits ascorbic acid synthesis (Robinson, Biosynthesis of this group of alkaloids appears to involve
1979) as well as plant cell division and cell elongation (Mar- p-coumaric acid andlor cinnamic acid, although a number
tin, 1987). This alkaloid also inhibits DNA polymerase, pro- of related compounds apparently can serve as precursors.
tein synthesis, Rauscher virus NIHl3T3, and herpes simplex The proposed pathways of origin are similar to those for
virus (Wink, 1993). crinine (27), an AmaryUidaceae alkaloid. Both phenylala-
Narciclasine (22) halts protein synthesis by blockage of nine and tyrosine, when administered to Sceletium strictum,
peptide-bond formation on the 60-S ribosomal subunit. The gave rise to labeled mesembrine (28) and mesembrenol (29).
effect is specific for eucaryotic cells. Narciclasine inhibits Radioactive label from DL-[2-14C] and DL-[3- 14C]phenyl-
Rauscher virus NIHl3T3 (Wink, 1993). Lycorine (8) blocks alanine was not incorporated into mesembrine, in agreement
mitosis in the broad bean (Vidajaba). The mechanism ap- with the general proposal of biosynthesis. Ring-labeled
pears to be related to inhibition of protein synthesis (Suffness pheylalanine was incorporated, as was label from [2-14C]tyr-
and Cordell, 1985). Pretazettine (15) has been used in combi- osine. Norbelladine (5) and its two O-methyl derivatives also
nation with DNA-binding and alkylating agents in the treat- were incorporated, but a number of experiments demon-
ment of the Rauscher leukemia virus (Cordell, 1981; Martin, strated that these compounds are not involved in the pathway
1987). This alkaloid inhibits purified RNA-dependent DNA leading to Mesembryanthenum and Sceletium alkaloids. Ty-
polymerase (reverse transcriptase) from avian myelo- rosine is converted to tyramine and then to N-methyltyra-
624 Alkaloids Derived from Tyrosine and Phenylalanine

mine (30) during the biosynthesis of mesembrine alkaloids CEFIIALOTAXUS AI'ID HOIIIOERYTHRIl'IA
(Fig. 33.8). Furthermore. the aromatic ring and the CH2 -N ALKALOIDS
side chain is incorporated as an intact unit. Incorporation
of phenylalanine has been shown to involve conversion to The gymnospermous genus Cephalotaxus (Cephalotaxa-
cinnamic acid and to p-coumaric acid (31). Reduction prod- ceae) (the only genus in the family) is native to Asia and is
ucts of the latter compound may be involved in additional generally considered to have eight species (Huang and Xue.
steps (Jeffs. 1981). 1984). Several of these species are cultivated as ornamental
A major pathway to mesembrenol (29) by cinnamic acid. shrubs in various parts of the world. Alkaloids from species
p-coumaric acid (31). and the corresponding dihydro-p- of Cephalotaxus have antitumor activity. Of the compounds
coumaric acid involves sceletenone (32). an efficient precur- present. about 20 are unique to the genus (such as com-
sor for mesembrenol (29) (Fig. 33.8). This suggests that hy- pounds (33) and (34) and belong to an unusual structural
droxylation of the aromatic ring occurs at a late stage of type. whereas another 10 are homoerythrina alkaloids (Fig.
biosynthesis. 33.9). Homoerythrina alkaloids are found in other groups

~
OH ~C02H

"" HOV NH2

HO~

U.: :,. .
phenylalanine I I h
¥
/" tyrosine

C02H ..-J' hi"" ~0~H3

~ I .,/
~ 1:
C02H NHCH3 HO " '"
::,... (31) (30) ---

clnnamicacld _0 ~N'CH

0"
~ OG~h
70
OO~"'/.
"".;::

I / - . ,__
o . N)
~

N --
'CH
~

I N' CH
%

3
3

~.'CH3 3 0 \
H • 0 . 0

C-:'bo
~, ~~'
CH3
I
:'bo
H
o~~ _
I
CH3
H
RO 0

seeletenone (32)
J

f~' r~~'
:'bo
IH
CH3
:·bOH
I
CH3
H

mesembrine (28) memembrenone mesembrenol (29) Scelelium~

Fig. 33.8. Biogenesis of Mesembryanthemum and Sceletium alkaloids (modified from Jeffs. 1981; used with pennission oftbe copyright owner,
Academic Press, New York).
 Alkaloids Derived from Tyrosine and Phenylalanine 625

HO~
o
<o
CH,o '" 1 NH

I'"
CH,O .&
OH

schelhammeridine (39) (38) schelhammerine

harringtonine (33) deoxyharringtonine (36) cephalotaxine (35)

< o
OH H'....

~····"rrO
OCH,
, HO~~OCH
H2'
homoharringtonine (34) isoharringtonine (37)

Fig. 33.9. Cephalotaxus and homoerythrina alkaloids.

of plants such as Schelhammera (Liliaceae) and Phelline A biogenetic pathway has been suggested (Fig. 33.10) (Hud-
(Phellinaceae or sometimes Aquifoliaceae) (Pusset et aI., licky et aI., 1987).
1989). More than 20 homoerythrina alkaloids are known. Cephalotaxine (35) occurs esterified to a series of some-
Homoerythrina alkaloids usually make up only a very minor what unusual hindered acids. In deoxyharringtonine (36), L-
proportion of the alkaloids present in most Cephalotaxus leucine appears to be the precursor of the acidic moiety as
species, although, in the species Cephalotaxus wilsoniana, indicated by labeling studies. The following pathway has
these alkaloids are major constituents (Dyke and Quessy, been suggested (Fig. 33.11) (Dyke and Quessy, 1981). 13C_
1981; Powell et aI., 1972a). NMR data for several Cephalotaxus alkaloids have been re-
Homoharringtonine (33) is an inhibitor of chloroquine- ported (Weisleder et al., 1980).
resistant Plasmodium jalciparum in vitro and P. yoelii in Callus cultures of Cephalotaxus harringtonii produce
mice (Borris and Schaeffer, 1992). Both homoharringtonine cephalotaxine (35), and the antitumor esters harringtonine
and harringtonine (34) inhibit protein synthesis (Wink, (33), isoharringtonine (37), and homoharringtonine (34) in
1993). both callus tissue and the medium. Deoxyharringtonine (36)
was found in the medium but not in the callus (Delfel, 1980;
Delfel and Rothfus, 1977; Spec. Per. Rep., 1975, 1979).
Biosynthesis of CephaJotaxus Alkaloids
Biosynthesis of Homoerythrina Alkaloids
Biosynthetic studies of Cephalotaxus alkaloids indicate
that ring A of the cephalotaxine molecule is derived from Homoerythrina alkaloids that occur in the genus Cephalo-
tyrosine, and rings C and D are derived from phenylalanine. taxus appear to be derived from phenylalanine and tyrosine
626 Alkaloids Derived from Tyrosine and Phenylalanine

o •

OR OR
\
b'-..
0 o •
0
0
(
0
(
0 <O~I
o ""
N.( 0
...- be~id O. II
rearrangement
o OR
o
cephalotaxlne (35)

Fig. 33.10. Proposed biogenetic pathway for the synthesis of cephalotaxine (modified from Huang and Xue, 1984; used with pennission of the
copyright owner, Academic Press, Orlando. FL).

by way of a phenethyJisoquinoline precursor such as (38) There are about eight species in the only genus of the family,
(this type of intennediate also is involved in the fonnation Cephalotaxus (Huang and Xue, 1984). Some workers have
of Cephalotaxus alkaloids) (Dyke and Quessy, 1981). The considered the family to he closely related to the Taxaceae,
intennediacy of such a compound has been demonstrated whereas others have even placed the family in its own order,
for schelhammeridine (39). Lahel from ring-Iaheled tyrosine the order Cephalotaxales.
is incorporated almost entirely into the A ring. Phenylalanine The genus Taxus (Taxaceae) is noted for the presence
is incorporated into rings C and D. Lahel from C-I (ofphe- of diterpenes and diterpene alkaloids such as taxine (see
nylalanine) appears mostly at C-8 of the alkaloid. Chapters 22 and 36). There is a complete dichotomy in the
types of alkaloids present in the two groups. These data do
Systematic Value of Cepbalotaxus and not support union of the two families.
Homoerthrina Alkaloids
Antitumor Activity
The relationships of the gymnospennous family Cephalo- Only the four esters harringtonine (33), homoharring-
taxaceae have heen viewed in a number of ways in the past. tonine (34), isoharringtonine (37), and deoxyharringtonine

OH

YLCO'~ ~O C~ ytCO'H --+ YlCO'H _

NH, I II CO,H CO,H
leucine

yxCO'H --+ yyCO,:... '-- ~ ~CO'H _ '-- ~ ~CO'H

HO CO,H I O~CO'H T ~ T ~ .
~ homoleucine

-
OH
the chain can be
extended further by
a similar series of !'{CO'H
reactions
CO,H
Fig. 33.11. Proposed biogenesis of the acid portion of Cephalotaxus alkaloids (modified from Huang and Xue. 1984; used with pennission of the
copyright owner. Academic Press. Orlando, FL).
 Alkaloids Derived from Tyrosine and Phenylalanine 627

(36) show significant antitumor activity (Cordell, 1978; HUDLICKY, L., L. D. KWART, and J. W. REED, Synthesis ofCephalo-
Powell et al., 1969, I 972b, 1974). Alkaloids of this type taxine alkaloids, in Alkaloids: Chemical and Biological Per-
generally inhibit the synthesis of DNA and protein synthesis spectives, Vol. 5 (S. W. Pelletier, ed.), 639-690, Wiley, New
(Blasko and Cordell, 1988; Wink, 1993). These compounds York, 1987.
significantly prolong the life spans of mice with lymphocyte JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary
leukemia. Harringtonine is effective in treating both acute compounds to the seed-eating larvae of the bruchid beetle Callo-
sobruchus maculatus, Phytochemistry, 16, 223-227 (1977).
and chronic myelocytic leukemia in human subjects (Spe-
cialist Periodic Reports, Alkaloids 1975, 1979). JEFFS, P. W., Sceletium alkaloids, in The Alkaloids, Vol. 19 (R. H.
F. Manske and R. G. A. Rodrigo, eds.), 1-80, Academic Press,
New York, 1981.
LEETE, E., The biosynthesis of alkaloids, Specialist Periodic Re-
REFERENCES ports, Biosynthesis, 102-223, The Royal Society of Chemistry,
London, 1983.
BLASKO, G. and G. A. CORDELL, Recent developments in the chem- MARTIN, S. F., The Amaryllidaceae alkaloids, in The Alkaloids,
istry of plant-derived anticancer agents, in Economic and Me- Vol. 30 (A. Brossi, ed.), 252-376, Academic Press, New York,
dicinal Plant Research, Vol. 2 (H. Wagner, H. Hikino, and N. 1987.
R. Farnsworth, eds.), 119-191, Academic Press, London, 1988. POWELL, R. G., R. V. MADRIGAL, C. R. SMTIH, JR., and K. L.
BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from MIKOLAJCZAK, Alkaloids of Cephalotaxus harringtonii var. dru·
plants, in Phytochemical Resources for Medicine and Agricul- pacea, J. Org. Chern., 39, 676-680 (1974).
ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum POWELL, R. G., K. L. MIKOLAJCZAK, D. WEISLEDER, and C. R.
Press, New York, 1992. SMITH, JR., Alkaloids of Cephalataxus wi/saniana, Phytochem-
BOYE, O. and A. BROSSJ, Tropolonic Colchicum alkaloids and a110 istry, 11, 3317-3320 (1972a).
congeners, in The Alkaloids, Vol. 41 (A. Brossi and G. A. POWELL, R. G., D. WEISLEDER, and C. R. SMITIl, JR., Antitumor
Cordell, eds.), 125-176, Academic Press, New York, 1992. alkaloids from Cephalotaxus harringtonii: Structure and activ-
CAPRARO, H. and A. BROSSI, Tropolonic Colchicum alkaloids, in ity, J. Phann. Sci., 61, 1227-1230 (1972b).
The Alkaloids, Vol. 23 (A. Brossi, ed.), 1-70, Academic Press, POWELL, R. G., D. WEISLEDER, C. R. SMITH, JR., and I. A. WOLFF,
New York, 1984. Structure of cephalotaxine and related alkaloids, Tetrahedron
CORDELL, G. A., Anticancer agents from plants, in Progress in Lett., 4081--4084 (1969).
Phytochemistry, Vol. 5 (L. Reinhold, J. B. Harborne, and T. PUSSET, J., S. LA BARRE, N. LANGLOIS, and J. HAMON, Alkaloids of
Swain, eds.), 273-316, Pergamon, London, 1978. Phelline comasa var. robusta, Phytochemistry, 28, 1298-1300
CORDELL, O. A., Introduction to Alkaloids, A Biogenetic Approach, (1989).
Wiley-Interscience, New York, 1981. ROBINSON, T., The evolutionary ecology of alkaloids, in, Herbi-
CRABB, T. A, Nuclear magnetic resonance of alkaloids, in Annual vores. Their Interaction with Secondary Plant Metabolites. (C.
Reports on NMR Spectroscopy, Vol. \3 (G. A. Webb, ed.), A. Rosenthal and D. H. Janzen, eds.) 413-448, Academic Press,
59-2\0, Academic Press, London, 1982. New York, 1979.
CRONQUIST, A., An Integrated System for the Classification of Sl>ECIALlST PERIODIC REPORTS, Alkaloids, Vol. 5 Chemical Society,
Flowering Plants, Columbia University Press, New York, 1981. London, 1975.
DELFEL, N. E., Alkaloid distribution and catabolism in Cephalo· SPECIALIST PERIODIC REpORTS, Alkaloids, Vol. 9, Chemical Society,
taxus harringtonia, Phytochemistry 19, 403-408 (1980). London, 1979.
DELFEL, N. E. and J. A. ROlHFUS, Antitumor alkaloids in callus SUFFNESS, M. and G. A. CORDELL, Antitumor alkaloids, in The
cultures of Cephalotaxus harringtonia, Phytochemistry, 16, Alkaloids, Vol. 25 (A. Brossi, ed.), 3-355, Academic Press,
1595-1598 (1977). New York, 1985.
DYKE, S. F. and S. N. QUESSY, Erythrina and related alkaloids, in ToJO, E., The homoaporphine alkaloids, J. Nat. Prod., 52,909-921
The Alkaloids, Vol. 18 (R. H. F. Manske and R. G. A. Rodrigo, (1989).
eds.), 1-98, Academic Press, New York, 1981. WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and Metabo-
FUGANTI, C, The AmaryJlidaceae alkaloids, in The Alkaloids, Vol. lism in Plants, Plenum Press, New York, 1978.
15 (R. H. F. Manske, ed.), 83-164, Academic Press, New York, WEISLEDER, D., R. G. POWELL, and C. R. SMITH, JR., Carbon-13
1975. nuclear magnetic resonance spectroscopy of Cephalotaxus alka-
GElSSMAN, T. A. and D. H. G. CROUT, Organic Chemistry of Sec- loids, Org. Magn. Res., 13, 114--1\5 (1980).
ondary Plant Metabolism, Freeman Cooper, San Francisco, SCHLEIFFER, H., Narcotic Plants of the Old World, Lubrecht, Monti-
1969. cello, NY, 1979.
HUANG, L. and Z. XUE, Cephalotaxus alkaloids in The Alkaloids, WINK, M., Allelochemical properties or the raison d'etre of alka-
Vol. 23 (A. Brossi, ed.), 157-226, Academic Press, New York, loids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), 1-118,
1984. Academic Press, New York, 1993.
34
Indole Alkaloids

Introduction leqnin et aI., 1993; Verpoorte et aI., 1991). Several types of

The Biosynthetic Pathway Leading to Key Intennediates alkaloids have an indole nucleus included in the strncture;
Strictosidine (Isovincoside) the ones considered in this chapter are those based on the
Subsequent Reactions of Strictosidine condensation of secologanin with trYptamine.
Classification and Strnctoral Types of Indole Alkaloids Methods for the chromatographic separation and purifica-
Fonnation of the Major Indole Alkaloid Types tion of indole alkaloids (TilJequin et aI., 1993; van der Heij-
Summary of Pathways Leading to Major Types of den et al., 1987a, 1987b, 1988) and both the 'H- and 13C_
Monoterpene-Derived Indole Alkaloids NMR (nuclear magnetic resonance) spectra of indole alka-
Chemosystematic and Phylogenetic Application loids have been reviewed (Cordell, 1979; Crabb, 1982; TiI-
Monoterpenoid Indole Alkaloids of Phannacologic lequin et aI., 1993). The results of feeding stodies leading to
Significance the fonnation of indole alkaloids have been reviewed (Leete,
Vincamine 1983).
Ajmalicine and Related Compounds
Ajmaline
Yohimbine
11m BIOSYl'lTHE11C PATHWAY LEADIl'IQ
Reserpine
TO KEY II'ITERMEDIATES
Picralima Alkaloids
StrYchnine and Its Relatives
Biogenesis Indole alkaloids are derived mechanistically in a manner
Curares from Strychnos Species similar to that discussed for ipecacuanha alkaloids in Chapter
Olivacine and Ellipticine 32. Schiff-base fonnation occurs between a carbonyl com-
Bisindole Alkaloids pound and an amine, but in this case, the amine is tryptamine
Catharanthus Alkaloids (I) (instead of 3,4-dihydroxyphenylethyl amine), and the
Gelsemium Alkaloids carbonyl-containing terpenoid unit is secologanin (2). The
Alkaloids from Tabernanthe iboga stereochemistrY of the condensation products is identical to
Camptothecine that of the ipecacuanha alkaloid series. The initial product
Biogenesis of the condensation of secologanin (2) and tryptamine (1)
Distribution of Camptothecine Among Higher Plants in most, if not all, plants that produce monoterpenoid indole
Biological Activity alkaloids is strictosidine (3) (Fig. 34.1) (fonnerly known as
Cinchona Alkaloids isovincoside).
Biogenesis
Biological and Medicinal Activity strictosidine (l!iovincoside)
Cinchophylline and Related Alkaloids
References Although there has been considerable debate conceming
the stereochemistrY of the initial precursor of indole alka-
loids, it is now finnly established that all naturally occurring
INTRODUCTION
monoterpenoid indole alkaloids are derived from strictosid-
Indole alkaloids (at least 4100 known) are among the most ine (3) (Battersby et al., 1978; Brown et aI., 1978; Scott,
numerous and complex of all secondary compounds (TiI- 1981; Stiickigt and Zenk, 1977). This alkaloidal glycoside

628
 Indole Alkaloids 629

0~
H'0. H O-glucosyl
O-glucosyl

CH30 2C
H.p'·

CH30 2C
: :,. . °
secologonin (2) strictosidine (3) viocoside (4)

11
3 :::,...2 (1_0) 'C02CH3
OH 0

(1.O)··X·- OH
'. I 4
··(1.0)
2109
-<:::,3. 3
5,9- H-Ia~lled
71 •• (1.0)
o ( 5) _
-HO
7105 H 0 secologamD
HOCH 2 IC02H
•
Ito I both (1.0)
!
10geraniol O-glucosyl
[4-3H,2-14C]_(3R,4R)_mevalooate [4-3H,2-14C]-(3R,4S)-menlonate, no tritium label introduced

X
11
C02CH3
.... OH ::::"'2
(1.0),.3. 4
'<:::: H'(I.O)
• 3 ,
4 2 -+-6 H 3 _ 3,7-3H-labeUed
HOCH2 I C02H 71 secologanin
•
10 9 .(1.0) loganin (11) O-gluoosyl
[2-3H,4-14C]-(2R,3R)-mevalonate •

-
H lto21
3to17
Sto15
O-glucosyl 7 to3
9islostlnajmalaclne
lito earbomelhoxy

CH30 2C
11 CH,O;C
..cologaoin (2) 'IImalicioe (7)
Fig, 34.1. Origin of monoterpenoid indole alkaloids.

possesses 3-(S) or 3-0l stereochemistry (Kapil and Brown, ole. The specific glucosidase occurs in the cytosol (Deus-
1979; Cordell, 1981). It was demonstrated to he the precursor Neumann and Zenk, 1984). This enzyme has been isolated
by Stiickigt and Zenk (1977), and subsequently confirmed from Catharanthus roseus plant tissue cultures at more than
by Battersby et aI. (1978) and Brown et al. (1978) in work 500 times the level that occurs in the plant itself (Hutchinson,
with cell-free systems from Catharanthus roseus (Apocyna- 1986). The cDNA clone for strictosidine synthase from Rau-
ceae). Strictosidine is accumulated by Rhazya stricta (Apo- volfia serpentina has heen sequenced and its expression in
cynaceae), the source from which it was first isolated (Smith, Escherichia coli examined (Kutchan et al., 1988). A similar
1968). clone from Catharanthus roseus has been introduced into
When tryptamine and secologanin were incubated with Nicotiana tabacum plants along with the CaMV 35S pro-
an enzyme preparation from a Catharanthus cell culture in moter. These plants produced levels of enzyme 3-22 times
the presence of a f3-glucosidase inhibitor (D-d-gluconolac- greater than Catharanthus roseus plants (McKnight et at,
tone), only strictosidine (3) was formed. The reaction clearly 1991). Furthermore, evidence was found for the production
was enzyme dependent. The enzyme, strictosidine synthase, of strictosidine.
a single polypeptide with 30,000 MW (Hampp and Zenk, Strictosidine (3), and not the 3f3-epimer vincoside (4),
1988; Stiickigt and Zenk, 1977), is found in the plant vacu- is the precursor of a numher of indole alkaloids, including
630 Indole Alkaloids

yohimbine (5) (Fig. 34.11), camptothecine (6) (Fig. 34.18), retention of tritium from loganin labeled at positions I, 5,
ajmalicine (7), serpentine (8) (Fig. 34.2), vindoline (9) (Fig. 7, and 8 occurs in positions 3, 15, 19 and 21 of Cory-
34.3), catharanthine (10), reserpiline, and mitragynine. Al- nantM-Strychnos-type alkaloids, respectively. For exam-
though it has been suggested that vincoside (4) is the precur- ple, [1-3H]-loganin (11) is incorporated into ajmalicine (or
sor of certain 3(3-CorynantM alkaloids, it has been shown class I, see Fig. 34.5) with retention of tritium at C-2!. Thus,
that these compounds also are derived from strictosidine any mechanism invoked to explain the formation of ajmali-
(Kapil and Brown, 1979; Riiffer et aI., 1978). cine (7) must explain the retention of tritium at both positions
C-21 and C-3 (Fig. 34.1). An enzyme preparation from Cath-
Subsequent Reactions of Sbictosidlne aranthus roseus has been shown to convert tryptamine (1)
and secologanin (2) into ajmalicine (7), 19-epi-ajmalicine
The formation of the monoterpenoid indole alkaloid aj- (12) and tetrahydroalstonine (13) in the presence ofNADPH
malicine (7) and its isomers from tryptamine (1) and secolo- (Griiger, 1980). The initial step is viewed as hydrolysis by (3-
ganin (2) involves the formation of four key intermediates glucosidase, followed by opening of the hemiacetal to yield a
(Fig. 34.2) (Hutchinson, 1986). Ajmalicine represents the dialdehyde (14). A (3-glucosidase highly specific for stricto-
first major stage in the synthesis of most major groups of sidine has been isolated from a number of indole-alkaloid-
monoterpenoid indole alkaloids (see below). Stereoselective producing plants. Attempts to trap the dialdehyde have been

I
strictosidine (3) (14)
(isovincoside)

CK,O,C

O'K
4,21-dehydrocorynantbeine
aldehyde (15)

~
-.;---
NAD'

OK
4,21-dehydrogeissoscbiziae (18)
(a) OK
Fig. 34.2 (a & b). Proposed biogenesis of ajmalicine and serpentine (modified from Hutchinson. 1986 and Stockigt. 1980; used with pennission of the
copyright owners, the Royal Society of Chemistry, Cambridge and Academic Press, London, respectively).
 Indole Alkaloids 631

CH,O,C
OH

4,21.dehydrogeissoschizine (18) cathenamine (16)

CH,O,C

ajmalicine (7) CH,O,C

~
~ NADPH tetrahydroalstonine (13)

serpentine (8) 19-epi-ajmalicine (12)

(b)

Fig. 34.2. (continued)

unsuccessful (Blasko and Cordell, 1990). Such an intermedi- (8) accumulates inside the vacuoles against a concentration
ate should recyclize and dehydrate to produce 4,21-dehydro- gradient, and the uptake system is specific for alkaloids in-
corynantheine aldehyde (15). However, subsequent experi- digenous to the plant from which the vacuoles have been
ments do not support the intermediacy of this aldehyde (15). isolated (Deus-Neumann and Zenk, 1984; Zenk et al., 1985).
Cathenamine (20,21-dehydroajmalicine) (16), in its iminium This uptake requires energy and probably ATP. In other
form (17), has been established to be an intermediate in the experiments, vindoline (9) (Fig. 34.3) did not occur as a
formation of tetrahydroalstonine (13) (StOckigt, 1980), and salt within the vacuoles and could exchange with excess
an NADPH-dependent enzyme that specifically hydrogen- vindoline in the medium (Zenk et al., 1977). Ajmalicine (7)
ates the iminium form of cathenamine (17) at C-21, called represents the first major branching point in the synthesis
cathenamine reductase, has been isolated (StOckigt, 1980). of other groups of alkaloids. These alkaloids are sometimes
This enzyme has a molecular weight of about 81,000 called heteroyohimbine types (Stockigt, 1980).
(Hemscheidt and Zenk, 1985). Strictosidine (3) is converted Proposed biogenetic pathways and mechanisms by which
into the same compounds (Fig. 34.2). In the absence of relatively simple indole alkaloids such as ajmalicine are con-
NADPH, cathenamine (16) is accumulated (Fig. 34.2) (Atta- verted into the subgroups of Corynanthti-Strychnos-type al-
ur-Rahman and Basha, 1983; Groger, 1980). kaloids are reviewed by Atta-ur-Rahman and Basha (1983).
Other cell suspension cultures of Catharanthus roseus
synthesize and accumulate substantial amounts of both ser- Classification and structural Types
pentine (8) and ajmalicine (7). Serpentine (8), the principal of Indole Alkaloids
alkaloid of some cell cultures of Catharanthus roseus, is
derived from ajmalicine (7); in contrast to some other indole The most important classification schemes are based on
alkaloids, serpentine is located inside the vacuole (Deus- biosynthetic relationships of the alkaloids involved. In 1971,
Neumann and Zenk, 1984; Zenk et aI., 1985). Serpentine Kompis et al. proposed a new approach to the classification
632 Indole Alkaloids

i
""'"
17 IS

18
ajmalicine (7) secodine (29) vincamine (35)
Class 1 Class 2 Class 3

(f-r\1
~N~
COlCH)

OH
vindoline (9) fruticosine catharanthine (10)

Class 3 Class 4 Class 5

Fig. 34.3. Biogenetic classification of indole alkaloids (Cordell, 1981; used with pennission of the author; Kompis et al., 1971).

of indole alkaloids that will be used in this work. These loid is derived from 4,21-dehydrogeissoschizine (18), now
workers divided monoterpene-derived indole alkaloids into known to be the Ime intermediate. This last alkaloid is con-
five classes, and, further, divided each class into subclasses. verted to preakuammicine (19) and akuammicine (20). Prea-
The main classes are subdivided as shown in Fig. 34.3 [based kuammicine (19) is subsequently converted to stemmade-
on a condensed version prepared by Cordell (1981)]. The nine (21) and alkaloids of classes 2, 3,4, and 5 (Fig. 34.5).
rationale for this system is shown in Fig. 34.4. Importantly, In feeding studies, it was shown that [O-methyl-3H,Ar-
the numbering system used is internally consistent. The sys- 3H]geissoschizine (22) can serve as a precursor for class 1,
tem of numbering was devised by Taylor and Le Men (1965) 3, and 5 alkaloids (Groger, 1980). Geissoschizine (22) also
and is now used by almost all workers in the field. Carbon can be converted to an intermediate related to preakuammic-
3 of one skeleton is carbon 3 of any other skeleton within ine (19) and stemmadenine (21). Geissoschizine has been
the monoterpenoid portion of the alkaloids. shown to be incorporated into ajmalicine (7), serpentine (8),
A bewildering number of subgroups of monoterpene-de- akuammicine (20) stemmadenine (21), the Aspidosperma al-
rived indole alkaloids occur [see, for example, the discussion kaloid vindoline (9) and coronaridine (17) (Fig. 34.7)
in Kisakiirek et al. (1983) and Cordell (1981)]. Because of (Groger, 1980; Hutchinson, 1986). Although geissoschizine
the vague lines delineating these groups and because of the (22) appears to be an important compound in indole alkaloid
resultant confusion among those lacking intimacy with this pathways, this alkaloid probably is not an intermediate; it
group of complex alkaloids, several classification schemes
appears to enter the pathway by an NADP+-dependent reac-
have been proposed. In the past, many alkaloids discussed
tion (Hutchinson, 1986; Stockigt, 1980).
below as Group 1 (Type A) were sometimes referred to as
Preakuammicine (19) serves as an efficient precursor of
"Corynanthe-Strychnos alkaloids," those of Group 3 (Type
akuammicine (20) (a class 1 Strychnos alkaloid) and vindo-
B) as "Aspidosperma alkaloids," and those of Group 5
line (9) (an Aspidosperma alkaloid). When labeled stemma-
(Type C) as "iboga alkaloids." To further complicate things,
denine (21) (a Strychnos alkaloid) (class 1) was fed to Cath-
older literature is oriented toward discussion of the alkaloids
of a particular genus. As will be observed, many genera aranthus roseus plants, labeled tabersonine (24) (class 1),
contain divergent groups of monoterpene indole alkaloids vindoline (9) (class 3), and catharanthine (10) (class 5) were
as well as other types of alkaloids. Several plants are known produced. Thus, preakuammacine (19) and stemmadenine
to contain representatives of more than one of the five groups (21) serve as key intermediates of both class 3 and 5 alka-
outlined above. For example, Catharanthus rose us contains loids. It has been proposed that incorporation of stemmade-
members of classes 1,2,3, and 5. nine (21) into vindoline (9) and catharanthine (10) proceeds
via dehydrosecodine (25). A Diels-Alder reaction could lead
Formation of the M~or Indole Alkaloid to the formation of tabersonine (24) and catharanthine (10),
Types respectively (Fig. 34.5).
Ajmalicine (7) was formerly considered a key intermedi- Catharanthus raseus and Stemmadenia pubescens are
ate in the synthesis of many classes of alkaloids. This alka- most unusual in that each of these two species contain four
 Indole Alkaloid, 633

R
I
NH
b
- - . . Class 1, e.g., ajmalicine (7)

17 15
14
---. Class 3
e.g., vincamine (35)
19
18

Class 2, e.g., secodine (29)

3 18

14

15 14

18
\
Class 5
e.g., catharanthine (10)

Class 3
/ 18

e.g., vindoline (9)

Class 4
fruticosine
Fig. 34A. Biogenetic classification of indole alkaloids (Cordell, 1981; used with pennission of the author; Taylor and Le Men. 1965).
634 Indole Alkaloids

CH30 1
.strictosidine (3) 4,21-debydrogeissoschizine (18) 0 . . . . " 8jmalicine (7)

+
Class 1

geissoschizine (22)

~_~~~_ ~<?
I IL.,~
H
CO, CH3
VHN~0 HOCH, CO,CH3
HN " ~
CO,CH3
stemmadenine (21)
N
/
!
debydrosecodine (25)
ClassZ

,~~~~
akuammicine (20) catharanthine (10) tabersonine (24)
CO,CH3

C.... S Class 3

Fig. 34.5. Fonnation of other major indole alkaloid types.

major types of indole alkaloids. Becanse of the diversity the intermediacy of dehydrosecodine (25), a class 2 monoter-
of alkaloids produced in various cultures and the medicinal pene-derived indole alkaloid (Fig. 34.6).
importance of several of these, Catharanthus roseus has When seeds of Catharanthus roseus are germinated in
been the plant of choice for biosynthetic studies. In one set the dark, the young seedlings first accumulate tabersonine
of experiments with Catharanthus roseus seedlings, geis- (24). In the subsequent 5-10 days, the cotyledons accumu-
soschizine (22) and preakuammacine (19) were detected in late vindoline (9) and its inunediate precursors (Balsevich
seedlings within 28-40 h, the Strychnos alkaloids stemma- et aI., 1986). Transfer of 5-day-old seedlings to the light
denine (21) and akuammicine (20), and the Aspidosperma results in the rapid loss of vindoline precursors, gradual loss
alkaloid (classes 3 and 5) tabersonine were detected after of tabersonine (24), and accumulation of vindoline (9).
40-50 h of growth, whereas the iboga alkaloid (class 5) Assuming that tabersonine (24) is a direct precursor of
catharanthine (10) was formed after 100 h. vindoline (9) (this assumption is based on whole plant feed-
These feeding experiments seem to suggest that As- ing data), tabersonine first appears to be oxygenated to yield
pidosperma-Hunteria alkaloids (classes 3 and 5) arise by 16-hydroxytabersonine (26). That compound is then hydrox-
prior formation of Corynanthi and then Strychnos alkaloids ylated to yield 16-methoxytabersonine (27). The next inter-
(class 1) (Atta-ur-Rahman and Basha, 1983). mediate produced is 16-methoxy-2,3-dihydro-3-dihydroxy-
Preakuammicine (19) and stemmadenine (21) (both class N(I)-methyltabersonine (i.e., desacetoxyvindoline) (28),
1) are converted to tabersonine (24) (class 3), probably via which is, in turn, converted to desacetylvindoline and vindo-
 Indole Alkaloids 63S

line (9) (Balsevich et aI., 1986). The scheme suggested by When seeds of Catharanthus roseus are germinated,
Blasko and Cordell (1990) probably is not correct. iboga alkaloids do not appear for 100-160 h. Only class 1
Both antipodes of several Aspidosperma and Hunteria (CorynantM and Strychnos) alkaloids are found during that
alkaloids are known to occur, but it is uncommon to find time. Vindoline (9) (a class 3 or Aspidosperma alkaloid)
more than one in a single plant. does not appear until 200 h after germination (Atta-ur-Rah-
Iboga alkaloids occur only in the Apocynaceae and are man and Basha, 1983).
most common in the tribe Tabemaemontana. This group of Within the iboga alkaloids, two antipodal series are pro-
alkaloids is derived by the same biogenetic pathway as the duced. (+ )-Catharanthine (10), (+ )-ibogarnine (30), and
class 3 (Aspidosperma-Hunteria) alkaloids, at least in the ( + )-coronaridine (23) belong to one, whereas (- )-coro-
early stages. Strictosidine (3), geissoschizine (22), stemma- naridine (31) and (- )-ibogamine (32) belong to the other
denine (21), 16,17-dihydrosecodin-17 -01, secodine (29) (Fig. (Alta-ur-Rahman and Basha, 1983). The occurrence of both
34.3), and tabersonine (24) all are incorporated into iboga ( + ) and ( - ) forms of both series of alkaloids [such as ( + )-
alkaloids, which lie at the end of the biosynthetic pathway quebrachamine (33) and ( - )-quebrachamine (34)] suggests
(Fig. 34.7). On one branch, class 1 (Strychnos alkaloids) that the pathway is closely linked and possibly occurs at the
give rise to class 3 (Aspidosperma) alkaloids and, on an- stage of cyclization of a secodine precursor (Atta-ur-Rahman
other, class 5 (iboga) alkaloids. A common secodine inter- and Basha, 1983).
mediate appears to be involved. Although plants of Voacanga africana normally produce

C\:qOv --
"" I HN I
COaCH3

dehyd....ecodine (25)

~I~
VHN\=~~
22C01CH3
COaCH3
tabersonine (24)

HO
~
HN ,;?"
"" I ~CH3-0
HI~ --
~HI
,;?
I
"" N
I
CH3
""
COaCH3
--

COaCH3
16-methoxytabersonine (27)
16-hydroxytahersonine (26)

CH3-O
~
I
HI
,;?

"" N
OH
. OH
~
CH3-O
I i
CH3 COaCH3
vindoline (9)
desatetylvindoline (28)

Fig. 34.6. Proposed biosynthesis of vindoline (modified from DeLuca et aI., 1986; used with pennission of the copyright owner, Gustav Fischer
Verlag, Stuttgart).
636 Indole Alkaloids

18

catharanthine (10) (+)·ibogamine (30) (-)·ibogamine (32)

(l---i;~
~HN'~

17 ~15
(+)·coranaridine (23) (-)·quebrachamine (34) (+)-quebrachamine (33) 18

Fig. 34.7. Representative iboga or class 5 alkaloids.

iboga-type alkaloids, cell cultures produce a much simpler ganin component (ebuman, plumeran, and ibogan types) can
pattern, mostly alkaloids of the class I (Aspidosperma) type occur with either absolute configuration. These skeletal
(Ellis, 1988). types were subdivided according to increasing "chemical
complexity" compared to the basic type of each group. Skel-
Summary of Pathways Leading to Migor etal types with a rearranged secologanin moiety (ebuman,
Types of "'onoterpene-Derlved Indole plumeran, and ibogan types) occur exclusively in the
Alkaloids subfamily Plumerioideae of the Apocynaceae. Alkaloids of
the aspidosperman and strychnan types are found only in
As observed above, there are numerous subgroups of mo- the Apocynaceae and Loganiaceae. Alkaloids of the cory-
noterpene-derived indole alkaloids. Unfortunately, various nanthean, vincosan, and vallesiachotaman types are found
authors have not always agreed or been consistent in placing in all three families, although those alkaloids of the corynan-
monoterpene-derived alkaloids into subgroups. Many thean type that are found in all three families were simply
subgroups and the distributional patterns of alkaloids be- derived representatives of the group; more complex alka-
longing to those groups are discussed and reviewed in Kisa- loids were restricted in distribution.
kiirek et al. (1983) and in Atta-ur-Rahman and Basha (1983) The corynanthean type represents the most extensive
(Fig. 34.8). class of indole alkaloids. At least 41 skeletal variations of
this type are known and these alkaloids are distributed
widely in all three families. Vincosan alkaloids apparently
CHE"'OSYSTElIIADC Al'ID PlIYLOGEl'IEDC are of two independent biosynthetic origins. One type occurs
AFFLICADOI'l in the Apocynaceae and a second occurs primarily in the
Rubiaceae, but also are found in some plants of the Logania-
Data from alkaloid chemistry and biochemistry have been ceae. Strychnan alkaloids are extremely numerous; II skele-
applied to a chemosystematic stody of three closely related tal types are recognized. Many occur in the Apocynaceae,
families: Apocynaceae, Loganiaceae, and Rubiaceae (Kisa- but the majority of skeletal variations are encountered in the
kiirek et aI., 1983). In this stody, eight types of indole alka- Loganiaceae. As preViously mentioned, the ebuman,
loids were utilized: aspidospermatan, corynanthean, ebur- plumeran, and ibogan types are only found in one SUbfamily
nan, ibogan, plumeran, strychnan, vallesiachotaman, and of the Apocynaceae; that is, only plants of the Apocynaceae
vincosan. Those skeletal types with a non-rearranged skele- have the ability to rearrange the secologanin moiety of these
ton have the same absolute configuration as secologanin it- alkaloids. The authors of this stody conclude that the Rubia-
self. On the other hand, alkaloids with a rearranged secolo- ceae are comparable to the Apocynaceae in regard to their
 Indole Alkaloids 637

ability to produce indole alkaloids with modified structures, nized have been placed in this group. Among these are Anar-
and that both families are evolutionarily more developed tia, Bonafusia, Capuronetta, Conopharyngia, Ervatamia,
than the Loganiaceae. Gabunia, Hazunta, Pagiantha, Pantiaca, Peschiera, Rejoua,
Within the family Apocynaceae, only four of seven tribes Sarcopharyngia, Stenosolen, and a number of less well-
contain indole alkaloids. These are the Carisseae, known genera (Danieli and Palmisano, 1986; Leeuwenberg,
Plumerieae, Rauvolfieae, and Tabemaemontaneae. All 1980). Only two of the eight genera studied do not contain
major skeletal types occur in the Carisseae. Within this alkaloids of the ibogan type. Otherwise, iboga alkaloids
group, the simpler alkaloids are more widely distributed than occur only in Alstonia, Catharanthus (Plumerieae), and Mel-
the more derived types. Alkaloids of the most complicated odinus (Carisseae). Plumeran alkaloids are distributed
skeletal type are found only in one species of Hunteria. The widely within the tribe. Some of these are known only from
Tabemaemontaneae represent a rather uniform tribe in com- the Tabemaemontaneae. The Plumerieae probably are the
parison to the Plumerieae. The genus Tabernaemontana is most thoroughly studied tribe of the Apocynaceae. Alkaloids
quite complex; many other genera that were formerly recog- of all skeletal types are known to occur in members of this

slriclosidine (3) corynantheine vallesiachotamine

stridosidine group corynantheine group vaUesiachotamine group

adifoline talbotine stemmadenine (21)

adifoline group talbotine group stemmadenine group
CHlOH
COl CH3

H
CHlOH
mavacurine cinchonamine (70) sarpagine
mavacurine group cinchonamine group sarpagine group
H

~ perakine
H ,
CHO
H
i
corynine
akuammiline
perakine group picraJine group corynine group
<a>
Fig. 34.8 (a-d). The major classes of iridoid derived indole alkaloids and representatives of the major subgroups of each (subgroups based on Atta-ur-
Rahman and Basha. 1983; modified and used with pennission of the copyright owner, Oxford University Press, Oxford),
638 Indole Alkaloids

cvgg
HO

H I I N
.•• OH"" HN
H. 0
H Ii

ajmaline (37)
peraksine rauvoxine
ajmaline group
peraksine group oxindole group

CO,CH,

condylocarpine akuammicine (20)

pseudoxindole group condylocarpine group akuammicine group

Q
~
~HNy
CO,CH,

strychnine (45)
secodine (29)
strychnine group
(b) Class 2

Fig. 34.8. (continued)

group. Corynanthean and plumeran alkaloids are the most loganin moiety as well as quinoline and isoquinoline alka-
commonly encountered. The Plumerieae have the greatest loids. Isoquinoline alkaloids (such as emetine) also are found
variety of each of these two types. The genera Catharanthus in Pogonopus (Rondeletieae), Tocovena (Gardenieae), some
and Vinca have many affinities with the tribe Tabemaemon- members of the Rubioideae, Psychotria and Cephaelis of
taneae, but are often placed in the Plumerieae. With the ex- the Psychotrieae, Bothriospora (Hamelieae), Borreria,
ception of the ibogan types, all other major stmctural types Spermacoce, and Richardsonia (Spermacoceae), and Manet-
occur in the Rauvolfieae. tia (Hedyotideae). Within the Hillioideae,Hillia species con-
According to the classification of Leeuwenberg (1980), tain only emetine-type alkaloids.
the Rubiaceae may be divided into four subfamilies: the Only two tribes of the Loganiaceae, the Gelsemieae and
Cinchonoideae, Guettardoideae, Hillioideae, and Rubi- Strychneae, contain indole alkaloids (Bisset, 1980; Leeu-
oideae. Altogether, about 7000 species and 500 genera make wenberg, 1980). Corynanthean-type alkaloids are the sole
up this family (Hemingway and Phillipson, 1980). Onlyal- type known from the Gelsemieae. The genus Gardneria of
kaloids with a non-rearranged secologanin portion occur in the Strychneae contains only corynanthean-type alkaloids.
these plants. Alkaloids of the corynanthean type are the most Members of the genus Strychnos (about 200 species) contain
common. Most indole alkaloids of the comyanthean, vinco- aspidospermotan, corynanthean, strychnosan, vallesiacho-
san, and vallesiachotaman types occur in the tribes maman, and vincosan types. Strychnosan types are the most
NaucIeeae and Cinchoneae of the subfamily Cinchonideae. abundant (11 skeletal variations) (Kisakiirek et aI., 1983).
The tribe Cinchoneae of the subfamily Cinchonoideae is the Although the majority of monoterpene-derived indole al-
only group to contain alkaloids with a non-rearranged seco- kaloids occur in the Apocynaceae, Loganiaceae, and Rubia-
 Indole Alkaloids 639

~059
H
, I 07
H~~'
N

""'HN 07, CH,·O HN HN

. ::,... HN "'"
I ~
........ CO,CH,
CO,CH,

(+)-quebrachamine (33) vincaditTormlne schizophylline

quebrachamine group aspidospermine group schizophylline group

~
H

(~:7,
H

o N
j N 07,
::,... HN
o A

vincamine (35) schizozygine tuboxinine

vincamine group schizozygine group vindolinine group

aspidofractinine kopsine vincatine

(0)
pleiocarpine group kopslne group oxindole group

fruticosine catharanthine (10) rupicoline

catharanthine group pseudooxlndole group
(d)
Class 4 Class 5

Fig. 34.8. (continued)

ceae, indole alkaloids also have been reported from the Alan- 1II01'l0TERPEl'IOID Il'IDOLE ALKALOIDS OF
giaceae, Annonaceae, Euphorbiaceae, Icacinaceae, and PHAKJIIACOLOGICAL SIGl'IIFICAl'ICE
Sapotaceae (Grager, 1980). Reports from the Annonaceae,
Euphorbiaceae, and Sapotaceae should be confinned, as Many of the alkaloids of this series have pronounced physio-
other alkaloids are known from those families, and the pres- logical properties in animals, and some are of great medici-
ence of indole alkaloids would be quite unusual in a chemo- nal importance (Cordell, 1981). The pharmacology and cer-
systematic sense. Further, it is difficult to distinguish many tain other biological properties of alkaloids from the genus
specimens of the Sapotaceae and Apocynaceae which could Tabernaemontana (Apocynaceae) have been reviewed
lead to misidentification of materials. (Danieli and Palmisano, 1986).
640 Indole Alkaloids

Vincamine pal alkaloid is yohimbine (5) (Fig. 34.11). The yohimban

system contains three asymmetric carbon atoms: C-3, C-15,
Eburnamine-vincamine alkaloids (about 70 in number)
and C-20. Only compounds with an a-configuration at C-
occur in a number of genera of the Apocynaceae (Lounasmaa 15 are known in nature (Atta-ur-Rahman and Basha, 1983).
and Tolvanen, 1992). One of the best known representatives This aIkaIoid also has been isolated from Amsonia, As-
of these groups of alkaloids, ( + )-vincamine (35) (Fig. 34.3), pidosperma, Catharanthus, Rauvolfia, and Vallesia (all
the major aIkaIoid of Vinca minor, is used to treat headache Apocynaceae), Ladenbergia (Rubiaceae), and Gelsemium
and vertigo. This compound has central nervous system ac- and Strychnos (Loganiaceae) and has been reported from the
tion and produces a fall in blood pressure. Its principal activ- Euphorbiaceae (Alchornea).
ity is to moderate cerebral vasodilation (Oanieli and Palmi- Yohimbine is an a-adrenergic blocking agent and a sero-
sano, 1986; Lounasmaa and Tolvanen, 1992; Neuss, 1980). tonin antagonist. It depletes catecholamines from the mouse
This alkaloid has proven useful in treatment of patients with brain (Neuss, 1980). Yohimbine derivatives exhibit hypoten-
advanced arteriosclerosis. Vincamine has an LOso i.v. in sive and cardiostimulant activities (Cordell, 1981; Szantay
mouse of75 mglkg and quenches singlet oxygen. Vincamine et aI., 1986).
is a feeding deterrent to polyphagous larvae of Syntomis Yohimbine (5) is a feeding deterrent to Phormia, Syn-
(Lepidoptera) and to bees (Wink, 1993). tomis larvae, and bees and is toxic to Lemna (Wink, 1993).

JijmaJicine and Related Compounds Reserpine

Ajmalicine (7) was first isolated from yohimbe bark and, The genus Rauvolfia (about 150 species from the tropics
later, from Rauvolfia species. This compound is one of the and subtropics) has long been used medicinally. Authenti-
principal alkaloids of Catharanthus roseus. Ajmalicine is cated reports of the use of R. serpentina as a medicine date
found in members of both the Apocynaceae and Rubiaceae. back to 1000 B.C. The hypotensive activity of the drug was
reported in 1933. As indigenous supplies became depleted
Approximately 3600 kg are produced each year at a market
in India, the development of R. vomitoria, an African spe-
price of about $2000lkg (Verpoorte et aI., 1991).
cies, and R. tetraphylla, a Central American species, was
Ajmalicine has been used for the treatment of circulatory
carried out (Cordell, 1981; Tyler et aI., 1981). Because the
disorders and 10-methoxyajmalicine (44) (Fig. 34.9) ex-
root of Rauvolfia serpentina contains more than 50 aIkaIoids,
hibits hypotensive and vasodilator actions (Cordell, 1981;
it is not surprising that the crude drug differs in activity from
Neuss, 1980; Verpoorte et aI., 1991).
isolated reserpine (39) (Fig. 34.11) (Tyler et aI., 1981). The
A number of species of Alstonia (Apocynaceae) are used
crude drug contains from 0.15% to 0.20% aIkaIoids.
in traditional medicine throughout Southeastern Asia for the
Tissue and cell cultures of Rauvolfia serpentina synthe-
treatment of malaria and dysentery (Phillipson et aI., 1993).
s~ze a plethora of alkaloids, dominated by yohimbine (5),
Alstonine (36) (Fig. 34.9) is used for the treatment of dysen-
a)maline (37), and ajmalidine, which differed from that of
tery (Phillipson and O'Neill, 1987). Examination of several
any plant part and from undifferentiated cultures (Ellis,
of the aIkaIoids from Alstonia angustifolia revealed that they
1988).
had relatively low activity against Plasmodium [alciparum
The central nervous system effects of reserpine are due to
and Entamoeba histolytica (Phillipson et aI., 1993; Wright
catecholamine and 5-hydruxytryptamine depletion in the
et aI., 1995). Ajmalicine (7) is a feeding deterrent to polypha-
brain. This alkaloid acts to produce sedation and tranquiliza-
gous Syntomis larvae (Lepidoptera) (Wink, 1993).
tion and is useful in the treatment of hypertension (Cordell,
1981; Neuss, 1980; Szantay et aI., 1986). Reserpine (39)
Jijmaline causes depletion of both 5-hydroxytryptamine and norepi-
Ajmaline (37) is derived from the sarpagine alkaloid po- nephrine (Neuss, 1980), is a stimulator of prolactin release (it
Iyneuridine aldehyde (Fig. 34.10). This alkaloid has power- may be carcinogenic in women) (Cordell, 1981; Mann, 1987),
ful coronary dilating and antiarrhythmic effects. Neoajma- and inhibits oxidative phosphorylation and uptake ofCa2+, se-
line, a derivative of ajmaline, is a central nervous system rotonin, and dopamine (Robinson, 1979). This aIkaIoid also
stimulant, a vasodilator, and uterine stimulant. This com- quenches singlet oxygen and inhibits noradrenaline transport.
pound lowers blood pressure, and, in high doses, causes Reserpine is antimicrobial against Gram-negative bacteria
death by respiratory arrest. Although ajmaline has been used and cytotoxic to Walker 256 carcinosarcoma. The oral LOso
to treat heart ailments, it is currently not used because of in Agelaius is 100 mglkg (Wink, 1993). Incorporation ofre-
toxicity problems (Tillequin et aI., 1993). serpine at 0.1 % into artificial diets is totally lethal to the larvae
ofCallosobruchus maculatus (Janzen et aI., 1977). This alka-
loid is a feeding deterrent to polyphagous Syntomis larvae at
Yohimbine
the 1% level in the diet (Wink, 1993).
The bark of the tree Corynanthe yohimbe (Rubiaceae),
indigenous to the Cameroons and the Congo Republic (for- PicraJima alkaloids
merly French Congo), has been used in the treatment of Several Picralima aIkaIoids also have interesting pharma-
arteriosclerosis and is said to be an aphrodisiac. The princi- cologic activity. Aknamrnine (40) augments the hyperten-
 Indole Alkaloids 641

akuammine (40) akuammidine (41)

cimiciphytine, R=CH3 (42)

norclmiciphydne, R=H (43)
CH.... O

10-methoxyajmBliclne (44) alstonine (36)

Fig. 34.9. Bioactive monoterpenoid indole alkaloids.

sive effects of adrenaline and has local anesthetic action tory, when the seeds of this plant were introduced into Ger-
almost equal to cocaine. The seeds of Picralima nitida (Apo- many as a rat poison. These seeds contain about 1.5-5%
cynaceae) have been used by natives of West Africa as an alkaloids, consisting mainly of strychnine (45) and brucine
antipyretic. Akuammidine (41) is about three times as active (10,n-dimethoxystrychnine) (46) (Fig. 34.12) (Tyler et a1.,
as cocaine as a local anesthetic (Fig. 34.9) (Cordell, 1981). 1981). To humans, these alkaloids have an intensely bitter
Another alkaloid, akuammine (40), has been isolated from taste; most persons can detect 1 part strychnine to 500,000
Picralima species. parts water.
An extract of Haplophyton crooksii (Apocynaceae) is Alkaloids of this type belong to class I and have an un-
said to be an effective poison for such insects as cockroaches, rearranged secologanin skeleton, but have a C-12-C-16
flies, mosquitoes, fleas, and lice (Correll and Johnston, bond (Tillequin et a1., 1993).
1970). This plant contains yohimbine and norcimiphytine Both strychnine and brucine (as well as other optically
(Mroue and Alam, 1988). Cimiphytine (42) and norcimiphy- active alkaloids) are used to effect the resolution of racemic
tine (43) are two minor, lactonic alkaloids isolated from mixtures of acids in synthetic organic chemistry.
Haplophyton cimicidum (Fig. 34.10) (Specialist Periodic Re- Strychnine inhibits growth of the radicle of Lepidium and
ports 1981). is toxic to Lemna (Wink, 1993). Strychnine (45) and brucine
(46) are feeding deterrents to polyphagous Syntomis, Pieris,
Strychnine and Its Relatives and Bombyx larvae (Lepidoptera) and a variety of other or-
The poisonous properties of Strychnos nux-vomica (Lo- ganisms. Both alkaloids are insecticidal to bees (Wink,
ganiaceae) have been known in Europe since the 16th cen- 1993).
642 Indo/e A/ka/oids

CH,02C

striclosidine (3) 4,21·dehydrogeissoschizine (18) OH polyneuridine aldehyde vinorine

H06

18
19

vomilinene norajmaline ajmaline (37)

Fig. 34.10. Biosynthesis of ajmaline (modified from Herbert, 1985, 1988; used with permission of the copyright owner, The Royal Chemical Society,
Cambridge).

Strychnine (Fig. 34.12) excites all portions of the central Brucine (46), the dimethoxy derivative of strychnine, has
nervous system and binds to the glycine receptor (Robinson, an oral LOso in rat of 1 mg/kg. Both strychnine and brucine
1979). The alkaloid is a powerful convulsant; death results quench singlet oxygen, bind to glycine receptors, and inhibit
from asphyxia. As little as 60-90 mg may be fatal to humans; muscle lactate dehydrugenase (Wink, 1993).
the LOso i.v. in rat is 0.9 mg/kg (Wink, 1993). Strychnine
currently has no therapeutic uses in Western medicine (Cor· Biogenesis
dell, 1981; Tyler et aI., 1981). Incorporation of strychnine
at 0.1 % into artificial diets is totally lethal to the larvae of Feeding studies have demonstrated that both geissoschi·
Callosobruchus maculatus (Janzen et aI., 1977). zine (22), probably as 4,21·dehydrugeissoschizine (18), and

CH,02C

yohimbine (5)

12 H
o
I8~OCH'
o ~ I
~ OCH,
OCH,
reserpine (39)
Fig. 34.11. Yohimbine and reserpine (modified from Cordell, 1981; used with pennission of the author),
 Indole Alkaloids 643

OH

striclosidine (3) 4,21-dehydrogeissoschizine

~71
;:". HN

CHO -
I
HO J H+

QC-CHz-COOSCOA

strychnine (42)

~'\
U~V.) / 0 CHO HO

diaboline Wieland-Gumlich aldehyde (44) vomicine

Fig. 34.12. Biogenesis of stryclurine (Heimberger and Scott, 1973; modified and used with pennission of the copyright owner, the Royal Society of
ChemislIy, Cambridge).

the Wieland-Gumlich aldehyde (47) are involved in the bio- The most potent curarizing alkaloids have little effect on
synthetic pathway leading to strychnine (Fig. 34.12) (Bisset, blood pressure (in cats), with some exceptions. Toxiferine
1980; Kapil and Brown, 1979). (48) at a dose of 2 mg intravenously produced paralysis for
50-160 min in a 70-kg human and is probably the most
Curares from Strychnos Species useful specific curarizing agent. The LDwo of toxiferine i.p.
Most curares derived from Strychnos species in South is 0.03 mglkg. Toxiferine binds to the acetylcholine receptor
America are known as calabash curares because they are (Wink, 1993). There is no depression of blood pressure, no
stored in a calabash or small gourd. Curares of this type are histamine liberation, and an absence of bronchoconstrlction.
regarded as the most paralytic natural or synthetic neuromus- The diallyl his-nor-derivative has more reproducible activity
cular blocking agents. At least seven alkaloids have been and is shorter-acting (Cordell, 1981).
isolated that are more active than d-tubocurarine (see Chap- The dimeric curare alkaloids are derived from monomeric
ter 32). For example, C-toxiferine (48) is about 10 times as units such as Wieland-Guntlich aldehyde (47) (Fig. 34.13)
active, and C-alkaloid G (49) and E (50) are 100 times more (Bisset, 1980), a compound first derived in 1932 from degra-
active, being active at I fLg per kg (Cordell, 1981). dation of strychnine.
644 Indole Alkaloids

OH
Wieland·Gumlich aldehyde (47)
(Nb-methyl, curacurine Vll)

HO
CHO
16-dihydronorfluorocurarine

venecurine (51)
caracurine V

OH
HO HO

C·alkaloid E (50) C·alkaloid G (49)

Fig. 34.13. Biogenesis of C-toxiferine I and similar alkaloids (modified from Cordell, 1981; used by pennission of the author).

CH,-O

olivacine (52) ellipticine (53) 9-methoxyellipticine (54)

Fig. 34.14. Olivacine, ellipticine, and 9-methoxyeIlipticine.

 Indole Alkaloids 64S

Although alkaloids with curarizing activity usually are their efficacy as anticancer agents (Fig. 34.15) (Cordell,
dimers, a monomeric alkaloid with curarizing activity, vene- 1978; Cordell and Saxton, 1981; Endo et al. 1988). Because
curine (51), has been isolated from the curare prepared by these alkaloids occur at extremely low levels in the plant,
the Hoti tribe of Venezuela (Quetin-Leclercq et al., 1989). enonnous quantities are required for commercial production.
Nearly 500 kg of plant material are needed to produce I g
Olivacine and Ellipticine of the active alkaloids (Tyler et al., 1981). Vincristine occurs
at 0.0003% by weight in the source plant. Vinblastine also
Olivacine (52) occurs in the bark of several Brazilian
is known from Catharanthus ovalis, C. longifolius, and C.
trees of the genera Aspidosperma and Tabernaemontana
(Apocynaceae); ellipticine (53) and 9-methoxyellipticine
trichophyllus (Blasko and Cordell, 1990). These antimitotic
agents inhibit cancer cell growth and are the treatment of
(54) have been isolated from species of Ochrosia (Apocyna-
choice for several types of leukemia and Hodgkin's disease
ceae) (Neuss, 1980). Similar alkaloids have been isolated
(Miura et al., 1987). The pharmacology and therapeutic use
from Strychnos dinklagei (Gribble, 1990). Both ellipticine
of these alkaloids has been reviewed (McConnack, 1990;
and 9-methoxyellipticine (54) have been isolated at low lev-
Neuss and Neuss, 1990). Synthetic strategies leading to the
els from callus cultures of Ochrosia elliptica (Gribble,
monomers vindoline and catharanthine and routes to couple
1990).
Olivacine (52) and ellipticine (53) are antileukemic them have been developed (Cordell, 1981; Miura et al.,
agents in mice, and derivatives of ellipticine are used clini- 1987).
cally in Europe (Fig. 34.14) (Cordell, 1978; Gribble, 1990). Vincristine (56) and vinblastine (57) bind to and dimerize
The principal mechanism of action is related to the planar tobulin, and inhibit protein synthesis and DNA-dependent
structures of this group of alkaloids. Ellipticine and its deriv- RNA polymerase. Both inhibit intracellular transport. Vin-
atives appear to inhibit topoisomerase IT; they also are muta- blastine (57) inhibits growth of Trypanosoma cruzi, the caus-
genic and clastogenic at the tk locus of mouse lymphoma ative agent of Chagas' disease. The LDso i.p. in mouse of
cells. The major mechanism of action appears to be chromo- vinblastine is 9.5 mg/kg; that of vincristine is 5.2 mg/kg
somal cleavage (Borris and Schaeffer, 1992; Gribble, 1990). (Wink, 1993).
These alkaloids inhibit DNA synthesis by intercalation into Although the seeds of Catharanthus roseus lack alka-
DNA. 9-Methoxyellipticine and ellipticine inhibit mitochon- loids, young seedlings contain several dozen alkaloids.
drial respiration and cytochrome c oxidase (Wiuk, 1993). These alkaloids disappear after about 3 weeks and reappear
Both ellipticine and olivacine are active against cultures at 8 weeks (Robinson, 1974); in the intennediate period,
of Crithidiafasciculata (a flagellate) and Trypanosoma cruzi alkaloids are absent from the plants.
(the causative agent of Chagas' disease) (Gilbert, 1977). EI- The products of cell culture of Catharanthus roseus are
Iipticine has an LDso i.v. in mouse of 19-22 mg/kg (Wink, many and varied (Ellis, 1988); bisindole alkaloids and other
1993). products from cultures of this plant have been reviewed
(Blasko and Cordell, 1990). Some lines fail to produce any
Bisindole Alkaloids of the alkaloids nonnally found in the whole plant. Ajmali-
cine (7) and serpentine (8) (class I or Corynanthe-type alka-
Numerous bisindole alkaloids, fonned by dimerization of loids) often are the principal alkaloids produced in Catha-
monomeric monoterpenoid-derived indoles, have been iso- ranthus roseus tissue cultures (Blasko and Cordell, 1990;
lated. Probably the best source of these alkaloids is Cathara- Ellis, 1988; Knobloch et al. 1982, Kurz et al. 1980). Taber-
nthus roseus; bisindole alkaloids from this source have been sonine (24) (a class 3 or Aspidosperma alkaloid) and cathara-
tabulated (Blasko and Cordell, 1990). Several of these alka- nthine (10) (a class 5 or iboga alkaloid) have been found in
loids antitomor activity (Blasko and Cordell, 1988); most relatively low yields in tissue cultures (DeLuca et al. 1986).
important in this regard are vincristine (55) and vinblastine Vindoline (9) (a class 3 or Aspidosperma alkaloid), a major
(56) from Catharanthus roseus (Fig. 34.15). Certain other alkaloid in Catharanthus roseus plant material, has never
dimeric indole alkaloids known as curares have paralytic been found in plant tissue cultures. The pathways leading
effects in animals (see the subsection Curares from to vindoline appear to occur in the shoots of Catharanthus
Strychnos species, above). roseus plants, but are not known to occur in roots (DeLuca
Nuclear magnetic resonance and mass-spectral data for et al., 1986).
bisindole alkaloids have been reviewed (Blasko and Cordell, Both vindoline (9) and catharanthine (10) are of great
1990). importance, as coupling of these two compounds (possibly
via an iminium intennediate) leads to 3',4'-anhydrovinblas-
Catbarantbus Alkaloids tine (57), a direct precursor to both vinblastine (57) and
Two clinically useful bisindole alkaloids, vincristine (55) vincristine (56) (Fig. 34.15) (Blasko and Cordell, 1990;
and vinblastine (56), are among the most expensive of all Stuart et ai., 1978). Enzymes from cell-free systems of Cath-
plant-derived drugs (as much as $23,000 per gram), because aranthus roseus have been demonstrated to carry out this
of their low abundance in the source plant Catharanthus coupling (Endo et al., 1988). The reaction also can be cata-
roseus (fonnerly placed in the Vinca by some workers) and lyzed by horseradish peroxidase (Goodbody et al., 1988) or
646 Indole Alkaloids

vindoline (9)

tabersonine (24)

r~ COlCH,

catbaranthine (10)

CH,O

iminium intermediate

OH
/ CH,O
COlCH,

3' ,4'·aohydrovinblastine (57)

vincristine (55), R = CHO

<a)
vinblastine (56), R = CH,

(b)

Fig. 34.15 (a & b). Proposed biosynthesis of vincristine and vinblastine (modified from Goodbody et aI., 1988; used with pennission of the copyright
owner, Georg Thieme Verlag, Stuttgart).

carried out synthetically (Kutney et al., 1976). Others have mon in the southeastern United States and has been involved
suggested that vinblastine (57) and vincristine (56) are not in poisoning of children. The major alkaloid is gelsemine
found in cell cultures because these cultures do not synthe- (58) (Fig. 34.16), but another alkaloid, gelsemicine (59)
size vindoline (9) (Goodbody et al., 1988). (MLD 0.05-0.06 mg/kg in rabbits), probably is responsible
for most of the toxicity. Gelsemine modulates glycine neuro-
Gelsemium Alkaloids chemical activity (Wink, 1993). A similar species, G. eleg-
ans, occurs in China and has been used in traditional medi-
A number of highly toxic alkaloids is found in members cine (Liu and Lu, 1988). A crude mixture of Gelsemium
of the genus Gelsemium (Loganiaceae). One species, Gelse- alkaloids has been used as an analgesic and antispasmodic
mium sempervirens or yellow jessamine, is extremely COffi- drug. Strictosidine is converted to gelsemine (58) in G. sem-
 Indole Alkaloids 647

CO~~:~~CH'OH
9" H
I I N

--
---+ "" HN H --

"'" "'"

strictosidine (3) akuammidine (41) koumidine

(koumicine)

vobasindiol / anhydrovobasindiol koumine

gelsenidine
humantienine type

CH,-O

gelsemicine (59)
Fig. 34.16. Proposed biosynthesis of gelsemine (Liu and Lu, 1988; modified and used with permission of the copyright owner, Academic Press,
Orlando, FL).

pervirens in 0.47% yield. Structurally similar alkaloids use of plant extracts enables the hunters to remain motionless
apparently occur in the related genus Mostuea (Bisset, for as long as 2 days while retaining mental alertness (Dan-
1980). ieli and Palmisano, 1986; Thompson, 1974).
The principal alkaloid, ibogaine (50) (lO-methoxyibo-
Alkaloids from Tabemantbe /boga gamine), was first obtained at the tum of the century and is
The roots of the West African plant Tabernanthe iboga regarded as the principal hallucinogenic agent of Tabernan-
(Apocynaceae) are used to combat fatigure, sleep, and hun- the iboga root. Several iboga alkaloids are central nervous
ger (central nervous system stimulation), and are associated system stimulants, as evidenced by their antagonism to reser-
with secret societies and religious practice (Schultes and pine catalepsy. Ibogaine (60) is a cholinesterase inhibitor
Hofmann, 1973). lboga, when used in small quantities, is a (Schultes and Hofmann, 1973). A number of alkaloids of
stimulant used to combat fatigue, whereas, in larger quan- this type cause hypotension and bradycardia. Ibogaline (61)
tities, it is hallucinogenic (Schleiffer, 1979). In Gabon, is the most active of these (Fig. 34.17).
where this practice is widespread, iboga may be derived from Iboga alkaloids are found in other genera [e.g., Conopha-
either Tabernanthe iboga or from T. subsesselis. Although ryngia, Ervatamia, Hazunta, Melodinus, Pagiantha, Pan-
the roots of the plant are usually used, the leaves have more daca, Tabernaemontanae, and Voacanga (Apocynaceae).
pronounced physiological activity (Raymond-Hamet, 1941). Catharanthine (10) possesses hypoglycaernic activity
Extracts of this plant also are used while stalking game; the (Neuss, 1980).
648 Indole Alkaloids

o
CH,O
CH'0'O:W~N
"'" I IN"", I I
HN HN
·...'H '. . H

ibogaine (60) ibogamine (30) iboluteine

(10-methoxyibogamine)

CH'°lrl?F _
CH,O ~
IIN
rn,o--J,,~
"'" HN
"H

tabernanthine
ibogaline (61)

Fig. 34.17. Alkaloids from Tabernanthe iboga.

Camptotbecine dell, 1985; Wall et al., 1966). Camptothecine is a potent

cytotoxic agent that acts by inhibiting nucleic acid synthesis.
Camptothecine (6) was fIrst isolated from the Chinese
In certain cell lines, DNA and RNA synthesis are inhibited
tree Camptotheca acuminata (Nyssaceae). Extracts of this
reversibly about 50% by 5 J.LM camptothecine (Cordell,
plant were shown to possess very high antitumor activity
1981). Camptothecine exerts its action on DNA topoisomer-
(Cai and Hutchinson, 1983; Cordell, 1978; Suffness and Cor-
ase I, an enzyme that relaxes supercoiled DNA. IO-Hydroxy-
dell, 1985). Derivatives of camptothecine are now in clinical
camptothecine (64) has a better therapeutic index than camp-
trials in the United States.
tothecine and has been subjected to clinical study. This
Camptothecine (6) does not give positive alkaloid tests
compound is used widely in China for treatment of liver
with Dragendorf's reagent or other similar reagents. Further,
cancer, leukemia, and gastric cancer (Blasko and Cordell,
this alkaloid does not form salts with mineral acids (Cai and
1988). The 20(R,S)-9-amino derivative of camptothecine
Hutchinson, 1983).
has been found effective in treating immunodefIcient mice
Biogenesis carrying three different lines of human colon cancer without
toxic effects (Kinghorn, 1992).
Feeding experiments have demonstrated that strictosidine Camptothecine (6) inhibits replication of DNA vimses
(3), tryptamine (1), and secologanin (2) all serve as precur- such as adenovims, vaccinia vims, and herpes vims, but has
sors for camptothecin (6) (Fig. 34.18) (Cai and Hutchinson, no effect on RNA vimses. Camptothecine inhibits 45 S
1983; Kapil and Brown, 1979). Feeding studies with [5- rRNA transcription (Wiuk, 1993) and is a strong inhibitor
14C,14-3H]strictosamide (63) indicate that this compound is of n\lcleic acid synthesis in mammalian cells (Blasko and
incorporated efficiently into camptothecine. Cordell, 1988). IO-Methoxycamptothecine (65) is abo\lt
eight times more active against herpes vims than campto-
Distribution of Camptotbecine Among thecine (Cai and H\ltchinson, 1983).
Higher Plants Camptothecine exhibits selective plant growth inhibition.
In addition to Camptotheca acuminata (Nyssaceae), cam- The growth of tobacco and com were retarded, whereas no
ptothecine (6) has been found in Nothapodytes (Mappia) effect was noted on beans and sorgh\lm when a spray of 1
joetida and Merilliodendron megacarpum (Ieacinaceae), X 10-4 ppm was applied. Growth inhibition appeared to
Ophiorrhiza mungos (Rubiaceae), and Ervatamia heyneana be limited to the rneristomatic areas of the plant (Cai and
(Apocynaceae) (Cordell, 1981). This distribution is of con- Hutchinson, 1983).
siderable interest to plant systematists and phylogenists, as Camptothecine (6) has been \lsed topically for the treat-
it reflects the same dichotomy seen in the distribution of ment of psoriasis (Cai and Hutchinson, 1983).
other types of monoterpene iridoids.
Cinchona Alkaloids
Biological Activity Alkaloids from plants of the genus Cinchona (Rubiaceae)
Plant extracts containing camptothecine (6) were shown have been used in Western medicine for more than 300 years.
to possess very high antitumor activity (Suffness and Cor- The use of Cinchona bark to treat malaria was first recorded
 Indole Alkaloids 649

in 1633; South American Indians do not appear to have used Biogenesis

it for this purpose. Quinine (66), isolated from this plant,
Despite the fact that quinine (66) and related alkaloids
was the first effective drug to treat Plasmodium falciparum
have quinoline skeleta; biogenetically, they are indole ~a­
and related organisms (Phillipson and Wright, 1991). As the
loids. Radioactivity from labeled loganin (11) and stnctosld-
medicinal properties of quinine became known, substantial
ine (3) is incorporated into quinine. The indole nitrogen be-
supply problems from South America developed and cultiva-
comes the quinoline nitrogen (Fig. 34.19). Loganin (11),
tion of Cinchona was initiated in other parts of the world.
corynantheal (69), and probably cinchonarnine (70) have
The Dutch introduced Cinchona into Java and, today, that
been shown to be key intermediates in the biosynthesis of
island is the world's major supplier. Before World War n,
qninine (66), cinchonine (67), and cinchonidine (68) (Kapil
quinine (66) was the major drug used to control malaria.
and Brown, 1979).
When the Japanese occupied the Dutch Indies, a major
Feeding experiments with doubly-labeled [1-3H2 , 1-
search was initiated to find substitutes. Several antimalarial
14C]tryptamine (1) showed that 50% of the tritium at C-I
drugs were developed and are still used.
of tryptamine was lost during the formation of Cinchona
At least 40 alkaloids have been isolated from various
alkaloids. This indicates that the cleavage between C-5 and
species of Cinchona (Cordell, 1981; Tyler et aI., 1981). An
N-4 in corynaltheal (69) is a stereospecific process. The ring
average yield of alkaloids from the bark is about 6-7%,
expansion to the quinoline nucleus must occur via opening
although up to 18% is occasionally found (Tyler et aI., 1~8.1).
of the N-containing ring and recyclization of (68) to furnish
Although first isolated in 1820, the structure of qmrune
cinchonidinone (71), which has been determined tu be a
was determined in 1909, and the first synthesis was carried
natural product in Cinchona ledgeriana (Kapil and Brown,
out in 1944 (Cordell, 1981). Quinine is used as a standard
1979). The specific incurporation of [11-3H21cinchonidinone
for bitterness.
(71) in cinchonine (67) and quinine (66) has been observed
Despite extensive searches, quinine and related alkaloids
(Kapil and Brown, 1979).
have been found only in Cinchona and a few closely related
The structure of guettardine (72), isolated from Guettarda
genera of the Rubiaceae, especially the genus Remijia.
heterosepala, suggests that this alkaloid may represent an
Because of the value and the widespread use of Cinchona
intermediate between the Corynanthe and cinchonamine
alkaloids, many attempts to produce alkaloids in plant tissue
types of alkaloids (Herbert, 1986). If so, cl~avage of the .13-
cultures and cell cultures from this group of plants have been
carboline system must precede the formation of the qum-
made (Verpoorte et aI., 1991). Trees require about 10 years
uclidine ring (Fig. 34.20) (Herbert, 1986).
before they can be harvested. Cell cultures of Cinchona ledg-
eriana and C. succirubra produce largely cinchonine (67)
Biological and JIIedicinal Activity
and cinchonidine (68), although small quantities of quinine
and quinidine were produced. Only small amounts of total Quinine forms a complex with haemin that seems to be
alkaloids were produced (Ellis, 1988). an essential part of the receptor for both quinine and chlo-

O-glucosyl
strictGsidine (3) strictosamlde (63) camptotheclne (6)

HO CH,-O

10-hydroxycamptothecine (64) lO.methoxycamptothecine (65)

Fig. 34.18. Biogenesis of camptothecine (modified from eai and Hutchinson, 1983; used with permission of the copyright owner, Academic Press,
Orlando. FL).
650 Indole Alkaloids

(Jc(1:' COzH
OH
I I
'>-. HN
NH
z
N

;5
tryptophan
+

CHO ~ O-gIU-:::-

'>-. 0
CH30 Z corynantheal (69) cinchonamine (70)

seoologanin (2)

quinamine

u
quinine (66) cinchonidine (68) cinchonine (67)

Fig. 34.19. Biogenesis of quinine (modified from Leete, 1969; modified and used with pennission of the copyright owner, the American Chemical
Society, Washington, DC),

roquine, Quinoline antimalarials are toxic preferentially to larval food consumption by Pieris brassicae by approxi-
the mature stages of the parasite (Borris and Schaeffer, mately 75% (Levinson, 1976).
1992). Certain insects that feed on Cinchona species appear to
Quinine (66), cinchonidine (68), cinchonine (67), and accumulate the alkaloids in their bodies. For example, a Hel-
quinidine (73) are toxic to Cinchona cells in culture and to opeltis species and Attacus atlas both accumulate cinchonine
Lemna. Cinchonine and quinidine inhibit growth of the radi- (67) or cinchonine-like alkaloids (Waller and Nowacki,
cle of Lepidium and Lactuca (Wink, 1993). 1978).
Several alkaloids of Cinchona species [e.g., quinine (66), Qninine (66) is toxic to many bacteria and unicellular
quinidine (73), cinchonidine (68), and cinchonine (67)] serve organisms. This alkaloid is active against Trypanosoma
as feeding deterrents to Syntomis larvae (Lepidoptera), bees, cruzi (Wink, 1993). Quinine is a local anesthetic and has
Leptinotarsa (Coleoptera), Locusta, Agelaius, and a variety cardiovascular effects, oxytocic effects, curarizing effects,
of other organisms (Wink, 1993). The presence of quinine and an analgesic effect comparable to salicylate. Quinine is
sulfate or chloride at 4 X 10-7 or 1.5 X 10-6 M reduced not commonly used alone as an antimalarial today (Cordell,
 Indole Alkaloids 651

OH 1981}. This alkaloid has the ability to supress. but not cure.
vivax malaria. However. quinine has regained importance
in the treattnent of chloroquine-resistant falciparum malaria
(Tyler et al .• 1981).
Quinine (66) intercalates with DNA. modulates ion chan-
nels. and inhibits glucose response in chemosensory cells
(Wink. 1993). Cinchonidine (68) has an LDso i.p. in rat of
206 mglkg; cinchonine (67) an LDso i.p. in rat of 152 mglkg;
quinidine (64) has an LDso Lv. in rat of 30 mglkg; and qui-
nine has an LDso p.o. (per os. oral) in Agelaius of 100 mglkg.
OH All of these organisms produce growth inhibition in Plasmo-
dium Jalciparum (Wink. 1993). Cinchonidine inhibits the
guettardine (70) potato X virus (Wink. 1993).
Quinidine (73). another alkaloid found in Cinchona. is
Fig. 34.20. Guettardine.
used to treat certain heart conditions; in partiCUlar. this alka-
loid is used to inhibit auricular fibrillation (Fig. 34.19) (Cor-
dell. 1981; Tyler et aI.. 1981).

(75) CHO

OH
(76)
4'
usambarenslne (77)
CH,-O

CH,-O

OH

tubulosine (74) OH
3,4·dihydroxyusambarenslne (78)

Fig. 34.21. Cinchopbylline alkaloids.

652 Indole Alkaloids

Cinchophylline and Related Alkaloids ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum
Press, New York, 1992.
The cinchophyllines, a series of alkaloids that structurally BROWN, R. T., J. LEONARD, and S. K. SLEIGH, The role of strictosid-
resemble emetine, cephaeline, and other isoquinoline alka- ine in monoterpenoid indole alkaloid biosynthesis, Phytochem-
loids, except that they are formed from tryptamine and seco- iotty, 17, 899-900 (1978).
loganin, are found in the leaves of certain Cinchona species CAl, J. and C. R. HUTCHINSON, Camptothecine, in The Alkaloids,
(Fig. 34.21). Somewhat similar alkaloids occur in Ochrosia Vol. 21 (A. Brossi, ed.), 101-138, Academic Press, New York,
and Dyera species (Apocynaceae) and in Strychnos species 1983.
(Loganiaceae) (Borris and Schaeffer, 1992; Phillipson et al., CORDELL, G. A., The biosynthesis of indole aIkaIoids, Lloydia, 37,
1993). Another structurally related alkaloid, tubulosine (74), 219-298 (1974).
has been isolated from a Pogonopus species (Rubiaceae) CORDELL, O. A .. Anticancer agents from plants, in Progress in
(see Chapter 32) (Hemingway and Phillipson, 1980). Phytochemistty Vol. 5 (L. Reinhold, J. B. Harbome, and T.
Strychnos usambarensis (Loganiaceae), the source of several Swain, eds.), 273-316, Pergamon, London, 1978.
of these alkaloids, is used as an arrow poison by the Ban- CORDELL, O. A., The Aspidosperma alkaloids, in The Alkaloids,
yarnbo tribe on the border of Tanzania and Rwanda (Phil- Vol. 17 (R. H. F. Manske andR. G.A. Rodrigo, eds.), 200-384,
lipson and Wright, 1991). These alkaloids probablY arise Academic Press, New York, 1979.
via precursors such as (75) and (76) that are derived from CORDELL, G. A., Introduction to Alkaloids, A Biogenetic Approach,
strictosidine (3) (Bisset, 1980). Wiley-Interscience, New York, 1981.
Several cinchophylline and related alkaloids have pro- CORDELL, G. A. and J. E. SAXTON, Bisindole aIkaIoids, in The
nounced arnebicidal and cytoxic activity. The alkaloids Alkaloids, Vol. 20 (R. H. F. Manske and R. G. A. Rodrigo,
usarnbarensine (77) and 3',4'-dihydroxynsarnbarensine (78) eds.), 3-296, Academic Press, New York, 1981.
from Strychnos usambarensis have ICso values against a CoRRELL, D. S. and M. C. JOHNSTON, Manual of the Vascular Plants
multiple-drog-resistant strain of Plasmodium Jalciparum of of Texas, Texas Research Foundation, Renner, 1970.
0.38 and 0.01 IJoglml, respectively (phillipson and Wright, CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual
1991). Usurnbarensine (77) has an ICso value of 1.40 IJoglml Reports on NMR Spectroscopy, Vol. 13 (G. A. Webb, ed.),
against Giardia intestinalis, 3',4'-dihydroxynsarnbarensine 60-210, Academic Press, London, 1982.
(78) of 0.023 IJoglml against Plasmodium Jalciparum, and DANIELl, B.and G. PALMISANO, Alkaloids from Tabernaemontana,
usarnbarine (79) 1.02 IJog/m1 against Entamoeba histolytica in The Alkaloids, Vol. 27 (A. Brossi, ed.), 1-130, Academic
(compared to emetine with 2.04 IJog/ml) (Pbillipson et al., Press, New York, 1986.
1993). DELUCA, V., J. BALSEVICH, R. T. TvLBR, U. EILERT, B. D. PANCHUK,
and W. G. W. KURZ, Biosynthesis of indole alkaloids: DEvelop-
mental regnlation of the biosynthetic pathway from tahersonine
to vindoline in Catharanthus roseus, J. Plant Physiol., 125,
REFERENCES 147-156 (1986).
DEUS-NEUMANN, B. and M. H. ZENK, A highly sensitive aIkaIoid
ATIA-UR-RAHMAN and A. BASHA, Biosynthesis of Indole Alka- uptake system in vacuoles of higher plants, Planta, 162,
loids, Clarendon Press, Oxford, 1983. 250-260 (1984).
BALSEVICH, J., V. DELUCA, and W. G. W. KURZ, Altered alkaloid ELus, B. E., Natural products from plant tissue culture, Nat. Prod.
pattern in dark grown seedlings of Catharanthus roseus. The Rep., 5, 581-612 (1988).
isolation and characterization of 4-desacetoxyvindoline: A ENro, T.. A. GooDBODY, J. VUKOVIC. and M. M!SAWA, Enzymes
novel indole alkaloid and proposed precursor of vindoIine, Het- from Catharanthus roseus cell suspension cultures that couple
erocycles 24, 2415-2421 (1986). vindoline and catbaranthine to form 3',4'-anhydrovinblastine.
BATIERSBY, A. R, N. G. LEWIS, and 1. M. 'fIpPETI, The basic Phytochemistty, 27, 2147-2149 (1988).
glucosides related to the biosynthesis of indole and ipecac alka- GEISSMAN, T. A. and D. H. G. CROUT, Organic Chemistty of Sec-
loids, Tetrabedron Lett., 4849-4852 (1978). ondary Plant Metabolism, Freeman Cooper, San Francisco,
BISSET, N. G., Alkaloids of the Loganiaceae, in Indole and Bioge- 1969.
netically Related Alkaloids (1. D. Phillipson and M. H. Zenk, GaBERT, B., Natural product derivatives in tropical insect and para-
eds.), 27-61, Academic Press, London, 1980. site control, in Natural Products and the Protection of Plants (G.
BLASKO, G. and G. A. CORDELL, Recent developments in the chem- B. Marini-Bett610, ed.), 225-252, Elsevier, Amsterdam, 1977.
istry of plant-derived anticancer agents, in Economic and Me- GOODBODY, A. E., T. ENDo, 1. VUKOVIC, J. P. KUI'NBY, L. S. L.
dicinal Plant Research, Vol. 2 (H. Wagner, H. Hikino, and N. CHOI, and M. MIsAWA. Enzymic coupling of catharanthine and
R. Farnsworth, eds.), 119-191, Academic Press, London, 1988. vindoline to form 3',4'-anhydrovinblastine by horseradish per-
BLASKO. G. and G. A. CORDELL, Isolation. structure elucidation, oxidase, Planta Medica, 54, 136-140 (1988).
and biosynthesis of the indole alkaloids of Catharanthus, in GRIBBLE, G. W., Synthesis and antitumor activity of ellipticine
The Alkaloids, Vol. 37 (A. Brossi and M. Suffness, eds.), 1-76, alkaloids and related compounds, in The Alkaloids, Vol. 39 (A.
Academic Press, New York, 1990. Brossi, ed.), 239-352, Academic Press, New York, 1990.
BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from GROOER, D., Alkaloids derived from tryptophan and anthranilic
plants, in Phytochemical Resources for Medicine and Agricul- acid, in Secondary Plant Products (E. A. Ben and B. V. Charl-
 Indole Alkaloids 653

wood, eds.), Encyclopedia of Plant Physiology, N. S., Vo!' 8, series. The biogenetic approach, Helv. Chim. Acta, 59.
128-159, Springer-Verlag, Berlin, 1980. 2858-2882 (1976).
HAMPP, N. and M. H. ZBNK, Homogeneous strictosidine synthase LEETE, E., Biosynthesis of quinine and related alkaloids, Ace.
from cell suspension cultures of Rauvolfia serpentina, Phyto- Chem. Res., 2,59-64 (1969).
chemistry, 27, 3811-3815 (1988). LEETB, E., The biosynthesis of alkaloids, Specialist Periodic Re-
lIEIMBERGER, S. I. and A. I. Scorr, Biosynthesis of strychnine, J. ports. Biosynthesis, Vo!. 7, 102-223, The Royal Society Chem-
Chern. Soc., Chern. Commun., 217-218 (1973). istry, Cambridge, 1983.
HEMmGWAY, s. R. and J. D. PinLLIPSON, Alkaloids of the Rubia- LEEUWENBERG, A. J. M., The taxonomic position of some genera in
ceae, in Indole and Biogenetically Related Alkaloids (J. D. Phil- the Loganiaceae, Apocynaceae, and Rubiacea.e, related families
lipson and M. H. Zenk, eds.), 63-90, Academic Press, London, which contain indole alkaloids, in Indole and Biogenetically
1980. Related Alkaloids (J. D. Phillipson and M. H. Zenk, eds.), 1-10,
HEMSCHEIDT, T. and M. H. ZENK, Partial purification and character- Academic Press, London, 1980.
ization of a NADPH dependent tetrahydroalstonine synthase LEVINSON, H. Z., The defensive role of alkaloids in insects and
from Catharanthus roseus cell suspension cultures, Plant Cell plants, Experientia, 32, 408-411 (1976).
Rep., 4, 216-219 (1985). Lru, Z. and R. Lu, Gelsemium alkaloids, in The Alkaloids, Vol.
HERBERT, R. B., The biosynthesis of plant alkaloids and nitrogenous 33 (A. Brossi, ed.), 84-140, Academic Press, New York, 1988.
microbial metabolites, Nat. Prod. Rep., 2, 163-179 (1985). LoUNASMAA, M. and A. TOLVANEN, Ebumamine-vincamine alka-
HERBERT, R. B., The biosynthesis of plant alkaloids and nitrogenous loids, in The Alkaloids, Vo!' 42 (G. A. Cordell, ed.), 1-116,
microbial metabolites, Nat. Prod. Rep., 3, 185-203 (1986). Academic Press, New York, 1992.
HERBERT, R. B.o The biosynthesis of plant alkaloids and nitrogenous MANN, J., Secondary Metabolism, Oxford University Press, Ox-
microbial metabolites, Nat. Prod. Rep., 5, 523-539 (1988). ford, 1978; 2nd edition, 1987.
HUTCHlNSON, C. R., Biological methods for studying the biosyn- McCORMACK, J. J., Pharmacology of antitumor bisindole alkaloids
thesis of natural products, Nat. Prod. Rep., 4, 133-152 (1986). from Catharanthus, in The Alkaloids, Vo!. 37 (A. Brossi and
JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary M. Suffness, eds.), 205-228, Academic Press, New York, 1990.
compounds to the seed-eating larvae of the bruchid beetle Callo- McKNIGHT, T. D., D. R. BERGEY, R. J. BURNETT, and C. J. NESSLER,
sobruchus maculatus, Phytochemistry, 16, 223-227 (1977). Expression of enzymatically active and correctly targeted strict-
KAPIL, R. S. and R. T. BROWN, Monoterpene alkaloid glycosides, osidine synthase in transgenic tobacco plants, Planta, 185,
in The Alkaloids, Vo!' 17 (R. H. F. Manske and R. G. A. Ro- 148-152 (1991).
drigo, eds.), 546-588, Academic Press, New York, 1979. MlliRA, Y., K. HIRATA, and N. KURANO, Isolation of vinblastine in
KINGHORN, A. D., Plants as sources of medicinally and phannaceut- callus culture with differentiated roots of Catharanthus roseus
ically important compounds, in Phytochemical Resources for (L). G. Don, Agric. Bio!. Chern., 51, 611-614 (1987).
Medicine and Agriculture (H. N. Nigg and D. S. Seigler, eds.), MROUE, M. and M. ALAM, Chemical investigation of alkaloids of
75-95, Plenum Press, New York, 1992. Hap/ophyton crooksii, Phytochemical Society of North Amer-
KIsAKiiREK, M. V., A. J. M. LEEUWENEERG, and M. HEsSE, A che- ica, Abstracts, Annual Meeting, 1988.
motaxonomic investigation of the plant families of Apocyna- NEUss, N., The spectrum of biological activities of indole alkaloids,
ceac, Loganiaceae, and Rubiaceae by their indole alkaloid con- in Indole and Biogenetically Related Alkaloids (J. D. Phillipson
tent, in Alkaloids: Chemical and Biological Perspectives, Vo!' 1 and M. H. Zenk, eds.), 294-313, Academic Press, London,
(S. W. Pelletier, ed.), 211-376, Wiley-Interscience, New York, 1980.
1983. NEUSS, N. and M. N. NEUSS, Therapeutic use ofbisindole alkaloids
KoMPIs, I., M. HEsSE, and H. SCHMID, An approach to the bioge- from Catharanthus, in The Alkaloids, Vo!' 37 (A. Brossi and
netic classification of indole alkaloids, Lloydia, 34, 269-291 M. Suffness, eds.), 229-240, Academic Press, New York, 1990.
(1971). PinLLIPSON, J. D. and M. J. O'NEILL, Antimalarial and amoebicidal
KNOBLOCH, K., G. BAST, and J. BERLIN, Medium- and light-induced natural products, in Biologically Active Natural Products (K.
formation of serpentine and anthocyanins in cell suspension Hostettmann and P. J. Lea, eds.), 49-64, Clarendon, Oxford,
cultures of Catharanthus roseus, Phytochemistry, 21, 591-594 1987.
(1982). i'BIu.IPSON, J. D. and C. W. WRIGHT, Can ethnopharmacology con-
KURZ, W. G. W., K. B. CHATSON, F. CONSTABEL, J. P. KUTNBY, tribute to the development of antimalarial agents? J. Ethnophar-
L. S. L. ClIm, P. KOLODZlEJCZYK, S. K. SLEIGH, K. L. STBWART, maco!.,32, 155-165 (1991).
and B. R. WORTH, Alkaloid production in Catharanthus roseus PinLLIPSON, 1. D., C. W. WRIGHT, G. C. KIRBy, and D. C. WARH-
cell cultures: britial studies on cell lines and their alkaloid con- URST, Tropical plants as sources of antiprotozoal agents, in Phy-
tent, Phytochemistry, 19, 2583-2587 (1980). tochemical Potential of Tropical Plants (K. R. Downum, J. T.
KUTCHAN, T. M., N. HAMPp, F. LoTrsPEICH, K. BEYREUTBR, and Romeo and H. A. Stafford, eds.), 1-40, Plenum Press, New
M. H. ZENK, The cDNA clone for strictosidine synthase from York,1993.
Rauvolfia serpentina, FEBS Lett., 237, 40-44 (1988). QUETIN-LBCLBRCQ, J., R. WARIN, N. G. BISSET, and L. ANGBNOT,
KUTNBY, J. P., T. HIBINo, E. JAHNOBN, T. aKUTAN!, A. H. RAT- Venecurine, an indole alkaloid from curare, Phytochemistry,
CLIFFE, A. M. ThBAsURYWALA, and S. WUNDERLY, Total synthe- 28, 2221-2223 (1989).
sis of indole and dihydroindole alkaloids. IX. Studies on the RAYMOND-HAMBT, M., L'!BOGA, drogue detatigante mal connue,
synthesis of bisindole alkaloids in the vinblastine-vincristine Bull. Acad. Nat. MOd., 124, 243-255 (1941).
6S4 Indole Alkaloids

ROBINSON, T., Metabolism and function of alkaloids in plants, Sci- M. Dey and J. B. Harborne, eds.), 309-371, Academic Press,
ence, 184, 430-435 (1974). London, 1993.
ROBINSON, T., THE EVOLUTIONARY ECOLOGY OF ALKALOIDS, IN Her- TYLER, V. E., L. R. BRADY, and J. E. ROBBERS, Pharmacoguosy,
bivores. Their Interaction with Secondary Plant Metabolism (G. 8th ed., Lea and Febiger, Philadelphia, 1981.
A. Rosenthal and D. H. Janzen, eds.), 413-448, Academic VAN DER HEDDEN, R., P. 1. LAMPING, P. P. OlIT, R. WUNSMA, and
Press, New York, 1979. R. VERPOORTE, High-performance liquid chromatographic de-
RilFFBR, M., N. NAGAKURA, and M. H. ZENK, Strictosidine, the termination of indole alkaloids in a suspension culture of Taber-
common precursor for monoterpenoid indole alkaloids with 3a naemontana divaricata, J. Cbrornatogr., 396, 287-295 (1987a).
and 313 configuration, Tetrahedron Lett., 1593-1596 (1978). VAN DER HEUDBN, R., A. HERMANS-LoKKERBOL, R. VBRPOORTE,
SCHULTES, R. E. and A. HOFMANN, The Botany and Chemistry of and A. BAERHEIM-SVENDSEN, Pharmacoguostical studies of Ta-
Hallucinogens, CC Thomas, Springfield, IL, 1973. bernaemontana species. XX. Ion-pair droplet countercurrent
SCHLBIFFER, H., Narcotic Plants of the Old World, Lubrecht and chromatography of indole alkaloids from suspension cultures,
Cramer, Monticello, NY, 1979. J. Cbromatogr., 396, 410-415 (1987b).
SCOTI, A. I., Indole alkaloid biosynthesis, Heterocycles, 15, VAN DBR HEUDEN, R., A. HERMANs-LoKKERBOL, J. P. J. DE KOOL,

1257-1274 (1981). P. J. LAMPING, P. A. A. HARKEs, and R. VBRPOORTE, Accumula-

tion of indole alkaloids in a suspension culture of Tabernaemon-
SMITH, G. N., STRIcroSIDINE: A key intennediate in the biogenesis
of indole alkaloids, Chern. Commun. 912-914 (1968). . tana divaricata, Planta Medica, 44, 393-397 (1988).
VBRPOORTB, R., R. VAN DER HEIJDEN, W. M. VAN GULIK. and H.
SPECIALIST PERIODIC REPoRTS, Alkaloids, Vol. la, Chemistry Soci-
J. G. TEN HOOPEN, Plant biotechnology for the production of
ety, London, 1981.
alkaloids: Present status and prospects, in The Alkaloids, Vol.
STiicKIGT, J., The biosynthesis of heteroyohimbioe-type alkaloids, 40 (A. Brossi, ed.), 1-188, Academic Press, New York, 1991.
in Indole and Biogenetically Related Alkaloids (J. D. Phillipson
WAu., M. E., M. C. WAN!, C. E. COOK, K. H. PALMER, A. T.
and M. H. Zeuk, eds.), 113-141, Academic Press, London,
McI'HAIL, and G. A. SIMS, Plant antitumor agents. I. The isola-
1980.
tion and structure of camptothecin, a novel alkaloidal leukemia
STiicKIGT, J. and M. H. ZENK, Strictosidioe (isoviocoside): The key and tumor inhibitor from Camptotheca acuminata, 1. Am.
intennediate in the enzymatic fonnation of monoterpene indole Chern. Soc., 88, 3888-3890 (1966).
alkaloids, J. Chern. Soc., Chern. Commun. 646-648 (1977).
WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and Metabo-
SWART, K. L., J. P. KUTNEY, T. HONDA, and B. R. WORTH, Studies lism in Plants, Plenum Press, New York, 1978.
on the biosynthesis of bisindole alkaloids. The fmal stages in
WINK. M., Allelochemical properties or the raison d'etre of alka-
biosynthesis of vinblastine, leurosine, and catharine. Heterocy-
loids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), 1-118,
cles, 9, 1391-1395 (1978).
Academic Press, New York, 1993.
SUFFNESS, M. and G. A. CORDBU., Antitumor alkaloids, in The
WRlGHT, C. W., D. AliEN, J. D. PHILLIPSON, G. C. KIRBy, D.
Alkaloids, Vol. 25 (A. Brossi, ed.), 3-356, Academic Press,
C. WARHURST, G. MASSIOT, and L. LE MEN-OUVIER, Alstonia
New York, 1985.
species: are they effective in malaria treatment? 1. Ethnophar-
SzANrAY, C., G. BLASKO, K. HoNTY, and G. DORNYBI, Corynanth- macol., 40, 41-45 (1995).
eioe, yohimbine, and related alkaloids, in The Alkaloids, Vol. ZeNK, M. H., H. EL-SHAGI, H. ARENS, J. STiicKIGT, E. W. WEILER,
27 (A. Brossi, ed.), 131-269, Academic Press, New York, 1986. and B. DEUS, Fonnation of the indole alkaloids serpentine and
TAYLOR, W. I. and J. LE MEN, A mtiform numbering system for ajmalicine in cell suspension cultures of Catharanthus roseus,
iodole alkaloids, Experientia, 21, 508-510 (1965). in Plant Tissue Culture and its Biotechnological Application
THOMPSON, H., Fear and Loathiog: On the Campaigu Trail, Popular (W. Barz, E. Reiohard, and M. H. Zeuk, eds.), 27-43, Springer-
Library, New York, 1974. Verlag, Berlin, 1977.
'i'Iu.EQUIN, F., S. MICHEL, and E. SEGUIN, Tryptamine-derived in- ZBNK, M. H., M. RilFFBR, M. AMANN, and B. DEUS-NEUMANN,
dole alkaloids, in Alkaloids and Sulphur Compounds (P. G. Benzylisoquinoline biosynthesis by cultivated plant cells and
Waterman, ed.), Vol. 8 of Methods in Plant Biochemistry (P. isolated enzymes, J. Nat. Prod., 48, 725-738 (1985).
35
Ergot and Other Indole Alkaloids

Introduction ceae, Sterculiaceae, and Verbenaceae are summarized in the

Ergot Alkaloids present chapter.
Biosynthesis
Biosynthesis of Peptide-Containing Ergot Alkaloids
Distribution of Ergot Alkaloids
Ergotism ERGOT ALKALOIDS
Biological Properties
Endophytes Indole alkaloids involving a hemiterpene unit have long been
Lysergic Acid Diethylamide known as mycotoxins and pharmaceutical products. Al-
Medicinal Properties though these alkaloids are best known from the fungal genus
Commercial Production of Ergot Alkaloids Claviceps, they also are found in several genera of the higher
Harmane or [:I-Carboline Alkaloids plant family Convolvulaceae. ReCently, ergot alkaloids also
Biosynthesis have been found in certain groups of endophytic fungi (An-
Distribution of Harmane Alkaloids tkowiak and Antkowiak, 1991). Ergot alkaloids may be di-
Biological Activity vided into three major groups: clavine alkaloids, simple lys-
Biosynthetically Related Alkaloids ergic acid alkyl amides, and peptide derivatives of lysergic
Cantbin-6-one and Related Alkaloids acid amide. Two of the most important ergot aIkaloids are
Carbazole Alkaloids lysergic acid (1) and agroclavine (2). Alkaloids of the
Alkaloids of Physostigma venenosum agroclavine type have a CH20H or methyl group at C-8 and
Alkaloids of the Calycanthaceae lysergic acid types have a carboxyl or carboxamide group
Polyindolenines at that center (Fig. 35.1) (Antkowiak and Antkowiak, 1991;
Other Alkaloids with Indole Skeleta Geissman and Crout, 1969; Mann, 1987). Free lysergic acid
References usually is not accumulated by Claviceps species, although
the simple amide ergine (3) has been isolated from Claviceps
paspa/i and from Ololiuqui, a hallucinogenic preparation
INTRODVcnOI'l from seeds of Ipomoea violacea and Rivea corymbosa (An-
tkowiak and Antkowiak, 1991). Cell-free extracts from
Many aIkaloids contain an indole nucleus as a part of their Claviceps species that synthesize clavine aIkaloids have
structure. Monoterpene-derived indole aIkaloids (see Chap- been obtained.
ter 34) are the most important of these compounds, but sev- Analytical methods for the detection and analysis of ergot
eral other significant types of indole aIkaloids exist. Ergot alkaloids by high-perfonnance liquid chromatography
alkaloids fonned from tryptamine and a mevalonate-derived (HPLC) and tandem mass spectrometry (Yates and Powell,
hemiterpene unit are of medical importance. Others, such as 1988), and both the IH_ and I3C-NMR (nuclear magnetic
harmane aIkaloids, physostigmine and a series of structurally resonance) spectra for these compounds have been reviewed
similar aIkaloids, Calycanthus alkaloids (Calycanthaceae) (Crabb, 1982). The results of feeding studies leading to the
and related compounds (principally from the Rubiaceae), fonnation of ergot alkaloids have been tabulated (Leete,
and relatively minor types of aIkaloids from the Ascelpiada- 1983).

IiSS
656 Ergot and Other Indole Alkaloids

17 H
Ho2e

agroclavine (2) lysergic acid (1) ergotamine (9)

HO HO

elymoclavlne (5) chanoclavine-J (7) 4-dimethylallyl- ergine (3)

tryptophan (4)

Fig. 35.1. Lysergic acid and agroclavine.

Biosynthesis indicates that a ZoE (cis-trans) isomerization must occur

in the formation of chanoclavine I (Floss, 1976, 1980). By
The biosynthesis of ergot alkaloids is initiated by prenyla- feeding 4-(-y;y-dimethylallyltryptophan)-(Z- 14CH3 ) and de-
tion of tryptophan with dimethylallyl pyrophosphate grading chanoclavine I (7), agroclavine (2), and elymoclav-
(DMAPP) to yield 4-dimethylallyltryptophane (DMAT) (4) ine (5), it was established that a ZoE isomerization occurs
(Fig. 35.2) (Floss, 1980). This alkylation step is believed to
(Floss, 1976). Additional evidence indicates that methyl
be rate limiting, and the enzyme that catalyzes this reaction,
group hydroxylation occurs before the closure of ring C,
DMAT synthetase, has been purified more than 60 times
formation of an epoxide may be involved in this oxidation
(Maier and Groger, 1976; Waller and Dermer, 1981). The
(GrOger, 1978). Chanoclavine I (7) is oxidized to the corre-
enzyme appears to be specific for L-tryptophan and is inhib-
sponding aldehyde (8) and this is cyclized to produce
ited by agroclavine (2) and elymoclavine (5) the end prod-
agroclavine (2). Another ZoE isomerization· is involved at
ucts of the reaction.
this stage. This rearrangement involves addition to the dou-
Mixed-function oxidase enzymes from ergot are capable
ble bond followed by elimination of the original vinyl proton
of converting DMAT (3) to the corresponding hydroxy de-
rivative (HODMAT) (6) have been studied (Maier and (Fig. 35.3). However, in subsequent addition steps, the en-
Groger, 1976; Waller and Dermer, 1981). zyme-proton complex (that now bears the original vinyl
Cyclization of a 4-dimethylallyltryptophan (4) intermedi- proton) can be added across the double bond of another mol-
ate yields chanoclavine (7). Many of the same mixtures of ecule of the aldehyde (8). This "recycling" of protons has
enzymes also produced chanoclavine I (7), the precursor for been confirmed by double-labeling experiments (Floss,
several ergot alkaloids with two rings as well as the agroclav- 1980). If ttitium is incorporated into the hydroxymethyl
ine and lysergic acid series. The synthesis of rings C and D group of chanoclavine I (7), half of the label is lost in the
has been studied with multiply-labeled mevalonic acid and subsequent formation of tetracyclic ergolines.
related precursors in work done primarily by Floss (1976, Agroclavine is converted into elymoclavine (5) and ulti-
1980). The reaction is mediated by the chanoclavine-I cy- mately into lysergic acid (1). An enzyme preparation that
clase enzyme complex. The enzyme is reasonably specific, catalyzes the aerobic conversion of agroclavine to ely-
requires ATP, a pyridine nucleotide, and Mg2+ for activity. moclavine (in addition to several other types of activity) has
Several cyclases of this type probably exist: one produces been studied. Feeding experiments with mevalonate-3-D3
agroclavine (2) and another elymoclavine (5) (Waller and established that both methylene hydrogens at C-17 of cha-
Dermer, 1981). Mevalonate-2- 14C specifically labels the C- noclavine I (7), but only one (at C-7) of elymoclavine (5)
17 methyl group of chanoclavine I (7). The pro-4R hydrogen comes from C-3 of mevalonate. The hydrogen that remains
of mevalonate is retained during alkaloid formation. This was shown to be the pro-7S hydrogen.
 Ergot and Other Indole Alkaloids 657

Biosynthesis of Peptide-Containing diate Iysergyltripeptide is linked to the enzyme and subse-

Ergot Alkaloids quently cyclized (Floss, 1980).

The structure of ergotamine (9) is a peptide alkaloid in

Distribution of Ergot Alkaloids
which a lysergic acid moiety is linked to alanine, phenylal-
anine, and proline. The manner of attachment of the amino Although best known from Claviceps species, these alka-
acids is not clear. Neither dipeptides or tripeptides are loids also are known to occur in other fungi, including Peni-
added in the synthesis. Lysergylalanine does not appear cillium, Aspergillus, and Rhizopus species. Ergot alkaloids
to be incorporated as an intact unit. Although the amino are also known to occur in certain higher plants, notably
acids were incorporated, the peptides appear to have been in the seeds of Argyreia, Convolvulus, Cuscuta, Ipomoea,
broken down and the component amino acids used. A Operculina, Rivea, Strictocarpia, and Turbina (Convolvula-
crucial step seems to be the hydroxylation of the amino ceae), but also in the leaves, stems, and roots of some species
acid directly linked to lysergic acid (Groger, 1978). Other (Amor-Prats, personal communication). Ololiuqui is a hallu-
major problems involve the formation of the cyclic struc- cinogenic drug used by the Indians of Mexico, made from
ture of the final product, ergotamine. Floss et aJ. (1974) Ipomoea violacea and Rivea corymbosa. These seeds contain
proposed that peptide chain formation takes place in a elymoclavine (5), chanoclavine-I (7), and lysergic acid
concerted fashion on a mUltienzyme complex. The interme- amide (10), the principal hallucinogenic constituent of ololi-

~ 'criNH2C02H

opp

~ I ~
~ NH

4-dimethylallyl. N.methyl-4-dimethyl-
tryptophan (4) allyltryptophan

o
HO

hydroxydimethylallyl chanoclavine-I (7) chanoclavine-I

-tryptophan (6) aldehyde (8)

CH H
HN' 3 H02C~1
N/CH3
H H
~

-+ ---+

c? I ~
~ NH

agroclavine (2) elymoclavine (5) lysergic acid (1)

Fig. 35.2. Biogenesis of lysergic acid (modified from Floss, 1980).

658 Ergot and Other Indole Alkaloids

-
H H

enzX

--
X-enz

--
Fig. 35.3. Isomerization of the double bond in the conversion of chanoclavine I to ergolines (modified from Floss, 1976; used with pennission of the
copyright owner, Elsevier Science Ltd., The Boulevard, Langford Lane, Kidlington OXS 1GB, UK).

uqui (Fig. 35.4). In general, the yield of ergot alkaloids from Ergotism is still an important disease of domestic animals.
higher plants is much less than from fungi. Cattle that feed on infested fields are regularly poisoned by
the sclerotia of Claviceps (Thomas and Basset, 1970).
Ergotism
Biological Properties
Ergot has been responsible for poisoning of both human
and domestic animals for thousands of years. Accounts are Ergocryptine (11) has an LOso Lv. in rabbit of 1.1 mg/kg,
known from the ancient Chinese, Greeks, Romans, and oth- ergocornine (12) of 1.2 mgikg, ergometrine (13) in mouse
ers. Ergot was not recognized as a fungus until 1764 by von of 0.15 mg/kg, and ergotamine (9) of 62 mg/kg (Fig. 35.5)
Munchhausen. In humans, poisoning usually occurred by (Yates et aI., 1989).
consumption of bread prepared from infested grain, typically Ergonovine (13), ergotamine (9), ergocryptine (11),
rye. In the Aquitane region of France in A.D. 944, about half agroclavine (1), and elymoclavine (5) reduced larval weights
of the popUlation died of ergot poisoning. Ergotism was a and amount of com leaf disks eaten by larvae of the fall
problem in Europe through much of the Middle Ages, but armyworm, Spodoptera /rugiperda, compared to controls
it has been largely eliminated by better grain-handling tech- (Clay and Cheplick, 1989). Ergocryptine (11), ergometrine
niques, inspection of grain, and pooling of grain supplies. (13), and ergotamine (9) are feeding deterrents to Syntomis
Some alkaloids of ergot possess potent vasoconstricting larvae (Lepidoptera) or are toxic to Oncopeltus (Hemiptera)
abilities; these were largely responsible for one of the effects (Wink, 1993).
of ergotism called SI. Anthony's Fire, a gangrenous condi-
tion of the extremities which often resulted in loss of limbs. Endophytes
Other related alkaloids produced delirium and hallucina- Although many plants are infected by fungal parasites,
tions, and, in more severe cases, convulsions. others are infested with endophytic fungi. These fungi live

o NH,

HO

lysergic acid (1) lysergic acid amide (10) lysergic acid diethylamide (14) 8~hydroxylysergic acid (15)
(LSD)

Fig. 35.4. Biologically active ergot aIkaloids.

 Ergot and Other Indole Alkaloids 659

'/:~~ "/:,l~
0 0

H H 0 0 H H 0 0
H H

~ b
ergotamlne (9) ergocryptine (11)

,/:f~
0 0

H H 0 0
H

ergocornine (12) ergometrine (13)

(ergonovine)

FIg. 35.5. Peptide-containing ergot alkaloids.

entirely within the leaf and stem tissues of the plant. Endo- of tall fescue, however, are of an unusual pyrrolizidine type
phytes of grasses (Poaceae) and sedges (Cyperaceae) are (Clay, 1988).
especially common and important (Clay, 1989). The fungi Infected individuals of several grass species are more re-
are members of the family Clavicipitaceae (Ascomycetes), sistant to a number of insect herbivores (Clay, 1988). Fur-
tribe Balansiae. Five genera are especially common: Atkinso- ther, seeds of infected species are more resistant to herbivore
nella, Balansia, Balansiopsis, Epichloi!, and Myriogen- attack (Cheplick and Clay, 1988). Lysergic acid amide (10),
osporae. A few species are epiphytic; however, none pro- isolysergic acid amide, 8-hydroxylysergic acid (15), ergono-
duce the haustoria common to parasitic fungi. Most of these vine, chanoclavine 1(7), andN-fonnylloline all were isolated
fungi produce fruiting bodies on the leaves or aborted inflo- from Stipa robusta infected with an Acremonium endophyte
rescences of host species, although others grow into the re- (Petroski et al., 1992; Powell and Petruski, 1992).
productive tissues of the plant and are maternally transmit-
ted. Many endophytes are transmitted through seeds of their Lysergic Acid Diethylamide
hosts. As is true for Claviceps infections of grasses, poison-
Lysergic acid diethylamide (LSD = Lysergsiiurediethy-
ing of herbivores by endophytes is well known. The cause
lamid) (14) was first syuthesized by A. Hoffman in 1943.
of many livestock poisoning problems of this narure was
This compound, which is not naturally occurting, is well
long mysterious, as obvious infection was not readily seen.
known for its hallucinogenic effects. These effects are proba-
Poisoning by tall fescue (Festuca arundinacea), orchard
bly due to an inhibition of a basic brain stem mechanism that
grass (Dactylis glomeratus), perennial ryegrass (Lotium per-
integrates sensory input. LSD apparently occupies serotonin
enne), annual ryegrass (Lotium tementulum), and species of
receptor sites and potentiates noradrenaline systems (Cor-
Stipa and Metica (often called "sleepy" or "drunk" grass)
dell, 1981).
is now known to involve endophytes (Clay, 1989; Petroski
et al., 1992; Powell and Petroski, 1992; Powell, and Yates
Medicinal Properties
1988).
Many endophytic fungi produce alkaloids; uninfected Ergot alkaloids are adrenergic blocking agents. These
host plants lack these compounds. The alkaloids of most of compounds also inhibit lactation in animals by inhibition
these grasses (presumably produced by the endophytes) are of prolactin release, a mammalian hormone responsible for
similar to those of Claviceps species described above. Those mammary and milk production (Fliickiger, 1980; Stadler and
660 Ergot and Other Indole Alkaloids

Stiitz, 1975). As prolactin is necessary for induction and tetrahydroharman are all incorporated into harman. These
growth of certain tumors in mice, ergocryptine (11) and er- results suggest that a second pathway involving N-acetyl-
gocornine (12) significantly reduced the size and incidence tryptamine is involved in the formation of harmane alkaloids
of these tumors. Ergometrine (13) and ergotamine (9) are (Fig. 35.6).
used as uterine-contracting agents following childbirth.
These uses are based on the well-established vasoconstric- Distribution of Hannane Alkaloids
tion action of these alkaloids. Both are amides of lysergic
acid (Cordell, 1981). Harmane or f3-carboline alkaloids have been reported
Jivaro women seem to control family size by determining throughout the family Rubiaceae (Hemingway and Phil-
the time they become pregnant. They do this by ingesting lipson, 1980). These alkaloids also occur in the Acanthaceae
infusions of Cyperus species at specified times. Women also (Adhatoda), Apocynaceae (Aspidosperma, Alstonia, Pi-
use a decoction made from the fungus Balansia cyperi to crasma), Eleagnaceae (Eleagnus), Fabaceae (Petalostylis,
aid in parturition or postpartum contractions and to reduce Gastrolobium), Loganiaceae (Strychnos), Malpighiaceae
bleeding following childbirth. In both instances, the presence (Banisteriopsis caapi), Passifloraceae (Passiflora), Ranun-
of fungi related to ergot may be involved (Lewis, 1992). culaceae (TiullictrumJoetidum), Rutaceae (Murraya, Clau-
seno), Simaroubaceae (Ailanthus), Solanaceae (Vestia), and
Commerdal Production of Ergot Alkaloids Zygophyllaceae (Peganum) (Cordell, 1979; Faini et aI.,
1978; Husson, 1985; Joshi et aI., 1977; Schiff, 1987).
Ergot has been used medicinally since the 1500s. Most Many of these plants also are known to contain other
of that used is from cultivated ergot. About 12,000 kg per alkaloids; reports of f3-carboline alkaloids should be con-
year is produced for a total value of about $50 million. Ergot firmed in some cases. Species of Ailanthus also contain can-
alkaloids are produced parasitically on rye, in saprophytic thine derivatives (see below) (Cordell et aI., 1978).
culture (usually with Claviceps paspali), or by partial or
total chemical synthesis. BiologiCal Activity
Harmane alkaloids inhibit membrane transport and act at
HARIIIAI'IE OK ~·CAKBOLII'III ALKALOIDS serotonin receptors (Robinson, 1979). Harmine (16) inhibits
monoamine oxidases and binds to DNA. Harmaline (17)
inhibits (Na+,K+)-ATPase, Na+ transport, and monoamine
Alkaloids based on the tetrahydro-f3-carboline system are
not common, but occur in a number of families. Several of oxidase (Wink, 1993).
these harmane or f3-carboline alkaloids occur in Peganum Alkaloids of this type are responsible for the hallucino-
iulrmala (Zygophyllaceae). AIta-ur-Rahman and Basha, genic activity of a number of plants. Among these are Pega-
1983. num harmala (Zygophyllaceae) and Banisteriopsis caapi or
Heterotrophic cultures of Peganum iulrmala accumulate (ayahuasca or yage) (Malpighiaceae), a plant used as a hallu-
about 0.1 % of their dry weight as harmine (16), harmaline cinogenic drink by Indians in Amazonian and Andean South
(17), harmol (18), and harmalol (19) (Berlin and Sasse, 1988; America. At oral doses of 4 mg/kg, harmaline (17) produces
Ellis, 1988). psychotomimetic activity in humans. High intravenous doses
The results of feeding studies leading to the formation of of harmine (16) also produce hallucinogenic effects (Neuss,
harmane alkaloids have been tabulated (Leete, 1983). 1980); harmine has an LDso i.v. in mouse of 38 mglkg
(Wink, 1993). The seeds of Peganum iulrmala are 2-3%
alkaloids. Certain alkaloids of this group have pronounced
Biosynthesis
hypotensive activity (Husson, 1985; Nuess, 1980).
The formation of harmine (16) is similar to the formation Harmine (16), harmol (18), and harmaline (17) are active
of pellotine (see Chapter 32), except that a tryptamine pre- against the epimastigotes of Trypanosoma cruzi at 1.9-5
cursor is utilized (Fig. 35.6). Tryptophan has been shown to fl.g/ml (Borris and Schaeffer, 1992; Phillipson and Wright,
be an effective precursor of eleagnine (20) and harmane (21) 1991). Harmaline (17) shows activity against Leishmania
in Eleagnus angustifolia and Passiflora edulis. The incorpo- mexicana amazonensis at 24 fl.g/ml, whereas 6-methoxy-N-
ration of [f3_ 14C- 1SN]_tryptamine into harmane (21) with methyl-l,2,3,4-tetrahydro-f3-carboline (24) from Nectandra
retention of isotopic ratio in Peganum iulrmala (Zygophylla- megapotamica (Lauraceae) inhibits the growth of Crithidia
ceae) suggests that a three-carbon pyruvate unit undergoes Jasciculata. 4-Methoxy-I-vinyl-f3-carboline (25) and the
condensation with tryptamine and subsequent cyclization. corresponding 6-hydroxy derivative (26) from Picrasmia ja-
Also, in the formation of eleagnine in Eleagnus angustifolia, vanica (Simaroubaceae) inhibit multidrug-resistantPlasmo-
the additional two carbon atoms come from pyruvic acid dium Jalciparum in vitro (Fig. 35.7) (Borris and Schaeffer,
(Fig. 35.6). The key intermediate (22) serves as a more effi- 1992).
cient precursor than either tryptophan or pyruvic acid (Hus- Harmaline (17), harman (21), norharman (27), and har-
son, 1985). However, in Passiflora edulis, N-acetyltrypto- mine (16) elicit phototoxicity in larvae of Trichoplusia (Lep-
phan, N-acetyltryptamine, harmalan (23), and idoptera), and serve as feeding deterrents to larvae of the
 Ergo/ and Other Indole Alkaloids 661

~H:­
~N) ~
~-
~~)'\~~
I 'CO,H

(Ir-llH
~NH")<:'
s:cY
"""
-- (In()HI
NH
I
0
N
~NHI'

e:y""
CO2"
(22) eleagnine (20)
/ harmalan (23)

I I N
CH,-O """ NH 0

harmine (16)

Fig. 35.6. Biogenesis of harrnane.

CH,-O
N=:C? ~
I

harmaline (17)
NH
I
h
N

harman (21)
Q:O
~
I NH
I
~
N

norharman (27)
h

~
~
~
I I N I I N
HO ~ NH h HO ~ NH h

harmol (18) harmalol (19)

OCH,
CH,-O
CH'-On9 n
4 ;, ?'

7 ~
I NH
I N'
'CH
~
8 1 '

6~methoxy-N-methyl-l,2,3,4- 4-methoxy-l-vinyl- 6-methoxy-l-vinyl-

tetrahydroxy-p-carboline (24) p -carboline (25) p-carboline (26)

Fig. 35.7. Biologically active hannane alkaloids.

662 Ergot and Other Indole Alkaloids

borreverine (29) isoborreverine borrerine (28)

Fig. 35.8. Borrerrine and related compounds (modified from Saxton. 1981; used with pennission of the copyright owner. the Royal Chemical Society,
Cambridge).

polyphagous lepidopteran Syntomis (Wink, 1993). Both har- occur in the Leitoeriaceae (K. Readel and D. Seigler, unpub-
mine (16) and harmalol (19) are reputed to bind to insect lished data).
synapses (Wink, 1993). Some of the plants that contain canthin-6-one (30) and
its relatives are used medicinally. I-Methoxycanthin-6-one
BiosyntbeticaUy Related AIkaloids (31) is found in Hannoa klaineana (Simaroubaceae) roots.
This plant is used to treat fever and intestinal diseases in
Borrerine (28), a compound probably analogous in syn- Zaire (Lumonadio and Vanhaelen, 1984). Brucea javanica
thesis to lophocereine but belonging to the (3-carboline series is used to treat malaria and dysentery as well as being an
of alkaloids, has been found in Borreria verticil/ata (Rubia- insecticide. Another simaroubaceous plant, Eurycoma longi-
ceae), along with a dimeric compound borreverine (29) (Fig. folia, also is used to treat dysentery, glandular swelling, per-
35.8). This alkaloid has also been found in Flindersia four- sistent fever, and tertian malaria (Kardono et al., 1991). The
nieri (Rutaceae) (Cordell and Saxton, 1981; Saxton, 1981). major alkaloids of this plant were 9-methoxycanthin-6-one
(32), 9-hydroxycanthin-6-one, and the corresponding N-ox-
ides (Kardono et al., 1991). The canthin-6-one derivatives
CAl'I11UN·6·0NE Al'ID RELATED ALKALOIDS were active against both KB and P-388 cell lines (Kardono
et al., 1991).
Canthin-6-one (30) and a series of related alkaloids are found Cultores of Ailanthus altissima and Brucea javanica pro-
in one species of Pentaceras, several species of Zanthoxylum duced canthin-6-one (30) and 1-methoxycanthin-6-one (31)
(Rutaceae), and several species of the Simaroubaceae (Fig. as well as a number of minor, related alkaloids (Ellis, 1988;
35.9) (Waterman and Khalid, 1981). These alkaloids also Liu et al., 1990).
have been reported from the Malvaceae, Caryophyllaceae, 1-Methoxycanthin-6-one (31), lO-hydroxycanthin-6-one
and Amaranthaceae (Tillequin, 1993). Similar compounds (33), and lO-methoxycanthin-6-one are cytoxic against car-

'59"
""
I
o
N
I
.&
-<
N

'59
""
1

o
N
IO~:'
.&
-<

canthinoo6..one (30) I-methoxycanthin-6-one (31)

CH,-O
~"" ""
1

o
N
1

.&
-<
N

10-hydroxycanthin-6-one (33) 9-methoxycanthin-6-one (32)

Fig. 35.9_ Canthin-6-one and related alkaloids.

 Ergot and Other Indole Alkaloids 663

"'" 000"'"
CCJY
OH

:::,...
I NH
I ~:::,...
I I ~
OCH, NH

1·methoxy·3·hydroxymethylcarbazole (37) girinimhin (34) 3-methylcarbazole (36) carbazole (35)

(koenoline)

Fig. 35.10. Carbazole alkaloids.

cinoma of the nasopharynx (KB) (Liu et al., 1990; Wink, ity (Chakraborty and Roy, 1991). Oxidative demethylation
1993). also appears to occur.
Koenoline (l-methoxy-3-hydroxymethylcarbazole) (37)
has been isolated from the root bark of Murraya koenigii, a
tree widely used in India to flavor foodstuffs (Fiebig et aI.,
CARBAZOLE ALKALOIDS 1985). This alkaloid has cytotoxic activity toward KB cells.

Carbazole alkaloids are found in the genera Clausena, Gly-

cosmis, Micromelum, and Murraya of the Rutaceae. There ALKALOIDS OF P/lYSOSTIG/I1A VEl'IElYOSUM
is a single report from the genus Ekebergia of the Meliaceae
(Bhattacharyya and Chakraborty, 1987; Chakraborty and The calabar bean (the seed of Physostigma venenosum, Fa-
Roy, 1991). baceae) was formerly used for "trial by ordeal" in Eastern
Girinimbin (34) (Fig. 35.10) is a common alkaloid in Nigeria in an area known as the Calabar Coast (Holmstedt,
several species of Murraya sect. Bergera (Kong et al., 1972). By this method, the guilt or innocence of the accused
1988a). The distribution of these alkaloids and 3-prenylin- is determined by whether that person survives after ingesting
doles and 8-prenylated coumarins strongly supports division extracts or concoctions prepared from the plant. The seeds
of this group into two genera, Murraya and Bergera (Kong of the calabar bean contain a number of indole alkaloids
et al., 1986, 1988a,b) (see Chapter 31). derived from tryptamine and S-adenosyl methionine. A con-
The isolation of several 3-methylcarbazole derivatives certed ring closure and nucleophilic attack on S-adenosyl-
from higher plants and the isolation of carbazole (35) shows methionine appears reasonable, but neither this mechanism
that the aromatic methyl group can be eliminated from the nor the origin of the carbamate group have been well studied
key compound 3-methylcarbazole (36) via oxidation to the (Fig. 35.11).
alcohol, the aldehyde, and, subsequently, the acid functional- Physostigmine (38), with a pyrrolo[2,3b lindole system,

(O~CH'_S'-

I I NH
/ ~ NV 2

(Jc)'1:
CH3
C02H
CH'NHCO-O~

I I NH
~ NH 2
~NAN)
I I
tryptophan CH] CH]

(O~ (o~CH3/ physostigmine (38)

/ U~) NH
~H2--
~
I
NHNH

Fig. 35.11. Biogenesis of physostigmine (modified from Geissman and Crout, 1969),
664 Ergot and Other Indole Alkaloids

the principal active ingredient from the seed of Physostigma Somewhat similar alkaloids have been isolated from skin
venenosum, Fabaceae, is a reversible cholinesterase inhibitor secretions of Australian frogs (~aly et al., 1990).
used in the treatment of glaucoma. It is employed in ophthal-
mology after administration of atropine to return the pupil to
its normal size (Neuss, 1980; Tillequin, 1993). This alkaloid ALKALOIDS OF THE CALYCAm1IACEAE
causes accumulation of acetylcholine and continuous stimu-
lation of the cholinergic receptors. As is true for most other A number of highly complex, multinuclear indole alkaloids
cholinesterase inhibitors, physostigmine also stimulates se- (indolenines) are found in the family Calycanthaceae (Ca/y-
cretions such as saliva and perspiration and increases tone canthus and Chimonanthus) (Cordell and Saxton, 1981).
and peristalsis of the gastrointestinal tract (Cordell, 1981). This family is closely related to several benzylisoquinoline-
Physostigmine has an oral LOs. in mouse of 4.5 mg/kg alkaloid-producing families, but lacks the characteristic al-
(Wink,1993). kaloids of that group (see Chapter 32). Similar compounds
Physostigmine is a feeding deterrent to polyphagous Syn- have been found subsequently in the genera Bhesa (Celastra-
tomis larvae (Wink, 1993). ceae) and Palicourea (Rubiaceae). Both calycanthioe (39)

O:::():
I I
CO.H

QJ()HCH~ ~-
7

~N~ NHCH,
~ NH.~
NH

tryptophan

calycantbine (39)

ofI})
CH,
I H

",... HN i N
HI
CH,
meso-chimonanthine meso-calycanthine
Fig. 35.12. Biogenesis of Calycantbaceae alkaloids (modified from Geissman and Crout, 1969),
 Ergot and Other Indole Alkaloids 665

CH, CH,

NNI:
~
,... I """'--_L-_
N N
CH, CH,
folicanlhine (40) qnadrigemine B quadrigemine A (42)
(+ diastereomer)

psycholridine (11)
hodgkinsine (41)
Fig. 35.13. Complex oligomers of tryptophan-derived precursors.

and chimonanthine also occur in Palicourea lentileri (Fig. iaceae) contains such compounds as hodgkinsine (41), in
35.12) (Hemingway and Phillipson, 1980). which three tryptophan units are involved, and quadrigemi-
Labeled tryptophan is incorporated into calycanthine (39) nine A (42). The latter contains four tryptophan units linked
and folicanthine (40) in Calycanthus florida (Cordell and into a tetrameric structure (Fig. 35.13). Psychotridine (43),
Saxton, 1981). the major alkaloid of Psychotria beccarioides, is a pentamer
based on similar precursors (Cordell and Saxton, 1981). In
POLYIl'IDOLENIl'IES addition to the simple harman-type alkaloids, species of the
genus Borreria (Rubiaceae) also contain dimeric forms of
Alkaloids similar to those of the Calycanthaceae have been tryptamine-derived alkaloids (Hemingway and Phillipson,
found in several other plants. Hodgkinsoniafrutescens (Rub- 1980).

OCH,
~10Wo
olf "
melosatin A (44) melosatin B (45) cryptolepine (47)

trichotomine (46) trichotomine G.

Fig. 35.14. Miscellaneous indole alkaloids.

666 Ergot and Other Indole Alkaloids

Ol1IER ALKALOIDS WITH Il'IDOLE SKELETA constituents of Ailanthus excelsa (Simaroubaceae), Lloydia, 41,
166-168 (1978).
Melochia tomentosa, a tumorigenic plant of the Sterculia- CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual
ceae, has been shown to contain the alkaloids melosatin A Reports on NMR Spectroscopy, Vol. 13 (G. A. Webb, ed.),
and B (44, 45) that contain an indole nucleus and are quite 60-210, Academic Press, London, 1982.
unusual in structure (Kapadia et al., 1977). DALY,1. W.. H. M. GARRAFFO, L. K. PANNELL, T. F. SPANOE, C.
The bright blue color of fruits of Clerodendron trichoto- SBVBRJNl, and V. ERsPAMER, Alkaloids from Australian frogs
mum (Verbenaceae) is due to the presence of bisindole alka- (Myobatrachidae): Pseudophrynamines and pumiliotoxins, J.
loids such as trichotomine (46). The blue color of these alka- Nat. Prod., 53, 407-421 (1990).
loids is uncommonly encountered in nature (Fig. 35.14) ELLIS, B. E., Natural products from plant tissue culture, Nat. Prod.
(Cordell and Saxton, 1981). Rep .. 5, 581-612 (1988).
Cryptolepine (47) from the roots of Cryptolepis sanguin- FAINI, F., M. CASTILLO, and R. TORRES, A new j3-carboline alkaloid
olenta (Asciepiadaceae), is highly active against Plasmo- from Vestia Iycioides, Phytochemistry, 17, 338 (1978).
dium Jalciparum. The in vitro IC50 value for this alkaloid FIEBIG, M., 1. M. PBzzUTO, D. SOEJARTO, and A. D. KINoHORN,
is 0.03 11g!m1, compared to 0.07 l1g!ml for chloroquine, a Koenoline, a further cytotoxic carbazole alkaloid from Murraya
koenigii, Phytochemistry, 24, 3041-3043 (1985).
common antimalarial drug (phillipson et al., 1993). Extracts
of the roots of the plant also are used to treat patients in FLoss, H. G., Biosynthesis of ergot alkaloids and related com-
pounds, Tetrahedron, 32, 873-912 (1976).
some Southeast Asian countries.
FLoss, H. G., The biosynthesis of ergot alkaloids (or the story of
the unexpected), in Indole and Biogenetically Related Alkaloids
(J. D. Phillipson and M. H. Zeok, eds.), 249-270, Academic
REFERENCES Press, London, 1980.
FLoss, H., M. TSCHANG-LlN, H. KOBEL, and P. STADLER, On the
ANTKOWIAK, R. and W. A. ANTKOWIAK, Alkaloids from mush- biosynthesis of peptide ergot alkaloids, Experientia, 30,
rooms, in The Alkaloids, Vol. 40 (A. Brossi, ed.), 189-340, 1369-1370 (1974).
Academic Press, New York, 1991.
FLUCKIGER, E., Recent advances in ergot phannacology. in Indole
AITA-UR-RAHMAN and A. BASHA, Biosynthesis of Indole Alka-
and Biogenetically Related Alkaloids (1. D. Phillipson and M.
loids, Clarendon Press, Oxford, 1983. H. Zeok, eds.), 285-291, Academic Press, London, 1980.
BERLIN, J. and F. SASSB, I3-Carboline and indole a1kylamines, in GEJSSMAN, T. A. and D. H. G. CROUT, Organic Chemistry of Sec-
Cell Colture and Somatic Cell Genetics of Plants, Vol. 5 (1. K.
ondary Plant Metabolism, Freeman Cooper. San Francisco,
Vasil, ed.), 357-369, Academic Press, Orlando, FL, 1988.
1969.
BHAITACHARYYA, P. and D. P. CHAKRABORTY, Carbazole alkaloids,
GROOER, D., Ergot alkaloids-Recent advances in chemistry and
Fortschr. Chern. Org. Naturst., 57, 159-210 (1987).
biochemistry, Federation of European Microbiology Societies
BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from Symposium Series 5 (R. Hutter, ed.), 201-217, Academic Press,
plants, in Phytochemical Resources for Medicine and Agricul- London, 1978.
ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum
HEMINGWAY, S. R. and J. D. l'Hn.LIPSON, Alkaloids of the Rubia-
Press, New York, 1992.
ceae, in Indole and Biogenetically Related Alkaloids (J. D. Phil-
CHAKRABORTY, D. P. and S. RoY, Carbazole alkaloids. m. Fortschr. lipson and M. H. Zeok, eds.), 63-90, Academic Press, London,
Chern. Org. Naturst., 57,71-152 (1991). 1980.
CHEPLICR, G. P. and K. CLAy, Acquired chemical defences in HOl.MSTEDT, B., The ordeal bean of old Calabar: The pageant of
grasses: The role of fungal endophytes, Oikos, 52, 309-318 Physostigma venenosum in medicine, in Plants in the Develop-
(1988). ment of Modem Medicine (T. Swain, ed.), 303-360, Harvard
CLAY, K., Fungal endophytes of grasses: A defensive mutualism University Press, Cambridge, 1972.
between plants and fungi, Ecology, 69, 10-16 (1988). HUSSON, H., Simple indole alkaloids including l3-carbolines and
CLAY, K., Trespassers will be poisoned, Nat. Hist., 8-14 (Septem- carbazoles, in The Alkaloids, Vol. 26 (A. Brossi, ed.), I-53,
ber 1989). Academic Press, 1985.
CLAY, K. and G. P. CHEPLICK, Effect of ergot alkaloids from fungal 10sm, B. S., V. N. KAMAT, and D. H. GAWAD, Some l3-carboline
endophyte-infected grasses on fall armyworm (Spodoptera alkaloids of Ailanthus malabarica, Heterocycles, 7, 193-200
/rugiperda), J. Chern. Ecol., 15, 169-182 (1989). (1977).
CORDELL, G. A., The Aspidosperma alkaloids, in The Alkaloids, KAPADIA, G. J., Y. N. SHUKLA, B. K. CHOWDHURY, S. P. BASAl<,
Vol. 17 (R. H. F. Manske and R. G. A. Rodrigo, eds.), 200-384, H. M. FALES, and E. A. SOKOLSKl, Phenylpentylisatins: A novel
Academic Press, New York, 1979. class of alkaloids from Melochia tomentosa, 1. Chem. Soc.,
CORDELL, G. A., Introduction to Alkaloids, A Biogenetic Approach, Chern. Commun., 535-536 (1977).
Wiley, New York, 1981. KAwONO, L. B. S., C. K. ANGERHOFER, S. TSAURI, K. PADMAWl-
CORDELL, G. A. and J. E. SAXTON, Bisindole alkaloids, in The NATA, J. M. PBzzUTO, and A. D. KINGHORN, Cytotoxic and anti-
Alkaloids, Vol. 20 (R. H. F. Manske and R. G. A. Rodrigo, malarial constituents of the roots of Eurycoma longifolia, J. Nat.
eds.), 3-296, Academic Press, New York, 1981. Prod., 54, 1360-1367 (1991).
CORDELL, G. A., M. OGURA, and N. R. FARNSWORTH, Alkaloid KONG, Y. P. P. BUT, K. No, K. CIIBNG, K. CHANG, K. M. WONG,
 Ergot and Other Indole Alkaloids 667

A. I. GRAY, and P. G. WATERMAN, The biochemical systematics ROBINSON, T., The evolutionary ecology of alkaloids, in Herbi-
of Merrillia,' in relation to Murraya, the Clauseneae, and the vores. Their Interaction with Secondary Plant Metabolites (G.
Aurantioideae, Biochem. Sys!. Ecol. 16, 47-50 (1988a). A. Rosenthal and D. H. Janzen, eds.) 413--448, Academic Press,
KONG, Y., P. P. BUT, K. NG, Q. Lt, F. CHENG, and P. G. WATERMAN, New York, 1979.
Micromelum: A key genus in the chemosystematics of the SAXTON, J. E., Indole alkaloids, Specialist Periodic Reports, Alka-
Clauseneae, Biochem. Syst. Bcol., 16, 485-489 (1988b). loids, Vol. 10, 141-210, The Royal Society of Chemistry Cam-
KONG, Y., K. CHENG, K. NG, P. P. BUT, Q. Lt, S. Yu, H. CHANG, bridge, (1981).
R. C. CAMBm, T. KiNOSHITA, W. KAN, and P. G. WATERMAN, SCHIFF, P. L., JR., The Thalictrum alkaloids: Chemistry and phar-
A chemotaxonomic division of Murraya based on the distribu~ macology, in Alkaloids: Chemical and Biological Perspectives,
tion of the alkaloids yuehchukene and girinimbine, Biochem. Vol. 5 (S. W. Pelletier, ed.), 271-637, Wiley, New York, 1987.
Sys!. Bcol., 14, 491-497 (1986). SPECIALIST PERIODIC REPoRTS, Alkaloids, Vol. 5 Chemical Society,
LEETE, E., The biosynthesis of alkaloids, Specialist Periodic Re- London, 1975.
ports Vol. 7, Biosynthesis, 102-223, The Royal Society of
SPECIALIST PeRIODIC REPoRTS, Alkaloids, Vol. 8, Chemical Society,
Chemistry, Cambridge, 1983.
London, 1981.
LEWIS, W. H., Plants used medically by indigenous peoples, in
Phytochemical Resources for Medicine and Agriculture (H. N. STADLER, P. A. and P. STiiTz, The ergot alkaloids, in The Alkaloids,
Nigg and D. S. Seigler, eds.), 33-74, Plenum Press, New York, Vol. 15 (R. H. F. Manske, ed.), 1-40, Academic Press, New
1992. York,1975.
LlU, K. C., Y. Sm-LIN, M. F. ROBERTS, and J. D. PHILLIPSON, Can- THOMAS, R. and R. A. BASSETI', The biosynthesis of ergot alkaloids,
thin~6-one alkaloids from cell suspension cultures of Brucea in Progress in Phytochemistry Vol. 3 (L. Reinhold and Y.
javanica, Phytochemistry, 29, 141-143 (1990). Liwschitz, eds.), 47-111, Pergamon, New York, 1970.
LUMONADtO, L. and M. VANBALEN, Indole alkaloids from Hannoa T!LLBQUlN, F., S. MICHEL, and E. SEGUlN, Tryptamine-derived in-
klaineana roots, Phytochemistry, 23, 453--455 (1984). dole alkaloids, in Alkaloids and Sulphur Compounds (P. G.
MAmR, W. and D. GROOER, Dimethylallyltryptopban-Synthetase Waterman, ed.), Vol. 8 of Methods in Plant Biochemistry (P.
in Clavieeps, Biochem. Physiol. Pflanzen, 170, 9-15 (1976). M. Dey and J. B. Hatborne, eds.), 309-371, Academic Press,
London, 1993.
MANN, J., Secondary Metabolism, Oxford University Press, Ox-
ford, 1978; 2nd edition, 1987. WALLER, G. R. and O. C. DERMER, Enzymology of alkaloidmetabo-
NEUSS, N., The spectrum of biological activities of indole alkaloids, lism in plants and microorganisms, in Secondary Plant Products
(E. E. Conn, ed.), Vol. 7 of The Biochemistry of Plants (p. K.
in Indole and Biogenetically Related Alkaloids (1. D. Phillipson
and M. H. Zeuk, eds.), 293-313, Academic Press, London, Stumpf and E. E. Conn, eds.), 317--402, Academic Press, New
1980. York,I981.
PeTROSKI, R. J., R. G. POWELL, and K. CLAY, Alkaloids of Stipa WATERMAN, P. G. and S. A. KHALm, The biochemical systematics
robusta (sleepygrass) infected with an Aeremonium endophyte, of Fagaropsis angolensis and its significance in the Rutales,
Nat. Toxins, 1, 84-88 (1992). Biochem. Sys!. Bcol., 9, 45-51 (1981).
PBn..LlPSON, J. D. and C. W. WRIGHT, Antiprotozoal agents from WINK, M., Allelochemical properties or the raison d'elre of alka-
plant sources, Planta Medica, 57, S53-S59 (1991). loids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), 1-118,
PBn..LlPSON, J. D., C. W. WRIGHT, G. C. KIRBy, and D. C. WAR- Academic Press, New York, 1993.
HURST, Tropical plants as sow-ces of antiprotozoal agents, in YATES, S. G. and R. G. POWELL, Analysis of ergopeptioe alkaloids
Phytochemical Potential of Tropical Plants (K. R. Downum, J. in endophyte-infected tall fescue, J. Agric. Food Chern., 36,
T. Romeo, and H. A. Stafford, ods.), 1-40, Plenum Press, New 337-340 (1988).
York, 1993. YATES, S. G., J. C. FENSTER, and R. J. BARTELT, Assay of tall
POWELL, R. G. and R. J. PeTROSKI, Alkaloid toxins in endophyte- fescue seed extracts, fractions, and alkaloids using the large
infected grasses, Nat. Toxins, I, 163-170 (1992). milkweed bug, J. Agric. Food Chern., 37, 354-357 (1989).
36
Alkaloids of Terpenoid Origin
(excepting indole alkaloids and ergot
alkaloids)

Introduction cally arise via the pathways leading to the various groups
Monoterpenoid Alkaloids of terpenes.
Biological Activity Some do not consider these compounds to be alkaloids,
Medical Uses of Monoterpenoid Alkaloids but prefer to call them "pseudoalkaloids" (Hegnauer, 1964;
Sesquiterpenoid Alkaloids Roddick, 1980). Many of these compounds are related
Dendrobium Alkaloids closely to non-uitrogenous plant metabolites, and, biosyn-
Nuphar and Castor Alkaloids thetically, are more related to those groups of terpenes from
Alkaloids of the Celastraceae and Hippocrateaceae which they are derived than to many groups of alkaloids
Khat (Gross et al., 1985). However, because of the many proper-
Miscellaneous Types of Sesquiterpenoid Alkaloids ties they share with other alkaloids, such a distinction will
Diterpenoid Alkaloids not be followed in this work.
Diterpenoid Alkaloids Derived from Kaurene Many spectral data for terpene-derived alkaloids have
Alkaloids from the Garryaceae been reviewed (Pelletier and Mody, 1979, 1981). Both the
Alkaloids of Delphinium and Aconitum 'H- and I3C-NMR spectra for several diterpene alkaloids
Erythrophleum Alkaloids have been discussed (Crabb, 1982). The results of feeding
Taxus Alkaloids studies leading to the synthesis of several alkaloids of this
Steroid and Triterpenoid Alkaloids group have been tabulated (Leete, 1983).
Solanum Alkaloids
Biosynthesis
Biological Activity of Solanum Alkaloids MONOTERPENOID ALKALOIDS
Solanum Alkaloids in Potatoes
Teratogenic Properties A number of important alkaloids are derived from several
Veratrum Alkaloids structural types of monoterpenes. Some involve substitution
Biosynthesis of nitrogen into ordinary monoterpene structures. Among
Biological Activity these compounds are simple monoterpene alkaloids in which
Steroidal Alkaloids in the Apocynaceae uitrogen is added to an existing monoterpene and more com-
Buxus Alkaloids plex structures, in which secologanin serves as a carbonyl
Daphniphy//um Alkaloids compound much as do the precursors for isoquinoline and
References benzylisoquinoline alkaloids. Alkaloids involving condensa-
tion of secologanin and tryptamine units are treated in Chap-
ter 34; those involving condensation of secologanin and tyra-
mine-derived units are included in Chapter 32. For
INTRODUCTION information concerning the precursor secologanin, the reader
should consult Chapters 19 and 20.
A large number of alkaloids are based on terpenoid struc- Both the 'H- and I3C_NMR spectra for several of the
tures. In most instances, the source of the nitrogen is not groups of alkaloids presented below have been reviewed
established, but the remaining portions of the molecule typi- (Crabb, 1982).

668
 Alkaloids of Telpenoid Origin 669

A number of alkaloids are known in which nitrogen is opicroside (6) (a monoterpenoid compound, but not an alka-
incorporated into monoterpenes. The source of the nitrogen loid), the proposed precursor of gentianine.
is not known. These alkaloids possess relatively simple
structures. Several common alkaloids of this type are derived 8iological Activity
from secologanin (1) or other monoterpenoid precursors by Flowers of Penstemon whippleanus (Scrophulariaceae)
condensation with ammonia (either on workup or, perhaps, serve as a host for the plume moth, Amblyptilia (Platyptilia)
in the plant itself). Others are derived from loganin (2), seco- mica. These flowers contain (+ )-boschniakine (10) and
loganin, or other precursors by largely unknown reactions other iridoid-monoterpene-derived alkaloids (McCoy and
(Mann, 1987). Several monoterpene alkaloids are thought Stermitz, 1983). Another host of this moth, Castilleja rhexi-
to be artifacts of isolation. For example, gentianine (3), bud- folia (Scrophulariaceae) contains rhexifoline (11), an iridoid
damine (4), and cantleyine (5) often are not found in the monoterpene alkaloid, as the major alkaloid of the seeds and
original plant materials when these are carefully investigated flowers. The moths and adults of the insect contain rhexifo-
(Fig. 36.1). Gentiopicroside (6) probably is the usual precur- line (Roby and Stermitz, 1984). Rhexifoline (11) also can
sor of gentianine (Cordell, 1981; Fodor and Colasanti, 1985). arise in some instances from the presence of ammonia in
Aucubin (7) is likely the precursor of buddamine (Stermitz workup from penstemonoside, but this possibility was spe-
and Harris, 1985). cifically precluded in this study by use of bicarbonate in
The biosynthesis of simple monoterpenes has not been place of ammonia in the workup.
studied extensively. Acetate, mevalonate, and geranylpyro- Actinidine (8) is found in the defensive secretion of the
phosphate are incorporated into actinidine. However, neither Australian cocktail ant, lridomyrmex nitidiceps (Fodor and
loganin (2) nor actinidine (8) serves as a precursor for 13- Colasanti, 1985). This compound is alleged to be attractive
skytanthine (9) or its derivatives in Tecoma stans (Bigno- to cats.
niaceae) (Fig. 36.2). These compounds appear to be derived
from an intermediate preceding loganin in the biosynthetic Medical Uses of Monoterpenoid AIkaloids
pathway. There has been much interest in gentianaceous plants be-
Both loganin and loganic acid are incorporated into genti- cause of their widespread use in folk medicine in Europe.

R
~ r-gIUCOSYI

- o~
. 0
NH,
,;? .&

N
o 0

gentianine (3)

a-
gentiopicroside (6)

W
OH
··,OCH'
f'
NH

"--0
\ i
HO
N

buddamine (4) cantleyine (5)

CH,O,C
o ':::H
~
H'···
:::,..
"'O~o~ OH
oHO~

secologanin (1)
HO
OH

'ow"' O-glucosyl

Joganin (2)

Fig. 36.1. Fonnation of simple monoterpenoid alkaloids.

670 Alkaloids of Terpenoid Origin

{]-- N
,>(11.<, N
I
'8- N
I
CH, CH,
actinidine (8) tecostidine hydroxyskytanthlne-n p-skytanthine (9)

~ ·X2:c
o -"H
OHC, ) : ' ) - - CH,O.C

'C.J N
I P'

""
H

w-·.· --
N

(+)-bosehniakine (10) rhexlfoline (11) gentionavine

H0Y'il
W
CHO
~
CHO HODJ"'N+
I
tr I an alkaloid of
Vakrlartllo!Ji<lrtIllls
(12)

"'"
NH
,

Fig. 36.2. Some representative monoterpene alkaloids.

At the beginning of the 1900s, at least 20 Gentiana spp. were as a tonic (Cordell, 1981). The principal alkaloid of this
commonly used. Gentianine (3), a widely studied alkaloid, is drug is dendrobine (13) (Fig. 36.3). This alkaloid is from a
not particularly toxic and, in low doses, exhibits a central cadalane skeleton, involving copaborneol (14), in a similar
nervous system stimulant action. Gentianine also exerts hy- manner to tutin and coriamyrtin (see Chapter 21). All three
potensive, anti-inflammatory, and muscular relaxant actions. parts of the molecule were labeled almost equally by mevalo-
The reputed tonic effects of gentian are probably due to the nate.
hypotensive and muscle relaxant actions. Dendrobine (13) lowers blood pressure and has weak an-
Valeriana officinalis is used widely in Europe as a seda- tipyretic and analgesic action. This alkaloid modulates gly-
tive. ill addition to iridoid monoterpenes, this plant contains cine neurochemical activity (Wink, 1993).
several monoterpenoid alkaloids, such as (12).
lfupbar and Castor Alkaloids
Sesquiterpenoid Alkaloids
The rhizomes of Nuphar japonicum and N. luteum
Several classes of sesquiterpene alkaloids have been re- (Nymphaeaceae) contain sesquiterpene-derived alkaloids
ported; these do not appear to be closely related biosyntheti- that possess a quinolizidine nucleus (Howard and Michael,
cally. Important among these alkaloids are those of Nuphar 1986). These compounds are based on a CIS skeleton of
and Dendrobium (Orchidaceae), and the Celastraceae and which part has been elaborated into a 3-substituted furan
Hippocrateaceae. About 130 sesquiterpene alkaloids are (Howard and Michael, 1986) (Fig. 36.4).
known (Verpoorte et al., 1991). The original structure of ( - )-deoxynupharidine (IS) was
not correct and much of the earlier literature concerning
Dendrobium Alkaloids these alkaloids contains erroneous structures. The compound
has a IR,4R,7R,9aR configuration, as indicated in Fig. 36.4
The genus Dendrobium (about 1200 species) is the largest (Howard and Michael, 1986). Similar alkaloids have been
genus of the family Orchidaceae [the largest plant family, isolated from the scent glands of the Canadian beaver, Cas-
with 20-25,000 species (Dressler, 1981»). Many of these tor fiber (Fig. 36.4). The oil of this gland or castoreum is
species contain alkaloids and some contain sesquiterpene used as a fixative in the perfumery industry. (- )-Deoxynu-
picrotoxins (see Chapter 21). A Chinese drug "Chin-Shih- pharidine (IS) is one of the components of the complex mix-
Hu," derived from the orchid Dendrobium nobile, is used ture of compounds found in this oil. Although it is not clear,
 T
T -- ~ T
T

TT
TT
# --
T

T
pT T
Alkaloids of Terpenoid Origin

#
__
671

T
from 4- 14C_mevalonate
T from S.lHz-mevalonate

copaborneol (14)
dendrobine (13)

FIg. 36.3. Biogenesis of dendrobine (modified from Cordell, 1981; used with permission of the author).

the beaver probably obtains the alkaloids from food plants have not been extensively stndied until comparatively re-
of the genus Nuphar (Howard and Michael, 1986). cently. These alkaloids most commonly occur in the seeds
of Euonymus europaeus, Tripterygium wilfordii, and in
Alkaloids of the Celastraceae leaves of khat, Catha edulis (16-20) (Fig. 36.5).
and Hippocrateaceae A series of sesquiterpene evoninoate alkaloids have been
isolated from the liana Hippocratea excelsa (Hippo-
Although members of the Celastraceae have been known
crateaceae) from Mexico (Mata, 1993; Mata et aI., 1990).
to contain aIkaIoids for at least 40 years, these compounds
This plant, called "cancerina," is used medicinally to treat
skin ailments and for its pesticidal properties. The main alka-
loid, hippocrateine I (21), was cytotoxic in the 9 PS system
(EDso 1.85 X 10- 1 flog/mI) (Mata et aI., 1990). This plant
also contains small amounts of the antitnmor triterpene
tingenone (see Chapter 23).
The presence of complex sesquiterpenoid alkaloids in
both the Celastraceae and Hippocrateaceae confirms the
close relationship between these families.

Khat
(+)-nupbaridine (-)-deoxynupharidine (15) Sesquiterpenoid alkaloids similar to those of other celas-
"traceous plants occur in leaves of khat (Brenneisen and El-
Sohly, 1992; Crombie, 1980) but do not appear to be respon-
sible for the effect of chewing fresh khat (Szendrei, 1980).
The active compound is a simple amine, cathinone (see
Chapter 28). A series of complex sesquiterpenoid cathedulin
aIkaIoids has been characterized from khat (Crombie et aI.,
1990).

Jlliscellaneous Types of Sesquiterpenoid

Alkaloids
castoramine l-epideoxynupharidine
Other sesquiterpenoid aIkaIoids are found in Fabiana im-
Fig. 36.4. Alkaloids from Nuphar and Castor. bricata (Solanaceae) [fabianine (22)] and in Pogostemon pa-
672 Alkaloids of Terpenoid Origin

OAc 0

,.o~ oA
OAc 0

Ac.o~O\Acr
O~
~, N
rn,

0
HO
OH
o
OAc OAc 0\Jy~
I 0,.,
"
(:~c
..
maytoline (16)

(OAC
:'('" % :~Ok
/ hippocraleineI(21) ~)
A.,.o~O\AC!
lAC 0 HO 10H 0 OAc

:~ ~
~.

o 0 0
o
01.0 OAc
o 0 OH CH,OH ~O
evoninol (18) I "" H

N H OH
evonine (19)
evoninic acid (20) 0

Fig. 36.S. Sesquiterpene alkaloids from the Celastraceae.

tchouli (Lamiaceae) [epiguaipyridine (23)] (Fig. 36.6). Other DlTBKPEI'IOID ALKALOIDS

sesquiterpene alkaloids from Gaillardia pulchella (As-
teraceae) [pulchellidine (24), neopulchellidine (25), and pul- Several types of aIka10ids are based on diterpenoid struc-
chellin (26) (Fig. 36.6)] have anti-inflammatory activity tures. Chief among these are aIka10ids derived from kaurene,
(Strunz and Findlay, 1985). those ultimately derived from macrocycIic precursors, and

H9JJOH
, 0

~.",O 0
HO~H
OHO
• H °
!
OH
H

pulcheJlidine (24)

~
neopuIchellidine (25) u1cheJlin(26)

17,

~ ~I
"'-N

N 'H

OH

epigualpyridine (23) fabianine (22)

Fig. 36.6. Some miscellaneous types of sesquiterpene alkaloids and pulchellin.

 Alkaloids of Terpenoid Origin 673

diterpene 19-carboxylic acid derivatives (Benn, 1993). pene-derived alkaloids. The family of plants is found primar-
About 640 diterpene alkaloids are known (Verpoorte et aI., ily in the southwestern United States and Mexico. The veat-
1991). chine (29) skeleton of Garrya alkaloids incorporates a
kaurene (30) skeleton without rearrangement (Fig. 36.7)
Diterpenoid Alkaloids Derived
(Geissman and Crout, 1969). Two of the principal alkaloids
from Kaurene
of these plants, garryine (31) and veatchine (29), are known
Oiterpene alkaloids derived from kaurene are known to be toxic. The LOso of veatchine hydrochloride in mice is
from several families. These compounds are found in the 12.5 mg/kg.
Ranunculaceae (Aconitum and Delphinium), Garryaceae
(Garrya), Asteraceae (Inula royaleana), Escalloniaceae
(Anopterus), and Rosaceae (Spiraeajaponica) (Benn, 1993; Alkaloids of Delphinium and Aconitum
Pelletier, 1992). In contrast to many other types of alkaloids,
Plants of the genera Delphinium and Aconitum have long
these diterpenoid alkaloids do not have a pronounced bitter
been recognized as poisonous (Cutler, 1992). Extracts of the
taste. Many (especially the aconitines) produce a tingling
roots and leaves of Aconitum species were used in ancient
sensation on the tongue, but in view of the extreme toxicity
times as poisons, but also for treatment of illnesses such as
of diterpene alkaloids, tasting these compounds is not recom-
neuralgia, hypertension, gout, and rheumatism. In the past,
mended. Two to five milligrams of several C l9 diterpene
preparations of Delpinium and Aconitum also have been used
esters is sufficient to be lethal in humans; aconitine (27)
medicinally as cardiotonics, febrifuges, sedatives, and ano-
and pseudoaconitine (28) are highly toxic to man (Cordell,
1981). Aconitine has an L0 50 i.v. in mouse of 0.17 mg/kg dynes (Benn, 1993; Benn and Jacyno, 1983). Alkaloids of
(Wink, 1993). Plant material containing alkaloids from Del- both genera have been used in Chinese traditional medicine
phinium, Aconitum (Ranunculaceae), and Spiraea (Rosa- (Han et al., 1988). Arrow poisons made from Aconitum spe-
ceae) have been used in Chinese traditional medicine (Han cies formerly were used by the inhabitants of Hokkaido and,
et aI., 1988). Aconitine (27) also inhibits growth of the radi- possibly, still are used by other peoples of Siberia and
cle in Lepidium seedlings (Wink, 1993). Alaska. The active ingredients of these arrow poisons are a
Four basic skeletal types of kaurene-derived diterpene series of diterpene alkaloids (Bisset, 1976). Aconitum spe-
alkaloids are known: the atisine, heteratisine, Iycoctonine, cies were used in Europe for trial by ordeal.
and veatchine types. Two main structural types of diterpene alkaloids are
found in these genera. One group includes a series of com-
Alkaloids from the Garryaceae paratively simple and relatively nontoxic amino alcohols and
A number of species of the genus Garrya, the only genus esters modeled on a Czo skeleton. These alkaloids are not
of the family Garryaceae, contain relatively simple diter- extensively oxygenated and contain, at most, one methoxyl

(+kaurene (30) veatcbine (29)

&9r o
garryine (31)

aconitine (27) pseudaconitine (28)

Fig. 36.7. Kaurene and veatchine.

674 Alkaloids ofTerpenoid Origin

group. Most of these alkaloids possess a veatchine (29) or Larger doses produced tachycardia and pronounced cardiac
an atisine (32) skeleton (Fig. 36.8). irregularities, finally culminating in cardiac arrest. Respira-
The second group is based on a C 19 , instead of a C20 tory failure appeared to precede heart failure (Benn and Ja-
skeleton. Compounds of this group are usually of an aconi- cyno, 1983). Electrophysiological studies have shown that
tine (27) or Iycoctonine (33) structural type (Fig. 36.8). aconitine prolongs depolarization after stimulation of a nerve
These compounds are more heavily substituted than those of and that this depolarization is blocked by tetrodotoxin. Thus,
the C20 series, and, usually, two of the alcohols are esterified, aconitine (27) appears to bind to the nerve-cell membrane
typically one with acetic acid, and the other with an aromatic in such a way as to prevent nonnal closing of sodium chan-
acid, such as benzoic or veratric acid. Lycoctonine (33) has nels (Benn and Jacyno, 1983). The LOs. of aconitine (27)
an LOs. i.p. in mouse of 350 mglkg (Wink, 1993). in rabbits is 0.04-0.5 mg/k:g.
Of these C 19 esterified alkaloids, most attention has been A second alkaloid, mesaconitine (34), also is widely dis-
paid to aconitine (27), the best known compound, because tributed in this group of plants, and predominates in some
it is extremely poisonous. One must regard many early re- Aconitum species. It has similar properties to aconitine. Mes-
ports with caution, however, as aconitine is extremely diffi- aconitine (34) has an LOso s.c. in mouse of 0.2 mg/kg (Wink,
cult to isolate and purify. Subcutaneous administration of 1993).
aconitine hydrobromide to cats produced bradycardia. Removal of the 8-acetate group of aconitine produces a

12
12
17

18
veatcbine skeleton atisine skeJeton

18

17

atisine (32)

OH

aconitine (27) Iyeoctonine (33)

Fig.36.8. Veatchine and atisine skeletons; aconitine and lycoctonine.

 Alkaloids o/Terpenoid Origin 675

~,"_ _ ."OCH3
OH
OCH3

,
, OH

aronine (36)

OH
~,-{5o methyllycaconitine (38)

OH

mesaconine (37)

OH

HO

mesaconitine (34)
Fig. 36.9. Aconine, benzaconine, and mesacorune.

compound, benzaconine (35), about 300 times less active As observed above, two types of activity, sometimes
than aconitine (27) (Fig. 36.9). Mesaconine (37) also belongs called aconitiform and curariform activity, are involved.
to this series. These compounds appear to exhibit neurotoxicity by inter-
Complete saponification of aconitine produces an alkan- acting with membranes in such a way as to hold sodium
olamine, aconine (36), that lacks the cardiac effects of aconi- channels open, following an action potential, so that pro-
tine but has antiarrhythmic properties. The compound has longed depolarization results. This activity is similar to that
curare-like effects on neuromuscular transmission, often re- of other natorallipophilic neurotoxins such as batrachotoxin
sulting in death from respiratory paralysis (Benn and Jacyno, (a steroidal alkaloid), veratrldine (a steroidal alkaloid), and
1983). grayanotoxin I (a nonalkaloidal diterpene). All four seem to
Methyllycaconitine (38) occurs in numerous Delphinium act at the same receptor site. As grayanotoxin I is not an
species. The hydroiodide of this alkaloid shows curare-like alkaloid, it has been suggested that the nitrogen may not be
properties greater than those of aconitine (27) or mesaconi- a necessary structural featore for this type of activity. Also,
tine (34). This compound is a potent, nondepolarizing neuro- as previously mentioned, the ester function of these com-
muscular poison, acting as a competitive inhibitor of acetyl- pounds is very important for activity. These ester groups
choline at nicotinic receptors in the same way as may play a role in the orientation of the molecules on the
tobocurarine (Benn and Jacyno, 1983). Methyllycaconitine receptor site (Benn and Jacyno, 1983).
(38) has an LDsoi.v. in mouseof5-18 mg/kg (Wink, 1993). Inspection of models indicates that methyllycaconitine
676 Alkaloids of Terpenoid Origin

(38) closely resembles ( + )-tubocurarine with regards to the Taxus Alkaloids

disposition of many nitrogen and oxygen atoms and would
fit the "curariform" template (Benn and Jacyno, 1983). Yews (Taxus spp., Taxaceae) produce a series of alka-
loids such as spicataxine (40) and taxine-B (41) that are
highly toxic (Blechert and Guenard, 1990) (Fig. 36.10). Be-
Erythropbleum Alkaloids cause of interest in taxol (42), the alkaloid chemistry of most
Species of the genus Erythrophleum (Fabaceae: Caesal- Taxus species and Austrotaxus spicata has now been investi-
pinioideae) are often highly toxic. As few as two leaves gated (Blechert and Guenard, 1990).
of Erythrophleum chlorostachys, a plant native to northern Taxus alkaloids are derived from macrocyclic diterpenoid
Australia, have been reported to be poisonous to a goat and precursors. These diterpenes are derived from macrocyclic
have produced poisoning of goats, sheep, horses, and cattle. precursors (Blechert and Guenard, 1990) (see Chapter 22).
In Africa, Erythrophleum guineense has been used as a trial- Four major structural types of Taxus diterpenes exist (Fig.
by-ordeal plant. 36.11).
Many species of this genus contain N-alkylarninoethyl Taxine I and II are extremely toxic. This activity may
esters of diterpene 19-carboxylic acids that possess cardiac arise because these alkaloids are effective calcium antago-
activity similar to digitalis glycosides [such as cassaine nists (Blechert and Guenard, 1990).
(39)]. Many of these alkaloids display potent cytotoxic activ- Taxol (42) is highly cytotoxic and has broad antitumor
ity (Fig. 36.10) (Suffness and Cordell, 1985). activity. This alkaloid acts by a mechanism different than
Erythrophleum alkaloids have cardiac activity (they in- any other antitumor drug. At concentrations completely in-
crease contractile strength) and a very strong local anesthetic hibitory to cell division, taxol had no significant effects on
action. Although the latter action is more potent than co- DNA, RNA, or protein synthesis, but BeLa cells were
caine, it is accompanied by intense irtitation at the site of blocked in late G 2 and/or M phase. In contrast to other anti-
admirtistration. Cassaine (39) inhibits (Na+, K+)-A TPase mitotic agents such as colchicine, maytansine, vincristine,
(Wink, 1993). When the double bond is saturated, almost all or podophyllotoxin, taxol (42) was found to promote assem-
biological activity disappears. Erythrophleum bark is used as bly of calf-brain microtubules, and the microtubules so pro-
a cancer remedy in Africa (Suffness and Cordell, 1985). duced were resistant to depolymerization by cold or CaC!,.

ODO
OCH,CH,N(CH,), spicataxine (40)

HO l,-o
H OCOCH,

~:-~' »~
~ rn,"/rn'd"" !: O~o
H ~o I I

o
HOP' OH OCOCH,
o i OCOC,Hs
HO OH ~
taxol (42)
taxine B (41
OCOCH,

~JtMo-~
'T ~O OH

cephalomannine (43)
HO !
OCOC,Hs
OCOCH
3

Fig. 36.10. Cassaine. taxine I and n, and taxol.

 Alkaloids of Terpenoid Origin 677

~ OPP

Fig. 36.11. Four major structural types of Taxus diterpenes (Blechert and Guenard, 1990; modified and used with peImission of the copyright owner,
Academic Press, Orlando, FL).

This is a unique mode for mitotic inhibitors (Suffness and batrachotoxin (45)] occur in animals (Fig. 36.12) (Daly et
Cordell, 1985). Two major problems are involved, however. al., 1993; Habermehl, 1967; Witkop and Gossinger, 1983).
The compound has extremely low solubility and occurs in Batrachotoxin occurs in the skins of the poison-dart frog
very low yields in the plant (Suffness and Cordell, 1985). Phyllobates. The LD50 on subcutaneous injection in mice is
Although the most common source of taxol is Taxus brev- about 40 ng; the lethal dose in man is estimated to be 200
ifolia, this alkaloid, as well as related alkaloids, is encoun- Ilg (Wink, 1993). The frog is so toxic that Indians only
tered in the needles of several species of Taxus (Witherup scrape the dart across the frog's back. Batrachotoxin is an
et al., 1990). important tool for the study of voltage-dependent sodium
Another family of gymnospermous plants that often has channels (Daly et al., 1993).
been considered closely related, the Cephalotaxaceae, do not
contain taxine alkaloids, but contain other structural types,
including homoerythrina alkaloids (see Chapter 33).
SOLAlVlJ/fI ALKALOIDS
The alkaloid cephalomannine (43) has been isolated from
Taxus wallichiana (the compound name reflects the fact that
plant source was originally considered to be a Cephalotaxus Steroidal alkaloids are widespread in the genus Solanum
species) (Miller et aI., 1981; Powell et al., 1979). This alka- (about 1400 species) and related genera of the Solanaceae
loid has antitumor activity in KB cell lines. (0' Arcy, 1979, 1986; Gross et al., 1985; Prelog and Jeger,
1953, 1960; Sato, 1970; Schreiber, 1968). Many plants of
this group have economic importance; among them are to-
mato (Lycopersicon esculentum), potato (Solanum tubero-
smROID AND TKITEKPEI'IOID ALKALOIDS sum), buffalo bur (Solanum rostratum), and eggplant (Sola-
num melongena). Steroidal alkaloids also occur in plants of
the genus Veratrum and related genera in the Liliaceae. All
At least 450 steroid and triterpenoid alkaloids have been told, steroidal alkaloids have been isolated from nearly 500
reported (Verpoorte et al., 1991). Most steroidal alkaloids species of plants from these 2 families. Five major structural
occur in a relatively small number of genera (Cestrum, Cy- types are known: the spirosolanes [solasodine (46)], solani-
phomandra, Lycopersicon, Nicotiana, and Solanum) of the danes [solanidine (49)], 22,26-epiminocholestanes [verazine
family Solanaceae and of the family Liliaceae (Fritillaria, (47), and etioline (48)], a-epiminocyclohemiacetals [solano-
Veratrum, and Zygadenus). Related alkaloids are known capsine (51)], and 3-aminospirostans [juribidine (50)]. An-
from the Apocynaceae (Funtumia, Holarrhena, and Ma- other large group of steroid-derived alkaloids have altered
louetia) and Buxaceae. Alkaloids based on triterpenoid C 27 skeleta based on cholestane. These are discussed in the
structures are much less common. Probably the best known subsection "Veratrum alkaloids," below. Most of the alka-
ones are found in plants of the Buxaceae (especially Buxus, loids occur as glycosides (Fig. 36.13) (Gross et aI., 1985;
Pachysandra, andSarcococca) (Gross et aI., 1985; Roddick, Schreiber, 1979).
1980, 1986). The co-occurrence of cuscohygrine (see Chapter 30) and
Other steroidal alkaloids [such as samandarine (44) and certain Solanum alkaloids has been reported for several spe-
678 Alkaloids o/Terpenoid Origin

~ NH

.amandarin. (44) batrachotoxin (45)

Fig. 36.12. Samandarine and batrachotoxin.

cies of Solanum and Cyphomandra (Ripperger and Cycloartenol and cholesterol serve as precursors for to-
Schreiber, 1981). matidine (52), solasodine (46), solanidine (49), soIanocap-
Techniques for the isolation and purification of steroid sine (51), and many other Solanum alkaloids. Cholest-4-en-
alkaloids and the instrumental methods for characterization 3-one (53) (Fig. 36.4) and 26-hydroxycholesterol were
(NMR and MS) have been reviewed (Atta-ur-Rahman and shown to be the first products of metabolism. The (25R)
Choudhary, 1993). (54) and (25S) (55) stereoisomers of 26-hydroxycholesterol
are converted stereospecifically into soladulcidine (56) and
Biosynthesis tomatidine (52), respectively (Gross et al., 1985).
Spirosolanes. The spirosolanes, such as tomatidine
Glycoalkaloids appear to be synthesized in aereal por- (52) and solasodine (46), have the basic nitrogen incorpo-
tions of Solanum species and not in roots, as in the case of rated into an oxaazaspirane unit that includes the original
tropane alkaloids of many solanaceous species (Waller and steroidal side chain (Figs. 36.13 and 36.14). A biogenetic
Nowacki, 1978). scheme has been proposed (Fig. 36.15).
The above-mentioned types of Solanum alkaloids are de- (25R)-Sapogenins can be formed from precursors such
rived from the same pathways as most other steroids in as (25R)-26-hYdroxycholesterol (54) or (25R)-50l-furostan-
plants. For more information on the formation of the various 3(3,26-diol (58). 10 precursor 58, the tetrahydrofuran ring
precursors involved, see Chapters 23 and 24. The biosyn- involving oxygenation at C-16 has already been formed. The
thesis and metabolism of the C27 sapogenins and alkaloids biosynthesis of (25S)-sapogenins differs from that of (25R)-
are closely related (Ripperger and Schreiber, 1981; Gross et sapogenins, because a different pro-chiral terminal group
ai., 1985). Acetate and mevalonate are incorporated into all (i.e., compound 54 versus compound 55) is involved, and
types of steroid alkaloids. not because of stereochemical differences in reduction of an

~~HN~
H,N
HO
HO '"
solacongestidine (6S) solanacapsine (51) solasodin. (46)

HO
H,N

juribldine (50) solanidine (49) procevin. (90)

Fig. 36.13. Representatives of the major structural types of steroid alkaloids (modified from Gross et at., 1985).
 Alkaloids oj Terpenoid Origin 679

HO
tomatidine (52) tomatine (69)
H R =P-lycotetraosyl
H

HO
ligogenin (9) cholest-4-en-3-one (53)
H H

17

HO

HO

<al solanidine (49) solacongestidine (65)

Fig. 36.14 (8 & b). Some Solanum alkaloids and their precursors.
680 Alkaloids oj Terpenoid Origin

HO HO
(2SR)-26-hydroxycholesterol (54)
H
H

(2SS)-Sa-furoslan-3p,26-diol (59)
HO
(2SS)-5a-cholestan-3p.26-diol (55)
~NH, H

~
H
iH
~ I m-OH

HO
HO~ H

26-aminodihydrodiosgenin (63) 16P-hydroxy-5a-choleslanol (60)

o

HO

(b) 16p.26-dihydroxy-5cx-choleslanol (61) 16P-hydroxy-22-oxo-5cx-cholestanol (62)

Fig. 36.14_ (continued)

unsaturated A24 intermediate. Synthesis of tomatidine (52) The C-27 methyl group of (25R)-solasidine (46) is de-
in Lycopersicon pimpinellifolium appears to utilize (25S)- rived from the C-2 group of mevalonate. whereas the C-27
5a-cholestan-3f3.26-diol (55). but not (25S)-5a-furostan- methyl group of (25S)-tomatidine (52) is derived from the
3f3.26-diol (59). C-3 group of mevalonate. This indicates that hydrogenation
20-Hydroxycholesterol is converted into both tigogenin of the A24 intermediate occurs by addition of hydrogen to
(64) and solasodine (46). 16f3~Hydroxy-5a-cholestanol (60). the 24 si.25 si face (Fig. 36.16).
16f3.26-dihydroxy-5a-cholestanol (61). and 16f3-hydroxy- Solanidine alkaloids. The second group of Solanum
22-oxo-5a-cholestanol (62) are convertible into spirostanes alkaloids. the solanidine alkaloids. also occur as glycoaIka-
[such as tigogenin (64)]. but not into spirosolanes (such as loids. An example of this type is solanidine (49).
52 and 46). The 16-hydroxy group appears to be introduced A biogenetic scheme for solanidine (49) has been pro-
after formation of the oxaazaspirane unit (ring F) (Fig. posed (Fig. 36.17) (Cordell, 1981; Gross et aI., 1985). The
36.15). amino acid arginine has been shown to be the source of
Introduction of the amino group occurs at an early stage. the nitrogen of solanidine (49) in Veratrum grandiflorum
This is supported by the incorporation of 26-aminodihydro- (Ripperger and Schreiber, 1981).
diosgenin (63) into solasodine (46) in Digitalis lanata (Gross The biosynthesis of solanidine (49) involves the intyrme-
et aI .• 1985). diates verazine (47) and etioline (66), and loss of the 1613-
 Alkaloids oj Telpenoid Origin 681

cholesterol (99) 25 Rand 25S (54, 55) 25R 25R and 25S
/<"25S)~dOrmantNinone spirostanes H
/ / tigogenin (64)

,~~~,~~
1) --- ~o

w'.
22S 25S
22,26~epiminocholestanes 25 R and25S (22R, 25R)-solasodine (46)
(25R)-26-aminocholesterol (68) spirosolanes
solacongestidine (65)
verazine (47)

......
/
~ '.N:::::1" jjDO...
2S ' H

w·····
N N l
~ 25 ~ ~j): HN -+ N ..z.;".

o "'",OH
-OH

,
solanidine (49)

\
25R (25R)-solal1oridine (78) (25S)~teinemine solanidine type alkaloids

teo
(25S)-etioline (66)

-- ~O!~HNN1.
N
(25R)-solanacapsine (51)
(25R)~solacasine "~ ct-epiminocyclohemiacetals
~O~""""
.... '?'
"0 ----",
OCH3 OH

Fig. 36.15. Proposed biosynthetic scheme for the origin of steroid alkaloids with unaltered ~7 skeletons (modified from Gross et aL, 1985).

hydrogen atom (Gross et aI., 1985). Rubijervine (67) is de-

rived from verazine (47), but not etioline (66) in the rhizome
of Veratrum grandiflorum (Gross et aI., 1985).
22,26-Epiminocholestanes. Alkaloids with this struc-
ture are intennediates in the biosynthesis of spirosolane, so-
lanidine, a-epiminocyclohemiacetal, and 3-aminospirostane
alkaloids. (25R)-26-Aminocholesterol (68) is precursor to
the 22,26-epiminocholestanes. The biosynthesis of verazine
H (47), etioline (66), and solacongestidine (65) is discussed

. .
above.

~
N
I
," 3-Aminospirostane alkaloids. Alkaloids of this group,
such as jurubidine (50) (Fig. 36.13) are probably synthesized
analogously to the corresponding spirostanols (e.g., tigo-
(2SR)-spirosolane
(2SS)-spirosolane genin). The 3-amino group is most likely introduced by
• from C-3 of mevalonate
transamination late in the biosynthesis (Gross et aI., 1985) .
• from C-2 of mevalonate

Fig. 36.16. Stereospecific reduction of the .6,24 intennediate in the Biological Activity of Solanum Alkaloids
fonnation of spirosolanes (modified from Gross et al., 1985).
",-Tomatine (69) (Fig. 36.18) has a half-life of about 6
days in tomato plants. This alkaloid inhibits the growth of
several Gram-positive and Gram-negative bacteria and ani-
682 Alkaloids oj Terpenoid Origin

no
dormantinol dormantinone

In :~n
~ ~ .... OH ........

--
no '" - no '" no.

~-r
rubijervine (67)
(l2a-hydroxysolanidine)

N~I
no
solanidine (49)
- no ""
etioline (66)
-
"'on ......

no
verazine (47)

Fig. 36.17. Proposed biogenesis of solanidine (modified from Cordell, 1981; used with permission of the author).

mal pathogenic fungi. a-Tomatine and demissine (70) have isozymes. This alkaloid has an LOso i.p. in rat of 84 mglkg
antifeeding effects for the Leptinotarsa decimlineata (Colo- (Wink, 1993). a-Chaconine also disrupts biomembranes by
rado potato beetle), the two-striped grasshopper, and the to- cholesterol binding. A 70: 30 mixtore of a-chaconine and
mato fruit worm (Juvik and Stevens, 1982; Juvik et al., 1982; a-solanine exhibited maximal synergistic effects in destabi-
Levinson, 1976; Roddick, 1986). Incorporation of either a- lization of cell membranes (Roddick et al., 1988). a-Chacon-
tomatine (69) or demissine (70) into a diet of potato leaves ine is a feeding deterrent to Clwristoneura (Lepidoptera)
at 0.1-0.5% makes the diet unpalatable to Leptinotarsa de- (Wink, 1993).
cemlineata larvae (0.3% for adults), whereas similar The potato glycoalkaloids leptine I (73) and demissine
amounts of solanine (71) do not. The adults normally feed (70) are among the most active inhibitors of human plasma
in the presence of up to 0.6% solanine (71) and a-chaconine cholinesterase (Beier and Nigg, 1992). a-Chaconine (72),
(72) (Levinson, 1976). The foliage of cultivated tomatoes commersonine, solamargine (79), solanine (71), and solani-
contains significantly lower levels of a-tomatine (69) than dine (49) all possess this activity (Wink, 1993).
the fruit and foliage of some wild Lycopersicon species Many steroidal alkaloids are repellent or antifeedant to
(Juvik and Stevens, 1982). Although a-tomatine (69) is cor- insects. This may be attributable largely to the membrane
related with mortality of Helicoverpa zea, this compound destabilizing effects of the alkaloids (Roddick et al., 1988).
does not occur at high enough levels to explain resistance Demissidine (76), solacauline (75), soladulcine (74), sola-
in many accessions of Lycopersicon species; other chemical nine (71), and tomatidine (52) are phagorepellent to Leptino-
factors such as 2-tridecanone may be involved (Barbour and tarsa decimlineata (Colorado potato beetle). a-Chaconine
Kennedy, 1991; Juvik et al., 1982). (72), solanidine (49), solanine (71), and tomatidine (52) are
a-Tomatine probably is toxic because it complexes with feeding deterrents to Clwristoneura (Wink, 1993).
3-J3-hydroxy steroids and alters membrane permeability. A variety of steroidal alkaloids have antifungal activity
Pure a-tomatine (69) has an LO so i. v. in rat of 18 mglkg (Clark and Hufford, 1992; Wink, 1993). In tomato and in
but is not very toxic orally (I glkg). potato, glycosidic alkaloids play a role in inhibition of the
Solanine (71) is a mitotic poison (Cordell, 1981) with an growth of fungi. Glycoalkaloids, such as a-tomatine (69)
LOso i.p. in mouse of 42 mg/kg (Wink, 1993). However, and solanine (71), are hydrolyzed by J3-glucosidases and the
2.8 mglkg is toxic to humans (Roddick, 1986). aglycones react with membrane sterols and cause damage
a-Chaconine (72) is a potent inhibitor of cholinesterase to the cell membrane of the fungos (Nahrstedt, 1979). A
 Alkaloids of Terpenoid Origin 683

highly active alkaloid, solacongestidine (65) exhibited mini- not derived from steroid alkaloids. The adults do not appear
mal inhibitory concentration (MIC) values of 0.4-0.8 ",g/ml to be able to sequester these compounds, and their toxicity
and verazine (47) at 3.1 ",g/ml for Candida albicans and is due to the presence of pyrrolizidine alkaloids (Drummond,
Trichophyton rubrum. Solacongestidine was active against 1986).
several other fungal species, but essentially inactive against As mentioned above, demissine and several other steroid
Aspergillus species tested (Clark and Bufford, 1992). Sola- alkaloids are repellent or toxic to the Colorado potato beetle.
casine (77), soladulcidine (56), solafloridine (78), solamar- Races of this insect are adapted to several Solanum species,
gine (79), solanidine (49), solanine (71), solanocapsine (51), including S. eleagnifolium, S. rostratum, and S. tuberosum,
solasodine (46), solasonine, tomatidenol, and tomatilladine and the insect has a recognized ability to adapt to new combi-
all exhibit varying degrees of antifungal activity (Wink, nations of steroid alkaloids (Barbome, 1986).
1993). Solanum alkaloids, especially those of the 22,26-epimino-
Although the larvae of many ithomiine butterflies feed cholestane types have been converted to pregnane deriva-
on solanaceous plants and must be adapted to the presence tives and used as sources of contraceptive and anti-inflam-
of steroid alkaloids, the distasteful properties of adults are matoty drugs (Bradley et al., 1979; Schreiber, 1979).

ORO_glucose

°HOH~~O'~O OH-:'....
OHOO~
RO D.xyl... ~O HO OR
",·tomatine (69) HOHO OH
R = p-.ycotet....y.
D-Rluoose OB Iycotetraose
H

OH

HO""""\ _0 0 OH
';o~L< _0
D-gluc~e ~~~O
D-gaJadose
o n-solanine (71)
L.~OH
'O~OH
L-rhamnose 0 OB

D.xyl.., ~OHD':""
OHO OOH
HO~O HO'L< ~O 0

O~O~A ?~
HOHO~ 0-..1 . -

D-glucose OH

OH

O~O :::,..

HO --r---...OJ O~ D-gI",."
",-chaconlne (72)

~ OH
L-rhamnose on o. OB
OB L-rhamnose
(a)

Fig. 36.18 (8 & b). Biologically active steroid alkaloids.

684 Alkaloids of Terpenoid Origin

H vated in western Bolivia because they are especially drought

and frost tolerant. The Aymara Indians that cultivate them
are relatively tolerant of the bitter taste present. Further,
these people consume clay along with the potatoes that ab-
sorbs much of the mixture of Solanum alkaloids. Geophagy
probably was an essential technique for solving the original
impasse these secondary compounds posed to the domestica-
R = O-rha(1-2)-O-rha(1-4)-glc tion process (Johns, 1985, 1990).

a-solamargine (79)
Teratogenic Properties
Many plants that contain Solanum alkaloids are responsi-
ble for the poisoning of livestock and humans. Potatoes in-
fected by the fungus Phytophthora infestans have been re-
ported to be responsible for an abnormal incidence of
congenital defects in humans and other animals. These re-
ports have prompted many studies into the nature of these
HO
problems. Of the alkaloids tested, demissine (70), 22-isode-
demissidine (76) missine, and a-tomatine (69) have been shown to be inactive.
Solasodine (46) had weak teratogenic activity, but
5a,22j3H,25j3H-solanidan-3j3-01 (81) or 22j3H,25aH-so-
lanid-5-en-3j3-01 (82) possessed strong teratogenic activity
(Cordell, 1981; Keeler, 1986).

VERATRlIM ALKALOIDS

solacasine (77) The alkaloids of Veratrum spp. (LiJiaceae) have long been
recognized as useful in medicine. In addition to the steroid
alkaloids found in this genus, structurally modified com-
pounds of two basic types are encountered: the jerveratrum
alkaloids and the ceveratrum alkaloids. Examples of the for-
mer class are jervine (83) and veratrarnine (84) (Fig. 36.19)
(Cordell, 1981; Jeger and Prelog, 1960a; Tomko and Vot-
icky, 1973); examples of the latter are protoveratrine A and
(b) (X--soladulcine (74) (L-rham., D-xyl, D-glu, D-gal) B (85 and 86) (Greenhill and Grayshan, 1992).
A simpler group of compounds with the cevane skeleton
Fig. 36.18. (continued)
occurs in the genus Fritillaria (Liliaceae). Medicinally, the
most important alkaloids of Veratrum are the polyesters of
Solanum Alkaloids in Potatoes protoverine (87) that have hypotensive properties.

It has long been known that "greened" potatoes (those Biosynthesis

that have been exposed to the sun), or potatoes exposed to
certain fungi, produce Solanum alkaloids and are somewhat 12-Hydroxysolanidine (epirubijervine) (88) may serve as
toxic. Both people and livestock have died from eating po- a precursor for a C-nor-D-homo-steroid derivative (91). Pos-
tato vines, sprouts, peels, or green and cull potatoes. Potato sibly 12j3-hydroxyteinemine (92) may serve as a precursor
breeders in the United States attempt to keep the a-solanine for veratramine (84). Reductive cleavage of the C(16)-N
(71) content of potato cultivars below 200 floglg fresh weight bond yields the skeleton of the jerveratrum alkaloids (Fig.
(Beier and Nigg, 1992). Fried potato peels contain as much 36.20) (Gross et al., 1985). Isorubijervine (89) (Fig. 36.21),
as 1400 flog/g a-solanine and a-chaconine (72), whereas the a solanidine alkaloid with a leaving group at C-1B, can form
recommended upper safety limit is 200 flog/g (Beier and a C(18)-N bond. Reduction of the resulting quaternary salt
Nigg, 1992). by cleavage of the C(16)-N bond yields the heterocyclic
Certain species of potatoes that are cultivated in Andean skeleton of the ceveratrum alkaloids. The alkaloid procevine
regions of South America are also known to contain larger (90) has been isolated from Veratrum grandiflorum (Gross
amounts of these alkaloids than are found in the usual Sola- et al., 1985). As the configuration at C-22 is opposite that of
num tuberosum cultivars. Plants of S. X ajanhuiri are culti- most naturally occurting ceveratrum alkaloids, anrakorinine
 Alkaloids o/Terpenoid Origin 68S

HO
27
veratramine (84)

J+1lo OH
HO

protoveratrine A, R=H (85) protoverine (87)

protoveratrine B, R=OH (86)

CH3-O ~I o
:::::,..
OCH3
OH
H

veralridine (94) H

HO
cyclopamine (96) cyclopsine (97)

Fig. 36.19. Veratrum alkaloids.

(93) may serve as a more likely precursor for these alkaloids Alkaloids of Veratrum are highly toxic to livestock. Con-
(Gross et al., 1985). sumption of these compounds results in teratological effects,
especially in sheep (Keeler, 1986). One of the most bizarre
Biological Activity effects is the production of lambs with one eye or "cyclop-
Veratrum alkaloids cause repetitive discharges of nerve ism." Under some circumstances, this deformity may affect
cells, apparently by delaying repolarization (possibly by ac- as many as 25% of the lambs produced (Keeler, 1986). The
tion on lipid components and an ATPase in the cell mem- three most active alkaloids are jervine (83), cyclopamine
brane) (Levinson, 1976). Veratridine (94) binds to the so- (96), and cyclopsine (97) (Keeler, 1986). Jervine has an LDso
dium channel and keeps it permanently open and active i.v. in mouse of 9.3 mg/kg and veratradine (94) of 1.4 mg/kg;
(Greenbill and Grayshan, 1992). Protoveratrines A and B rubijervine (67) in rat of 70 mg/kg; and protoveratrine a
inbibit inactivation of Na+ channels. Cevadine (95) and pro- lethal dose in rabbit of 0.1 mg/kg (Wink, 1993).
toveratrines A and B (85, 86) depolarize membranes (Wink, Many of this group of alkaloids possess antifungal prop-
1993). erties. Among the active compounds are cevadine (95), iso-
686 Alkaloids ojTerpenoid Origin

HO HO

(91)
I (not known to
epirubijervlne (88) .. occur naturally)
(12.hydroxysolanidine)

&n ~
;
~ H 7' •....•",QHN
f ~
7' !
H 0

HO ~
H
1 ! ~ HO ~ (not known to
occur naturally)

HO
~
U~.,~th
~
H H

jervine (83)
HO

/ veratramine (84)
/'

HO
Fig. 36.20. Proposed biogenesis of jerveratrum alkaloids (modified from Cordell. 1981; used with permission of the author).

rubijervine (89), jervine (83), protoveratrine A and B Sorrn, 1967, Jeger & Prelog, 1960). Most important among
(85,86), pseudojervine, rubijervine (67), veratramine (84), these are Funtumia, Holarrhena, and Maluoetia. Similar al-
veratridine (94), and verazine (47) (Wink, 1993). kaloids occur in the family Buxaceae (Pachysandra and Sar-
These alkaloids are widely used today in the treatment cocca). Alkaloids such as holaphylline (98) are based on
of hypertension. Medicinally, the most important alkaloids the 5a-pregnane skeleton and appear to be derived from
of Veratrum are the polyesters of protoverine (87) that have cholesterol (99) and pregnenolone (100) (Fig, 36,22) (Gross
hypotensive properties, Plants from Veratrum and Fritillaria et ai., 1985). A series ofbiosynthetic routes leading to major
have been used in Chinese traditional medicine (Han et ai., subgroups of these alkaloids has been proposed (Gross et
1988), ai., 1985),
Veratrum plant material has also been used as an insecti- Holarrhena antidysenteriea (Apocynaceae), an Indian
cide (Greenhill and Grayshan, 1992). Protoveratrine B (86)
plant, is used for treatment of dysentery. The steroidal alka-
is a feeding deterrent to Syntomis larvae; veratrine (94) is a
loid conessine (101) was the main antiamebic constituent
feeding deterrent for Sehistoeerca (Wink, 1993).
(Borris and Schaeffer, 1992), The same compound occurs
in Wrightia tomentosa, Another Indian plant, Chonemorpho
STEROIDAL ALKALOIDS IN THE
fragrans (Apocynaceae), afforded the alkaloid chonemor-
APOCYNACEAE
phine (102) which had an MIC of 25 fLg/ml in vitro (Borris
In addition to the indole alkaloids of the Apocynaceae, a and Schaeffer, 1992).
number of genera possess alkaloids based on a steroidal Conessine (101) is active against Candida albieons at
structure (Alta-ur-Rahman and Muzaffar, 1988; Cerny and a concentration of 100 fLg/ml (Clark and Hufford, 1992),
 Alkaloids of Terpenoid Origin 687

HO
skioooomenioe

Fig. 36.21. Proposed biogenesis of ceveratrum alkaloids (modified from Gross et aI., 1985).

HO

cholesterol (99) holophyllamine conessine (101)

chonemorphine (102) pregnenolone (100) bolaphylline (98)

Fig. 36.22. Alkaloids with a ~I-carbon skeleton and possible precursors.

688 Alkaloids oj Terpenoid Origin

~"'''''N#HCH3 NHCH,
..."H.

l
...."OH
"
H
.~ D
·""OH .... "OH

CH,-NH CH,-NH \~. CH,-NH

cyclovirobuxine (l05) buxamine-G (104)

cycIobuxine·D (103)
o carbon from C-2 of mevalonic acid (buxenine-G)
• 4-pro-R H of mevalonic acid

Fig. 36.23. Buxus alkaloids (modified from Gross et aI., 1985).

CH'02C~~
~
HO
N
. . . j" '" ......"..,,',--,..' /
i OCOCH,
OCOCH 3
.,
yuzunmme

Fig. 36.24. Proposed biogenesis of daphniphylline (modified from Cordell, 1981; used with pennission of the author) and some representative
Daphniphyllum alkaloids.
 Alkaloids of Terpenoid Origin 689

Conessine (101) is phagorepellent to Pieris, Bombyx, Ly- cynaceae and Buxaceae. in The Alkaloids, Vol. 32 (A. Brossi,
rnantria, and Dysdercus (Wink, 1993). ed.), 79-240, Academic Press, New York, 1988.
BARBOUR, J. D. and G. G. KENNEDY, Role of steroidal glycoalkaloid
a-tomatine in host-plant resistance of tomato to Colorado po-
tato beetle, 1. Chern. Bcol., 17, 989-1005 (1991).
BUXUS ALKALOIDS
BBlER, R. C. and H. N. NIGG, Natural toxicants in foods, in Phyto-
chemical Resources for Medicine and Agriculture (H. N. Nigg
The genera Buxus and Pachysandra of the family Buxaceae and D. S. Seigler, eds.), 247-376, Plenum Press, New York,
contain a series of steroidal alkaIoids similar to those of 1992.
HolarrhelUl (Apocynaceae) (Atta-ur-Rahman and Muzaffar, BBNN, M. H., Diterpene alkaloids, in Alkaloids and Sulphur Com-
1988; Brown, 1970; Cerny and Sorn 1967; Tomko and Vot- pounds (P. G. Waterman, ed.), Vol. 8 of Methods in Plant Bio-
icky, 1973); in addition, these plants contain a series of tri- chemistry (p. M. Dey and J. B. Harbome, eds.), 373-419, Aca-
terpenoid alkaloids commonly known as the Buxus alka- demic Press, London, 1993.
loids. These are typified by cyclobuxine-D (103), based on BBNN, M. H. and J. M. JACYNO, The toxicology and pharmacology
a cycloartenol skeleton (Fig. 36.23). The six terminal carbon of diterpenoid alkaloids, in Alkaloids. Chemical and Biological
atoms have been removed and nitrogen functions are at- Perspectives, Vol. I (S. W. Pelletier, ed.),451-472, Wiley, New
York,1983.
tached at C-3 and C-20. Buxenine-G (104) probably results
from cleavage of the cyclopropyl ring to generate a seven BISSET, N. G., Hunting poisons of the North Pacific region, Lloydia,
39, 87-124 (1976).
membered Bring.
Incorporation of [2_ 14C,(4R)-4-3HIl mevalonic acid BLECHERT, S. and D. GUENARD, Taxus alkaloids, in The Alkaloids,
Vol. 39 (A. Brossi, ed.), 195-238, Academic Press, New York,
(MVA) into cyclovirobuxine (105) and cyclobuxine-D (103)
1990.
in shoots of Buxus sempervirens (Buxaceae) gave 3H: I'C
BORRIS, R. P. and J. M. SCHAEFFER, Antiparasitic agents from
ratios of -3.4 and 3.3, consistent with the labeling pattern
plants, in Phytochemical Resources for Medicine and Agricul-
shown and the involvement of ketone groups at C-3 and C- ture (H. N. Nigg and D. S. Seigler, eds.), 117-158, Plenum
20 in the biosynthetic pathway. Oxidative removal of the Press, New York, 1992.
4a-methyl group of cyclobuxine-D (103) also is evident BRADLEY, V., D. 1. COu.tNS, F. W. EASTWOOD, M. C. IRVINE, J.
from these ratios (Fig. 36.23) (Gross et aI., 1985). M. SWAN, and D. E. SYMON, Distribution of steroidal alkaloids
in Australian species of Solanum in The Biology and Taxonomy
of the Solanaceae O. G. Hawkes, R. N. Lester, and A. D. Skeld-
DAPlI1YlPlIl'LWM ALKALOIDS ing, eds.), Linnear Society of London Symposium Series 7,
203-209, Academic Press, London, 1979.
BRENNEISEN, R. and M. A. ELSmR.Y, Socio-economic poisons:
The genus Daphniphyllum (Daphniphyllaceae) is the source Khat, the natural amphetamine, in Phytochemical Resources for
of complex triterpenoid alkaloids. Daphniphyllum rnacropo- Medicine and Agriculture (H. N. Nigg and D. S. Seigler, eds.),
dum, a tall tree from Japan, has been particularly studied 97-116, Plenum Press, New York, 1992.
(Yamamura, 1986; YamamUTa and Hirata, 1975). At least BROWN, K. S., JR., Abnormal steroidal alkaloids, in Chemistry of
19 alkaloids have been isolated from the bark, leaves, and the Alkaloids (S. W. Pelletier, ed.), 631-667, Van Nostrand
fruit of this plant. DaphniphyIIine (106) and codaphniphyl- Reinhold, New York, 1970.
line (107) are the two major alkaloids of the bark (Fig. CERNy, V. and F. SORM, Steroid alkaloids: Alkaloids of Apocyna-
36.24). ceae and Buxaceae, in The Alkaloids, Vol. 9 (R. H. F. Manske,
Labeled squalene was incorporated into both alkaloids ed.), 305-426, Academic Press, New York, 1967.
and degradation studies indicated derivation from six meva- CLARK, A. M. and C. H. HUFFORD, Antifungal alkaloids, in The
lonate units. A possible biosynthetic scheme has been pro- Alkaloids, Vol. 42 (G. A. Cordell, ed.), 117-150, Academic
posed (Fig. 36.24) (Cordell, 1981; Yamamura, 1986). Press, New York, 1992.
Plants of this genus have long been known to produce CORDELL, G. A., Introduction to Alkaloids: A Biogenetic Approach,
mortality in livestock in Japan (Yamamura, 1986). D. ma- Wiley-Interscience, New York, 1981.
cropodum has been used as a vermicide and asthma remedy CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual
(Cordell, 1981). Reports on NMR Spectroscopy, Vol. 13 (G. A. Webb, ed.),
60-210, Academic Press, London, 1982.
CROMBIE, L., The cathedulin alkaloids, Bull. Narco!., 32, 37-50
(1980).
REFERENCES CROMBIE, L., W. M. L. CROMBIE, and D. A. WHITING, Alkaloids
of khat (Catha edulis), in The Alkaloids, Vol. 39 (A. Brossi,
ATIA-UR-RAHMAN and M. I. CHOUDHARY, Steroidal alkaloids, in ed.), 139-164, Academic Press, New York, 1990.
Alkaloids and Sulphur Compounds (P. G. Waterman, ed.), Vol. CUlLER, H. G., An historical perspective of ancient poisons, in
8 of Methods in Plant Biochemistry (P. M. Dey and J. B. Har- Phytochemical Resources for Medicine and Agriculture (H. N.
borne, eds.), 473-510, Academic Press, London, 1993. Nigg and D. S. Seigler, eds.), 1-13, Plenum Press, New York,
ATI'A-UR-RAHMAN and A. MUZAFFAR, Steroidal alkaloids of Apo- 1992.
690 Alkaloids oj Terpeno;d Origin

DALY, J. W., H. M. GARRAFFO, and T. F. SPANOE, Amphibian JUVIK,J. A .. M. J. BERLINGER, T. BEN-DAVID, and J. RUDICH, Resis-
alkaloids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), tance among accessions of the genera Lycopersicon and Sola-
185-288, Academic Press, New York, 1993. num to four of the main insect pests of tomato in Israel, Phyto-
D'ARCY, W. G., The classification of the Solanaceae, in The Biol- parasitica, 10, 145-156 (1982).
ogy and Taxonomy of the Solanaceae (1. G. Hawkes, R. N. KEBLER, R. F., Teratology of steroidal alkaloids, in Alkaloids:
Lester, and A. D. Skelding, eds.), Linnear Society of London Chemical and Biological Perspectives, Vol. 4 (S. W. Pelletier,
Symposium Series 7, 3-47, Academic Press, London, 1979. ed.), 389-425, John Wiley & Sons, New York, 1986.
D'ARcY, W. G. (ed.), Solanaceae. Biology and Systematics, Co- KUPCHAN, S. M. and A. W. By, Steroid a1ka1oids: The Veratrum
lumbia University Press, New York, 1986. group, in The Alkaloids, Vol. to (R. H. F. Manske, ed.),
DREsSLER, R. L., The Orchids: Natural History and Classification, 193-286, Academic Press, New York, 1968.
Harvard University Press, Cambridge, MA, 1981. LEETE, E., The biosynthesis of alkaloids, Specialist Periodic Re-
DRUMMOND, B. A., m, Coevolution of ithomiine butterflies and ports, Biosynthesis, 102-223, The Royal Society of Chemistry,
solanaceous plants, in Solanaceae. Biology and Systematics (W. Cambridge, 1983.
G. D'Arcy, ed.), 307-327, Columbia University Press, New LEVINSON, H. Z., The defensive role of alkaloids in insects and
York,1986. plants, Experientia, 32, 408-411 (1976).
FODOR, G. and B. COLASANTI, The pyridine and piperidine alka- MANN, J., Secondary Metabolism, Oxford University Press, Ox-
loids: Chemistry and pharmacology, in Alkaloids: Chemistry ford, 1978; 2nd edition 1987.
and pharmacology, in Alkaloids: Chemical and Biological Per- MATA, R., Chemical studies and biological aspects of some Mexi-
spectives, Vol. 3 (S. W. Pelletier, ed.), 1-90, Wiley, New York, can plants used in traditional medicine, in Phytochemical Poten-
1985. tial of Tropical Plants (K. R. Downum, J. T. Romeo, and H. A.
GEISSMAN, T. A. and D. H. G. CROUT, Organic Chemistry of Sec- Stafford, eds.), 41-64, Plenum Press New York, 1993.
ondary Metabolism, Freeman Cooper, San Francisco, 1969. MATA, R., F. CALZADA, E. DiAz, and R. A. TOSCANO, Sesquiterpene
GREENHn..L, J. V. and P. ORAYSHAN, The cevane group of Veratrum evoninoate alkaloids from Hippocratea acelsa, PIanta Medica,
alkaloids, in The Alkaloids, Vol. 41 (A. Brossi and G. A. Cor- 56, 517 (1990).
dell, eds.), 177-237, Academic Press, New York, 1992. McCoy, J. W. and F. R. STBRMITZ, Alkaloids from Castilleja mini-
GROSS, D., H. R. SCHiiTrE, and K. SCHREWER, Isoprenoid alkaloids,
ata and Penstemon whippleanus, two host species for the plume
moth,Amblyptilia (Platyptilia) mica, J. Nat Prod., 46, 902-907
in Biochemistry of Alkaloids (K. Mothes, H. R. Schutte, and M.
(1983).
Luckner, ods.), 354-384, VCH Veriagsgeselischaft, Weinheim,
1985. MILLER, R. W., R. G. POWBU-, C. R. SMITH, JR., E. ARNOLD, and
J. CLARDY, Antileukemic a1ka1oids from Taxus wallichiana
HABERMllHL, G., The steroid a1ka1oids: The Salamandra group, in
Zucc., J. Org. Chem., 46,1469-1474 (1981).
The Alkaloids, Vol. 9 (R. H. F. Manske, ed.), 427-440, Aca-
demic Press, New York, 1967. NAHRSTBDT, A., Chemische Waffen bei hoheren Pflanzen, Pharma-
zie in unserer Zeit., 8, 129-138 (1979).
HAN, G., Y. CHEN, and X. LlANG, Distribution of a1ka1oids in tradi-
tional Chinese medicinal plants, in The Alkaloids, Vol. 32 (A.
i'Bu.ETlER, S. W., Studies in the chemistry of natural products:
Brossi, ed.), 241-270, Academic Press, New York, 1988.
Rearrangement reactions of diterpenoid and norditerpenoid al-
kaloids, J. Nat. Prod., 55, 1-24 (1992).
HARBORNB, J. B., Systematic significance of variations in defense
i'Bu.ETlER, S. W. and N. V. Mooy, The structure and synthesis of
chemistry, in Solanaceae. Biology and Systematics (W. G.
C l9-diterpenoid a1ka1oids, in The Alkaloids, Vol. 17 (R. H. F.
D'Arcy, ed.), 328-344, Columbia University Press, New York,
Manske and R. G. A. Rodrigo, ods.), 2-104, Academic Press,
1986.
New York, 1979.
HBoNAUBR, R., Chemotaxonomie der Pflanzen, Vol. 3, Birkhliuser
i'ELLETlER, S. W. and N. V. MODY, The chemistry of C 20-diterpen-
Verlag, Baael, 1964.
oid a1ka1oids, in The Alkaloids, Vol. 18 (R. H. F. Manske and
HOWARD, A. S. and J. P. MICHAEL, Simple indolizidine and quino- R. G. A. Rodrigo, eds.), 100-216, Academic Press, New York,
lizidine a1ka1oids, in The Alkaloids, Vol. 28 (A. Brossi, ed.), 1981.
183-308, Academic Press, New York, 1986.
POWELL, R. G., R. W. MnLER, and C. R. SMITH, JR., Cephaloman-
JEGER, O. and V. i'RELoo, Steroid alkaloids. Veratrum group, in nine; a new antitumour alkaloid from Cephalotaxus mannii,
The Alkaloids, Vol. 7 (R. H. F. Manske, ed.), 364-418, Aca- Chern. Commun., 102-104 (1979).
demic Press, New York, 1960a.
i'RELoo, V. and O. JEGBR, The chemistry of Solanum and Veratrum
JEGER, O. and V. PREwo, Steroid a1ka1oids: The Holarrhena group, alkaloids, in The Alkaloids, Vol. 3 (R. H. F. Manske and H. L.
in The Alkaloids, Vol. 7 (R. H. F. Manske, ed.), 319-342, Holmes eds.), 248-312, Academic Press, New York, 1953.
Academic Press, New York, 1960b. PREwo, V. and O. JEGBR, Steroid alkaloids. The Solanum group,
JOHNS, T. A., Chemical ecology of the Aymara ofWestem Bolivia: in The Alkaloids, Vol. 7 (R. H. F. Manske, ed.), 343-363,
Selection for glycoalkaloids in the Solanum X ajanhuiri domes- Academic Press, New York, 1960.
tication process, Ph.D. dissertation, University of Michigan, RIPPERGER, H. and K. SCHREIBER, Solanum steroid alkaloids, in
Ann Arbor, 1985 (Diss. Abs. DA8512440). The Alkaloids, Vol. 19 (R. H. F. Manske and R. G. A. Rodrigo,
JOHNS, T. A., With Bitler Herbs They Shall Eat It, University of eds.), 81-192, Academic Press, New York, 1981.
Arizona, Tucson, 1990. ROBY, M. R. and F. R. STBRMITZ, Pyrrolizidine and pyridine mono-
JUVlK, J. A. and M. A. STEVENS, Inheritance of foliar a-tomatine terpene a1ka1oids from two Castilleja plant hosts of the plume
content in tomatoes, J. Am. Hort. Soc., 107,1061-1065 (1982). moth, Platyptilia pica, J. Nat. Prod., 47, 846-853 (1984).
 Alkaloids oj Terpenoid Origin 691

RODDICK, I. G., Isoprenoid alkaloids, in Secondary Plant Products Alkaloids, Vol. 25 (A. Brossi, ed.), 3-355, Academic Press,
(E. A. Bell and B. V. Charlwood, eds.), 167-184, Springer- New York, 1985.
Verlag, Berlin, 1980. SZENDRBI, K., The chemistry of khat, Bull. Narco!., 32, 5-35
RODDICK, J. G., Steroidal alkaloids of the Solanaceae, in Solana- (1980).
ceae. Biology and Systematics (W. G. D'Arcy, ed.), 201-222, TOMKO, J. and Z. VOTICKY. Steroid alkaloids: The Veratrum and
Columbia University Press, New York, 1986. Buxus groups, in The Alkaloids, Vol. 14 (R. H. F. Manske, ed.),
RODDICK,1. G., A. L. RIJNENBERG, and S. F. OSMAN, Synergistic 1-83, Academic Press, New York, 1973.
interaction between potato glycoalkaloids a-solanine and a- VERPOORTE, R., R. VAN DER HEnoEN, W. M. VAN GULlK, and H.
chaconine in relation to destabilization of cell membranes: Eco-
J. G. TEN HooPEN, Plant biotechnology for the production of
logical implications, 1. Chern. Bcol., 14, 889-902 (1988).
alkaloids: Present status and prospects, in The Alkaloids, Vol.
SATO, Y., Steroidal alkaloids, in Chemistry of the Alkaloids (S. 40 (A. Brossi, ed.), 1-188, Academic Press, New York, 1991.
W. Pelletier, ed.), 521-629, Van Nostrand Reinhold, New York,
WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and Metabo-
1970.
lism in Plants, Plenum Press, New York, 1978.
SCHREIBER, K., Steroid alkaloids: The Solanum group, in The Alka-
loids, Vol. 10 (R. H. F. Manske, ed.), 1-192, Academic Press, WINK, M., Allelochemical properties or the raison d'elre of alka-
loids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), 1-118,
New York, 1968.
Academic Press, New York, 1993.
SCHREIBER, K., The steroid alkaloids of Solanum, in The Biology
and Taxonomy of the Solanaceae (J. G. Hawkes, R. N. Lester, WITHERUP, K. M., S. A. LooK, M. W. STASKO, and G. M. CRAGG,
and A. D. Skelding, eds.), Linnear Society of London Sympo- Taxus spp. needles contain amounts of taxol comparable to the
sium Series 7, 193-202, Academic Press, London, 1979. bark of Taxus brevifolia: Analysis and isolation, J. Nat. Prod.,
SPECIAUST PERIODIC REPoRT, Alkaloids, Vol. 5, Chemical Society, 53, 1249-1255 (1990).
London, 1975. WITKOP, B. and E. GOSSINGER, Amphibian alkaloids, in The Alka-
STERMI1Z, F. R. and G. H. HAiuus, Reappraisal of the structore loids, Vol. 21 (A. Brossi, ed.), 139-254, Academic Press, New
for the alkaloid buddantin from Buddleya davidii, Tetrahedron York,1983.
Lett., 26, 5251-5252 (1985). Y AMAMURA, S., Daphniphy/lum alkaloids, in The Alkaloids, Vol.
STRUNZ, G. M. and J. A. FINDLAY, Pyridine and piperidine alka- 29 (A. Brossi, ed.), 265-286, Academic Press, New York, 1986.
loids, in The Alkaloids, Vol. 26 (A. Brossi, ed.), 89-184, Aca- Y AMAMURA, S. and Y. HIRATA, The Daphniphyllum alkaloids, in
demic Press, New York, 1985. The Alkaloids, Vol. 15 (R. H. F. Manske, ed.), 41-82, Aca-
SUfFNESS, M. and G. A. CORDELL, Antitumor alkaloids, in The demic Press, New York, 1975.
37
Miscellaneous Types of Alkaloids

Introduction Both the 'H- and 13C-NMR (nuclear magnetic resonance)

Hemiterpene Alkaloids spectra for several of the alkaloid groups below have been
Imidazole Alkaloids reviewed (Crabb, 1982). The results of feeding stodies lead-
Biosynthesis ing to the synthesis of several alkaloids of this group have
Other Sources of Imidazole Alkaloids been tabulated (Leete, 1983).
Lythraceae alkaloids
Maytansinoids
Antitumor Activity HEMITERPENE ALKALOIDS
Other Biological Activity
Muscarine Alkaloids Many groups of alkaloids possess five-carbon mevalonate-
Naphthylisoquinoline Alkaloids derived units as a part of the overall structure of the com-
Peptide Alkaloids pounds. In one previously discussed group, the ergot alka-
Purine or Xanthine Alkaloids loids, this mevalonate unit is attached to a tryptophan-de-
Biosynthesis rived portion and cyciized to form tricyclic and tetracyclic
Physiological Activity of Xanthine Derivatives alkaloids.
Pyrazine Alkaloids Another series of hemiterpene alkaloids is found in the
Securinega Alkaloids genus Alchornea (Euphorbiaceae). In these alkaloids (1-4)
Sesbania Alkaloids the hemiterpene uuit constitutes a major portion of the struc-
Betalains tore (Fig. 37.1) (Hart et al., 1970).
Biosynthesis
Chemosystematic Value
Betalains in Foods and Beverages IMIDAZOLE ALKALOIDS
Miscellaneous Alkaloids
Trigonelline and an Unusual Pyrrole Several alkaloids derived from histidine occur in plants.
Histronionicotoxins Probably the best known of these is pilocarpine (5), found
Huperzine A in species of Pilocarpus (Rutaceae) (Fig. 37.2). This com-
References pound is a peripheral stimulant of the parasympathetic sys-
tem and is used topically as a myotic to counteract the mydri-
atic effects of atropine and other parasympatholytic drugs.
Il'ITRODUCTION Pilocarpine is used for the treatment of glaucoma (as a
0.5-10% solution).
A number of miscellaneous types of alkaloids not easily Positively inotropic histamine-derived compounds have
assignable to other alkaloid classes, but that are of interest been isolated from members of the Cactaceae. N' ,N'-Dimeth-
because of their medical, drug, systematic, biosynthetic, or ylhistamine (6) from Echinocereus blanckii and dolicothe-
other properties, are surveyed in this chapter. Many of these line (7) from Dolichothele sphaerica have been studied. Dol-
are of limited distribution, whereas others, such as caffeine ichotheline has an EDso of 1.6 X 10-7 mollL (Figure 37.3)
(a xanthine base), are widely and sporadically distributed. (Wagner, 1988).

692
 Miscellaneous Types of Alkaloids 693

alchorneine (1) alchorneinone (2)

)(N'l
yTJ"""''\
N /
N~N\.,,"'"
~ V~I-H
r"'·····\.-N-/
OCH,
I
<
o

alchornine (4) isoalchorneine (3)

Fig. 37.1. Hemiterpene alkaloids from the genus Alchornea.

Biosynthesis baceae), Cynometra (Fabaceae), Cypholophus (Urticaceae),

Dendrobium (Orchidaceae), Dolichothele (Cactaceae),
Pilocarpine has been suggested to arise from condensa-
Glochidion (Euphorbiaceae), and Macrorungia (Acantha-
tion of an alcohol involved in the biosynthesis of histidine
ceae) (Maat and Beyerman, 1983). These include many
(as the phosphate) and acetoacetate (Fig. 37.2). A second
structural variants and may be of quite distinct origins.
path has also been suggested. There is little experimental
evidence to establish either pathway (Maat and Beyerman,
1983).
LYTHKACBAE ALKALOIDS
other Sources of Imidazole Alkaloids
About 45 alkaloids from the above-ground parts of members
Other imidazole alkaloids are known from Alchornea of the Lythraceae have been studied (Golebiewski and Wro-
(Euphorbiaceae), Casimiroa (Rutaceae), Cassia absus (Fa- bel, 1981). Several of these alkaloids contain quinolizidine

-+

pilocarpine (5)

Fig. 37.2.Proposed pathways for the biogenesis of pilocarpine (Maat and Beyennan. 1983, modified and used with pennission of the copyright owner,
Academic Press, Orlando, FL).
694 Miscellaneous Types of Alkaloids

serves as a precursor for much of the rest of the molecul.

[3- 14C]Phenylaianine is incorporated specifically into cry,
genine (8) (Fig. 37.3), and is the precursor for both of th
aromatic rings. Feeding experiments with other labeled ph.
nylalanine precursors established that two intact C6 -C3 unit
are incorporated into lactonic Lythraceae alkaloids, such a
N,N·dimetbylbistamine (6) dolicotheJine (7)
vertine (9) and verticillatine (10) (Fig. 37.4) (Golebiewsk
and Wrobel, 1981).
Fig. 37.3. Dolichotheline and cryogenine. The American Indians used plants of the genus Heimi.
to prepare intoxicating and stimulating beverages that pro
duce a mild psychosomirnetic effect. Plants of this genu:
ring systems. Some of the best known alkaloids have been
were commonly used medicinally (Golebiewski and Wrobel
isolated from the hallucinogenic plant Heimia salicifolia and
1981).
from crepe myrtle Lagerstroemia subcostata (Howard and
Michael, 1986).
The biosynthesis of Lythraceae alkaloids is not well JllAYTANSINOmS
understood. Lysine serves as a precursor for part of the mole-
cule; all carbons except the carboxyl group of lysine are A series of novel ansa-macrolides has been isolated from
incorporated. Further, the lysine-derived portion enters the plants of several genera of the Celastraceae, as well as othel
molecule via a symmetrical intermediate. Phenylalanine families. The macrolide alkaloid, maytansine (11), was first

O~

.o~ ~~~:~ ~~
CO,H 0 H
o _~~

OCH, OCH, OCH,

HO~i ~ '! O~l!

.~.
HO
l! H N
H
N
H
N

?' ?' 1
I"'"
.0 OCOCH, HO ""- I HO ""-
OCI;I, OCOCH, OCH, OCH,

(-)-Iasublne I demethyllasubine I

10-epidemethoxy· abresoline
(al abresoline

Fig. 37A (8 & b). Some representative Lythraceae alkaloids and a proposed biogenetic scheme for their fonnation (Golebiewski and Wf, 981;
modified and used with pennission of the copyright owner, Academic Press, Orlando. FL).
 Miscellaneous Types of Alkaloids 695

H H

-+-Iysine

OH OCH,

OCH, OH OCH, OH

Iythrine cryogenin. (8)

0 0

12

L3

15

I'

OCH, OH OCH,
OH

(b)
vertine (9) verticillatin. (10)

Fig. 37.4. (continued)

obtained from the fruits of May tenus ovatus (Fig. 37.5). Sim- Trewia nudiflora (Powell et al., 1981, 1983). Several include
ilar alkaloids have been obtained frum the stem bark of an ususual second macrocyclic ring attached to the macrocy-
Maytenus buchanan;; andM. serrata and lither plant sources. dic ring found in inost maytansinoids (Powell et aI., 1981,
Another celastraceous plant, Putterlickia verrucosa (Celas- 1982).
traceae), proved to be rich in maytansine (11); the plant The biosynthesis of these alkaloids has been little stodied.
contained as much as 12 mg/kg of maytansine. Maytansi- The origin of the various parts of the molecule has been
noids also have been isolated from Trewia nudiflora (Eu- suggested (Fig. 37.5). In general, the biosynthesis of ansa-
phorbiaceae) and Colubrina texensis (Rbamnaceae) (Reider mycin antibiotics of microbial origin and maytansanoids
and Roland, 1984; Smith and Powell, 1984). from higher plants are related (Smith and Powell, 1984).
Techniques for the isolation and purification and both 1H-
and 13C_NMR and mass spectrometry of these alkaloids have Antitumor Activity
been reviewed (Smith and Powell, 1984).
Of the compounds found in these plants, maytansine (11), Maytansine (11) has been stodied as an antitomor agent.
maytanprine (12), maytanvaline, and maytanbutacine Average ED,o values for these materials are in the range
showed antitomor activity, but most of an accompanying 10-4 _10- 6 ltg/mi. Maytansine (11) is highly active in a
series of structurally· related compounds did not (Fig. 37.5) variety of murine tomor systems including BI6 melanoma,
(Smith and Powell, 1984). The LDso s.c. in mouse of may- colon 26, Ll210 leukemia, Lewis lung carcinoma, and P-
tansine is 0.48 mg/kg (Wink, 1993). 388 leukemia (Smith and Powell, 1984). Several members
A complex series of maytansinoids has been isolated from of the maytansinoid family of alkaloids have antileukemic,
696 Miscellaneous Types of Alkaloids

CR,·O

maytanslne (11) maytanvaline (12)

CI o CI o

OCR,

maysenine N-methylmaysenine

(aJ actomycin

Fig. 37.5 (a & b). Maytansinoids (modified in part from Suffness and Cordell, 1985; used with pennission of the copyright owner, Academic Press,
Orlando. Fl.).

cytotoxic. antitubulin, and antintitotic properties (Cordell. trewiasine (14), and demethyltrewiasine (15) were excep-
1978; Suffness and Cordell, 1985). The antitumor activity tionally active in the PS, B1. and KB tumor lines (Powell
is mainly produced by the prevention of microtubule protein et al., 1981).
polymerization because of tubulin binding. Maytansine (11)
and vincristine have been shown to compete for the same Other Biological Activity
binding sites but utilize different binding mechanisms (Re-
ider and Roland. 1984). At 6 X 10-8 M, maytansine irrevers- Maytenus ilicifolia from southern South America has
ibly inhibits cell division in the eggs of sea urchins and clams been used as a menses inducer by indigenous people of Para-
and also irreversibly inhibits the in vitro polymerization of guay (Ahmed et al., 1981).
tubulin. This compound is 100 times more effective than Extracts of Trewia nudiflora (Euphorbiaceae) that contain
vincristine as an inhibitor of marine eggs. Maytansine inhib- maytansanoids have been shown tu have insect antifeedant
its DNA synthesis. but not RNA or protein synthesis. activity and produce several other effects in insects that may
Although maytansine has been studied in an extensive limit herbivory (powell et al .• 1982, 1983; Reider and Ro-
series of clinical trials, there does not appear to be sufficient land, 1984; Smith and Powell, 1984). Several maytansinoids
antitumor activity and the future of maytansine as an antican- were antifeedant to larvae of Ostrinia nubilalis, the Euro-
cer drug appears doubtful (Reider and Roland, 1984). pean com borer. whereas others significantly retarded devel-
The maytansinoid compounds trewiasine (13). dehydro- opment of the larvae (Madrigal et al., 1985).
 Miscellaneous Types of Alkaloids 697

CI

CH,-O

CI dehydrotrewiasine (14)

CH,-O

OCH,

Irewiasine (13)

(b) demethyltrewiasine (15)

Fig. 37.S. (continued)

Trewiasine (13), from the seeds of Trewia nudiflora (Eu- rhea, bronchoconstriction, hypotension, and vasodilation.
phorbiaceae), gives greater than 50% inhibition of radicle Amanita muscaria has been used by several indigenous
elongation with velvetleaf (Abutilon theophrasti) (Malva- groups for religious rites and as a hallucinogen.
ceae) (Powell and Spencer, 1988). Muscarine (16) is derived from glutamic acid and pyruvic
acid (Fig. 37.6) (Wang and Joullie, 1984). The carbons of
pyruvate are incorporated at positions 2 and 3 of the ring
!llVSCARll'lE ALKALOIDS and the methyl group at C-2. The carbons at position 2,3,
and 4 of glutamate were incorporated into positions 4 and
The alkaloid muscarine (16) is found in a number of fungi 5 of the ring of muscarine and at the methylene at C-5. The
of the genera Amanita, Clitocybe, and Inocybe (Fig. 37.6). carbons at positions 1 and 5 of glutamate are lost during
The amount of muscarine (16) present usually is quite low biosynthesis (Wang and Joullie, 1984).
(about 0.0003%). Fungi of the genera Clitocybe and Inocybe The action of muscarine (16) and acetylcholine on smooth
usually are responsible for' 'muscarine" poisoning. The best muscle is very similar; muscarine is a potent parasympatho-
known species of muscarine-containing alkaloids is un- mimetic drug (Antkowiak and Antkowiak, 1991). The term
doubtedly Amanita muscaria; however, much of the activity "muscarinic activity" is used to describe the direct periph-
of this species is attributable to ibotenic acid (17) and musci- eral action on cholinergic receptors in smooth muscles of
mol (18) (Antkowiak and Antkowiak, 1991; Kendrick, the gastrointestinal tract, eye, exocrine glands, and heart
1985). This attractive mushroom grows in most temperate (Wang and Joul!ie, 1984). Much early experimental work
climate areas of the world and occasionally is responsible for with muscarine is unreliable, as many of the materials used
human poisonings, although these are relatively uncommon were impure.
(Wang and Joullie, 1984). Symptoms of poisoning include The mushroom Amanita muscaria has a reputation for its
headaches, nausea, vomiting, salivation, lacrymation, diar- insecticidal properties. Pure muscarine, however, is devoid
698 Miscellaneous Types of Alkaloids

4~).CO.- H OH
HO,C 3 __1'\ _
H NH3+ HO.C~CO,- --+
H NH3+

(+)-(2S,3R,SS)-muscarine (16) musclmol (18) ibotenic acid (17)

J:J-
HO

"
J:) N+
,CH3 HO
N+
,CH3

--""'-"CH
o CH3 3 o I 'CH3
CH3

(+)-(2S,3R,SR)-allomuscarine (+)-(2S,3S,SR)-epimuscarlne

Fig. 37.6. Muscarine and related alkaloids (modified from Wang and Joullie, 1984; used with pennission of the copyright owner, Academic Press,
Orlando, FL).

of such properties. Compounds such as ibotenic acid (17) posed (Fig. 37.7) (Bringmann, 1986). Feeding studies with
and muscimol (18) appear to be responsible for much of this acetate suggest that these alkaloids are derived by a type of
activity, as well as the potent effect on the central nervous polyketide pathway.
system in mammals (Wang and Joullie, 1984). Both ibotenic Ancistrociadus abbreviatus is widely used in folk medi-
acid and muscimol bind to 'Y-aminobutyric acid and gluta- cine (Bringmann et al., 1990). Leaves of the Ancistrociadus
mate receptors (Antkowiak and Antkowiak, 1991). Ibotenic tectorius are used in Thailand to treat dysentery and malaria
acid is converted to muscimol upon drying (Kendrick, 1985). (Tantisewie and Ruchirawat, 1992).
The oral LDs. ofmuscimol in rat is 45 mgikg (Wink, 1993). Michellamine B (19), from Ancistrocladus korupensis
Muscimol (18) induces food aversion in Opossum (Wink, (formerly considered to be A. abbreviatus) has been shown
1993). to possess potent activity against mv-1 and mV-2
(Manfredi et al., 1991; Thomas and Gereau, 1993).

l'IAFHTIfYLlSOQUIi'IOLIl'IE ALKALOIDS PEPTIDE ALKALOIDS

An unusual group of perhaps 20 alkaloids is found in the A series of over 100 alkaloids with a peptide structure have
families Ancistrocladaceae and Dioncophyllaceae. The been isolated and characterized. In these compounds, a 10-
monotypic genus Ancistrociadus, Ancistrocladaceae, occurs or 12-membered (sometimes 13 or 14) ring spans the 1,3 or
in Africa and Asia (Bringmann, 1986). The genera Dionco- 1,4 positions of a benzene ring. These alkaloids are primarily
phyllum and Triphyophyllum are members of the Dionco- composed of simple amino acids. The basic properties are
phyllaceae, the only other plant family known to produce caused by an N-methyl or an NoN-dimethyl amine group of
this type of alkaloid. These plants are native to the Old World an N-terminal amino acid (Jouillie and Nut!, 1985;
tropics (Bringmann, 1986). The co-occurrence of certain al- Tschesche and Kaussman, 1975).
kaloids suggests a relationship between the two families Peptide alkaloids were first studied because of their thera-
(Bringmann et al., 1990). peutic, antibiotic properties. There is some evidence that
A biogenetic scheme for these alkaloids has been pro- peptide alkaloids may serve as ionophores in the plant and
 Miscellaneous Types of Alkaloids 699

from Ancistrocladus species from Triphyophyllum species

OCH,
(a) triphyophyttine isotriphyophyttine

OCH,

(b) michellamine B (19)

Fig. 37.7 (a & b). Proposed biogenetic scheme for naphthylisoquinoline alkaloids and michellamine B (Bringmann, 1986; modified and used with
pennission of the copyright owner, Academic Press, New York).
700 Miscellaneous Types of Alkaloids

are important in the absorption of nutrients from the soil and Phillipson, 1980). Cola (or kola) is derived from Stereu
(Joullie and Nutt, 1985). iaceae trees of the genus Cola (Stereuliaceae) which at
The bark of Scutia buxifolia (Rharnnaceae) contains scut- widely cultivated in West Africa, but also in other tropic,
ianine A, B (20), C, D, F (21), and G (22). Similar com- areas of the world. The cotyledons of the seed are the soorc
pounds have been isolated from the roots of Melochia tomen- of cola flavoring. The caffeine content may be as high a
tosa (23 and 24). A number of 14-membered-ring peptide 3.5%. A dried paste made from the seeds of Paullinia cupan,
alkaloids have been isolated from members of the genus (guararui) is the base of several beverages prepared in Brazil
Zizyphus (Rbarnnaceae) (Fig. 37.8). An aIkaIoid from this The beverage often contains 2.5-5% caffeine (29). Yerb.
family, ziziphin (25), blocks activity of the sugar (but not mare, flex paraguariensis, is widely used as a tea in southen
the salt) receptors of flies (Stiidler, 1984). South America. The dried leaves of this plant often contail
These compounds are particularly common in the Rbam- about 2% caffeine. The leaves of another species flex guay
naceae but also occur in the families Celastraceae, Euphor- usa, which contain 1.5-3.5% caffeine, are used daily bJ
biaceae, Hymenocardiaceae, Menispermaceae (Cocculus), Jivaro Indians (Lewis, 1992).
Pandaceae, Rubiaceae (Canthium, Oldenlant/ia), Stereulia- Tea is derived from the leaves of Camellia sinensi,
ceae (Melochia, Waltheria), and Urticaceae (Lagarias et al., (Theaceae), a plant native to eastern Asia. Different grade,
1979; Schmidt et al., 1985). Several structural types have and types of teas are prepared by various processes of bruis-
been recognized (Fig. 37.8) (Schmidt et al., 1985). ing, aging, and drying tea leaves. Significant chemical
Stems, leaves, and flowers of Bouvardia ternifolia (Rubi- changes occor during these procedures. Dried tea often con-
aceae) from Mexico were shown to contain cytotoxic and tains 1-4% caffeine (29) along with smaller amounts of
antitumor compounds. Fractionation revealed two cyclic theob.romine (31) and theophylline (31) (Cordell, 1981).
hexapeptides, bouvardin (26) and deoxybouvardin (27) (Fig. Cacao, from Theobroma cacao (and related species of
37.8) (Suffness and Cordell, 1985). The antitumor effects the Sterculiaceae), is derived from the seed, which is fer-
of these compounds were attributed to the ability of these mented and roasted. These seeds contain 0.9-3% theobro-
compounds to inhibit protein synthesis because of an effect mine (31) and the husks 0.2-3%. The genus Theobroma is
on the 80-S ribosome. Similar compounds have been found native to the New World but is extensively cultivated in
to be the antitumor principles of Rubia akane and R. cordi- several tropical areas, especially West Africa.
folia (Suffness and Cordell, 1985). Other purine and pyrimidine derivatives with side chains
Certain linear peptide aIkaIoids are known from marine have been found in plant extracts. Many of these are physio-
sponges (Schmidt et al., 1985). logically active in plants as cytokinins and are related struc-
turally and metabolically to zeatin (Horgan and Scott, 1991).
Other purine aIkaIoids occor in Banisteriopsis inebrians
(Malpighiaceae), Holarrhena mitis (Apocynaceae) [triacan-
PURINE OK XAN11IIl'IE ALKALOIDS thine (32)], Leontinus edodes (Tricholomataceae) [deoxyeri-
tadenine (33)], trans-zeatin (34) in Zea mays (Poaceae), and
Both purine and pyrimidine bases occur as constituent parts Lupinus angustifolius (Fabaceae) [lupinic acid (35)] (Atta-
of nucleic acids and plant products. In addition, a number or-Rahman and Choudhary, 1990). The uracil-derived amin'o
of free purine and pyrimidine bases have been isolated from acid willardiine (36) has been isolated from the seeds of
plants (Atta-or-Rahman and Choudhary, 1990; Brown, Acacia willardiana and other species of this genus (Brown,
1991; Wang, 1973). The pyrimidine glucoside vicine (28) 1991). The seeds of Lathyrus tingitanus contain another
was ftrst isolated. from seeds of Vicia species in 1870. How- amino acid, lathyrine (37), that is similar in structure (see
ever, purine bases appear to be much more common that Chapter 13) (Brown, 1991). The pyrimidine glucoside vicine
pyrimidine bases in nature. The most important purine alka- (28) is the causative agent of favism, a genetic defect in
loids are derived from the xanthine nucleus. Xanthine itself individuals that result in acute hemolytic anemia that can
has not been found naturally, but several of its N-aIkyl deriv- prove fatal. This problem is most common in those of Medi-
atives are of considerable importance. The most important terranean descent (Brown, 1991).
of these are 1,3,7-trimethylxanthine (caffeine) (29), 1,3-di- The fruits of Zizyphus jujuba (Rbarnnaceae) are con-
methylxanthine (theophylline) (30), and 3,7-dimethylxan- sumed directly and also used as a folk medicine in several
thine (theobromine) (31) (Fig. 37.9). These alkaloids are countries. This material, zizyphiJructus, contains the highest
major constituents of plants used as stimulating beverages levels of cyclic AMP (1100-500 nmoVg) known from either
by people throughout the world. plant or animal sourees (Hikino, 1985). This uriental plant
Coffee, Coffea arabica and Coffea canephora (Rubia- drug also contains high levels of cyclic GMP (30-60
ceae), is native to East Africa but widely cultivated in many nmoVg). Cyclic AMP may be responsible for the antiallergic
tropical countries. The seeds contain 1-2% caffeine com- activity of this material (Hikino, 1985).
plexed with chlorogenic acid. All parts of the coffee plant
contain caffeine, although the greatest concentration is in Biosynthesis
the frnits (Waller and Nowacki, 1978). Caffeine also occurs Cell-free enzyme systems capable of synthesizing caf-
in Genipa and Oldenlandia (both Rubiaceae) (Hemingway feine (29) and related xanthines have been prepared from
 Miscellaneous Types of Alkaloids 701

(b) deoxybouvardin (27)

Fig. 37.8 (a & b). Peptide alkaloids.

702 Miscellaneous Types of Alkaloids

o CH, o

CH31~)
o '-NJ--N
'"llO
o N
I
N

CH,
I
CH,
xanthine skeleton caffeine (29) theophylline (30) theobromine (3I)

Y NH,

(x);
NH, ( N:):NH
I);
N?' N
~N NH OH

N N ~CO'H
(aJ
triacanthine (32) deoxyeritadenine (33) trans-zeatin (34) lupinic acid (35)

o
1~ N NH,

~CO'H
willardiine (36) lathyrine (37) vicine (28)
o o 0yNH,

HO_N~
Ayo.gIUCOSYI
o
J-.... NH,
O~N)
HN , ...
OJ-.HN

convicine
HO~
HO OH
3·hydroxyuridine (39) saxitoxin (40)
(bJ

Fig. 37.9 (a & b). Xanthine and related alkaloids.

Camellia (Thea) sinensis (tea) and Coffea arabica (coffee) Physiological Activity of Xanthine
plants. Caffeine was fonned rapidly by extracts of green Derivatives
coffee berries, but little by more mature ones, and not at
all by seedlings. Biosynthesis of caffein proceeds from 7· Xanthine derivatives have a number of phannacological
methylxanthosine in the presence of an active purine nucleo- properties in common. Five major actions are observed: (I)
side phosphorylase or 7.methyl·,y"·nucleoside hydrolase. central nervous system and respiratory stimulation, (2) skele·
Methionine and S-adenosylmethionine serve as precursors tal muscle stimulation, (3) diuresis, (4) cardiac stimulation,
for the methyl groups of purine alkaloids. These act in the and (5) smooth·muscle relaxation. Caffeine (29) increases
presence of methyltransferases on 7·methylxanthine (38) central nervous system activity and its main effect is on the
and theobromine (31) to produce caffeine. A pathway for cerebral cortex, where it acts to produce clear thought and
the origin of these compounds in coffee and tea plants has reduce drowsiness and fatigue. The nonnal dose is 100-200
been proposed (Suzuki et aI., 1992; Waller and Denner, mg (Cordell, 1981). The oral LDso in mouse is 127-137
1981) (Fig. 37.10). mg/kg; the oral LD50 in rat for theobromine (31) is 950
 Miscellaneous Types of Alkaloids 703

HO-P-O~
I
?
OH
&>--i6:f).- i:\) -iYy) OANH NH
I
ribosyl
OANH N
oANJ-N
I
CH3

HO OH 7-methylxanthosine 7-methyl,anthine (38) 3,7-dimethylxanthine

(31) (theobromine)
adenosine monophosphate (AMP)

-),j::> -'l)=5
o

1,3-dimethylxanthine (30) I-methylxanthine 1,7-dimethylxanthine

caffeine (29)
(theophylline) (paraxanthine)

Fig. 37.10. Biogenesis of xanthine alkaloids (modified from Suzuki et aI., 1992; modified and used with permission of the copyright owner, Elsevier
Science Ltd., The Boulevard, Langford Lane, Kidlington OXS 1GB, UK).

mg/kg (Wink, 1993). Both caffeine and theophylline (30) (Nahrstedt, 1982). Monarch butterflies sequester pyrazines
inhibit cyclic AMP phosphodiesterase (Wink, 1993). that have a quinoline-like odor from their host plant, Asclep-
Incorporation of I % caffeine (29) or theobromine (31) ias curas.avica. It is thought that these compounds serve as
into artificial diets was totally lethal to the bruchid Calloso- warning odors. In a sense, they may be aposematic odors,
bruchus maculatus (Janzen et al., 1977). Caffeine has anti- as the insects are well protected with cardiac glycosides and
feedant properties toward Phormia, polyphagous Syntomis pyrrolizidine alkaloids. Other lepidoptera such as Zygaena
larvae (Lepidoptera), bees, Agelaius, Bombyx, and Ly- lonicerae and an Australia Amata species also contain these
mantria (Wink, 1993). compounds. One can smell the compounds from the last two
Caffeine is autotoxic to Coffea seedlings. Caffeine, the- insects simply by disturbing them (Harborne, 1987, 1989;
ophylline and theobromine inhibit growth of lettuce seed- Rothschild et al., 1984). The compounds are thought to arise
lings and certain other species of plants (Wink, 1993). from the host plants.
3-Hydroxyuridine (39), from plant material of Baillonella 3-Isobutyl-2-methoxypyrazine (41) is a major odor com-
toxisperma (Sapotaceae), inbibits growth of seedlings of sev- ponent of Capsicum annuum. Pyrazines that lack the 2-me-
eral plants and is responsible for the allelopathic effects ob- thoxyl substituent are partially responsible for the odor of
served. However, the compound did not inbibit growth of fermented and roasted coffee and cacao (Nahrstedt, 1982;
Zea mays (Ohigashi et al., 1989). Rothschild et al., 1984).
An unusual purine base, saxitoxin (40), has been isolated
from several marine dinoflagellates (Atta-ur-Rahman and
Choudhary, 1990). Saxitoxin inhibits Na+ channels. This
compound is considered to be the most toxin nonprotein SEClJIlIl'lEGA ALKALOIDS
compound known; it has an LD50 i.p. in mouse of 10 J.Lg/kg
(Wink, 1993). Securinine (42) is the major alkaloid of Securinega suffruti-
cosa and related species. These alkaloids only occur in the
Euphorbiaceae. Eight of the 13 carbon atoms are derived
PYKAZIl'IE ALKALOIDS from tyrosine; lysine serves to label other parts of the mole-
cule (Fig. 37.12).
Pyrazine alkaloids (Fig. 37.11) are known to occur in both Securinine nitrate is a central nervous system stimulant
,Iants and insects. These alkaloids serve as warning and similar to but less toxic than strychnine. This compound
!larm substances from lridomyrmex humilis ("driisena- raises blood pressure, stimulates respiration, and increases
neisen") and Ondontomachus species ("Stachelameisen") cardiac output (Cordell, 1981).
704 Miscellaneous Types of Alkaloids

3·propyl-2-mdhoxypyrazoline 3-butyl-2.methoxypyrazoline (41)

Capsicum annuum (Solanaceae)

Fig. 37.11. Pyrazine alkaloids.

G o
w
OH

CO,H
c8."~09~ CO,H HO CO,H

--+

~~- OP
securinine (42)

Fig. 37.12. Biogenesis of securinine (Leete, 1980; modified and used with permission of the copyright owner, Springer-Verlag, Berlin).
 Miscellaneous Types of Alkaloids 705

;600
~1u HN
o
HN
~
0,/'-..0
"
H
OH
H
OHl

0
0
~
HN OH
'"

o o 0
o
sesbanine (43) sesbanimide A (44) sesbanimide B (45) streptimidone (48)

OH

o
HN

o o
sesbanimide C (46) cycloheximide (47) streptovatacin (49)
(actidione)

Fig. 37.13. Alkaloids from Sesbania.

SESBA1'IlA ALKALOIDS glucoside, betanin (50), a representative of the most common

type. In addition to these betalain pigments, the betaxanthins,
A number of unusual alkaloids have been isolated from the in which proline appears to replace the cyclodopa-derived
seeds of Sesbania drummondii (Fabaceae, coffee bean, rattle portion, are encountered in many plants of this group. This
bush, or rattle box), a plant of the southeastern United States second type is usually yellow in color (Steglich and Strack,
(Fig. 37.13) (Powell and Smith, 1981; Powell et ai., 1976, 1990).
1984; Suffness and Cordell, 1985). Other species of this Betalain pigments in plants often were confused with an-
genus also appear to contain alkaloids. The seeds are poison- thocyanins until a number of investigators in the late 19th
ous to cattle, goats, and sheep. Although the alkaloid sesban- century recognized that betacyanins contain nitrogen. How-
ine (43) occurred in small amounts and was isolated as a ever, purification of these pigments was difficult and, despite
part of antitumor screens, the actual activity resided with study by many well-known chemists, betalains were not
an even more minor compound, sesbanimide (44), that was fully characterized until 1963 (Wyler et ai., 1963). The be-
finally isolated in 5 X 10-'% yield. Sesbanimide is the taxanthin, indicaxanthin (51), was characterized a year later
major cytotoxic and antileukemic compound in the plant. (Fig. 37.14) (Piatelli et ai., 1964).
Two additional antitumor compounds have been isolated, Recently, a number of betalain pigments, as well as re-
sesbanimide B (45) and C (46) (Powell et al., 1984). lated metabolites, have been reported from fungi such as
The structure of sesbanimide is similar to that of the anti- Amanita muscaria (Dopp and Musso, 1974). Muscaflavin
biotics cycloheximide (47), streptimidone (48), and strepto- (52), muscaaurin I (53), muscaaurin II (54), and similar com-
vitacin A (49), but, so far, all evidence suggests that these pounds now are known to occur in this species and several
alkaloids are plant products and not fungal metabolites (Suff- other fungi, especially those of the genera Amanita, Hygro-
ness and Cordell, 1985). cybe, and Hygrophorus (Antkowiak and Antkowiak, 1991;
Steglich and Strack, 1990).
Techniques for the isolation and purification and instru-
8ETALAINS mental methods for analysis of betacyanins have been re-
viewed (Strack et al., 1993).
Betalains are a group of alkaloidal pigments restricted to
8iosynthesis
about 10 families in the order Caryophyllales (formerly the
Centrospermae). These pigments occur in all plant parts, Labeling studies indicate that tyrosine gives rise to both
vary in color from yellow to red-violet, exist as zwitterions, parts of the molecule. However, little is known of the spe-
are water soluble, and are accumulated in the vacuole. Two cific enzymology (Steglich and Strack, 1990). The formation
main structural types and about 50 compounds have been of cyclodopa (56) from DOPA has been difficult to establish.
reported from plants (Strack et ai., 1993). The red color of Possibly, this occurs via a monohydroxylated precursor (Fig.
beets (Beta vulgaris) is due to the presence of the betalain 37.15) (Geissman and Crout, 1969).
706 Miscellaneous Types of Alkaloids

glucosyl-O
H

HO
, 10
CO"

11

I.

19 H .. ,.
CO,H
CO,H

£=<
betanin (SO) indicaxanthin (51)

00 """ o
HO"]N ,
"'"-
H - H

6 :5
HN+ CO,, CO,H
HN+ CO"
CHO

~~-
H H I
H H

HO,C"'- HN I CO,H
Ho,O"""" HN I CO,H
H
H
muscaaurin n (54) muscsaurin I (53) muscsflavin (52)
Fig. 37.14. Betalains, betaxanthins. and related pigments.

The presence of a series of structurally modified com- (Geissman and Crout, 1969; Fischer and Dreiding, 1972).
pounds derived from DOPA in both higher plants and fungi Stizolobic acid (55) occurs in Mucuna (Fabaceae); and mus-
has been established (Fig, 37.17). Betalamic acid (57) has caflavin has been found in several fungi (Fig. 37.17) (Mabry,
been isolated from a number of betalain-synthesizing species 1977).
and a biogenetic pathway has been proposed (Fig. 37.16) Most betalains and indicaxanthin (51) have a 2S configu'

H0:():J
HO
I :::,...
H,N CO,H
--+
~'<::::
u+
0
"".Q4"
H,N
--
CO,H
O~

HO~NH~CO'H -
DOPA

H0:(Q
I
HO:::"" NH CO,H
--
~
I
HO:::"" NH CO,H
4-

cyclodopa (56)

Fig. 37.15. Biogenesis of cyc1odopa (modified from Geissman and Crout, 1969).
 H0'fln
HO~
H.
H,W/A....CO,H
-
e· H'

,;7:1
"'"
OH

OH
Miscellaneous Types of Alkaloids

--
707

DOPA HOzCI>""" NHz

H

betanidin betanin (SO)

Fig. 37.16. Biogenesis of betanin (modified from Fischer and Dreiding. 1972; used with pennission of the copyright owner, Swiss Society of
Chemistry, Basel).

Hofu'~
: I
HO .."" H,N eO,H

/
b
DOPA

° ° HO

HO'cH~
O~NJe~
~
---' HO,e

HO ~ I
H,N eO,H
HO'~ :;=
H~H,N"~eO'H
/0
C

H'N-\'H

D
° ° eO,H

muscaflavin (52) stizolobic acid (55) betalamic acid (57)

Fig. 37.17. Cleavage of DOPA to produce muscaflavin, stizolobic acid, and betalamic acid (Mabry, 1977; modified and used with permission of the
copyright owner, Dr. T. J. Mabry),
708 Miscellaneou.f Types of Alkaloids

ration. Differences at C-15 have been encountered; both an members of the order (Rodman et al., 1984). If so, the ability
R and S series at that center are naturally occurring. to synthesize anthocyanins has been lost in most members
In the presence of a weak base, betacyanin and betaxan- of the order but has been regained in these two families.
thin pigments undergo exchange reactions in which the dihy- Two other families frequently allied with the order Caryo-
dropyridine portion of the pigment is transferred from one phyllales, the Polygonaceae and the Plumbaginaceae, are
amino acid to another. For example, in the presence of am- shown to be more distantly related (Rodman et al., 1984).
monium hydroxide and proline, betanin (50) yields indicax- To date, no plants are known that contain both types of
anthin (51). Similar in vivo conversions may occur in plants. pigments.
Betalains are synthesized by hairy root cultures of Beta
vulgaris. The alkaloids are not excreted into the medium Betalalns In Foods and Beverages
(Flores et al., 1987).
Adulteration of wines with juices of beets and of the froits
of Phytolaeea species has been known since at least 1860.
Chemosystematic: Value
The similarity of spectral properties of anthocyanins and
The distribution of betalains and anthocyanins in plants betalain pigments makes this somewhat difficult to detect.
is completely mutually exclusive. Betalains are found in the However, the pigments can readily be separated by electro-
families Aizoaceae, Amaranthaceae, Basellaceae, Cacta- phoresis and routine screening of many wines is conducted
ceae, Chenopodiaceae, Didieriaceae, Nyctaginaceae, Phyto- (Strack et al., 1993).
laccaceae, Portolacaceae, and Steguosperrnaceae. These Betanin from beets is used as a colorant in ice cream,
families, along with the families Caryophyllaceae and Mol- jam, and froit conserves (Strack et al., 1993).
luginaceae, are placed in the order Caryophyllales or, for-
merly, Centrospennae. Anthocyanins are known from the
last two families, but not from others in this order. Betalain JIIISCELLAl'IEOUS ALKALOIDS
pigments from plants of the Caryophyllales have been tabu-
lated (Steglich and Strack, 1990). Trigonelline and an Unusual Pyrrole
A number of small genera from several families of the
Caryophyllales have been segregated and some are consid- Trigonelline (59), N-methyloicotinic acid (Fig. 37.18), is
ered as families. However, the presence of these pigments capable of promoting specific G2 cellular arrest but this ac-
within the genera Agdestis, Dysphania, Gisekia, Halophy- tivity is dependent on the age of the organism. An unusually
tum, Heetorella, and Petiveria suggest that these plants substituted pyrrole (58), produced by older seedlings of
should be maintained in the order Caryophyllales. Nonethe- Pisurn sativum (Fabaceae), functions as an endogenous regu-
less, these data alone do not answer questions concerning lator of trigonelline-induced G2 arrest. The ED50 of exoge-
the rank at which they should be accepted. nously added (58) in the inhibition of trigonelline-induced
Some have suggested that the small families Batidaceae, cellular arrest in G2 is 5 X 10-7 M. This compound occurs
Gyrostemonaceae, and Theligonaceae should be included in in sufficient quantities to account for the inability of trigonel-
the Caryophyllales. The absence of betalains and special line to induce G2 arrest in older roots (Lynn et aI., 1987).
sieve-tube plastids containing protein inclusions from all
three families has been reported (Behnke and Turner, 1971; HistronlonlC:otoxins
Goldblatt et al., 1976; Mabry and Turner, 1964; Mabry et
A series of potent alkaloids were first isolated from den-
al., 1975). These rmdings suggest that these families are
drobatid frogs of western Colombia and northwestern Ecua-
not close relatives of the other betalain-containing families. dor, but are now known to be more widespread in distribu-
Although the Batidaceae lack anthocyanins, these com-
tion. These alkaloids affect at least three classes of channels
pounds are present in the Theligonaceae and Gyrostemona-
in nerve and muscle. The first two are receptor-regulated
ceae. channels, in particular the nicotinic acetylcholine receptor
It has also been suggested that the betalain- and anthocya-
channel. The histrionicotoxins are noncompetitive blockers
nin-producing families of this order developed from a com-
of this receptor-channel complex (Daly et al., 1993). The
mon ancestor before the widespread occurrence of floral pig-
second class of channels are the voltage-dependent sodium
ments in angiospenns (Mabry, 1976). All families of the
channels. Histrionicotoxins reduce conductances in a man-
Caryophyllales contain sieve-tube cells with proteinaceous
ner reminiscent of local anesthetics (Daly et al., 1993). De-
inclusions which are not encountered in other plant families.
spite these effects, these alkaloids have relatively low toxic-
However, because anthocyanins are known from most other ity (Daly et al., 1993).
groups of plants including gymnospenns and ferns, it seems
likely that the ancestors of the Caryophyllales also possessed
HuperzineA
anthocyanins.
Certain results of cladistic analyses suggest that the Mol- The club moss Huperezia serrata has long been used in
luginaceae and CaryophylJaceae are not closely related, but Chinese traditional medicine. An alkaloid from this plant,
are relatively advanced and derived from betalain-containing huperzine A (60) is one of the few drugs that appears to
 Miscellaneous Types of Alkaloids 709

~C02-
HOJJ---CHO

~N)
N

Cr"
o
I
CH3

(58) trigonelline (59) huperzlne A (60)

Fig. 37.18. Miscellaneous alkaloids.

have activity for treating Alzheimer's disease (Borman, Alkaloids, Vol. 43 (G. A. Cordell, ed.), 185-288, Academic
1993; Zhou et al., 1993). Clinical trials in China suggest that Press, New York, 1993.
the alkaloid improves memory and learning in aged individu- DOpp, H. and H. Musso, Chromatographic analysis of betalain
als with memory impairment and also improves function in pigments in toadstools and higher plants, Z. Naturforsch., 29c,
patients with myasthenia gravis. Huperzine A exhibits strong 640-642 (1974).
anticholinesterase activity (Zhou et al., 1993). FISCHER, N. and A. DRBIDING, Biosynthesis of betalaines. On the
cleavage of the aromatic ring during the enzymatic transfonna-
tion of Dopa into betalamic acid, Helv. Chim. Acta, 55,
649-658 (1972).
REFERENCES
FLORES, H. E., M. W. Hoy, and J. J. PICKARD, Secondary metabo-
lites from root cultures, Tibtech, 5, 64-69 (1987).
AHMED, M. S., H. H. S. FONG, D. D. SOBIARTO, R. H. DOBBERSTEIN,
GEISSMAN, T. A. and D. H. G. CROUT, Organic Chemistry of Sec-
D. P. WALLER, and R. MORENO-AzoRERO, High-performance
ondary Plant Metabolism, Freeman Cooper, San Francisco,
liquid separation and quantitation of maytansanoids in Mayte-
1969.
nus iIIicifolia, J. Chromatogr., 213, 340-344 (1981).
GOLDBLATI, P., J. W. NOWICKE, T. J. MABRY, and H. D. BEHNKE,
ANTKOWIAK, R. and W. Z. ANTKOWIAK, Alkaloids from mush-
Gyrostemonaceae: Status and affinity, Bot. Notiser, 129,
rooms, in The A1kaloids, Vol. 40 (A. Brossi, ed.), 189-340,
201-206 (1976).
Academic Press, New York, 1991.
GOLEBIEWSKI, W. M. and 1. T. WROBEL, The Lythraceae alkaloids,
ATIA-UR-RA>!MAN and M. I. CHOUDHARY, Purine alkaloids, in The
in The A1kaloids, Vol. 18 (R. H. F. Manske and R. G. A. Ro-
A1kaloids, Vol. 38 (A. Brossi, ed.), 225-323, Academic Press,
drigo, eds.), 263-323, Academic Press, New York, 1981.
New York, 1990.
HAREORNE, J. B., Chemical signals in the ecosystem, Ann. Bot.,
BEHNKE, H. D. and B. L. 'fuRNER, On specific sieve-tube plastids
60, 39-57 (1987).
in Caryophyllales, Taxon, 20, 731-737 (1971).
HARBORNE, J. B.o Recent advances in chemical ecology, Nat. Prod.
BORMAN, S., Natural product may be Alzheimer's therapeutic,
Rep., 7, 85-109 (1989).
Chern. Eng. News, 35-36 (September 20, 1993).
HART, N. K., S. R. JOHNS, J. A. LAMBERTON, and R. I. WILLING,
BRINGMANN, G., The naphthyl isoquinoline alkaloids, in The Alka-
Alkaloids of Alchornea javanensis (Euphorbiaceae). The isola-
loids, Vol. 29 (A. Brossi, ed.), 141-184, Academic Press, New
tion of hexahydroimidazo[I,2-aJpyrimidines and gnanidines,
York,1986.
Aust. J. Chern., 23, 1679-1693 (1970).
BRINGMANN, G., F. POKORNY, H. REUSCHER, D. LISCH, and L. A. HEMINGWAY, S. R. and J. D. PHiLLIPSON, A1kaloids of the Rubia-
AsS!, Novel Ancistrocladaceae and Dioncophyllaceae type ceae, in Indole and BiogeneticalJy Related Alkaloids (J. D. Phil-
naphthylisoquinoline alkaloids from Ancistrocladus abbrevia- lipson and M. H. Zenk, eds.), 63-90, Academic Press, London,
tus: A phylogenetic link between the two families? Planta Med- 1980.
ica, 56, 496-497 (1990).
H:naNo, H., Recent research on oriental medicinal plants, in Eco-
BROWN, E. G. a. Purines, pyrimidines, nuc1eosides and nuc1eotides. nomic and Medicinal Plant Research, Vol. I (H. Wagner, H.
in Amino acids, proteins, and nucleic acids (L. J. Rogers, ed.), Hikino, and N. R. Farnsworth, eds.), 53-85, Academic Press,
Vol. 5 of Methods in Plant Biochemistry (P. M. Dey and J. B. London, 1985.
Harbome, eds.), 53-90, Academic Press, London, 1991.
HORGAN, R. and I. Scorr, Cytokinins, in Amino Acids, Proteins,
CoRDELL, G. A., Anticancer agents from plants, in Progress in and Nucleic Acids (L. J. Rogers, ed.), vol. 5 of Methods in Plant
Phytochemistry Vol. 5 (L. Reinhold, J. B. Harbome, and T. Biochemistry (P. M. Dey and J. B. Harbome, eds.), 91-120,
Swain, ods.), 273-316, Pergamon, London, 1978. Academic Press, London, 1991.
CoRDELL, G. A., Introduction to A1kaloids: A Biogenetic Approach, HOWARD, A. S. and J. P. MICHAEL, Simple indolizidine and quino-
Wiley, New York, 1981. lizidine alkaloids, in The A1kaloids, Vol. 28 (A. Brossi, ed.),
CRABB, T. A., Nuclear magnetic resonance of alkaloids, in Annual 183-308, Academic Press, New York, 1986.
Reports on NMR Spectroscopy, Vol. 13 (G. A. Webb, ed.), JANZEN, D. H., H. B. JUSTER, and E. A. BELL, Toxicity of secondary
60-210, Academic Press, London, 1982. compounds to the seed-eating larvae of the bruchid beetle Callo-
DALY, J. W., H. M. GARRAFFO, and T. F. SPANOE, Amphibian sobruchus maculatus, Phytochemistry, 16, 223-227 (1977).
710 Miscellaneous Types of Alkaloids

JOULLlE, M. M. and R. M. NUTI, Cyclopeptide alkaloids, in Alka- velvetleaf (Abutilon theophrasti) germination as models for
loids: Chemical and Biological Perspectives, Vol. 3,113-168, new biorational herbicides, in BiologicallY Active Natural Prod-
Wiley, New York, 1985. ucts: Potential Use in Agriculture (H. G. Cutler, ed.), ACS Sym-
KENDRICK, B., The Fifth Kingdom, Mycologue Publications, Wa- posium Series 380, 211-232, American Chemical Society,
terloo, Ontario, Canada, 1985. Washington, DC, 1988.
LAGARIAS, J. c., D. GOFF, F. K. KLEIN, and R. RAPPOPORT, Cyclo- POWELL, R. G., C. R. SMITH, JR., and R. V. MADRlGAL, Antitumor
peptide alkaloids. Phenylcyclopeptines from the polymorphic activity of Sesbania vesicaria, S. punicea, and S. drummondii
species Ceanothus integerrimus, 1. Nat. Prod., 42, 220-227 seed extracts, Planta Medica, 30, 1-8 (1976).
(1979). POWELL, R. G., C. R. SMITH, JR., R. D. PLATTNER, and B. E. JONES,
LEETE, E., Alkaloids derived from ornithine, lysine and nicotinic Additional new maytansinoids from Trewia nudiflora: lO-Epi-
acid, in Secondary Plant Products (E. A. Bell and B. V. Charl- trewiasine and nortrewiasine, 1. Nat. Prod., 46, 660-666
wood eds.), 65-91, Springer-Verlag, Berlin, 1980. (1983).
LEETE, E., The biosynthesis of alkaloids, Specialist Periodic Re- POWELL, R. G., C. R. SMITH, JR., and D. WEISLEDER, Novel may tan-
ports, Biosynthesis, 102-223, The Royal Society of Chemistry, sinoid tumor inhibitors from Trewia nudiflora: Trewiasine, de-
London, 1983. hydrotrewiasine, and demethyltrewiasine, 1. Org. Chern., 46,
4398-4403 (1981).
LEWIS, W. R., Plants used medicinally by indigenous peoples, in
Phytochemical Resources for Medicine and Agriculture (H. N. POWELL, R. G., C. R. SMITH, JR., and D. WEISLEDER, Sesbanimide
Nigg and D. S. Seigler, eds.), 33-74, Plenum Press, New York, A and related tumor inhibitors from Sesbania drummondii:
Structure and chemistry, Phytochemistry, 23, 2789-2796
1992.
(1984).
LYNN, D. G., K. JAFFE, M. CORNWALL, and W. TRAMONTONO, Char-
POWELL, R. G., D. WEISLEDER, C. R. SMJTH, JR., 1. KOZLOWSKI,
acterization of an endogenous factor controlling the cell cycle
and W. K. ROHWEDDER, Treflorine, trenudine, and N-metbyl-
of complex tissues, J. Am. Chem. Soc., 109, 5858-5859 (1987).
trenudone: Novel maytansinoid tumor inhibitors containing two
MAAT, L. and R. C. BEYERMAN, The imidazole alkaloids, in The
fused macrocyclic rings, J. Am. Chern. Soc., 104, 4929-4934
Alkaloids, Vol. 22 (A. Brossi, ed.), 282-334, Academic Press,
(1982).
New York, 1983.
REIDER, P. J. and D. M. ROLAND, Maytansinoids, in The Alkaloids,
MABRY, T. J., Pigment dichotomy and DNA-RNA hybridization
Vol. 23 (A. Brossi, ed.), 71-156, Academic Press, New York,
data for centrospermous families, Plant Syst. Evo!., 126, 74-94
1984.
(1976).
RODMAN, 1. E., M. K. OLIVER, R. P. NAKAMURA, 1. U. MCCLAMMER,
MABRY, T. 1., The order Centrospennae, Ann. Mo. Bot. Gard., 64, JR., and A. H. BLEDSOE, A taxonomic analysis and revised clas-
210-220 (1977). sification of Centrospermae, Syst. Bot., 9, 297-323 (1984).
MABRY, T. 1. and B. L. TuRNER, Chemical investigations of the ROTHSCHILD, M., B. P. MOORE, and W. V. BROWN, Pyrazines as
Batidaceae, Taxon, 13, 197-200 (1964). warning odour components in the monarch butterfly, Danaus
MABRY, T. J., I. J. EIFERT, C. CHANG, H. MABRY, C. KIDD, and H. D. p/exippus, and in moths of the generaZygaena andAmata (Lepi-
BEHNKE, Theligonaceae: Pigment and ultrastructural evidence doptera), BioI. J. Linn. Soc., 23, 375-380 (1984).
which excludes it from the order Centrospennae, Biochem. Syst. SCHMIDT, U., A. LIEBERKNECHT, and E. HASLINGER, Peptide alka-
Ecol., 3, 53-55 (1975). loids, in The Alkaloids, Vol. 26 (A. Brossi, ed.), 299-326, Aca-
MADRIGAL, R. V., B. W. ZILKOWSKI, and C. R. SMITH, JR., Struc- demic Press, New York, 1985.
ture-activity relationships among maytansinoids in their effect SMITH, C. R., JR. and R. G. POWELL, Chemistry and pharmacology
on the European com borer, Ostrinia nubilalis (Hiibner), 1. Org. of rnaytansinoid alkaloids, in Alkaloids: Chemical and Biologi-
Chern., 11, 501-506 (1985). cal Perspectives (S. W. Pelletier, ed.), 149-204, Wiley, New
MANFREDI, K. P., 1. W. BLUNT, J. H. CARDELLINA n, J. B. McMA- York,1984.
HON, L. L. PANNELL, G. M. CRAGG, and M. R. BOYD, Novel STADLER, E., Contact chemoreception, in Chemical Ecology of In-
alkaloids from the tropical plant Ancistrocladus abbreviatus in- sects (W. J. Bell and R. T. Cardi, eds.), 3-35, Chapman &
hibit cell killing by HIV-I and HIV-2, 1. Med. Chern., 34, Hall, London, 1984.
3402-3405 (1991). STEGLICH, W. and D. SmACK, Betalains, in The Alkaloids, Vol.
NAHRSTEDT, A., Strukturelle Beziehungen zwischen pflanzlichen 39 (A. Brossi, ed.), 11-62, Academic Press, New York,
und tierischen Sekundarstoffen, Planta Medica, 44, 2-14 1990.
(1982). STRACK, D., W. SrnGUCH, and V. WRAY, Betalains, in Alkaloids
OHIGASHI, R., M. KAJI, M. SAKAKI, and K. KOSHIMIZU, 3-Rydrox- and Sulphur Compounds (P. G. Waterman, ed.), Vol., 8 ofMeth-
yuridine, an allelopathic factor of an African tree, Baillonella ods in Plant Biochemistry (P. M. Dey and J. B. Harborne, eds.),
toxisperma, Phytochemistry, 28, 1365-1368 (1989). 421-450, Academic Press, London, 1993.
PIATELLI, M., L. MINALE, and G. FROTA, Isolation, structure and SUFFNESS, M. and G. A. CORDELL, Antitumor alkaloids, in The
absolute configuration of indicaxanthin, Tetrahedron, 20, Alkaloids, Vol. 25 (A. Brossi, ed.), 3-356, Academic Press,
2325-2329 (1964). New York, 1985.
POWELL, R. G. and C. R. SMITH, JR., An investigation of the antitu- SUZUKI, T., H. ASHllIARA, and G. R. WALLER, Purine and purine
mor activity of Sesbania drummondii, J. Nat. Prod., 44, 86-90 alkaloid metabolism in Camellia and Coffea plants, Phytochem-
(1981). istry, 31, 2575-2584 (1992).
POWELL, R. G. and G. F. SPENCER, Phytochemical inhibitors of TANTISEWIE, B. and S. RUCHIRAWAT, Alkaloids from the plants of
 Miscellaneous Types of Alkaloids 711

Thailand, in The Alkaloids, Vol. 41 (A. Brossi and G. A. Cor- WALLER, G. R. and E. K. NOWACKI, Alkaloid Biology and Metabo-
dell, eds.), 1-40, Academic Press, New York, 1992. lism in Plants, Plenum Press, New York, 1978.
THOMAS, D. W. and R. E. GEREAU, Ancistrocladus korupensis (An- WANG, D., Purines and pyrimidines and their derivatives, in Phyto-
cistrocladaceae): A new species of liana from Cameroon, chemistry, Vol. 2 (L. P. Miller, ed.), 61-117, Van Nostrand
Novon, 7, 242-245 (1993). Reinhold, New York, 1973.
TSCHESCHE, R. and E. U. KAUSSMAN, The cyclopeptide alkaloids, WANG, P. and M. M. JOUI.LIE, Muscarine alkaloids, in The Alka-
in The AIka1oids, Vol. 15 (R. H. F. Manske, ed.), 165-206, loids, Vol. 23 (A. Brossi, ed.), 328-380, Academic Press, New
Academic Press, New York, 1975. York,1984.
WAGNER, H., Non-steroid, cardioactive plant constituents, in Eco- WINK, M., Allelochemical properties or the raison d'etre of alka-
nomic and Medicinal Plant Research, Vol. 2 (H. Wagner, H. loids, in The Alkaloids, Vol. 43 (G. A. Cordell, ed.), 1-118,
Hikino, and N. R. Farnsworth, eds.), 17-38, Academic Press, Academic Press, New York, 1993.
London, 1988. WYLER, H., T. J. MABRY, and A. S. DRBlDING, Zur Struktur des
WALLER, G. R. and O. C. DE1!MER, Enzymology of alkaloid metabo- Betanidins. 'Obor die Konstitution des Randenfarbstoffes Be-
lism in plants and microorganisms, in Secondary Plant Products lanin, Helv. Chim. Acta, 46, 1745-1748 (1963).
(E. E. Conn, ed.), Vol. 7 of The Biochemistry of Plants (P. K. ZHou, B., D. Zoo, M. HUANG, L. LIN, L. LIN, X. HAN, and G. A.
Stumpf and E. E. Conn, eds.), 317-402, Academic Press, New CORDELL, NMR assignments of huperzine A, serratinine and
York,I981. lucidioline, Phytochemistry, 34, 1425-1428 (1993).
Index

Abies 337, 338 acetic acid, 59, 514, 532, 674 actinomycin D, 568
Abies balsamea, 338, 383 acetoacetate, 19, 532, 539, 693 actinomycins, 568
Abies iasiocarpa, 338 acetoacetyl-CoA.313 Acu/eiferum, 287
abietanes. 405, 406. 412 acetoacetyl-CoA thiolase (AAeT), 313 acyl-ACP, 3, 21
abietic acid, 312, 412, 415 acetogenin, 56, 67, 74, 139 acyl-ACP thioesterase, 21
abiotic stress, 8 acetone, 206, 276, 535 acyl-ACP:glycerol-3-phosphate acyltransferase,
abortifacient, 404. 420 acetosyringone, 123 23
abortion, 227, 404 6-acetoxytoonacilin, 480. 484, acyl-ACP-l :lysophosphatidic acid
abrin. 231, 244 N-acetylanthranilic acid, 568, 570 acyltransferase, 23
Abrus fruticulosus, 463, 471 acetyl-ACP, 19 acylation, 23
Abrus precatorius, 244, 245, 463, 470 acetylcholine, 510, 511, 526, 528, 600, 609, acyl carrier protein (ACP), 3, 16, 19, 21, 23,
abrusosides A-D, 463, 470 664, 675, 697 40,41
abscisic acid, 3, 6, 33, 132, 145,367,500,501, acetylcholine receptors, 526, 528, 643 acyl-CoA, 3, 21, 57, 67
504,505 acetylcholinesterase, 599, 600 acyl-CoA alcohol transacylase, 5 j
( + )-(S)-abscisic acid. 500 acetyl-CoA, 6, 19-21,24,25,57,58, 121, acyl-CoA:lyso-phosphotidylcholine
abscission,leaf,415 313,531,532,539 acyltransferase, 26
absinthe, 346 acetyl-eoA carboxylase, 19 l-acyl-glycerol-3-phosphate,23
absinthin. 389 acetylenic carotenoids, see carotenoids, 3-0-acylingenol, 402
Abuta, 442, 588 acetylenic acyltransferases, 21, 23
Abuti/on theophra,., 84, 99, 309-311, 390, acetylenic compounds, 16, 42-50, 113. 136, Acyrthyosiphon, 559
697,711 222,324,381,387,420,514 Acyrthosiphon nipponicus, 363
Acacia, 53, 79, 193, 194, 218, 227, 233, 273, N-acetylglucosamine, 261 Acyrthosiphon pisU, 559
284,287,289,299 N-acetyl-D-glucosamine, 253 Acyrthyosiphon spartii, 559
Acacia berlandieri, 517 ,B-N-acetylglucosaminidase, 561 addictive properties, 523, 526, 588
Acacia catechu, 204, 260, 528 P.N-acetylhexosaminidase, 562 additives, food, 171,245
Acacia chiapensis, 289 3-acetyl-6-metboxybenzaldehyde, 125 Adenia, 285, 296
Acacia collinsii, 53 2'-acetylneriifolin, 468, 472 adenosine 5'-(a-D-glucopyranosyl
Acacia cornigera. 289 N-acetylphenylalanine, 238 pyrophosphate),249
Acacia jarnesiana. 289 O-acetyIserine, 219. 220 Adenostomafasiculatum, 125, 126
Acacia globulifera, 289
O-acetyl-L-serine, 219. 220 Adenostyles leucophylla, 552
Acacia hindsii, 289
N-acetyltryptamine, 660 S-adenosyhnethionine (SAM), 27, 45, 69, 186,
Acacia /caroo. 193 Achariaceae. 285, 287, 296 230, 238, 436, 508, 518, 531, 587, 598, 663,
Acacia mearnsii, 204, 206
Achillea millefolium, 88 664,702
Acacia melanoxylon, 79, 200
achiote, 499 S-adenosyl-L-methionine:3'-hydroxy-N-methyl-
Acacia senegal, 260
Achlya 444 (S)-coclaurine-4'-O-methyltransferase, 587
Acacia sieberiana. 284 S-adenosylmethionine:bergaptoIO-
Achlya ambisexualis, 444
Acacia sutherlandii, 284
Achlya heterosexuaIis, 444 methyltransferase, 133
Acacia willardiana. 700
Achras sapota, 318 , 321 S-adenosyl-L-methionine:(S)-scoulerine 0-9-
acacipetalin. 284
Acalypha indica, 286, 528, 529
Achryanthes, 442 methyltransferase, 598
acalyphin, 274, 286, 528 L-acofriose, 250, 251, 252 S-adenosylmethionine:xanthot9xol 0-
Acalyphoideae, 286 Acokanthera schimperi, 468 methyltransferase, 133
Acanthaceae, 101, 173, 361, 364, 365, 370, aconine, 675 adenovirus, 648
387,432,522,533,569,570,660,693 aconitiform activity, 675 adenylate cyclase, 420, 424
Acanthomyops cJaviger, 341 aconitine, 673-675 adenyltransferase inhibitor, 339
Acanthus mollis, WI, 105 Aconitum, 269, 673, 674 Adhatoda, 569, 570, 660
accumulation, vii, 1-9, 11, 13, 15, 19.25,29, Acorus calamus, 111, 129 Adhatoda vasica, 570
30,32,33-35,43,48,77,80,82,84-86, L-acovenose, 251, 252 adhesives, 12, 413
88, 92, 95, 115, 124, 132, 138, 143, 145, ACP transacyiase, 19 Adiantum, 174
151-153, 155, 156, 158, 166, 173-175, 184, ACP-track, 52 Adonis, 269
186, 195,200,217,229,249,253,256,257, Acraea, 289, 297 Adoxaceae, 360
261, 262, 264, 265, 270, 275, 294, 303, 326, Acraea horta, 289 ADP/ATP carrier protein, 420
337,339,368,377,502,535,557,558,565, Acremonium, 659 ADP-glucose, 162,249
566, 634, 664 Acrididae, 208, 316 adrenal gland, 439
Acer, 195, 197, 223, 232, 523 actidone alkaloids, 8, 12, 568, 570, 573-576, adrenaline, 591, 640
Acer pseudoplatanus, 242 610 adrenergic blocking agents, 659
Aceraceae, 195, 197,217,222,223,231,458, Acro/epiopsis assectella, 227, 459, 468, 470 a-adrenergic blocking agents, 640
523 Acronychia, 574 adrenergic drugs, 511, 522
Aceratium, 563 acronycine, 574 adrenolytic activity, 591
acetate, 19, 26, 31, 42, 45, 52, 56, 58, 59, 63, Actaea, 269 adults, 41, 99,109,111,113,124,221,223,
64,67,68,72-74,80,85, 113, 114, 118, actin filaments, 237 227,228,253,285,294,308,317,362,364,
139, 142, 152, 174, 182, 187,203,206,222, Actinidia po/ygama, 362, 366 366,384,418,441,447,550,551,552,669,
269,303,313,316,377,384,442,447,463, Actinidiaceae, 361, 362 682,683
465,466,471,507,543,669,678 actinidine, 362, 669 Aegilops, 101, 128
acetate-malonate pathway, 56, 65, 76, 80, actinidiolide, 362 Aegi/ops ovala, 118
85-87, 104, 121, 122, 148,531-534,539, Actinomycetes, 65, 244 aesculetin, 130
543, 561, 570, 571, 573 actinomycin A, 568 Aesculus californicus, 222

713
714 Index

Aesculus hippocas/anum, (horse chestnut), 231 alcohol. 16.30,31,37.38.44,51-55.58. 108. isopavine. isoquinoline, jervatrum. lupine.
Aesculus parviflora, 218, 222 109. ll5. 122, 315 mesembrine. Mesembryanthemaceae,
Afghanistan. 553 alcohol dehydrogenase, 599 monoterpene, monoterpene-iridoid.
aflatoxin, 5, 65, 66, 70 alcohols,long chain, 51 monopterpene-derived indole alkaloids,
aflatoxin Bit 420 aldehyde. 2. 3. 13. 16.30.31.44.51.58.63. morphinandienone, morphine, muscarine,
Africa. 29. 48, 77. 85.177.209.210.213.222. 109. 113. 122. 288. 324. 328. 339. 376. 507. naphthylisoquinoline, Nuphar, oxaporphine,
228. 245. 288. 361. 365. 379. 390. 395.402. 508.509.513.531.548.549.578-581.586, pavine, peptide, phenanthroindolizidine.
4ll. 425, 460. 468. 477. 478, 480. 481. 484. 587.600.610.630.631.656.663 phenanthroquinolizidine,
514. 522. 551. 553. 558. 588. 623. 640. 641. alder. European black. 83 phthalldeisoquinoline, Picralima, piperidine,
647.676. 698.700, 7ll alder, green, 451 polyamine, polyhydroxypiperidine,
(+ )-afzelechin-7-0-,B-D-apioside. 463 aldol condensation, 57. 63. 64. 142. 155.507. proaporphine, protoherine, protopine, purine,
Agalinis.78 531 pyranoquinoline, pyrazine, pyridine,
AgaUnis purpurea, 78, 452 aldol reductase, 588 pyrrolidine, pyrrolizidine,
agar, 260 aldopentoses, 248 pyrroloquinazoline, quinazoline,
Agaricales,46 aldose reductase. 588, 591. 599, 600 quinazolinocarboiine, quinoline, 4-quinolone,
Agatha. 413 aldoses, 247. 248. 249.262 quinolizidine, quinolone, rhoeadane,
agathic acid, 420 aldoxime. 275. 276. 303 rutacridones. sarpagine, Sceletium,
Aga/his australis, 405 Alectra, 392 secophthalideisoquinoline, Securinega,
Agauria salicifolia, 411 aleprolic acid, 26, 28 taxane, Taxus, tetrahydrobenzylisoquinoline,
Agavaceae. 231. 257. 457 Aleurites, 402, 403 tetrahydroprotoberberine,
Agave wightii, 457 Aleurites ford;;, 35, 244 tetrahydroisoquinoline, tropane,
Agdes/is, 708 Alexa, 253, 561 tropoloisoquinoline, Veratrum, and xanthine
Agelaius. 523. 528. 619. 640. 650. 651. 703 alfalfa. 31. 39. 53. 83. 167. 179. 181. 183. alkaloids, and C-alkaloid E. C-alkaloid G,
Agera/um houstonianum, 317. 384 189-192.261,270.393,457.459.460 glycoalkaloids, glycosidic alkaloids,
agglutination, 244 algarobilla, 195 protoalkaloids. pseudoalkaloids, saponic
aggregation pheromones. 35-37, 41. 111,324. algae. 4. 9. 26. 29, 32. 39. 46. 48. 56. 70. 96. alkaloids
342. 344, 383 114.130.142.172.242.246.254,265.266, alkaloid N-oxides. 507. 546, 547. 550, 552.
aginoside, 459 324. 340, 380. 381. 396. 419. 420. 425. 433. 553. 565. 662
Aglaia cordata, 480 435.486.488.492,495.497,510.515 alkamides. 514. 528
Aglaia adorata, 533 algae. blue-green, 4. 234. 239, 240. 246 aikaues. 30. 53. 54
aglycone. 5, 85. 124. 136. 151. 152. 158. 159. algae. brown. 32. 210. 260, 265. 420. 495. 497 alkenes. 30. 53
165-167. 170. 171. 173. 174. 177. 178.201. algae. green. 172,234.239.240.246,257. alkylation. 316. 549. 553. 656
N-alkylation, 316
267.273.274.287.288.296.302.303.361. 260.265.340.381.396.419.420.494.495,
497 O-alkylation.316
366. 441. 456. 459. 682
Allamanda cathartlca, 364, 365
agmatine, 531 algae. red, 29. 40. 260, 265. 340. 381. 442.
Agrimonia. 209 495.513.515 allamandin. 364. 365
allelochemics. 9
agrimony. 209 algal blooms. 239
allelopatbie compuunds. 26. 32. 70. 78. 79. 83.
Agrobac/erium tumefaciens. 47, 110, 123, 129, algicidal. 78. 145
85.99. 122. 125-127. 174, 310. 311,
520 alginate. 260
agrobactine, 521 alginic acid, 260 344-346.350.367.389.390.394.395,424.
451. 510. 523. 703. 7ll
agroclavine, 655-658 alimentary toxic aleukia, 378
allelopathy. vii. 38. 41, 78. 83. 85. 92, 106.
Agros/is/achys hookeri, 420, 423 alizarin, 65, 87. 89
109. 122, 125-127.300.309.324.344.345,
AIDS. 562 alkaline phosphatase, 208
350. 367. 376, 395. 396, 453. 454
Ailanthus, 660 alkaline picric acid, 273 allenic acids, 49
Ailanthus aitissima, 662 alkaloid biosynthesis, 2, 8, 16 aIlenic group, 49
Aizoaceae, 623, 708 alkaloid enzymology, 2 allenic linkage, 45
ajma1acine. 2. 10. 506. 628-632. 640. 645. 654 alkaloids derived from both phenylalanine and allergenic effects. 67. 78. 267. 269. 389
19-epi-ajmalicioe, 630 tyrosine, 617 alligator weed. 168
ajmalidine, 640 alkaloids. 2. 3, 6, 8-15.93.97. 102. 106. 107, alliin. 227
ajmaline, 640 138. 171. 188. 209. 240. 254. 270. 289. 309. meso-allitol. 264
A}uga. 418. 442 321.324.351.360.361.365.366.369.385. Allium. 226, 227. 459. 468. 588, 597
Ajuga remota, 418 398.412.428,441.472,477.506.507. Allium cepa, 34, 38
ajugarins, 418 509-513. 517, 520-523. 525. 528-533, 538. Allium porrum, 51
akee.222 539.542-571.573-588.591-601.603-617, allocryptopine, 600, 601
akoammicine, 632. 634 619-632.634-643.645-659.661-678. allomatatabiol, 362
akuanunidine, 641 680-686. 689-695. 697, 698. 700. 703. 705. D-allomethylose, 250
akuammine. 641 708-711; see also acridone, Ammyllidaceae, allomones.9. 10. 83. 110. 168. 300. 307. 324.
Alangiaceae. 360. 579, 581. 598. 612. 639 3-aminospirostane. aporpbine. 339-341.352.367.379.380,382,383,390.
alangiside, 611 Aspidosperma, Aspidosperma- Hunteria, 447.486,511
Alangium lamarckii. 611-613 azafluoranthene, benzophenanthridine, Allomyces arbuscula, 376
alanine. 220. 300. 302. 513 benzylisoquinoline, berberine, Allomyces javanicus, 376
alanine aminotransferase, 601 bisbenzyHsoquinoline, bisindole, Buxus, Allomyces macrogynus, 376, 396
L-alanine:aldehyde aminotransferase, 1 Calycanthus, carbazole, p-carboline. Allophylus cobbe. 285. 289
DL-alanine-DL-aspartine (DL-ala-DL-asp), 240 Ca/haranthus, Cephalotaxus. ceveratrum, allose. 247. 249
DL-alanine-DL-Ieucine (DL-ala-DL-leu), 240 clavine, Corynanthe. Cryp/ostyline, cularine, alloxanthin. 498
DL-alanine-DL-metbiooine (DL-ala-DL-met). dehydropyrrolizidine, dibenzopyrrocoline, allyl isothiocyanate, 266, 308, 309
240 dihydrofuroquinolone, diterpene, S-allylcysteine, 227
alantolactone, 390, 392 ebumamine-vincamine, 22,26- S-allylcysteine sulfoxide, 227
Alaska,673 epiminocholestane, ergot, peptide, Erythrina, allylglucosinolate. 303. 307. 308. 309
Alberta. 338 Erythrophleum, Eupomatia, evoninoate, allylic oxidation, 42, 43, 387
Albizia, 227, 228 furoquiooline. harman, hasubanan, allylic pyrophosphate. 315. 326. 370. 371, 393
Aiblzia amara, 521, 529 hemiterpene, homoerythrina, Hun/eria, S-allylmercaptocysteine, 227
albiziine, 227, 228 hydroxyindolizidine, iboga, ibogan, almond. 287. 290. 294. 415. 561
Alces alces, 207 imidazole, indole, indolizidine, Alnus, 363
Alchornea, 640, 692, 693 iodoJoquinazoline, ipecac, ipecacuanha, Alnus crispa, 451
 Index 715

Alnus glutinosa, 83 2-aminoadipic acid, 218, 226 /l-aroyrin, 432, 447, 452, 458
Aloe, 85-87, 91, 543 L-2-aminoadipic acid, 234 p.amyrin cyclase, 447
Aloe saponaria, 85 6-(L-aminoadipyl)-L-cysteinyl-D-valine, 234 anabasine, 507, 508, 510, 526, 528, 538, 539
aloesaponarin. 85, 86 L-2-amino-6-(amidino)hexanoic acid, 227 Anabasis aphylla, 557
alpinigenine. 603 p-aminobenzoate synthase, 97 Anacardiaceae, 67, 68, 88, 173, 195, 197,206,
Alstonia. 637, 640, 660 p-aminobenzoic acid, 97, 105 269,337,412,447
Alstonia angustifolia, 640 4-aminobutanal, 549 anacardic acid. 68, 74
alstonine, 640 y-aminobutyric acid (GABA), 221, 517, 588, Anacridium melanorhodon, 208, 212
Aistroemeria, 269 698 Anadenanthera peregrina, 514
Alstroemeriaceae, 269 2-aminobutyric acid, 513 anaferine, 539
Alternanthera philexeroides, 168 4-aminobutyric acid, 221 Anagallis, 444
Alternaria, 238 4-arylcoumarins, 187 anagyrine, 552, 558, 559
Alternaria alternata, 67 S-(3-amino-3-carboxypropy1)-8-methyl anabygrine, 539
Alternaria kikuchiana, 238 sulfoxime, 231 anal glands, 343
Alternaria mali, 238 l-aminocyclopropane-l-carboxylic acid (ACC), analgesia, 122,346,596,597,623,646,650,
Alternaria solani, 377, 397 230,231,233,238,297,289 670
altholactone, 139 l-aminocyclopropane-l-carboxylic acid anamafana, 36
L-altromethylose, 250 synthase (ACC-synthase), 230 Anamirta, 385
altrose, 249 2-amino-2-deoxy-D-glucose, 250 Anartia. 363, 637
aluminum, 171 26-aminodibydrodiosgenin, 680 Anastrepha suspensa, 340
Alyssum, 309 L-2-amino-6-guanidinohexanoic acid, 223 anatabine, 526, 538
Alzheimer's disease, 710 cis-l-amino-3-hYdroxymethYlcyclobutane-l- Ancistrocladaceae, 698, 710, 711
Amanita, 235, 237, 246, 697, 705 carboxylic acid, 222 Ancislrocladus, 698
Amanita muscaria, 35, 697, 705 2-amino-4-methylhexanoic acid, 222 Ancistrocladus abbreviatus, 698, 710, 711
Amanita pantherina, 235 2-amino4-methylenehex-4-enoic acid, 222 Ancistrocladus korupensis, 698, 711
Amanita phalloides, 235, 237 2-amino4-methylhex-4-enoic acid, 222 Ancistrocladus tectorius, 698
Amanita verna, 235 2-amino-4-methyl-6-hydroxyhex-4-enoic acid, andirobin, 476
Amanita virosa, 235 222 Andrachne, 610
Amanitaceae, 35 2-amino-5-methyl-6-hYdroxyhex-4-enoic acid, Andrena, 344, 381
a-amanitin, 237 222 androcymbine, 617
amanullin,237 L-2-aminooxy-3-phenylpropionic acid (AOPP), Androcymbium, 617
Amaranthaceae, 79, 168, 179,387,390,442, 195 Andrographis paniculata, 370, 375
458, 662, 708 a-amino-,8-propionic acid, 294 andromedane, 411
3-aminopropionitrile, 223, 226 Andropogon gerardii. 125
Amaranthus palmeri, 79, 376
,8-aminopropionifrile, 226 Andropogon nardus, 346
Amarathus retrojlexus, 390
( - )-a-aminopropiophenone, 522 anemia. acute hemolytic, 700
Amaryllidaceae, 231, 457, 510, 560, 617, 619,
aminopropyltransferases, 517 Anemia mexicana, 414. 424
620, 622-624, 627
3-aminospirostane alkaloids, 677, 681 Anemone, 269
Amaryllidaceae alka1oids, 617, 619, 620
Ammi majus, 133, 135, 138 Anemonel/a, 269
Amato, 703
Ammi visnaga, 137 anemonin. 269
amatoxins, 237
(R)-anunirin, 135 Anemonopsis, 269
Amazon, 588, 607
ammodendrine, 538 anesthesia. 36
arober,413 Ammodendron conolly;;, 538 anesthetic compounds, 136. 525, 535, 537,
Amblyptilia mica, 669 ararnonia, 220, 221, 253, 290, 669 607,641,650,676
Ambrosia, 47, 376, 387, 388, 392, 395 ammonium hydroxide, 708 anethole, 344
Ambrosia artemisiifolia, 376 amebiasis. 599 E-anethole, 113
Ambrosia chamissonis, 388 amoorastatin, 481 Anethum graveolens, 241
Ambrosia confertiflora, 388 Amorpha, 158, 186 angelic acid, 321, 322
Ambrosia cumanensis, 388 Amorpha fruticosa. 158 Angelica, 136
Ambrosia psilostachya, 387, 388 AMP phosphodiesterase, 267 angelicin, 135, 136
ambrosiol, 388 cAMP phosphodiesterase, 120 angina pectoris, 588
amebicidal, 559, 652 amphetamine, 522, 529 angiospenn, 12, 13, 19,50,92, 115, 142, 172,
amenorrhea, 570 amphibians, 510, 552, 560, 561 173, 176, 194,212,265,280,286,310,324,
amentoflavone, 173 amphipods, 340, 381, 420 365, 387, 401, 405, 442, 454, 510, 533, 544,
Ames assay (mutagenesis), 165 Amphypterygium adstringens, 67, 74 545,708
aroides, 36, 508, 513, 514, 517, 518, 528, 529, Ampithoe /ongimana, 420 angustifoline, 554, 555, 556, 558
576, 617, 655, 659 Amsonia. 640 (+ )-angustifoline, 554
amine oxidases, 509, 510, 531 AM-toxin I, 238 Angylocalyx, 253
aroines, 10,37, 105, 123,507,508,513-517, aroygdalin, 254, 277, 279, 280, 287-289, 297, anbaladine, 581
529, 530, 575, 579, 610, 666 299 anbalarnine, 579, 580, 581
amines, primary, 507, 509 (R)-aroygdalin, 277, 287, 297, 299 anhalanine, 581
amines, secondary, 507 amygdalin hydrolaae, 280 anhalonidine. 581
amines, simple, 507, 513-515, 521 aroygdalin 6"(4-hydroxybenzoate), 277 anhalonine, 581
amino acid metabolism, 508 amygdalin 6"[4-hydroxy-(E)-cinnarnate], 277 (3,6-anhydro-L-xylo-hexulono-l,4-lactone
amino acids, vii, 3,5, 6, 8, 13, 19, 28, 46, 94, Amygdaloideae, 264, 287 - hydxate, 265
97, 101, 102, 104, 105,207,209,216-232, a-amylases, 259 1,5-anbydro-D-mannitol, 262
234,237,241,244,274,277,290,294,300, /l-aroylases, 259 3',4'-anbydrovinblastine,- 645, 652
302,303,314,462,506,508,513,568,619, amyloglucosidase, 561 Aniba, 139
657,680,698,700,708,710 amyloidosis, 619 Aniba colo, 139
a-amino acids, 508 aroylopectin, 257, 259, 260 Aniba duckei, 528
D-amino acids, 234 aroylose, 257-260 Aniba pseudocoto, 139
L-amino acids, 215, 216, 234, 302 amylo-(l,4-> 1,6)-transglucosylase, 259 anibine, 528
amino acids, a-hydroxy1501 amyotrophic lateral sclerosis (ALS), 226. 294. animals, 3, 9, 10, 12, 13, 16, 17, 19, 24, 30,
amino acids, primary, 215 299 32,33,35,47,59,70,71,76,78, 80,91,
amino acids, sulfur-containing, 226. 227 a-aroyrin, 432, 447 94, 102, 106, 137, 165, 167, 176, 177, 179,
716 Index

187, 193,206-208,210,222,223,226,234, antibiotics, 4, 6, 35, 48, 56, 64, 65, 71, 77, ants, 35, 53, 54, 88, 124, 176,273,289,291,
235,239,241,242,244,245,252-254,261, 192, 209, 234, 235, 249, 253, 262, 309, 310, 340,341,343,344,361,362,380,382,419,
264,274,286,288-290,294,296,306,307, 389, 395, 415, 458, 481, 601, 695, 698, 705 447,452,514,538,540,542,552,563; see
317,324,329,338,339,341,349,361,365, antibiotics, macrolides, 65 also ant, Driisenameisen. fire ants,
378,390,420,427,433,435,437,439,442, antibiotics, macrocyclic. 65 Stachelameisen
445, 452, 456, 460, 462, 466, 479, 486, 498, antibiotics, ansa-macrolide, 694 anls, leaf cutter, 340, 380,452, 514
503,506,508,510,511,513,515,517,518, antibiotics, polypeptide, 601 ants, Pharaoh, 563
520, 533, 537, 538, 542, 546, 549, 552, 553, anticancer properties, 379, 423, 452, 453, Aphanamixis, 477
559, 561, 568, 581, 588, 597, 615, 618, 640, 481-483,564,576,619,627,645,652,696 Aphanamixis grandi/olia, 481
645, 658, 659, 677, 681, 682, 684 anti-carcinogenic properties, 210 aphanostatin. 481
animals, polyphagous, 380, 390, 437, 445, 591, anticholinergic drug, 537 Aphelandra squarrosa, 522
597,603,613,618,619,640,641,662,664, anti-cholinesterase activity, 709 Aphelenchoides besseyi. 419
703 anticoagulants, 132,261 aphids, 41, 53, 99, \05,262,269,270,308,
anisaldehyde, 122 anticonvulsants, 136, 591 340, 343, 349, 362, 363, 365, 366, 383, 468,
anise. 122 antidepressant, 148 525, 550, 552, 559, 562, 567
Anisomorpha buprestiodes, 341 antidiarrheal compounds, 599, 600 aphid, black hean, 363
Anisotes, 569, 570 antidotes, 179 aphid, cabbage, 308
annatto,499 antiexudative, 460 aphid, damson-hop, 363
Annona cherimolia. 67. 73 antifeedants, 49, 113, \18, 136, 138, 213, 267, aphid-host plant compatibility, 262
Annona muricata. 67 288, 316, 361, 365, 379, 380, 390, 395, 396, aphid. melon, 67
Annona reticulata. 585 398, 412, 415-418, 423-425, 472, 473, aphid, pea, 363
Annona squamosa, 67 478-481,483,484,570,573,591,623,682, aphid, peach-potato, 363
Annonaceae, 46, 59, 67, 74, 104, 105, 139, 696,703 aphid, turnip, 383, 393
290, 392, 581, 585, 588, 598, 609, 610, 614, antifertility compounds, 553. 566 aphid, vetch, 363
639 antifungal activity, 26, 35, 40, 47, 48, 64, 85, Aphis cylisorum, 559, 567
AnnonaIes, 337 \18, 145, 146, 148, 179, 183, 269, 309, 3\0, Aphis fahoe, 363
Annoniflorae. 337 339,392,414,447,458,459,510,515,517, Aphis nerii, 468
anodynes, 673 525, 558, 591, 599, 601, 603, 682, 683, 685 Aphis po/ygama, 362, 363
Anopterus. 673 antigenic effects. 244 aphmdisiac, 36, 382, 537, 550, 552, 640
anolexia, 364 antihepatotoxic compounds. 168 Apiaceae, 3, 26, 39, 42, 45-48, 73, 109, \11,
anrakorinine, 684 antiherbivore compounds, 447 122, 130, 134, 136, 137, 153,208,251,252,
ansamycin. 695 anti-mY activity, 209 328, 332, 337, 341, 346, 360, 387, 395,460,
ant, see also ants anti-HSV-I activity, 423 537, 542, 543, 6\0
ant, Australian cocktail, 669 antiinflammatory compounds, 85, 122,261, Apieae, 134
ant, thief, 533, 542, 552
392,420,460,564,601,606,670,672,683 apiforol,l64
antagonisms, 4; see also calcium antagonists. anti-itching effects. 346 apigeuin, 152, 158-160, 162, 164-166
serotonin antagonists antijuvenile honnone activity, 49, 317, 384 apiin.252
antenna pigments, 496
antileishmaniacal activity, 847 2R-,B-D-apio-D-furanosy1-(1-6)-,B-D-
antennae, 552
antileukemic activity, 364, 365, 482, 484, 533, glucopyranosyloxyphenylacetonitrile. 277
antenna! receptors, 309
615, 645, 694, 695, 705 apiose, 251, 252
anthelminthic compuods, 28, 69, 346, 479,
539,599 antimalarial compounds, 69, 392, 395, 482 D-apiose, 252
Anthemideae, 36, 46, 387 antimalarial effects, 569, 648, 649, 650, 651, Apis mellifera, 445
Anthemis altissima. 277 652,666 Apium, 146
Anthemis cairica. 277, 297 antimicrobial effects, 40. 50, 59. 67, 118, 137, apocarotenoids, 499
Anthem;! glycoside A. 277 346, 458, 574, 591, 599, 600, 601, 603, 606, Apocynaceae, 98, 319, 361, 364, 466, 468,
Anthemi! glycoside B, 277 640 469,506,510,511,521,549,615,629,
antheraxanthin, 496, 504 antimitotic effects, 120, 510, 645, 676, 696 635-641,645,647,648,652,653,660,668,
antheridia. 444 antimutagenic effects, 165 677, 686, 689, 690, 700
antheridiogens, 414 antimycotic effects, 140 apoplastic space, 288
antheridiol, 444 antineoplastic effects, 120 aporphine alkaloid biosynthesis, 588
antheridium, 414, 424, 444 anti-nutritional effects, 208 aporpbine alkaloids, 578, 588, 590-592, 594,
anthochlors. 174 antioxidant properties, 120, 164, 166,207,499 605,627
anthocyanidin, 151, 152, 162, 163, 167, antiplaque properties, 603 aposematic labelling, 364. 366. 466. 468, 470.
170-172,177, 191, 193, 200, 204, 206, 213 antiprotozoan effects, 392 498, 551, 564, 567, 703; see also warning
anthocyanin biosynthesis. 162. 163 antipyretic properties, 122, 136,641,670 coloration
anthocyanins, 8, 151, 152, 156, 158, 159, 162, antirheumatic effects, 122 apparency, 209, 212
163, 170, 171, 175, 177, 189, 190, 193,498, antirrhinoside. 358, 364 apple pomace, 260
653, 70S, 708, 709 Antirrhinum majus. 158, 189. 358 apples, 174, 2\0, 215, 238, 277, 290, 346, 381,
Anthonomis grandis. 35, 342, 542 antismoking properties, 538, 540 397
anthothecol, 480 antispasmodic effects, 137. 346, 646 apricots, 277, 290, 346
anthranilate synthase, 98 anti-termite properties, 460 Aquifoliaceae. 290. 625
anthranilic acid, 11, 94, 98, 99, 459, 507, Antitoxicum, 563 Aquilegia, 269
568-571, 575, 576 antitubulin, 696 Arabidopsis Ihaliana. 4, 41, 303. 310, 314
anthraquinone glycosides, 76, 89, 91 antitumor compounds, 67, 104, 120, 145, 149, arabin,310
anthraquinones, 8, 9, 56, 64, 65, 76, 80, 82, 209, 234, 235, 254, 261, 378, 393,402, 404, D-arabinitol, 264
85-89,91,92,94, 104, 177 420, 445, 452, 460, 468, 473, 481, 482, 511, L-arabinitol, 262, 264
anthrone dimers, 76, 91 546,553,559,564,568,574,576,581,591, O-a-L-arabinopyranosyl-(1-6)-/3-D-glucose,
anthrones, 76, 85, 88, 91 599, 601, 605, 606, 613, 623-625, 627, 645, 266
Anthyl/is l'uineraria, 184 648, 653, 654, 671, 676, 677, 695, 696, 700, arabinose, 172,248,249,456
antiamebic activity, 559, 599, 612, 686 705,711 L-arabinose. 248. 260, 456
D-antiarose, 250 antitussive effects. 460, 591, 596, 597, 603 Araceae, 111, 123, 287, 515, 6\0
antiarrhythmic effects, 600, 640, 675 antiulcer effects, 167, 460 arachidic acid, 17
antiarthritic compounds, 392 antiviral properties, 48, 120, 128, 192,209, arachidonic acid, 34. 209
antibacterial activity. 48, 85, 267, 339, 392, 2\0,261,468,482,510 Arachis hypogaea, 65. 145. 226
510,525,601 antiyeast effects, 603 Arachnoides standish;;, 253
 Index 717

Araliaceae, 42, 45, 46, 48, 134,337,360,459, Artemisia annua, 348. 392 Atalantia, 573
460, 569, 570, 610, 612 Artemisia californica. 345 Atelia herbert-smith;;, 222, 231
Aralidiaceae, 360 artemisia ketone. 348 Athalia rosae, 418
Araliiflorae, 337 Artemisia maritima, 392 atisine, 673, 674
Araliopsis, 570, 573 artemisinin. 392. 607 Atkinsonella. 659
Araucariaceae, 405. 412, 413 artemisyl skeleton, 346 ATP, 6, 9, 14, 19,21,23,81,97, 155, 166,
Arbacia. 420 arteriosclerosis, 640 226,315,588,603,631,656
Arbacia punctulata. 420 Arthrobacter globiformis, 483 ATPase, 601, 603, 660, 676, 685
arborine, 568, 569 arthropods, 13, 79, 324, 340, 349, 439, 479 atractylenolide I, 392
arbusculin-A, 389, 390 artichokes, 520 Atractylodis macrocephala, 392
arbutin, 77, 78. 104 Arum maculatum, 515 Atractylodis rhizoma, 392
Arbutus. 78 Arum nigrum, 515 atrioventricular conduction disturbances, 623
Arbutus unedo. 78 Arundo, 101 Atropa belladonna, 3, 534, 535, 537, 558
Archaebacteria, 401 aryl hydroxylation, 136 Atrophaneura aleinaus. 591
Arcthosiphon pisum, 363 aryl-sulfatase, 160, 188 atropine, 511, 535, 537, 664, 692
Arctiidae, 551 ,B-asarone, III Alta cephalotes, 340, 380, 452
Arctostaphylos, 78, 125, 126 E-asarone, 113, 136 AUa sexdens, 514
Arctostaphylos giandulosa, 125 ascaricide, 392 attack, 319, 326, 339, 340, 342, 370, 372, 377,
Arctostaphylos uva-urai. 209 Ascaris, 392 380,391,401,414,415,447,468,478
Arctoteae-Calenduleae, 387 Asclepiadaceae, 361,466,515,563,571,573, Attacus atlas, 650
Areca, 513, 528 655,666 attIactants, 339, 342-344, 350, 362, 363, 366,
Areca catechu. 528 Asclepias, 466 381,382,397,445
Arecaceae, 25, 458, 569 Asclepias curassavica, 466, 468, 471. 703 aubergenol, 377
arecoline, 511, 528 Asclepias labri/ormis, 466 aubergenone, 377
Argemone, 603, 605 Asclepias syriaca, 468 Aucubaceae, 360
Argemone mexicana. 603 Ascobolus furfuraceus, 71 aucubin, 358, 363, 669
Argemone platyceras, 587 ascochitine, 63 aucuparin, 148
argemonine. 605 Ascochyta/abae, 63 Aurantioideae, 571, 573, 575
argentilactone, 269 Ascochyta rabiei, 186 aureomycin, 65
Argenlina, 244, 380, 563 Ascomycetes, 659 auricular fibrillation, 651
arginase, 220, 221 ascorbic acid, 158, 159, 162,202,207,209, aurones, 151, 156, 173, 174, 177, 188
arginine, 215, 220, 221, 223, 226, 517, 521, 210,247,265,266,271,300,587,623 auroxanthin, 499
531,534,546,547,680 L-ascorbic acid, 247, 265, 271 AQstralia, 29, 79, 177,226,229,287,294,
L-arginine, 220, 239 Asia, 525, 574, 624, 640, 666, 698, 700 380, 389, 551, 561, 563, 574, 605, 615, 617,
arginine decarboxylase, 517. 531, 547, 548 asimicin, 67 664, 666, 669, 676, 703
arginyl-tRNA synthetase, 220 Asimina triloba. 67, 74 Austrobaileyaceae,610
Argyreia. 658 asirinin, 118 Austrotaxus spicata, 676
aril, 222, 223 asparagine, 215, 226, 230, 231, 290, 291 autonomic nervous system (ANS). 510
Arion rufus. 559 asparagus, 227 autumnaline, 617
aristolactams, 591 Asparagus adescendens, 459 auxin, 3, 53, 146, 165, 166, 190,306,389,483
(- )-aristolochene, 371, 372 Asparagus ojficinalis, 227 Avena coleoptile assay, 33, 146, 389, 510
5-epi-aristolochene cyclase, 372 asparanin B, 459 Avena sativa, 98, 230, 459
Aristolochia, 591, 613 Aspartame, 245, 246 avenacin, 459
Aristolochia argentina, 269 aspartate aminotransferases, 601 avenalumin I, 98
Aristolochia clematitis. 591 aspartic acid, 215-217, 220, 230, 290, 293, avenalumin n. 98
Aristolochia indica, 372 294,570 avenalumin m, 98
Aristolocbiaceae, 36, 269, 372, 387, 591, 609 aspartic ,B-semialdehyde, 518 avenalumins, 568
aristolochic acids, 591, 615, 616 3-aspartylsemialdebyde, 217 avenic acid, 230
Aristotelia, 563, 564 Aspergillus, 59, 63-65, 212, 234, 658, 683 avocado, 419
Arizona, 553, 582, 690 Aspergillus niger, 212, 414, 591 axivalin, 390
Armoracia lapathifolia, 303, 306 Asperula, 85 ayabuasca, 660
Armoraeia rusticana, 168 Aspbodelaceae, 87 8-azabicyclo[3.2.1]octane system, 533
anny wonn, 361, 379 asphyxia, 642 Azadirachta, 473, 476, 484
anny wonn, African, 361, 379, 380,415 Aspidiaceae, 544 Azadirachta indica, 473, 478, 479, 484
anny wonn, bertha, 308 Aspidosperma, 560, 632, 634-636, 640, 645, azadirachrin, 379, 479, 480, 483-485
anny wonn, fall, 145, 390, 479, 481, 658 652, 660, 666 azafluoranthene alkaloids, 588
anny wonn, southern, 228, 289, 390 Aspidosperma alkaloids, 632, 645 azetidine-2-carboxylic acid, 3, 222
anny wonn, yellow-striped, 390 Aspidosperma-Hunteria alkaloids, 634 azidirone, 479
Arnica, 392 aspidospennatan, 636
arogenate dehydratase, 101, 106 Aspilia, 48 baccharinoids, 379
arogenate dehydrogenase, 106 aspirin, 122, 123 Baccharis, 378. 395
arogenic acid, 101, 102 Asteraceae, 26, 36,42, 43, 45, 46, 48-50, 77, Baccharis cordifolia, 378
aroma, 324 88, 108, 109, 117, 125, 134, 158, 159, 168, Baccharis gaudichaudiana, 420
aromatic alkaloids, simple, 513 173-175, 177, 179,257,279,280,282,286, Baccharis megapolamica, 378, 395
aromatic compounds, 45, 94, 96, 508, 529 287,316,317,319,320,322,326,328,337, bacilli, Gram-positive, 600
aromatic ring, 45, 56, 57, 76, 80, 99, 106, 115, 345, 346, 374, 378, 380, 384, 386, 387, 390, Bacillus, 234, 253
151,187 392,393,395,396,405-407,412,413,415, Bacillus acidocaldarius, 26, 39
aromatization, 525 418,420,439,458,510,514,549,550,552, Bacillus brevis. 234
arrow poisons, 12, 187, 456, 463, 469, 470, 553, 559, 673 Bacillus polymyxa, 234
511,588,607,652,673 Astereae, 387 Baeillus subtilis, 69
Artabotrys uncinatus, 392 Asteridae, 277 backcrosses, 338
Artemia salina, 145 asthma, 137, 564, 576, 689 bacteria, viti, 2-4, 8, 9, 19, 22, 23, 27, 32, 49,
Artemisia absinthum, 333, 346. 389 Astragalus, 229, 253, 290, 299, 561 56,65,72,77,80,83,94,97-99, 101, 102,
artemisia alcohol, 348 Astragalus lentiginosus, 561 104, 106, 121, 122, 125, 134, 167, 193,210,
Artemisia, 332, 345, 346, 389, 552, 559 astringency, 206, 207, 209, 210, 346 218,234,235,237,239,240,244-246,249,
718 Index

252.253.259,270.273,286.290,306.312, bean, mung, 102, 108, 166. 222, 260 Berberidaceae, 1I8, 280, 282, 557, 598, 603,
313,324.339,401,402,460,481,483,484, bean, precatory, 244 609,610
486,488,490,494-496,498,510,517,520, bean, tonka, 130 Berberidales, 337
559,561,573,601,613,650,681 bean, winged, 290 Berberidopsis beckleri, 282, 296
bacteria, anaerobic, 22 bearberry, 209 berberine, 506. 511, 586, 598-600
bacteria, Gram negative, 603, 640, 681 beaver, 670 berberine alkaloids, 586, 597
bacteria, Gram positive, 69, 558, 603, 681 beaver, Canadian, 670 berberine bridge, 598
baikiain, 218 bebeerine, 606 "berberine-bridge" enzyme, 598
Bailfonella toxisperma, 703, 711 bee flowers, 176, 177 Berberis, 586, 587, 598
Baja California, 441 beech. 115 Berberis aggregata, 599
Balanites aegyptica, 460 beer, 70, 165,316,378; also see kaffir beer, Berberis canadensis, 587
Balansia, 659 root beer Berberis slolonifera, 241, 587
Balansia cyperi, 660 bees, 151, 176, 177,257,344,362,385,445, Berberis wilsoniae, 598, 613
Balansiae, 659 640,641,703; also see bee flowers, bumble bergamotene, 376
Balansiopsis, 659 bees, carpenter bees. euglossine bees, (+ )-(E)-endo-p-bergamoten-12-oic acid, 381,
Balfourodendron, 573 honeybees, solitary bees 394
Baliospermum, 403 bees, solitary, 381 bergamotin, 335, 336
Balsaminaceae, 82 beetle, Agasicles, 168 bergapten, 113, 133, 135-137
Balsamodendron pubescens, 475 beetle, blister, 382 bergaptol, 133
bamboo, 115, 607 beetle, bombardier, 79 Bergera, 663
bamboo shoots, 290 beetle, Colorado potato. 31, 39, 231, 682, 683, berry, see bearberry, blueberry, chinaberry,
bananas, 215, 228, 517 689 mulberry, raspberry ketone, raspberry,
Banisteriopsis argentea, 514 beetle. confused flour, 35 serendipity berries, strawberries
Banisteriopsis caapi, 660 beetle, cucumber, 445 Besseya alpina, 363
Banisteriopsis inebrians, 700 beetle, driedfruit, 36 Besseya plantaginea, 363
Banyambo tribe, 652 beetle, flour, 562 Beta vulgaris, 3. 222, 705. 708
I/I-baptigenin, 181 beetle, hide, 35 betacyanins, 423, 705
Baptisia, 188-190,229,558 beetle, Khapra, 35 betalnin, 171, 177, 506, 692, 705, 706, 708,
Baptisia australis, 554 beetle, ladybird, 363, 366, 542, 552 710.711
Baptisia leucophaea, 8, 558 beetle, Mexican bean, 67. 480 betalain glucoside, 705
Baptisia spherocarpa, 8 beetle, Mexican water, 439 betalamic acid, 706, 708
bark beetle, European elm, 83, 169 beetle, mustard, 309 betanin, 705, 708
bark beetle, hickory, 84 beetle, red flour, 35 betaxanthins, 705, 708
bark beetle, pine 343 beetle. rust-red flour, 460 betel,210
b",k beetles, 37, 72, 168,339 340,342,343 beetle, sawtoothed grain, 37 betel nut, 528
b",k, 30, 40, 67, 85, 86. 88,110.117,139, beetle, scarabaeid, 53 Betula alba. 447
174,193-195,206,209,244,252,319,342, beetle, soldier, 49, 50, 542 Betula pendula, 120, 129
343,353,403,405,413,447,480,528,561, beetle, striped cucumber, 67 Betula platyphyl/a, 121
608.640,645.649,663,676,689,691,695, beetle, western pine, 343 Betula resinijera, 451
700 beetles, see also bark beetles, bruchid beetles, Betulaceae, 197, 242
bark, willow, 122 chrysomelid beetles. diabrotid beetles, dung betulin, 447
bark, yohimbe, 640 beetles, flea beetles, beetle-pollinated beverages, 110, 136, 140, 364, 692, 694, 700,
b",ley. 15,59,70, 129,240,517,518,522, flowers, water beetles 708
523 beetles, dung, 515 beverages, intoxicating, 694
bartenders, 137 beetles, leaf, 444, 445, 447 beverages, stimulating, 694
base, see purine base, pyrimidine base, beets, 423, 705, 708 Beyeria, 407
tetrabase, Schiff-base, xanthine base Begonia, 444 d-beyerene, 408
Basellaceae, 708 Begoniaceae, 444 ent-beyerene, 407, 408
basidiomycetes, 45, 46, 48, 107 Belgium, 378 bhang, 336
Bataceae, 304 belladonna, 534, 535, 537 Bhesa, 664
batatasin I, 147 Bellardia trixago, 500 bibenzyl compounds, 136, 147, 148
batatasin III, 147 Beltian bodies, 289 bibenzyl synthase, 147, 149
batatasin IV, 147 Bentazon, 99 BMens, 47
Batidaceae, 708, 711 benzaconine, 675 BMens pilosa, 47
batrachotoxin, 675, 677 benzaldehyde, 581 biflavonoids, 151, 173
bats, 176, 177,344 benzoate hydroxylase, 104 big blue stem, 125
Battus philenor, 591 benzofurans, 312, 316, 317, 322 biglandulic acid, 269
Battus poiydamas, 591 benzoic acid, 121, 148,674 Bignoniaceae. 82, 86, 88, 114,337,361,362,
bay leaf, 340, 352 benzoic acid 2-hydroylase. 121 568,669
Bayer Company, 122 benzophenanthridine alkaloids, 578, 586, 600, bilharzia, 459
bean, viii, 26, 32, 437, 559 601,610.614,616 Billia, 223
bean, black, 167, 561 benzophenones, 136, 139, 148 binding, see cholesterol binding, HIV -1 cell
bean, broad, 48, 244, 414, 623 benzopyrans, 312, 316 binding, hormone binding sites
bean, calabar, 663 benzoquinone, 76, 77, 78, 80, 104, 124 bioassay, brine shrimp, 67, 145, 149
bean, castor, 22-24, 26, 244, 261, 369, 399, p-benzoquinone, 76, 77, 124 biochanin A, 186
407,414 benzoxazinone, 99 biogenetic isoprene rule, 312, 427
bean. coffee. 705 N-benzoylantbranilic acid, 568 Biomphalaria glabrata, 209
bean, dry, 261 benzyl acetone, 113,344 biosynthesis, see alkaloid, anthocyanin,
bean, dwarf, 414 benzyl benzoates, 140 aporphine, carotenoid, fatty acid, flavon-3,4-
bean, fava, viii benzyl isothiocyanate, 309 diol, flavonoid, iridoid monoterpene, lupine
bean, French, 156, 186 benzylglucosinolate, 287, 303, 310 alkaloid, monoterpene, polyamine,
beans, jack, 222, 244 benzyIisoquinoline alkaloids (BIQ), 2, 3, 12, polyketide, proanthocyanidin, protein,
bean, kidney, 241, 244 136,337,560,561,575,578,579,583-588, quinolizidine alkaloid, rubber, sesquiterpene,
bean, lima, viii, 287, 290, 295, 299. 341 591,594,597,598,605,607-610,613,614, squalene, stilbene, terpenoid, tetraterpene,
bean, mescal. 560 616, 652, 660, 664, 668 and triterpene biosynthesis
 Index 719

biosynthesis, channeled,S, 153,275,277,556 Bombyx, 341, 641, 689, 703 Brucea javanica, 482, 662, 667
biosynthesis, rubber, 319 Bombyx mori, 35, 36, 118, 341, 419 bruceantin, 482
biosynthetic pathways, vii, 2, 3, 6, 11, 12, 56, Bonajusia, 637 bruceoside A, 482
85,95, 179, 182,217,226 bone, 223. 437 bruchid beetles, 40, 132, 138,217,220,221,
birch, 8, 129 Boraginaceae, 3, 18,77,82,85, 108, 177,257, 223, 232, 245, 296, 523, 529, 544, 562, 565,
birch, Alaska paper, 451 285,290,294,336,337,452,510,547,549, 615,627,653,703,708
birch, Japanese 121 551-553 Bruchophagus roddi, 31
birch, Scandinavian, 120 borneol, 331, 345, 346, 451 bmcine, 641, 642
birch, white, 447 ( + )-boroeol, 335 Brugia pahangi, 479
birds, 175, 176, 179, 344, 361, 466, 486, 498, D-bomesitol, 264 Brugmansia, 537
511,517; see also bird-pollinated flowers, D-( - )-bornesitol, 264 BrUtifeisia, 537
hummingbird flowers, hummingbirds, L-bomesitol, 264 Brussel's sprouts, 306
ladybird beetles, red-winged blackbirds L-( +)-bornesitol, 264 Bryonia, 34, 35
bisabolenes, 374 bornyl acetate, 343, 344 bryophyllin B, 468
P.bisabolene, 376 ( - )-boroyl pyrophosphate, 331 Bryophyl/um, 468
y-bisabolene, 375 (+ )-bornyl pyrophosphate, 330, 333 Bryophyllum pinnatum, 468
a-( - )-bisabolol, 383, 384 ( - )-bomyl pyrophosphate cyclase, 330 bryophytes, 9, 114, 164, 324, 369, 500, 510
bisbenzylisoquinoline alkaloids, 605-607 d-bomyl pyrophosphate cyclase, 330 Bryum capillare. 179
bisindole alkaloids, 628, 645, 652-654, 666 I-hornyl pyrophosphate cyclase, 330 Buchenroedera, 549, 566
bisnaphthoquinones, 85 (+)-boroyl pyrophosphate syuthase, 330 buckwheat, 76, 162
bitter principles, 386, 389 Borrer;a, 611, 612, 638, 665 bud, see dormancy, bud
bitter taste, 171, 175,253,269,272,316,322, Borreria verticillata, 662 buddamine, 669
361,361,364,386,389,390,417,444,452, borrerine, 662 Buddleja davidii, 390, 397
460, 463, 466, 478, 482, 483, 511, 552, 641, borreverine, 662 Buddlejaceae, 361, 390
649, 673, 684 boschnaloside, 358 buddlejin A, 390
bitterbrush, 208 Boschniakia rossica, 361 buddlejin B, 390
bittersweet taste, 525 ( + )-boschniakine, 669 buddlejin C, 390
Bua orellana, 372, 499 boschnialactone, 361 buds, 7, 136, 335, 436
Bixaceae, 372 Bothriospora, 611, 612, 638 budwonn, cotton, 30, 481
bixin, 499 Bothrops atrox, 179 budwonn, tobacco, 169, 171, 220, 380, 415
black locust, 244 Botrytis, 48, 594, 601 bufadienolides, 466, 468
black rot disease, 72 Botrytis cinerea, 145 buffalo bur, 677
black spot disease, 67. 238 Botrytis tulipae, 269 Bufo marinus, 594
blackbirds, red-winged, 523, 619 Bouvardia ternifolia, 700 bufonid toad, 594
blackbrush, 208 bouvardin, 700 bufotenin, 514
blackcurrant, 346 Bowdichia nitida, 79 bugs, 285, 468; see also greenbugs, ladybugs,
blackwood, 79 Brachynus, 79 lygaeid bugs, mealybugs, milkweed bugs
B1akeslea, 501 bradycardia, 623, 647, 674 bulbocapnine, 590, 591
B1attela germanica, 136 brain, 597, 640, 659, 676 Bulbocodium, 617
bleomycin A2, 235 brain, mouse, 640 bulbs, 457, 472, 623
Blighia, 222, 223 Brassica, 306, 309, 310, 345, 363 a-bulnesene, 373, 376
Blighia sapida, 222 Brassica campestris, 306, 309 bumble bees, 362, 420
Blighia unijuga, 222 Brassicajuncea, 306, 309, 310 Bupleurum falcatum, 460
blindness, 603 Brassica napus, 30, 39, 306, 307, 311, 437 Burseraceae, 30, 337, 387, 412, 473, 475, 576
blisttning, 267 Brassica nigra, 309, 311 bush rats, 29
blocking agents, see a-adrenergic blocking Brassica oleracea, 51, 52, 190.306,309,310 Bushmeo of Namaqualand, 623
agents, adrenergic blocking agents, Brassica rapa, 306 Busse massaiensis, 222
cholinergic blOCking agents. ganglionic Brassicaceae, 4, 51, 52, 108, 168, 179,267, Z,E-l,3-butadiene hydroperoxide, 24
blocking agents, neuromuscular blocking 300,302,303-305,308,310,311,345,418, 2-butanone, 276
agents 437,444,466, 468, 521, 537 E-Ll2-butenoyl-S-ACP, 20
blood, 174,209,244,384,447,452,456,479 brassinolates, 444 butterflies, 10, 13, 123, 136-138, 177, 191,
blood brain barrier, 229, 597 brassinolide, 437, 454 192,265,297,307,309,310,341,344,350,
blood pressure, 517, 522, 528, 606, 640, 643, Brazil, 378, 390, 395, 588, 595, 610, 645, 700 361,466,470,535,550-552,559,564,566,
670,703 bread, 30, 658 591,711
blood sugar, 223 brefeldin A, 420 butterflies, danaid, 550, 551
bloodroo~ 601 Bretschneideraceae, 304, 310 butterflies, ithomiine, 535, 683
blossoms, see flowers brevicomin, 37, 343 butterflies, swallowtail, 591
blowfly, 67 exo-brevicomin, 343 butterfly, black swallowtail 136, 138,307
blue jays, 466 Brevicoryne brassicae, 308 butterfly, blue lycaenid, 559
blue mahoe, 77 brevilagin 1, 196 butterfly, buckeye, 364, 365
blueberries, 171 brevilagin 2. 196 butterfly, cabbage, 307, 309
bluegrass, 52, 54 British Columbia, 338 butterfly, checkerspot, 363
Boehmeria, 563 Briza spicata, 30,41 butterfly flowers. 177
D-boivinose, 250 broccoli, 306, 311 butterfly, hairstreak, 294
boldine, 591 bromohydroquinone, 340 butterfly, monarch, 383, 466, 471, 472, 551,
Bolivia, 114, 535, 684, 690 Bromus rigidus, 126 591,703
boll weevil, 35, 342, 380, 542 bronchial dilator, 137 butterfly, queen, 551
bollwonn, 49, 50, 169, 380 bronchoconstriction. 643, 697 butterfly, yellow, 623
bollworm, cotton, 415 bronchodilating effects, 522 butylated hydroxyanisole (BHA), 166
bollwonn, pink, 49, 50, 415 Brosimum, 134 butyrate, 152
Bomarea, 269 Broussonetia, 571 butyrospennol, 473, 476
Bombacaceae, 28, 77, 387 Broussonetia papyri/era, 145 butyrylcholinesterase, 600
Bombax,77 broussonin A, 145 butyryl-S-ACP, 19, 20
Bombus terrestris, 419 broussonin B, 145 Buxaceae, 677, 686, 689
bombykol, 36 Brucea antidysenterica, 482 buxenine-G, 689
720 Index

Buxus, 677, 689 caUinecdysone-B, 237 canthine derivatives, 660

Buxus alkaloids, 689 Callistephus chinensis, 158, 439 Canthium, 700
Buxus sempervirens, 689 Callitricaceae, 361 cantleyine, 669
callose, 247, 259 Capirona, 611
C 4 starter unit, 69 Callosobruchus chinensis, 460 CapparaIes, 304
Cs units, 326, 370 Callosobruchus maculatus, 5, 40, 132, 138, Capparidaceae, 302, 303, 308, 521
C6 -C, compounds, 106, 121, 122, 139, 148, 293,296,217,223,232,315,522,523,529, Capparis, 302
195,530,622 537,544,558,562,565,609,615,627,640, Capparis decidua, 521
C6 -C2 compounds, 106 642, 653, 703, 708 capric acids, 25
C6 -C3 compounds, 106, 109, 115, 121, 139, Callosobruchus phasecoli, 340 Caprifoliaceae, 173,355,361
148, 193, 195,613,694 Ca/oncoba echinata, 28 capsaicin, 110,517
C7 non-protein amino acids, 222 calophyUolide, 187 capsaicinoids, 517
C-17 side-chain cleavage, 439 Calophyllum inophyllum, 187 capsanthin, 486, 494, 504
C,racetylenes, 45 Calospilos miranda, 417 Capsella bursa-pastoris, 309
C 17-tenninal olefms, 44, 45 calotropin, 466, 468 Capsicum, 110,377,488,499,504,513,517,
C 20 isoprenyl ethers of glycerol, 401 Caltha, 269, 549 530
Cn-sterols, 437 Calvin cycle, 248 Capsicum annuum, 377, 494, 703
C 29 alkane, 52 Calycanthaceae, 610, 655, 664, 665 capsidiol, 373, 377, 517
C 29-sterols, 439 calycanthine, 664, 665 capsorubrin, 494
cabbage, 51, 52,171,306-309,520 Calycanthus alkaloids, 655 capsules, 593, 596
cabbage, Chinese, 132, 135 Calycanthus florida, 665 Capuronetta, 637
cabbage looper, 390 Calyceraceae, 361 Caragana, 215
cabbage, red. 171 CaJypogeia granulara, 368 Carapa procera, 480
Cacalia,77 Calypogeia tosana, 143 caraway, 47
cacao, 219, 700, 703 Camellia sinensis, 171, 195, 700, 702 carbamate group, 663
Cactaceae, 515, 522, 523, 578, 581, 692, 693, camels, 525 carbamoylputrescine, 531
708 Cameroons, 640 carbazole alkaloids, 575, 576, 655, 662, 663,
cacti, 148,441,453,523,581-583,615; see cammelinin, 310 666
cactophilic yeasts, coccus cacti Camoensia, 558 carbinolamines, 508
cactus, agria, 441, 582 Campanulaceae, 45, 46, 257, 458, 540 carbocation, 329, 374. 432
cactus, cina, 582 campesterol, 427, 435, 582 carbohydrate, 194,207,208.217,247-250,
cactus, organ pipe, 441, 582 camphene, 331, 345 252,254,260,261,265,324
cactus, peyote, 148, 151,522,523,581 ( - )-camphene, 331 carbohydrates, structural, 259
cactus, saguaro, 441, 582, 615 ( + )-camphene, 331 ,B-carboline alkaloid, 10, 514, 649, 655, 660
cactus. senita, 441, 582 d-camphene,331 2·carbomethoxy~4,4*bis(3-methyl*2-
cadalane skeleton, 670 I-camphene, 331 butenyloxy)chalcone, 168
cadaverine, 8,510,517,518,537.540,545, campho" 330, 331, 345, 346, 451 carbon dioxide, 6, 19,20,21,24,94,230,231,
554-556 ( - )-camphor, 345 261,290,313,328,329,336,435,488
cadinenes, 346, 375 ( + )-camphm, 335, 345 cis-2-(carboxycyclopropyl)glycine, 218
y-cadinene, 375, 381 Camptotheca acuminata, 648, 654 2-carboxy-4-hydroxy-l-tetralone, 82, 86
cadinol, 346 camptothecine, 628, 630, 648 trans-2-(2-carboxymethylcyclopropyl)glycine,
Caenocoris nerii, 468 camptothecine, 20(RS)-9-amino-derivative, 648 222
Caenorabditis elegans, 67 Canada, 322, 338 2-carboxy-4-oxotetraIone (COT). 81, 82, 86,
Caesaipinia, 194 canadine, 598, 600 87
Caesalpiniodeae, 86,105.179,197,221,228, ( - )-canadine, 598 3(3-carboxyphenyl)aIanine. 215
412, 543, 558, 676 canaline, 220, 221, 232 S·(2-carboxypropyl)cysteine, 226
caffeic acid, 108, 193, 518 Canavalia ensiformis, 220, 221, 244 carcinogenesis, 306
Z-caffeic acid, 113 canavanine, 220, 221, 231 carcinogenic compounds, 59, 65, 70, 110. 165,
E-caffeic acid, 113 L-canavanine, 215, 220, 221, 232 294,306,311,510,529,553,574,591,640
caffeine, 511, 692, 700. 702, 703 cancer, see also anticancer, antitumor, carcinoma, nasopharyngial, 606
caffeoyl acid, 104 carcinoma, leukemia, tumor carcinoma, squamous cell, 235
caffeoylputrescine, 518 cancer cell growth, 645 Cardamine cordi/olia, 309
3-caffeoylquinic acid, 171 cancer, esophageal, 210 cardenolides, 9, 309, 456, 462, 463, 465, 466,
Cakile, 305, 311 cancer, gastric, 601, 648 468-472
Cakile edentula, 305 cancer, human colon, 64& cardiac activity, 676
Cakile lanceolata, 305 cancer, liver, 553, 648 cardiac arrest, 674
Cakile maritima, 305 cancer metastasis, 562 cardiac effects, 675
Calabar Coast, 663 cancer remedy, 676 cardiac glycosides, 249, 250, 267, 433, 456,
calabash, 643 cancer, stomach, 606 463, 465, 466, 468-470, 552, 703
caiactin, 466 cancer, throat, 193,210 cardiac irregularities, 674
Calamintha ashei, 346, 451 cancerina, 671 cardiac muscle, 468
calcium, 260, 264, 402, 418, 437, 640 Candida albicans, 432, 588, 591. 683, 686 cardiac stimulation, 522, 640, 702
calcium antagonists, 676 candy, 462 cardiomuscular calcium transport, 418
calcium chloride, 676 Canellaceae, 379, 387 (S)-cardiospennin, 282, 285, 287, 289
Calendulae, 287 Cannabaceae, 316, 336, 522 (S)-cardiospermin p-hydroxybenzoate, 282,
California, 125, 128, 338, 345, 349, 389, 483, cannabidiol, 336 284,287
559 cannabidiolic acid, 336 (S)·cardiospennin p-hydroxycinnamate, 282.
California bay laurel, 340 canniabinol, 336 287
caliminthone, 346 cannabinolic acid, 336 (S)-cardiospermin sulfate, 282
Calindrinia ciliata, 126 Cannabis sativa, 336, 522 cardiotonic, 114, 392, 445, 462, 673
C-alkaloid E, 643 canteloupe, 468 cardiovascular effects, 650
C-alkaloid G. 643 Cantharellus cibarius, 496 cardon. 441, 582
C-alkylation, 316 cantharidin, 382 3-carene, 343
Callianthemum, 269 canthaxanthin, 498 caribou, 207
Callicarpa, 417 canthin·6-one, 655, 662, 667 Carica papaya, 287, 309
 Index 721

Caricaceae, 286, 287, 304 Castor jiber, 670 cell culture, colon 8, 517, 695
Carisseae, 637 castor oil, 35 cell culture, L-121O (lymphoid leukemia), 420,
canninic acid. 89 castoreum, 670 601,695
carnation, 156,231 .232 casuarictin, 197 cell culture, LE, 482
carnauba see wax, camauba, 52 Casuarina, 173 cell culture, LL, Lewis lung carcinoma, 482,
camegeine, 581, 582 Casuarinaceae, 173, 197,242,244 695
Carnegiea g;gantea. 441, 582 catabolism, 4, 5, 297 cell culture, murine P-3381ymphocytic
carob gum, 260 catalase, 79 leukemia, 482
carotene, 401 Catalpa, 361, 362 cell culture, murine tumor systems including
a-carotene, 486, 491, 492. 495 Catalpa oVata, 82 B16 melanoma, 695
p-carotene, 486, 491, 492, 494-497, 499, 504 Catalpa speciosa, 362 cell culture, plant, 1,2,8,39, 156, 179, 181
£-carotene. 486 catalpalactone, 82 cell culture, 9 PS system, 671
{-carotene, 490 catalpal,361-364 cell culture, Walker 256 carcinosarcoma, 640
(6'R)-p, ..carotene, 486 catalpanol, 82 cell cultures, parsley, 4, 156
(15Z)-{-carotene, 491 catalposide, 361-363 cell cultures, PS, B 1 and KB tumor lines, 696
,8-carotene dioxygenase. 497 catechin, 159, 171,200-204,206 cell cycle, 231
(3R,3'R)-p,p-carotene-3,3'-diol, 494 (+ )-catechin, 171, 193,202-204,206 cell cycle, 0, phase, 231, 520, 528
(3R,3'R,6'R)-,B,E-carotene-3,3'-diol.486 (+ )-(ZR,3S)-catechin, 201 cell cycle, G2 and/or M phase, 676
carotenoids, acetylenic. 495 (- )-(2S,3R)-catechin, 201 cell cycle, G2 cellular arrest, 708
carotenes, alicyclic, 492 ent-catechin, 201 cell cycle, G2 Factor, 231
carotenoid biosynthesis, 488. 496 catechin xyloside, 169 cell cytosol, 19
carotenoid pigments, 498 catechol, 67, 124 cell division, 520, 619, 623, 676, 696
carotenoids, 10, 171, 173, 177, 314, 322, 348, catechol structure, 67 cell free enzyme systems, see enzyme systems
367, 384, 486-488, 490-492, 494, 496-504 catecholamine, 517, 640 cell recognition, 261
carotenoids, acyclic, 491, 492 caterpillar, saddleback, 390 cell suspension cultures, 2, 13, 92, 93, 128,
carpenter bees, 177, 362 caterpillars, 29, 32, 137, 138,316,437,455; 138, 155, 158, 178, 179, 184,553,554,587,
Carpophilus hemipterus, 37 see also lepidopteran caterpillars, noctuid 600,601,631,652,653,654,667
carrageenan, 260 caterpillars cell wall fractions, 135
carrot, 26, 48, 72, 73, 113, 128, 129, 136, 328, Catha, 452 cell walls, 184,226,253,259-262,271,561,
399,486,490,492 Catha edulis, 210, 522, 671 613
Carthamus tinctorius, 48 catharanthine, 630, 632, 634, 635, 645, 647, cell walls, plant, 3, 8, 259-262, 270
Carum carvi, 47 652 cell-cell communication, 261
(-)-trans-carveol, 335 (+ )-catharanthine, 635 cellobiose, 254
carvone, 339, 340, 341, 346, 451 Catharanthus, 629, 637, 640
cells, 1-3, 8, 9, 13, 14, 18, 22, 32, 35, 51, 53,
d-carvone. 344 Catharanthus alkaloids, 628, 645
59,71,72,85,96, 110, 120, 128, 131, 138;
Catharanthus longijolius, 645
carvone oxide, 344 see also cancer
Catharanthus ovalis, 645
Carya, 82, 84 cells, cell free enzyme systems, chemosensory
Cotharanthus roseus, 2, 110, 241, 354, 355,
Carya illinoensis, 84 cells, epidermal cells, epithelial cells,
366, 506, 511, 629-632, 634, 635, 640, 645,
Caryedes brasiliensis, 220 extracellular enzymes, mv-1 cell binding,
652-654
Caryophyllaceae, 88, 156, 171, 458, 514, 568, leukemic cells, mesophyll cells, nerve-cell
Catharanthus trichophyllus, 645
662, 708, 709
cathartic, 35, 91, 364, 365, 402, 479 membrane, oil cells, cell culture, cell walls,
Caryophyllales, 197,705,708,709
caihenamine, 631 red blood cells, nelVe cells, secretory cells,
caryophyllanes, 375 cathenamine reductase, 631 sex cells, squamous cell carcinoma
caryophyllene, 342, 346, 371, 375, 379 cathinone, 522, 671 cells, chemosensory, 588, 651
p.caryophyllene, 371, 375 (S)-cathinone, 522 cells, HeLa, 676
caryopbyllene cyclase, 375 cats, 361, 362, 591, 643, 669, 674; see also cells, leukemic, 145
caryopbyllene epoxide. 379 civet cat cells, mouse lymphoma, tk locus, 645
caryophyllene oxide, 346, 380 cattle, 71, 91, 290, 317, 420, 437, 460, 540, cells, nerve, repetitive discharges, 685
Caryophyllidae, 171 553, 658, 676, 705 cellulose, 254, 257, 259, 260, 270, 272
Caryopteris.417 Caulanthus, 305 cembrene, 398, 401, 402, 413, 419
casbene, 401, 408, 414 Cauierpa, 380, 396, 419 Centaurea, 47
casbene synthase, 401, 402 Caulerpa ashmeadii, 380, 396 Centaureo solstidalis, 317, 322
cascara sagrada, 91 cauliflower, 306, 309, 311 Central America, 229, 298, 525, 640
Casimiroa, 693 Caulophyllum thalictroide, 557 central nervous system, 229, 510, 514, 522,
casimiroine, 571 Cavariella aegopogii, 340 596,640,642,647,670,698,702
cassaine, 406 , 412, 676 CD, see circular dichroism Centrospennae, 171,705,708,711
cassava, viii, 274, 276, 290, 294, 297 cedar leaf oil, 346 cephaeline, 610, 611, 652
Cassia, 85, 88, 91, 543 cedar, western red, 340 Cephaelis, 610-612, 638
Cassia absus, 693 Cedrela toona, 399 Cephaelis ipecacuanha, 610, 612
Cassia fistulosa, III cedrelone, 475, 479, 480 cepbalomannine, 677
Castanea sativa, 252 Celastraceae, 210, 240, 269, 270, 321, 452, cephalosporin C, 235
castanospennine, 254, 270, 560-562, 566 466, 470, 522, 525, 528, 549, 575, 664, 668, cephalosporins, 234
Castanospermum australe, 254, 561 670, 671, 694, 695, 700 Cephalosporium, 234, 235
castelanone, 482 celery, 48, 137, 556 Cephalotaxaceae, 617, 624, 626, 677
Cassia, 85, 88, 91, 543 celery root, 48 cephalotaxine, 625, 627
Castilla elastica, 319 cell culture, 3, 4, 8, 9, 67, 82, 86, 87, 117, Cephalotaxus, 617, 624-627, 677
Castilleja hispida, 552 132, 133, 143, 153, 156, 181, 182, 184, 186, Cephalotaxus alkaloids, 617, 624-627
Castilleja integra, 363 188, ZOO, 202, 209, 290, 506, 533, 554, 557, Cephalotaxus harringtonii, 625, 627
Castilleja lineoriijolia, 552, 559 558, 567, 586, 598, 601, 614, 616, 629, 631, Cephalotaxus wilsoniana, 625, 627
Castilleja miniata, 557 636, 640, 645, 646, 649, 653 Ceratiola ericoides, 125, 175,451
Castilleja occidentalis, 552 cell culture, Bl6 murine melanoma tissue ceratiolin, 175,451
Castilleja rhexi/olia, 552, 669 culture, 482 Ceratitis capitala, 31, 40, 381
Castilleja sulphurea, 552 cell culture, carcinoma of the nasophamyx Ceratocystis, 342
Castor, 668, 670 (KB), 380, 420, 444, 445, 452, 468, 479, Ceratocystis fimbriatum, 27, 40, 72, 135, 377
castor bean, see bean, castor 481, 606, 663 Ceratocystis u1mi, 72, 73, 237
722 Index

Ceratonia siliqua, 260 cherry laurel, 274 chromatography, centrifugal thin layer (C1LC),
Ceratophalus, 269 chestnut, 194, 252 457
CercidiphylIaceae, 197 chestnut, Moreton Bay, 561 chromatography, countercurrent, 1
Cercospora rosicola, 501 chestnut, Spanish, 195 chromatography, droplet countercurrent
cereal grains, 78, 98, 99, 256, 378, 518, 520, chewing gum, 321, 462 (DCCC),457
658 chicken, 35 chromatography, gas liqnid (OC or OLC), 54,
cerebral cortex, 346, 702 chickpea, 184, 186, 188 142,217,302,305,326,351
cerebral vasodilation, 640 chicle, 312, 318, 321 chromatography, gas liquid (FID), 457
cerin,447 childbirth, 660 chromatography, gas capillary, 326
Cervus eZaphus, 207 Chile, 388, 563 chromatography, gel permeation, 318
Cestreae, 537 chimonanthine, 665 chromatography, high performance (HPLC), I,
Cestrum, 677 chimpanzee, 48 105, 132, 138, 142, 187, 188, 191,212,217,
cevadine, 685 China, 210, 244, 306, 522, 606, 646, 648, 709 232,295,302,353,365,398,457,465,471,
cevane skeleton, 684 china aster, 158 487,514,655
ceveratrum alkaloids, 684 chinabeny, 479 chromatography, ion exchange, 302
a-chaconine, 682, 684 chio-shih-hu, 670 chromatography, liqnid (LC), I, 38, 132, 295
Chaenorrhinum, 522 Chisocheton paniculatus, 475 chromatography, paper (PC), I, 212, 217, 223,
Chaetocnema con/inis, 7 chitin, 247, 253, 260, 261, 270-272, 414 294,302,305,309
Chagas' disease, 479, 645 chitinase, 414 chromatography, thin layer (1LC), 1,38, 142,
chalcid,31 chives, 226 212,217,294,302,353,362,398,465,466,
chalcone 2,3-epoxide, 159 chloramphenicol, 234 514
chalcone glycosides, 178 Chiorelia fusca, 242 chromenes, 312, 316, 322
chalcone isomerase, 156, 158, 173 chlorofonn, 507 chromium ion «(Yi+), 209
chalcone reductase, 179 chlorogenic acid, 104, 108, 128, 171, 193,700 chromones, 477, 576
chalcone synthase, 142, 156, 179, 184 Chlorophyceae, 260 chromoplast stroma, 488
chalcones, 151, 156, 158, 159, 173, 174, 177, chlorophyll, 8, 171, 177, 207, 314, 398, 399, chromoplasts, 177,488,492,499,504
179, 181, 188, 190 414, 486, 488, 496, 498, 499 cluomoproteins, 496
Chamaecyparis, 337 chlorophyll a, 496 chromosomal cleavage, 645
Chamaecytisus hirsutus, 559 chlorophyll synthesis, 230 chromosomes, 4, 5
chamigrane, 381 chlorophyUide a, 399 chrysanthemic acid, 348, 430, 431
chamise, 125, 128 Chlorophyta, 396, 419, 425, 495 Chrysanthemum, 348, 436
Chamorros, 226, 294 Chiorophytum, 280 Chrysanthemum coronarium, 49
channa, 623 chloroplast membranes, 17, 18 chrysanthemyl alcohol, 348
channel, see nicotine acetylcholine receptor chloroplasts, 8, 9, 14, 17-19, 22, 23, 26, 82, chrysanthemyl skeleton, 346
channel, sodium channels, voltage-dependent 94,99, 105, 130, 166,241,313,314,488, chrysazin, 88
sodium channels, channeled biosynthesis, 492,494-499,518,520,554,556,557, chrysmanthemic acid, 348
ion channels chloroquine, 606, 607, 614, 625, 649, 650, 666 trans-cbrysanthemic acid, 348
chanoclavine cyclase, 3 chloroquine phosphate, 606 Chrysolina, 92
chanoclavine I, 656, 658, 659 chloroquine-resistant, 606, 607, 625, 651 Chrysolina brunsvicensis, 92
chanoclavine-I cyclase, 656 chloroquine-sensitive, 606, 607 Chrysolina quatirigemina, 92
chaparral, 78, 125, 126, 128, 345 chlorosis, 237, 238, 423 Chrysomela, 124
chaparrinone, 482 chocolate, 346, 462 chrysomelid beetles, 14, 84, 88, 124, 128, 363,
Chauliognathus, 542 Choisya (ernata, 573 468,552
Chauliognathus iecontei, 49 cholera, 77 chrysomelid, elm leaf, 88
chaulmoogric acid, 29, 36 (25S)-a-cholestau-3,B,26-diol, 680 Chrysomelidae, 289, 447, 453, 454, 472
chavicine, 539 5,B-cholestan-3,24-mone, 437 Ciuysomelina, 124, 363
chavicol, 113 cholestane, 677 Chrysopa septempunctata, 362
chayote, 31,40 7-cholestenol,582 Chrysopa siossonae, 53
chebulagic acid, 196 cholest-7-en-3,8-01-5,6-epoxide,442 chrysophsnol, 85, 88
cheese, 517 cholest-4-en-3-one, 678 Chrysopidae, 362, 366
cheilanthatriol, 422 cholesterol, 312, 313,427,428,433,435,437, Chusquea, 101
Cheilanthes farinosa, 422 439, 440, 442, 458-460, 462, 468, 470, 582, chymotrypsin, 245
Cheiranthus, 468 678,686 Cker, 181, 186
Cheiranthus X allion;;, 468 cholesterol binding, 682 Cker arietinum, 181
chelerythrine, 601, 603 chollc acid, 208, 213 Cichorieae, 387
chelidonine, 601, 603 choline, 208, 300, 510 Cichorium intybus, 439
chelidonine sulfate, 601 choline phosphotransferase, 23 Cicindelinae, 289
Chelidonium majus, 557, 600, 601 chOlinergic blocking agents, 510 Cicuta virosa, 47
chelilutine, 601 cholinergic drugs, 510, 511, 623, 664, 697 cider, 210
chelirubrine, 601 cholinesterase, 510, 549, 623, 647, 664, 682 cigarettes, 462, 526
Chelone giabra, 361 cholinesterase inhibitors, 537, 664 Cimifuga, 269
chemical ecology, 189 213, 231, 232, 271, chondriol, 48 cimiphytine, 641
296, 510, 529, 565, 709 chondrocoIe A, 340 Cinchona, 85,628, 648-652
chemical messages, 12, 338 chondrodendrine, 606 Cinchona ledgeriana, 649
chemosystematic studies, viii, 324, 337, 338, Chondrodendron, 511 Cinchona succirubra, 649
352,367,387,398,413,473,477,486,494 Chondrodendron tomentosum, 607 cinchonamine, 649
chemotaxis, 367, 376, 391, 392, 444 Chonemorpha fragrans, 686 Cinchoneae, 612, 638
Chenopodiaceae, 3, 88, 179. 222, 390, 458, chonemorphine, 686 Cinchonideae. 638
557, 581, 708 chorismate mutase, 101, 102, 104, 106 cinchonidine, 649-651
Chenopodium, 390, 442 chorismate synthetase, 97 cinchonidinone, 649
Chenopodium quinoa, 460 chorismic acid, 65, 80, 93-98, 101, 102, 106, cinchonine, 649-651
Chenopodium rubrum, 3, 557 122 Cinchonoideae, 612, 637
Chenopodium strictum, 390 Choristoneura, 682 cinchophylline, 628, 652
cherries, 37, 277, 290 Chortoicetes, 228 cineole, 331, 340
cheny, black, 277, 288, 298 chromatography, 367 1,4-cineole, 345
 Illdex 723

1,8-cineole, 331, 342, 344~346 CNS, see central nervous system colorin, 559
cinnamaldehyde, 110 CoA esters, 23, 109 Colubrina lexensis, 695
cinnamic acid, 3, 106~11O, 118, 121, 122, 126, CoA ligases, 109 columbamine, 598-600
128, 130-132, 138, 139, 147, 148, 152, 155, coadaptation, vii, 10,84, 177,208.209,307, columbianetin, 134
156, 165, 193, 196,203,509,515,529,533, 441,447,466,511,525 cotyledons, 700
564, 620, 624 coal, 53 Colymbetinae, 439
E-cinnamic acid, 106~ 109, 130 coca, 535 Combretaceae, 88, 147, 195, 197,533,610
cinnamic acid o-hydroxylase, 130 cocaine, 511, 531, 534, 535, 537. 544, 641. comfrey, 553
Cinnamomum camphora, 585 676 Commelilla, 146
Cinnamomum zeylanicum, 419, 424 cocarcinogenic diterpenes, see diterpenes. Commelilla communis, 171
cinnamon, 110 cocarcinogenic commersonine, 682
cinnamoyl-CoA oxidoreductase, 109 Coccinellidae, 542 Commiphora rostrata, 30, 40
cinnamyl acetate, 110 coccinellines, 542 communication, interplant, 33
cinnamyl alcohol dehydrogenase, 109, 115 cocculidine. 609 community succession, 209
cinnamyl alcohol, 109, 113 cocculine, 609 compartmentation, 9
cinnzeylanine, 418, 424 cocculolidine, 609 complexation, 170, 177, 207~209, 213
cinnzeylanol, 418, 424 Cocculus, 385, 608, 609, 700 concanavalin A, 244
circular dichroism (CD), 230 Cocculus carolinus, 609 condensation, 19, 21. 56-58, 65, 94, 96, 97.
Cirsium japonicum, 49 Cocculus trilobus, 609 117,315,326,327,348,351,359,369,396,
Cistaceae, 197 coccus cacti, 89 398,405,428,430,431,432,444,465,473,
citral, 340~343, 346 cochineal, 89 488; see also aldol condensation, Claisen
(E)-citra!, 343 Cochlearia, 537 condensation, Claisen-type condensation,
citric acid cycle, 3 Cochliobolus, 423 imino-aldol type~condensation, Mannich
citrinin, 63, 70 cocklebur, 389 condensation
citronella!, 342, 344, 346 cockroaches, 340, 640 condensed tannins, 8, 12, 156, 164, 193, 194,
citronellol, 342~344, 346 cockroaches, American, 343, 383 198,200,204,206,207-209,212
(S)-( - )-citronellol, 358 coclaurine, 586 condensed tannins, acid butanol assay, 212
Citrullus vulgaris, 499 (R,S)-coclaurine, 595 conessine, 686
Citrus, 158, 175,337,340,370,476,482-484, (S)~coclaurine, 586 confertiflorin, 388, 390, 392
499,573 cocoa, 165 Congo Republic, 640
Citrus aurantium, 34 coconut, 346 y-coniceine. 543, 544
Citrus paradisii, 175 coconut oil, 25 8-coniceine, 561
citrus waste, 260 codaphniphylline, 689 coniferin, 115, 117
civet cat, 37 codeine, 511, 593~597 Coniferopsida, 337
civetone,37 Codium.260 conifers, 198,324,337,338,342,343,352,
Cladophora, 260 codonocarpine. 521 398,510
Cladosporium, 415 Codonocarpus australis, 521 coniferyl alcohol, 115, 117, 123
Claisen condensation, 57, 64, 148, 155 Coelospermum, 85 coniine, 8, 531, 537, 543, 561
Claisen-type condensation, 57, 142 coenzyme Q, 76, 104, 125 Conium, 542, 543
clams, 696 coevolution, vii, 10, 12~14, 137,511 Conium maculatum, 3, 8, 543, 557
clastogenic effects, 645 coevolved, 10 Connaraceae, 231
Clausena, 660, 663 cofactor, 156, 158, 159. 162, 179.207,276, Conopharyngia, 637, 647
Claviceps, 3. 26, 315, 655, 658, 659 371,488,491,508,535,587 Conospermum, 82
Claviceps paspa/i, 655, 660 Coffea, 702, 703 Conradina canescens, 346, 352, 451
Clavicipitaceae, 659 Coffea arabica, 700, 702 Consolida, 269
clavine alkaloids, 655 Coffea canephora, 700 contraceptives, 376, 462, 683
clay, 684 coffee, 104, 165,219,346,700,702,703 Convallaria majus, 222
Cleistopholis patens, 588 coffee bean, see bean, coffee convergence, 12, 54
Clematis, 267 Coit, 101 Convolvulaceae, 41. 72, 251, 377, 533, 537,
C!ematopsis, 267 cola, 165 549, 559, 565, 655, 658
clerodanes, 406, 417, 418 Cola, 700 Convolvulus, 658
neo-clerodane, 417 colchicine, 507, 511, 617~619, 676 convulsions, 591, 607, 642, 658
ent-clerodanes, 418 (7S)-( - )-colchicine, 617 cooking, 36, 137, 245
clerodendrin A, 417 Colchicum, 617 copabomanes, 375
clerodendrin B, 417 Colchicum autumnale, 617, 619 copabomeol, 386, 670
Clerodendron, 417, 418 Coleogyne ramosissima, 208 (+ )-a-copaene, 381
Clerodendron infortunatum, 417 Coleoptera, 179,297,352,393,447,453,454, COPalfera, 226, 232, 413
CJerodendron trichotomum, 418, 666 472,650 copalic acid, 420
clinal variation, 8, 338 coieoptile, 33, 146,280 copalyl pyrophosphate, 408, 414
Clitocybe, 697 Coleus barbatus, 420, 426 copalyl pyrophosphate synthetase, 408
clones, 8, 288 Coleus forskohlii, 420 copigmentation, 171, 177
Clostridium perjringens, 261 coliolic acid, 26 co~pigments, 152
clove, 59, 110 collagen, 200, 206. 220 copper ethylacetoacetate, 294
clover, 35, 179, 179, 296 collards, 306 Coprosma, 85
clover, red, 72, 179, 186, 561 Colletes, 37, 344 Coptis, 269, 598
club moss, 540, 708 Col/etia, 610 Coplis japonica, 598, 600
Clusiaceae, 69, 88, 91, 148, 149, 173, 187, colletotrichin,415 Cordia, 77
197,209,412 Colletotrichum, 415 Cordia alliodora, 336, 337, 452
Cneoraceae, 134, 473, 475~477, 485, 575, 576 Colletotrichum coffeanum, 186 Coreopsis, 45
Cneorum tricoccum, 477 Colletotrichumfalcatum, 145 Coreopsis lanceolata, 49
Cnestis glabra, 231, 232 Colletotrichum lindemuthianum, 148, 184 coriamyrtin, 670
cnicin, 389 Collosobruchus maculatus, 619 coriander, 26
Cnicus benedictus, 389 CoIocasia alltiquorum, 26, 40 Coriandrum sativum, 26
Cnidosco!us texanus, 517, 529 Colombia, 607, 708 Coriaria arborea, 384
Cnidoscolus urens, 517 coloration, warning, 542 Coriariaceae, 197,244,384,610
724 Index

coriariin A, 210 cowpeas, 392 cultures, callus, 47, 326, 413, 473, 535, 586,
corilagin, 194, 209 Coxsackie, 210 587,601,610,625,627,645,653
( - )-coronaridine, 635 crambe, 306, 307 cultures, hairy root, 47, 353, 526, 528, 534,
( + )-coronaridine, 635 Crambe abyssinica. 306, 307 708
cork, 53, 447 Crassulaceae, 282, 290, 539 cultures, suspension, 2, 3, 9, 13, 93, 128, 135,
com (Zea mays), 53, 65, 105, 173, 192,290, crayfish,44O 326,353,366,368,452,457,473,553,554,
435, 648; also see maize Creatonos, 552 557,587,594,598,601,610,614,616,631,
com borer, 417 Creatonotos transiens, 552 654
com borer, Asiatic, 480 creosote bush, 120 Cunoniaceae, 197
com borer, European, 696, 711 crepe myrtle, 694 Cupressaceae, 46, 179,338
Comaceae, 197, 359-361 crepenynic acid, 42. 43 Cupressus macnabiana, 338
Cornales, 360 Crepis, 45 Cupressus sargentii, 338, 351
Comanae, 360 o-cresol, 124 cupric ion (CU2 +), 209
Comiflorae, 337, 360 p-cresol, 124 Curacao, 210
comin,355 Crithidia fasciculata, 645, 660 curare, 511, 607, 628, 643, 645
coronaridine, 632 crocein, 499 curare-like effects, 675
coronary artel}' flow, 588 crocin,254 curares, calabash, 643
coronary dilation, 640 Crocus sativus, 499 curares, tube, 605, 607
coronatine, 238 crops, 53, 219, 227, 300, 306-308, 317, 336, curarizing agent, 643
Coronilla, 290 366, 378, 389, 457, 482 curarizing effects, 650
Coronilla varia, 291 Crotalaria, 548, 549, 551-553 curcin,244
coronopilin, 388, 389 Crotalaria retusa, 551 curine, 606
corpora allata, 318, 384 Crotalarieae, 550, 566 Curtisiaceae, 360 ,
cortex, 320, 439 crotepoxide, 104 cuscohygrine, 531, 532-534, 539, 677
cortexone, 440 Croton, 403, 405, 420, 591, 610 Cuscuta, 658
corticosteroids, 439 Croton macrostachys, 104 Cuscuta platyloba, 559
cortisone, 462 croton oil, 402, 403, 425 Cuscuta refiem, 559
Corydalis, 605 Croton salutaris, 595 Cuscutaceae, 559
cOl}'nantheal, 649 Croton tiglium, 402 cutch,204
CorynantM alkaloids, 630-632, 634, 640, 645, Crotonoideae, 286 cuticle, 53, 53
crown gall, 110, 128 cutin, 51, 53, 54, 111
649
corynantheal, 649 crown rust, 98, 105 cutworm, variegated, 316, 479
corynanthean, 612, 636, 638 crownvetch, 291, 295 Cyamopsis tetragonolobus, 260
Corynocarpaceae, 290, 610 Cruciferae, see Brassicaceae cyanide determinations, 294; see also Feigl-
Corynocarpus, 290 crustaceans, 260, 437, 440 Anger method, Guignard test. picrate
cost, see metabolic cost method, sodium picrate paper
crustacyanin, 498
cyanide oxygenase, 290
Costa Rica, 220, 295 cryogenine, 694
cyanide-resistant respiration, 6
Costelytra zealandica, 179, 291 cryptaustoline. 605
cyanidin, 152, 167, 170, 172, 177, 190, 191,
costunolide, 391 cryptic marking, 550
193,200, 210
COT, see 2-carboxy-4-oxotetralone cryptocapsin, 494
cyanidin 3-0-g111coside, 171
cotinine, 526 Cryptocarya, 563, 565
{J-cyanoa1auine, 226, 289, 290
Cotoneaster, 499 Cryptocarya bowie;, 605 3-cyanoalanine, 223, 226, 231
cotlon, 3D, 34-36, 39, 78, 171,228,342,378, Cryptocarya massoia, 269
3-cyano-L-alanine, 215
380, 391, 395 cryptolepine, 666 ,B-cyanoa1auine synthase, 232, 287, 289
cotton boll weevil, 342 Cryptolepis sanguinolenta, 666 cyanobacteria, 17, 495
Cotton effect, 230 cryptopine, 594 cyanoformic acid, 231
cottonseed, 256 cryptopleurine, 564 cyanogenesis, 273, 274, 280, 287-289
cottonseed meal, 380 cryptostyline alkaloids, 581 cyanogenic glycosides, vii, 5, 6, 8, 9, 14, 273,
cottonseed oil, 35, 36 Cryptostylis.581 274, 276, 277, 280, 282, 285-290, 294, 295,
cotyledoos, 52, 128, 164, 174, 190,220,231, cryptotanshinone, 413 297,298,300,302,303,310,517,528; see
244,261,276,288,308,447,634 cryptowolline, 605 also pseudocyanogenic glycosides
cotylenins, 415 Ctenuchidae,551 cyanogenic glycosides, meta-substituted, 280
p-coomarate, 107, 108, 152 cubebanes, 375 cyanogens, cyclopentenoid, 289
p-coumarate:CoA ligase. 155 cubenol,381 cyanoginosin-LR, 239, 245
4-coumarate:CoA ligase, 109, 133 cucumber, 53, 346, 444, 472 cyanohydrin, 273, 274, 276, 280, 282, 290,
p-coumarate-3-hydroxylase, 108 Cucumis sativus, 444 295-297, 299
o-coumaric acid, 122, 126, 130, 132 cucurbic acid, 33, 79 cyanolipids, vii. 6, 273, 285, 287, 289. 294,
p-coomaric acid, 54,102,106-109,118,121, Cucurbita. 452. 488 297, 298, 321
122, 131, 132, 142, 147, 155, 156, 193, 195, Cucurbita maxima, 444, 452 cyanophoric compounds, 282
269, 509, 518, 624 Cucurbita pepo, 33, 39, 370 Cyathea spinulosa, 173
coumarin, 11, 113, 130-138, 179, 190,262, Cocorbitaceae, 31, 33, 229-232, 411, 444, Cybister tripunctatus, 440
387, 478, 575, 576 445, 447, 454, 463, 472 Cycadaceae, 226
coumarins, 8-prenylated, 663 cucurbitacin B, 444 cycads, 173, 226, 290, 294
Z-o-coumarinic acid, 130 cucurbitacin C, 444 Cycas, 226, 294
p-coumaroyltryptamine, 518 cucurbitacin E, 444 Cycas circinalis, 226
Coumarouna, 130 cucurbitacin E glucoside, 445 cycasin, 294, 296
p-coumaryl-CoA, 156 cllcurbitacin F, 444, 445 cyclamates. 462
coumaryl glycosides, 9 cucurbitacin I, 445 cyclase ll, 331
coumestan glycoside, 179 cucurbitacins, 309,427,444,445,447, cyclic AMP, 700
coumestans, lSI, 179 452-455 cyclic AMP phosphodiesterase, 703
coumestrol, 179, 191 lOa-cucurbita-5,24-dien-3p-ol, 444, 452 cyclic GMP, 700
couplings, see NMR couplings, oxidative cucurtiitine, 229 cyclilols, 247, 264, 271
coupling, radical coupling cularine alkaloids, 563, 605, 606, 610 cycloalliin, 227
courtship, 383, 551, 552 Culex pipiens, 402 cyc1oartenol, 427, 432, 433, 435, 436, 444,
cows, 35 cultures, see also cell cultures 463, 678, 689
 Index 725

cycloartenol cyclase, 432 Dalechampia spathulata, 344 dehydrochloroprepacifenol.381

cyc10buxine D, 689 damage, 7, 8, 31, 32, 91, 136 4,21-dehydrocorynantheine aldehyde, 631
cyclodopa, 705 damascenine. 568 dehydrocrepenynic acid, 42, 43
cycloeucalenol, 435, 436 ,B-damascenone, 500 dehydrodiconiferyl alcohol ,B-D-glycosides,
cycloeucalenol:obtusifoliol isomerase, 436 dambonitol, 265 llO
cyclohexene oxides, 104 danaidal, 551 dehydmgenation, !l7, !l8, 158, 508, 553
cyc1ohexylcarboxylic acid, CoA derivative, 26 danaidone, 551 7,8-dehydroiridotrial glucosides, 358
cyclooxygenase, 167 Danainae, 550, 551 dehydmiridodiol, 362
cyclopamine, 685 Da""us, 383, 466, 551 dehydromatricaria ester, 43, 49
cyclopentenoid rings, 273, 274, 285, 287 Danaus chrysippus, 552, 564 dehydropyrrolizidine alkaloids, 550
"cyclopism," 685 Danaus gilippus, 551 3-debydroquinate dehydratase, 97
(2-cyclopentenyl)g!ycine, 274, 276, 277, 285 Danaus plexippus, 383, 466, 471, 472, 551, 3-dehydroquinate synthase, 97
cyc1opentenylcarboxylic acid, 28 7ll 3-dehydroquinic acid, 96, 97
cyclopsine, 685 daphnanea, 402, 403 debYdrosanguinarine, 601
cyc1osativene, 381 Daphne genkwa, 404 dehydrosecodine, 634
cyclosporin A, 235 Daphne mezereum, 404 debydroshikimic acid. 95, 193
cyclovirobuxine, 689 Daphne odora, 419 3-dehydmshikimate, 97
D-cymarose, 250, 251, 465 daphnetoxin, 403 3-dehydroshikimate reductase, 97
p-<:ymene, 345, 346 Daphniphyllaceae, 361, 689 12,13-dehydrosparteine, 559
cymopol, 340 daphniphylline, 689 dehydrotestosterone, 440
Cymopolia barbata, 340 Daphniphyllum, 668, 689, 691 dehydrotrewiasine, 696, 711
Cymopterus watsoni, 136 Daphniphy/lum macropodum, 689 deidaclin, 285, 286
Cymmchum. 563 Datiscaceae. 244 Delia antiqua, 227
Cynarese, 387 Datura, 269, 532, 537, 545 Delia brassicae, 309
Cynometra, 693 Datura !erox, 534, 535 delirium, 537, 658
Cyperaceae, 337, 659 Datura innoxia, 534 delirium tremens, 537
Cyperus, 660 Datura stramonium, 534. 535, 537. 573 Delonix regia, 222
Cyperus iria, 384 Datureae, 537 delpbinidin, 152, 167, 170, 177
Cypholaphus, 693 Daucus carata, 3, 26, 48, 72, 73, 128, 136, delpbinidin 3-glucoside, 171
Cyphomandra, 677 328,557 Delphinium, 269, 673, 675
Cyphomandra betacea, 534 Davallia, 280 demethylation, also see oxidative
cysteine, 45, 217, 226, 232, 241, 290, 321, 513 Davallia trichomanoides, 287 demethylation
L-cysteine, 234, 303 Davidiaceae, 361 N-demethylation. 595
cysteine sulfoxides. S-substituted 226 DCCC, see chromatography, droplet demethylcoclaurine, 591
cysteine synthase, 220 countercurrent 3-demethylfamesyl pyrophosphate, 430
cystine, 215. 226 de novo origin, 9, 18 demethylpapaverine, 588
Cystoseiraceae, 210 de novo synthesis, 21 demethylsuberosin, 133, 137
cytisine, 556, 558, 559 dealkylation, 437 demethyltrewiasine. 696, 711
Cytisus praecox, 559 delmyol, 377 demethylvestitol, 184
Cytisus scoparius, 559 decanoic acid. 25 demissidine, 682
cytochalasin, 72 2-decanone, 30 demissine, 682-684
cytochalasin B, 72 decanortriterpenes, 476 denbrobine, 385, 375, 670
cytochrome bs, 22 decarboxylation, 45, 52, 53, 58, 77, 81, 87, 98, Denbrobium, 385
cytochrome oxidase, 288 142,222,223,226,229,253,315,360,435, Dendrobates, 560
cytochrome c oxidase, 645 507,508,513,514,517,531,537,580; also Dendrabiurn, 385, 560, 668, 670, 693
cytochrome P-450 dependent monooxygenases, see oxidative decarboxylation Dendrobium nobile, 670
133, 158, 159, 179, 182, 202, 276, 296, 303, I-decene-l,IO-dicarboxylic acid, 32 Dendroctonus, 37, 340, 342, 343
317,335,355,441,600; see also Decussocarpus gracilior, 173 Dendroctonus brevicomis, 343
monooxygenases de-epoxidation, 497 Dendroctonus frontalis, 343
cytokinins, llO, 700 deet,340 Dendrodoris limbata, 380
cytomegalovirus, 561 deer, white-tailed, 207, 390 dendmlssin, 382
cytoplasm, 8, 19, 107, ll5, 152, 153,261,313, defense, 6, 7, 8, 12-14, 30, 39, 50, 78, 79, dendroprimine, 560
314,318,319,483,488 124, 128, 129, 134, 137, 179, 183, 193, Dendrosius, 382
cytosol, 18, 19, 26, 106, 319, 368, 414, 430, 207-209,213,231,232,261,269,270,273, Dennettia tripetala, 290. 297
629 288-290, 296, 310 deodorization, 35
cytotoxic activity, 59, 67, 69, 74, 120. 149, defense, plant, 6, 7, 213, 288 I-deoxocucurbitacin, 444
209,267,316,338,380,389,393,415,420, defensive compounds. vii, 6, 7, 8, 33,41.79. 2-deoxy sugars, 250
444, 445, 452, 455, 468, 471, 472, 479, 481, 122, 124, 125, 209, 231, 269, 273, 288, 290, 6-deoxy-D-allose, 250
482,484,510,591,601,606,640,648,652, 291,293,310,322,339-343,353,362,367, 6-deoxy-L-altrose, 250
663, 666, 671, 676, 696, 700, 705 372,376,380,382,439,442,447,455,458; 3-deoxyanthocyanins, 164
see digestibility reducing substances deoxybouvardin, 700
Dactylis glomeratus, 659 defensive compounds, qUalitative, 7, 209 deoxychalcone. 158
Dacus dorsalis, 111, 128 defensive compounds, quantitative, 7, 209 deoxyeritadenine, 700
Daemonorops, 569 defensive secretion, 49, 500 6-deoxy-D-galactose, 250
daffodils, 430, 452, 510, 620, 622 defensive substances, digestibility-reducing, 6-deoxy-L-galactose, 251
daffodils, King Alfred, 620 207 6-deoxy-D-glucose, 251
daga, 336 deficiency, see fatty acid deficiency, human 6-deoxy-D-gulose, 250
Dahlia, 200 immunodeficiency virus (HN) deoxyhaningtonine. 625, 626
DAHP, see 3-deoxy-D-arabino-heptulosonic Degenariaceae, 610 3-deoxY-D-arabino-beptulosonic acid
acid 7-phospbate dehydration, 20, 44, 97, 315 7-phosphate (DAHP), 97
daidzein, 178, 181, 184, 191 dehydroabietic acid, 415, 420 2-deoxybexoses, 247, 250
Dalbergia, 78, 187 20,2I-dehydmajmalicine, 631 6-deoxybexoses, 247, 250
Dalbergia refusa, 78 dehydroaporphines, 588 deoxyloganic acid, 354, 355
dalbergiones, 187 dehydroascorbic acid, 265 epi-deoxyloganic acid, 354
dalbergiquinols, 187 dehydrochelerythrine, 601 8-epi-deoxyloganic acid, 358
726 Index

epi-deoxyloganin, 353, 358 diabrotid beetles, 447 digitonin, 468, 470

6-deoxy-L-mannose, 250, 251 1,4-diacetoxy-l.3-diene, 380,419 D-digitoxose, 250, 251, 465
6-deoxy-3-0-methyl-D-galactose, 251 Diacl'isia virginica, 390 diglycerides, 287, 437
6-deoxy-4-0-methyl-D-glucose, 251 diacylglycerol, 19,23.24,40 diglycerides, monogalactosyl, 17
6-deoxy-3-0-methyl-L-mannose, 250, 251 diacylglycerol acyltransferase, 24 digoxin, 468
6-deoxy-3-0-methyl-L-talose, 250, 251 Diadema, 340 dihydroactinidiolide, 362
( - )-deoxynupharidine, 670 Diadema antillarum, 420 dihydrobenzophenantbridine oxidase, 600, 616
2-deoxy-D-erythro-pentose, 250 Diadromus pulchellus, 227 dihydrobenzophenanthridines, 600, 60 I
4-deoxyphorbol, 402 diamides, 518 dihydrobiochanin A, 186
12-deoxyphorbol, 403 diamine oxidases, 509, 510, 554, 599 dihydrochalcone, 156, 174-176
deoxypodophyllotoxin, 120 diamines, 509, 510, 513, 515, 517, 518, 531, dihydrocinnamic acid, 109, 125, 147,451
5-deoxyproanthocyanidins, 201, 206 540 dihydro-m-coumaric acid, 147
2-deoxy-D-ribose, 250 2,4-diaminobutyric acid, 223, 226 dihydro-p-coumaric acid, 624
6-deoxy-L-talose, 251 a, y-diaminobutyric acid, 234 24,25-dihydro-9a,11a-epoxyparkeol, 444
dephosphorylation, 23 L-2,4-diaminobutyric acid, 226 dihydroflavonoI4-reductase, 162, 163,200
L-a, y-diaminobutyric acid, 226 dihydroflavonols, 151, 156, 158, 159, 162,
depolymerization, 676
2,6-diaminopimelic acid, 217 163,175,188,190,201
depressants, central nervous system, 539
2,3-rliaminopropionic acid. 223, 226 dihydrofuroquinolone alkaloids, 573
Depressaria pastinacella, 136, 138
a,,B-diaminopropionic acid, 226 dihydrokaempferol, 159, 162, 163
depression, 514, 517, 607, 643
dianthalexin, 568 (2R,3R)-dibydrokaempferol, 162
depression, bone marrow, 235
dianthrone, 85, 88 dihydrokawain, 140
depression, smooth muscle, 603
Dianthus, 568 8-Z-dihydromatricaria acid, 49
depside,70 Dianthus caryophyllus, 156
dermatitis, 336, 389, 394, 403, 424 dihydromethysticin, 140
diaphoresis, 528 dihydromyricetin, 163
Dermestes maculata, 35
Dermocype austroverwta, 91
6,6'-diapocarotenoic acid. methyl ester. 499 ( + )-dihydromyricetin, 200
diarrhea, 452, 599, 612, 697 dihydromyricetin reductase, 202
Derris, 186, 253 Diarthron vesiculosum, 404, 425
Derris elliptica, 543 dihydronepetalactone, 358. 362
dibenzopyrrocoline, 605 dihydroparthenolide, 390, 392
desacetoxyvindoline, 634, 652 dibenzopyrrocoline alkaloids, 605 dihydrophenanthrenes, 139, 140, 146, 147, 149
desacetylconfertiflorin, 388, 390, 392 S-(1,2-dicarboxyethyl)cysteine, 227 9,10-dihydrophenanthrenes, 147
desacetylipecoside, 611 Dicentra, 605 3,4-dibydroxypyridine, 220
desacetylisoipecoside, 611 Dichapetalaceae, 29 dihydroquercetin, 159, 163, 175
desacetylvindoline, 634 Dichapetalum, 29, 30 (+ )-dihydroquercetin, 175, 200
desaturase, 3, 21, 22, 26, 34, 43 Dichapetalum toxicarium, 29 (2R,3R)-dihydroquercetin, 159. 162
desaturation, 3, 21-23, 34, 35; also see fatty 1-(3 ,5-dichloro-2,6~dihydrox y-4-
dihydroquercetin 3-acetate, 175, 176
acid desaturation methoxyphenyl)-l-hexanone, 71
dihydroquercetin reductase, 200
desert, 52, 120, 125, 128 2,5-dichlorophenol, 125
dihydroquercetin 3-0-rhamnoside, 175
Des/onfainia, 444 Dichroa, 569
dibydrosanguinarine, 600, 601, 616
Desfontainiaceae, 444 dicotyledonous plants. 19,69,88, 108, 115,
16,17-dihydrosecodin-17-ol, 635
desmethoxyyangonin, 140 164, 172, 189, 194,242,286,324,458,510,
rlihydrostilbenes, 140
desoxymannojirimycin, 253 5l7,562
dihydroxyacetone phosphate (DHAP), 19
desoxynorjirimycin (ON]), 253, 254 dicoumarol, 132, 137
3,4-dihydroxybenza1dehyde, 620, 621, 622
desoxyvasicine, 570 Dicranum. 173
2,4-dihydroxy-I,4(2H)-benzoxazin-3-one
desoxyvasicinone, 570 dictamnine, 571, 573
Dictylomatoideae, 574
(DIBOA), 98, 99
desulfobenzylglucosinolate, 303 2,4~dibydroxy-1 ,4(2H)-benzoxazin-3 -one
detergent, 208, 428, 430, 432, 459, 490 dictyol-E, 420
(DIBOA) glucoside, 99, 101
deterrency, 603 dictyopterene A, 32
dictyopterene B, 32 (3R,3'S,5'R)-3.3'-dihydroxy-,8,K-caroten-6'-
deterrent, 173, 179,288,291,295,309,316, one, 494
339,340,350,361,380,382,390,395, Dictyopteris deticulata, 32. 39
Dictyostelium, 71, 74 16,8,26-dihydroxy-5a-cholestanol, 680
415-418,441, 451v 452, 468, 472, 479,511, 20,22-dihydroxycholesterol, 439
523,528,537,538,540,543,550,558,559, Dictyotales, 420
1,2-didehydrosparteinium ion, 556 3,4-dihydroxycinnamic acid, 118
562,582,588,591,597,600,603,613,618, ent-Ia,10j3-dihydroxy-9a,ISa-cyclo-20-
640,641,650,658,660,664,682,686 2,6-dideoxy-D-ribo-hexose, 250, 251
2,6-dideoxy-D-_l}'lo-hexose, 250, 251 norgibberell-16-ene-7,9-dioic acid 10,19-
detoxication, 5, 10, 136, 138, 186, 209, 288, lactone, 414
2,6-dideoxy-3-0-methyl-O-lyxo-hexose, 250,
290 7,2'-dihYdroxy-3,4' ~dimethoxyisoflavan, 184
251
detritus, 207 2,7-dihydroxy-3,4-dimethoxyphenanthrene, 148
2,6-dideoxy-3-0-methyl-D-ribo-hexose, 250,
Deutzia crenata, 355 2,7-dihydroxy-4,8-dimethoxyphenanthrene, 148
251
deutzioside, 355 2,3-dihydrodipicolinic acid, 217
2,6-dideoxy-3-0-methyl-D-xylo-hexose, 250,
development. 1,3,6,7,9, 10, 12, 14,40,46, (22S,25R,26R)-3/3,26-dihydroxy-22,26-epoxy-
251
71,78,92,98,111,122,135,171,196,207, 2,6-dideoxy-3-0-methyl-L-arabino-hexose, 251 6-oxo-Sa-cholest-7-ene, 463
234,235,238,239,259,275,287,288,297, Didieriaceae, 708 6a,7-dihydroxymaackiain, 186
298 Diels Alder reactions, 573 6a,7-dihydroxymedicarpin, 186
dextrins, 259 (3S,5R,6S,3'S,5'R,6'S)-5,6,5',6'-diepoxy- 2,4-dibydroxy-7-methoxy-l,4(2H)-benzoaxazin-
DHAP. see dihydroxyacetone phosphate 5,6,5',6'-tetrahydro-,6,j3-carotene-3,3'-diol, 3-one (DIMBOA), 94, 98, 99, 101, 104, 568
DHNA, see 1,4-dihydroxy-2-naphthoic acid 486 2,4-dihydroxy-7-methoxy-l,4(2H)-benzoaxazin-
dhurrin, 5, 276, 280, 282, 287, 288, 290, 296, diet, 16, 17,34,35,92,99,113,136,167, 176, 3-one (DIMBOA) glucoside, 99
299 191,193,207,208,212,217,220,223,228, 3,3'-dihydroxy-5-methoxybibenzyl, 148
(S)-dhurrin, 277, 282 245,254,261,289,291,306-308,310 7,4'-dihYdroxy-2'-methoxyisoflavan, 184
diabetes, 174 digalactosyl diglycerides, 17 2-(3,4-dihydroxy-S-methoxyphenyl)ethyIamine,
dhurrin j3-g1ucosidase, 282 1,6-digalloylglucose, 196 592,593
Diabrotica barberi, 113 D-diginose, 250, 251, 465 2R,5R-dihydroxymethyl-3R,4R-
Diabrotica undecimpunctata IlOwardii, 113, digitalis glycosides, 676 dihydroxypyrrolidine (DMDP), 253, 254
128 Digitalis, 85, 465, 468 erythro-9,10-dihydroxy-l-octadecanyl acetate,
Diabrotica virgifera virgifera, 113. 128 Digitalis lanata, 468, 680 35
Diabroticina, 447 D-digitalose, 251, 465 1,4-dihydroxy-2-naphthoate, 87
 Index 727

1,4-dibydroxy-2-naphthoic acid (DHNA), 81, Diospyros, 85 Z-13-docosenoic acid, 35

82,86,87 Diospyros diepenhorstii, 209, 212 2-doclecanone. 30
l,4-dihYdroxy-2-naphthoic acid synthetase, 81 dioxygenases, 158, 159, 186, 202 dogs, 609
12,13-dihydroxyoleic acid, 26 dipeptides. 657 Dolichoderus scabridus. 362
lO,16-dihydroxypalmitic acid, 53 diphosphomevalonate decarboxylase, 315 dolichodial, 358
3,4-dihydroxyphenethylamine (dopamine), 523, Diphysa robinioides, 145 dolicholactone. 358
579, 580, 628 Diploclisia, 442 dolichoprenols, 321
3,4-dihydroxY-P.phenethyl-O-P.D- Dip/odea pinea, 340 Dolicholhele, 693
glucopyranosyl-(I->3)-4-0-caffeoyl-P.D- Dip/adus, 420 Dolichothele sphaerica, 692
glucopyranoside, 108 Diplodus holbrooki, 420 dolichotheline, 692
3',4'-diliydroxyphenols.208 diploids, 4, 337 dopa, 587, 591, 705, 706
3,4-dihydroxyphenylacetaldehyde, 2, 3, 584, dipropyl sulfinate, 227 L-dopa (dihydroxyphenylalanine), 229
587 Dipsacaceae, 360, 361 dopa decarboxylyase, 515
3,4-dihydroxyphenylacetonitrile, 277 Dipsacales, 360, 361 dopamine, 2, 229, 510, 523, 535, 578-582,
3,4-dihydroxy-L-phenylalanine (L-dopa), 229, Dipterocarpaceae, 197,412 585-587. 591, 595, 610, 640
587 Dipteryx odorala, 130 dopamine receptors, 591
3,4-dihydroxyphenylpyruvic acid, 3, 587 disaccharide esters, 108 dopaminergic, 510
5,7-dihydroxyproanthocyanidins, 201 disaccharides, 247, 253, 254. 266, 270, 287 donnancy, bud, 500
5,8-dihydroxypsoralen, 133 Discaria, 610 dormancy, seed, 500, 501
2,7-dihydroxy-3,4,6-trimethoxy-9,10- disease, see Alzheimer's, blackrot, black spot, Dorslenia, 134
dihydrophenanthrene, 148 Chagas', Dutch elm, Hodgkin's, Lou Draba cuneifolium, 305
2,3-dihydroxy4,7,8-trimethoxyphenantltrene, Gehrig's, Parkinson's, rust, veno-occlusive. Draba platycarpa, 305
148 and yellow rice diseases Dragendorff's reagent (alkaloids), 547, 560,
2,7-dihydroxy-3,4,6-trimethoxyphenanthrene, Dislium, 228 648
148 dispersal, vii, 9, 35, 40, 149, 151, 152. 171 drimane, 379
3,4-dihydroxytyrarnine, 578, 579 dispersal, seed, vii Drosera, 63, 80
3',4'-dihydroxyusambarensine, 652 dissemination, 30 Droseraceae, 63, 80, 82, 197,297
la,25-dihydroxy-vitamin D, glycoside, 437 distasteful properties, 550 , 683 Drosophila, 441, 453, 454, 549. 582, 583, 615
3,4-di-O-isobutyryl-6-0-caprylsucrose, 269 distillation, 324 Drosophila melanogasler, 441
Dilleniaceae, 197 diterpene acids, 342 Drosophila mettleri, 441, 582
DIMBOA, see 2,4-dihydroxy-7-methoxy- diterpene alkaloids, 408, 626, 668, 672, 673, Drosophila mojavensis, 441. 582, 583
1,4(2H)-benzoaxazin-3-one 689,690 Drosophila nigrospiracula, 441, 582
dimeric aporpbines, 588 diterpene 19-carboxylic derivatives, 673 Drosophila pachea, 441, 582
2,6-dimethoxybenzoquinone, 78, 79 diterpenes, 77, 312, 337, 339, 341, 369, 370, drugs, see adrenergiC drugs, anticholinergic
2,6-dimethoxy-p-benzoquinone, 78 380,381,398,399,401-408,411-416. drugs, cholinergic drugs, drugs of abuse,
3,5-dimethoxy-4-hydroxycinnamic acid, 300 418-420,423-427,431,488,500,626 parasympatholytic drugs,
3,5-dimethoxy4-hydroxycinnamoylcholine, seco-diterpenes, 413 parasympathomimetic drugs, soporific drugs
302 diterpenes, acyclic, 399 drugs of abuse, 12,511,535,593
1O,l1-dimethoxystrychnine, 641 diterpenoids, B-secokaurene, 420 drununondin A, 69
di-O-methyl tubocurarine, 607 diterpenes, bicyclic, 404, 405 drummondin B, 69
dimethylallyl pyrophosphate (DMAPP), 312, diterpene 19-carboxylic acids, N- drmmuondin C, 69
315,316,326-329,335,348,369,370,430, a1kylaminoethyl esters, 674 DrUsenameisen. 703; also see ants
431,488,656 diterpenes, cocarcinogenic, 244, 402, 403, 413. Dryopleris, 68
dimethylallyl transferase, 3, 133,488 420,424 Drypetes, 304
4-dimethylallyltryptophan, 656 diterpenes, 5a,lOJ3-configuration, 398, 404, Duboisia, 528, 535, 537
4-()', ~imethylallyltryptophan), 656 405,412 Duboisia leichhardlii, 532, 534
1,4-dimethylazulene, 368 diterpenes, 5,B,lOa-configuration, 398, duckein, 528
N' ,N"-dimethylhistamine, 692 404-407,411,412 duckweed, 145
ID-di-O-methyl-chiro-inositol, Z64 diterpenes, grayanoid, 411, 412 Dufour's glands, 37, 563
1,3-di-O-methyl-myo-inositol, 265 diterpenes, macrocyclic, 414 dulse, 260
I-D-l,4-di-O-methyl-myo-inositol, 265 diterpenes, monocyclic, 401, 402 dung, 14,71
E-4,8-dimethyl-l,3,7-nonatriene, 341 diterpenes. tetracyclic, 401, 407 dunnione, 87
N,N-dimethyltryptamine, 511, 514 diterpenes, tricyclic, 398, 405-407 dust, 535, 552
1,3-dimethylxanthine,7oo diterpenoid precursors, macrocyclic, 676 Dutch eJm disease, 72
3,7-dimethylxanthine, 700 diuretic, 136, 392, 702 Dutch Indies, 649
a-dimorphecolic acid, 26 diurnal rhythm, 7, 510, 543, 553 duvatrienediol, 401, 414
dinoflagellates, 703 diversity. 3, viii, 6, 7, 10, 13 4.8,13-duvatrien-l,3-diol,414
dinosaurs, 511 divi-divi, 195 a-4,8,13-duvatrien-I,3-diol,414
Dioclea megacarpa, 220 djenkolic acid, 227 Dyera.652
2,3-lrans-3,4-trans-diol (2R,3S,4R)-( +)- Djibouti, 210 dyestuff, 85, 89, 267
3,4,5,7,3',4'-hexahydroxyflavan, 200 DMAT synthetase, see tryptophan Dysdercus, 689
Dioncophyllaceae, 698, 708 dimethylallyltransferase synthetase Dysdercus koenigii, 111
Dioncophyllum, 698 DNA,3-5, 14.26, 135, 137, 156, 237, 246, dysentery, 564, 570, 612, 640, 662, 686, 698
Dioseorea, 33,457,462,510,513,528 314,393,529,549,553,568,573,623,648. dysentery, amebic, 564, 612
Dioscorea balalas, 147 653,660 Dysoxylum, 607
Djoscorea composita, 241 eDNA, 156,280.295,314.629 dyspepsia, 364
Dioscorea delloidea, 457 T-DNA,123 Dysphania, 708
Dioscorea rotundata, 148 DNA intercalation, 573, 599, 601, 645, 651 Dytiscinae, 439
Dioscoreaceae, 147, 148,457,462,510,528, DNA polymerase, 521, 601, 623
537 DNA recombinant methods, 3, 10 earwonn, com, 99, 173, 192
Dioscoreophyllum cumminsii, 245 DNA, synthesis, 520, 613, 645, 648, 676, 696 earwonn, European com, 99
dioscorine, 528 DNA-dependent RNA polymerase, 645 Ebenaceae, 82, 85, 197
diosgenin, 457, 458, 462 ONJ, see desoxynojirimycin ebumamine-vincamine alkaloids, 640, 653
diosindigo A, 82, 85 dock, 76, 91 ebuman, 636
Diosmeae, 575, 576 l-docosanol, 53 Ecballium. 230
728 Index

Ecballium e/aterium, 437 Encelia ventorum, 316 epiJucumin 4"-p-(,B-D-glucopyranosyloxy)-(E)-

ecdysis, 419, 479, 484, endophytic fungi, 655, 658, 659 cinnamate, 277
ecdysone, 440-442, 454 endoplasmic reticulum, 18,21,23,26,51,313, epilucumin 4"-p-(,B-primeverosyloxy)-(E)-
a-ecdysone, 444 318,326, 368, 526, 528, 598 cinnamate, 277
,B-ecdysone, 440 endopolygalacturonase, 261 epilucumin, 279
ecdysteroid, 427, 440, 442, 452, 453, 455 a-l,4-D-endopo!ygalacturonic acid lyase, 262 epimastigotes, 660
echinacein, 36 endosperm, 22-24, 26, 39, 287, 288, 399 22,26-epiminocholestane type alkaloids, 677,
Echinocereus blanckii, 692 energy storage, 16, 17,32,591,497 681,683
Eclipta alba, 179 England, 190,210,363 a-epiminocyclohemiacetals, 677, 681
ecology, 3, 10, 12, \3,41, 128, 135, 177, 189, enkephalins, 597 epinephrine, 511, 522
311,510,544,550; see also chemical enrnein, 408, 419, 420 Epipactis palustris, 147
ecology enniatins, 238 epipassicoriacin, 285
Ecuador, 708 5-enolpyruvylshlkimate, 97 epiphytic, 659
edema, 378. 460 5-enolpyruvylshikimate-3-phosphate (EPSP), epiproacacipetalin, 282
edible oils, 16 97 (R)-epiproacacipetalin, 282
eggplant, 677 5-enolpyruvylshlkimate-3-phosphate synthase epirubijervine, 684
eggs, 37, 88, 124, 128, 179,220,291,340, (EPSP synthase), 97 epitetraphyllin B, 285, 296, 298
342,363,393,437,444,447,498,551,696 enoyl-ACP reductase, 20 epithelial cells, 324
Ehrlich's reagent, 547 Entamoeba histolytica, 392, 564, 612, 640, 652 epivolkenin, 285, 286, 296
eicosatetra-5,8,1l,14-enoic acid, 34 (S)-epilotaustralin, 282 epoxidation,431
Ekebergia, 663 (S)-epilucumin, 277, 279 epoxide, 282
Elaeocarpaceae. 444, 537, 543, 546, 563 Enterobacter aerogenes, 97, 105 epoxydon, 72
elaeocarpine, 563 enzyme-linked immunosorbent assay (ELISA), (8R,9R,10S)-9,10-epoxyheptadec-16-ene-4,6-
Elaeocarpus, 444. 543, 563, 565 1 diyne-8-ol. 49
Elaeocarpus dolichostylus, 444, 453 enzyme, nitrogenase, 290 24-ethoxyhopane, 431
elaidate,35 enzyme systems, cell free, 326, 328, 329, 331, Z-12,13-epoxyoleic acid, 27
elaiosome, 35, 40 373,407,488,587,592,629,645,655,700 Z-9,lO-epoxystearate,27
Eleagnaceae, 244, 660 enzymes, 1-10, 13, 19,21-24,42,58,78,94, (3S,5R,6R,3'S,5'R,6'Sj-5',6'-epoxy-5,6,5',6'-
eleagnine, 660 95,97,98,101,102,107-109,115,117, tetrahydro-,B,,B-carotene-3,5,3'-triol,486
Eleagnus, 660 120, 132, 135, 138, 153, 155, 156, 158, 159, 14,15-epoxyprieurianin, 481
Eleagnus angustifolia, 660 162-165,173,178,179,181,182,184,187, EPSP, see 5-enolpyruvylskikimate
electroantennogram, 31, 113 188, 190, 195, 196,200,203,207-209,212, EPSP synthase, see 5-enolpyruvylskikimate
electron carriers, 22, 76 220,221,227,230,231,241,252,257, synthase
electron microscopy, 318 260-262, 266, 269, 270, 273, 275, 276, 280, Equisetaceae, 521
electron transport chain, 166 287,288,290,294-296,300,302,303,311, Equisetum paiustre, 521
electrophoresis, 217, 708 313-315,319,321,322,326,327,329, equol, 179
electrophoretic separation, 587 331-333,335,348,351,369-373,375,371, Erantis, 269
elemanes, 374 389,396,401,408,414,424,425,428,430, eremanthine, 391
elemicin,111 432,436,444,447,452,453,455,459,471, Eremanthus, 390
Eleocharis microcarpa, 32 473,488,490,492,497,503,504,508-510, eremophilanes, 371, 374
a-eleostearic acid, 35 513,516,520,528,531,532,535,556,561, ergine, 655
elephantin, 393 586,587,594,598,599,600,614,616, ergocornine, 658, 660
Eleutherococcus senticosus, 462, 470 629-631, 648, 654, 656-658; see also ergocryptine, 658, 660
elicitors, 33, 39,133,134,138,142,178,179, "berberine·bridge" enzyme, enzyme-linked ergolines, 656
184, 186,261,262,270,271,372,377, 402, immunosorbent assay, rubber transferase ergometrine, 658, 660
414,600,601,6\3 enzyme ergonovine, 658, 659
elimination reaction, 107,327,329,349,370, enzymes, anabolic, 4 ergosterol, 442
431,621,656 enzymes, branching, 259 ergol, 26, 316, 568, 575, 655, 658-660, 666
Z-elimination, 22, 97 enzymes, catabolic,S, 7 ergot alkaloids, 511, 655-660, 666-668, 692
ELISA, see enzyme linked immunosorbent enzymes, debranching, 259 ergot alkaloids, peptide containing, 657
assay enzymes, extracellular, 209 ergotamine, 511, 656-658, 660
elk,207 enzymes, hydrolytic, 194, 319 ergotism, 655, 658
ellagic acid, 194,208,210 enzymes, peroxidase, 117 Ericaceae, 71, 78, 82, 88, 122, 195, 197, 361,
ellagitannins, 193-197,206,209,210,212, Epacridaceae, 361 408,411,413,554
213 Ephedra sinica, 522 Ericales, 361
ellipticine, 628, 645 Ephedraceae, 522 Ericanae, 360, 361
elm, 237 ephedrine, 511, 513, 522, 530 Eriobotrya japonica, 148
elongation, 3, 21, 23, 51, 52 epicatechin, 165,201 eriodictyol, 158, 162-164, 167, 175
elymoclavine, 656, 657, 658 ( - )-epicatechin, 202-204, 206 (2Sj-eriodictyol, 162, 164
Elymus, 101 ( + )-epicatechin, 172 erucic acid, 35
embelin,77 ( - )-(2R ,3R)-epicalechin, 201 Ervatamia, 637, 647
embryo, 288, 361, 411 (+ H2S,3S)-epicatechin, 201 Ervatamia heyneana, 648
emetic, 361, 466, 468, 479, 601 ent-epicatechin, 201 Erwinia carotovora, 262
emeline, 361, 511, 564, 579, 610, 612, 613, Epichloe, 659 erychroside, 468
638,652 epidermal cells, 9, 51, 52, 137, 155, 156, 166, erysimoside, 468
emodin, 64, 65, 85, 87, 91 191, 193, 282, 299, 510, 557 Erysimum, 468
emodin anthrone, 91 epievodone, 346,451 Erysimum cheiranthoides, 309,468
emodin 1-0-methyltransferase, 85 epigallocatechin, 172, 203 Erysiphe graminis, 288, 558
Empetraceae, 125, 175,451 ( - )-epigallocatechin, 172 erysodiene, 608
emulsin, 280. 287 ( + )-epigallocatechin, 172 erysodienone, 608
emulsin, almond, 561 epiguaipyridine, 672 (- )-(5S)-erysodienone, 608
enamine, 507, 553 epiheterodendrin, 282, 284, 288 erysopine, 608
encecaiin, 316, 317, 322 (R)-epiheterodendrin, 282 erythraline, 608
Encelia, 316, 322 Epilachna borealis, 445 Erythrina, 290, 294, 299, 560, 598, 608-610,
Encefia farinosa, 125 Epilachna varivestis, 288, 297 614,627
 Index 729

Erythrina alkaloids, 608. 609 Eumaeus ataia, 294 fagopyrin, 91

Erythrina christa-galli, 608 Euodia, 570, 573 Fagopyrum, 76
Erythrina flabelliformis, 609 Euonymus, 452 Fagopyrum esculentum, 162, 190,543
meso-erythritol, 262 Euonymus europaeus, 269, 270, 671 Fagus grandifolia, 117
erythrocytes, 459 eupahyssopin, 392 falcarindiol, 50, 113, 136
a-erythroidine, 608 Eupatorieae, 387, 510, 549, 550 falcarinol,48
,B-erythroidine, 608 eupatoriochromene, 317 falcarinone, 45
erythromycin, 65 Eupatorium jhanii, 500 familial Mediterranean fever, 619
erythromycin A, 65, 67 Eupatorium rugosum, 317, 321 a-farnesene, 381
Erythronium, 269 euphol, 447, 473 (E)-,B-farnesene, 383
Erythrophleum, 412, 676 apo-euphol, 473, 475, 480 (E,E)-a-famesene, 383
Erythrophleum alkaloids, 668, 676 LJ1-euphol, 473, 476 (Z,E)-a-famesene, 383
Erythrophleum chlorostachys, 676 Euphorbia, 197,319,337,344,385,398,402, farnesol, 344, 370, 380-383, 396
Erythrophleum guineense, 676 403 E-farnesol, 370, 501
erythrose, 249 Euphorbia iagascae, 26, 27 (E,E)-farnesol, 381, 382
erythrose-4-phosphate, 94, 96, 97 Euphorbia lathyrus, 229 Z,E-farnesol, 370
Erythroxylaceae, 412, 533-535, 537 Euphorbia tirucalli, 402 2-E-6-E-farnesol, 370
Erythroxy!um, 412, 532, 533, 534 Euphorbiaceae, 26, 35, 45, 46, 50, 69, 88, 173, 2-Z-6-E-farnesol, 370
Erythroxylum coca, 532, 533, 535 197,229,240,244,245,253,269,276,282, famesyl acetate, 381
Escalloniaceae, 359, 361, 673 286,288,299,304,319,321,337,344,385, farnesyl diphosphate, see famesyl
Escherichia coli, 81,96,97, 101. 104, 106, 398,401-407,412-414,420,423,424,426, pyrophosphate
156,245,517,629 444,466,510,513,517,537,542,549,554, farnesyl diphosphate synthase, 319
Eschscholtzia, 605 575,581,592,595,610,639,640,692-697, farnesyl octanoate, 381
Eschscholtzia californica, 33, 290, 296, 600, 700, 703, 709 farnesyl pyrophosphate (FPP), 315, 319, 321,
601,616 Euphoria longan, 222 322,326,327,335,351,367,369,370-373,
Eschscholtzia tenuifolia, 2. 587, 616 euphoric effect, 597 394,396,428,430,431,455,488
( - )-eschscholtzine, 605 Euphydryas, 361, 363, 366 2-cis-6-trans-farnesyl pyrophosphate, 370
esculetin, 130, 132 Euphydryas anicia, 363 E,E-farnesyl pyrophosphate, 321, 367, 369,
essence d'iris, 452 Euphydryas chalcedona, 363 370,375,377,387
essence, 324 Euphydryas cynthia, 363 E,E-farnesyl pyrophosphate synthetase, 369
essential oil, 30-32, 42, 45, 47, 52, 59, 109, Euphydryas phaeton, 361 famesyl pyrophosphate synthetase. 326, 327.
lll, 122, 130,324, 326, 332, 337-344, 346, Eupomatia, 591 351,368-370,396,399,488
349,351,367,368,370,381,384,388,398, Eupomatia alkaloids, 591, 616 2-E-6-E-famesyl pyrophosphate, 370, 374, 375
452,499,516,568,575 Eupomatiaceae, 591, 609 2-trans~6-trans-farnesyl pyrophosphate, 370
estradiol, 439, 440 Euproctis subfiava, 417 2-Z-6-E-famesyl pyrophosphate, 367, 370
17,B-estradiol, 439 Eurema, 623 famesyl transferase, 414, 488
estragole, 113, 128 Eurema hecabe mandarino, 265, 623 fatigue, 647, 702
estrogen, 179 Europe, 37, 83, 91, 92, 138, 169, 184,210, Fatoua, 134
estrogenic effect, 70, 439 233,235,237,244,246,297,340,349,351, fatty acid biosynthesis, 19,21.40.56
estrone, 436, 439, 440 363,364,383,393,417,426,452,471,537, fatty acid deficiency, 35
ethanol, 206, 217 563,610,641,645,658,669,673 fatty acid, desaturation, 54
ether, 535 Eurycoma longifofia, 662 fatty acid elongation, 4
Ethiopia, 459 eusiderin, 121 fatty acid metabolism, 339, 343
ethylamine, 513 Euxylophora, 569, 570, 573 fatty acid synthase, 56, 156
ethylene, 3, 6, 73, 231-233, 238, 289, 297, Evodia, see Euodia fatty acids, vii, 3, 16-19,21-29,31,32.
520 evodiamine, 570 34-43,45,51-53,55,56,58,67,68,109,
ethylene glycol, 262 evolution, vii, 1, 5-7. 9-14, 32, 160. 135, 190, 113,223,324,339,340,343,383,402,441,
ethylmalonate, 57 246,304 499,515,582; see also fluorofatty acids,
5-ethyl-2-methoxyphenol. 124 evoninoate alkaloids, 671, 690 fatty acid deficiency
etiolated, 275, 277, 282 Excoecaria, 403 anteiso-fatty acids, 25
etioline, 680, 681 excretion, 5, 7, 70,165,208.245,288,413 E-fatty acids, 35
eucalyptol, 346 expectorant, 460, 601 iso-fatty acids, 25
Eucalyptus, 69, 140, 204 expression, 3, 9, 14, 33 trans-fatty acids, 35
eucalyptus, 194,212 exudate, 30, 40, 125 fatty acids, acetylenic, 35, 42, 43
Eucalyptus camaidulensis, 345 eye-spot, 495 fatty acids, conjugated unsaturated, 18, 26
Eucalyptus citriodora, 345 fatty acids, cycIopentenoid, 26-29, 36
Eucalyptus robusta, 69 F.b.ce.e, 29, 31, 46, 79, 85, 86, 88, 91, HI, fatty acids, essential, 35
Eucalyptus sideroxylon, 142 123, 130, 134, 140, 142, 145, 149, 158, 159, fatty acids. hydroxy, 16.24,26,27,35,51, 53
Euclea, 85 168,173-175,178,179,183-188,190,197, fatty acids, 3-acetoxy, 35
Eucommiaceae, 321. 361 206,215,218-223,226-229,231,244,246, fatty acids, cycIopropanoid, 16,27,35
Eucommiales, 360 253,254,257,260,261,267,280,282,286, fatty acids, cyclopropenoid, 16, 17.27,35
eudesmane, 372. 374. 377 287,290,298,405,412,413,456,458,460, fatty acids, dihydroxy, 26, 27
eudesmanolides, 387, 388, 392 462,463,466,510,514,515,517,521,528, fatty acids, epoxy, 16, 27
(+ )-eudesma-4(14)-7(ll)-dien-8-one, 392 538,543,549,552-554,557-559,561,562, fatty acids, long chain, 51-53
eugeniin, 197 564,566,569,570,581,598,608,610,660, fatty acids, odd carbon numbers, 24
eugenol. 110, 113, 344 663, 664, 676, 693, 700, 705, 706, 708, 709 fatty acids, primary, 17, 18,25
eugenone, 58, 59 Fabiana imbricata, 671 fatty acids, saturated, 16, 41
Euglena, 495 fabianine, 671 fatty acids, unsaturated, 16, 18,21,22,24,34,
Euglenophyceae, 495 FAD, 432, 508 35,36,40,41,402
Eug/ossa, 344 Fagaceae, 195, 197,252,264 fatty acids, ..:19-unsaturated C 18• 4
euglossine bees, 344, 352 Fagara, 569, 570, 573-575, 610 fatty acids, unusual, 16, 25, 26
eukaryotic organisms, 4, 8, 22, 23, 234, 244, y-fagarine, 573 fatty acyl-CoA reductase, 51
433,623 fagaronine, 601, 615 fatty alcohols, 51
eukaryotic pathway, 16, 18, 19,23 Fagaropsis, 575, 577, 610, 667 fatty aldehydes, 51
Euiaema, 344 Fagaropsis angolensis, 576 favin,244
730 Index

favism, 700 flavone synthase n, 158 forb,207

feathers, 486, 498 flavone synthases, 158 formamide, 290
febrifuge, 569, 673 flavones,4, 151, 152, 156, 158-160, 162, fonnic acid, 99, 294, 343, 570
febrifugine, 569 164-166,171,174,177,178,189,191 Formica, 343
fecal pellets, 460 flavonoid biosynthesis, 151, 153, 173, 186 fonnononetin, 178, 179, 181, 186
Fedia cornucopiae, 353 flavonoid sulfates, 159 2~formylantbraquinones, 88
feeding activity, 35 flavonoids, 4, 5, 8, 9, 11, 14, 56, 102, 108, 4-formyl-8-hydroxyquinoline, 571
feeding stimulants, 35, 168, 174, 265, 308, 136, 139, 142, 149, 151-153, 155, 156, N-formylloline, 659
310,350,363,418 158-160, 162, 164-169, 172, 173, 175, 177, forskolin, 420, 426
Feigl Anger method, 273, 286, 290, 294 179,184,187-191,193,194,198,200,210, Forsythia intermedia, 117, 129
Felidae, 362 212, 213, 261, 387, 420, 498, 499, 533, 558, Forsythia suspensa, 117
( - )~endo~fenchol cyclase, 332 575 founder effect, 389
fenchone, 345 flavonoids, A-ring, 151, 179 Fouquieraceae, 197, 360, 361, 365, 458
fennel,332 f1avonoids, B-ring, 151, 152, 174, 178, 202 fragrances, 12
fenugreek, 528 flavonoids, C-ring, 151 frsgrances, floral, 326, 352, 709
Ferdinandusa, 611 flavonol glycosides, 4, lSI, 167, 168 France, 210, 658
fermented products, 700, 703 flavonol sulfate, 159 Frankia, 83
fern allies, 164,442 f1avonols,4, 151-153, 156, 158-160, frsss, 136, 343, 361, 552
femane, 431 164-167, 175, 177, 189, 190 free radical, 580
ferns, 9, 40, 68, 164, 172-174, 194,253,254, flavoprotein, 280, 431 French Congo, 640
279,286,318,324,376,3%,414,422,424, flavoring agents, viii, 12, 109, 324, 346, 349, friedelin,447
431,442,462,497,510,533,543 506,700 Frilillaria, 677, 684, 686
fern, bracken, 376, 396, 414, 442, 454 flavoxanthin, 499 frogs, 466, 560, 564, 664, 666, 677
ferric ions (Fe+ + +),209,543 flavylium cation, 171 frogs, dendrobatid, 560, 708
ferrous ion (Fe++), 158, 159, 162, 166. 202, flax, 274, 276, 282, 288, 290 frontalin, 37, 343
266,598 flea beetle, 168,308,445 fructans, 247, 257
ferruginol,413 flea beetle, sweet potato, 7 {3-D-fructofuranosyl O-a-D-galactopyranosyl-
ferulate, 152 fleas, 641 (1-6)-a-D-glucopyranoside, 256
ferulic acid, 54, 107, 108, 113, 115, 118, 122, flexilin, 380, 419 fructosans, 257
126,131,132,517 flies, 177, 289, 344, 441, 515, 582, 641, 700; fructose, 247-250, 253, 256
feruloylputrescine, 517 also see blowfly, fruit fly, houseflies, sawfly, D-fructose, 247, 248, 250, 256
Festuca arundinacea, 659 Spanish fly; see fly below D-fructose 6-phosphate, 248
fever, 619, 662 Flindersia /ournieri, 662 fructose-l,6-bis(phosphate),252
fiber, 336 flindersine, 571, 573 fructose-I,6-bis(phosphat)ase, 520
Ficus, 134, 563 Flindersioideae, 571, 573, 575, 576 frui~ 7-9, 30, 31, 35,47,59,77,94, 110, 121,
Ficus carica, 137 Florida, 191, 245, 270, 294, 346, 349, 350, 127, 128, 152, 171, 175, 177, 194,203,206,
fig, 137 352, 365, 394, 3%, 423, 453, 504 210,220,223,229,231,237,238,244,256,
filamentous fungi, 517 flour, 30, 41 265,269,272,277,279,280,285,287-290,
Filipendula ulmaria, 209 Flourensia dentala, 317 297,305,320,324,337,346,362,381,397,
filixic acid, 69 Flourensia ilicifolia, 317 402, 403, 420, 445, 457,459, 460, 463, 472,
fire, 126, 127, 345 flowers, 7, 8, 26, 30, 111, 136, 152, 153, 156, 473, 476, 480, 482, 483, 486, 490, 492, 494,
rife ant, 35, 383, 397, 533, 542, 543 159, 162-164, 170, 171, 173-177, 188-190, 498,499,504,517,535,537,558,567,570,
fir, Douglas, 202, 257, 338 207,231,232,256,269,299,306,320,324, 611, 659, 666, 689, 695, 700; see also dried
fir, balsam, 338, 383 328,335,344,348,362,381,382,385,386, fruit beetle, fruit flies, grapefruit, miracle
fir, alpine, 338 392,396,411,412,439,457,459,471,472, fruit, oriental fruit moth, passion fruit, pome
firs,338 486,494,498,499,511,515,516,552,558, fruits, tomato fruit worm
Fischer projections, 246 608,669,700; see also bee flowers, fruit conserves, 708
fisetin,166 cauliflower, sunflower moth, flowering fruit flies, 582
fisetinidol-4a-ol, 206 plants, safflower, sun flower, wall flower fruit fly, Caribbean, 340
fish, 12, 32, 39, 79, 85, 340, 348, 380, 381, flowers. beetle-pollinated, 177 fruit fly, European cherry, 37
390, 393, 419, 420, 439, 456, 460, 486, 498 flowers, bird-pollinated, 176, 177 fruit fly, Mediterranean, 31, 340, 381
fish poisons, 12, 460 flowers, hummingbird, 177 fruit fly, oriental, 111
fitness, increase, 6 fluoroacetic acid, 29 fruit juices, 165
fixative, 670 fluorofatty acids, 29 fruits, pome, 381
Flacourtiaceae, 28, 282, 285, 287, 296, 298, tv-fluorofatty acids, 29 Frullania, 389
418,522 tv-fluorooctadec-Z-9-enoic acid, 29 fucose, 247, 250, 260
flagellate, 645 lrJ-fluorooleic acid. 29 D-fucose, 249, 250, 465
flatus, 261 lrJ-fluoropalmitic acids, 29 L-fucose, 247, 250
flavan-3,4-diol biosynthesis, 200 fluridone, 501 a-fucosidase, 561
flavan-3,4-diols, 162, 163, 193, 200-204, 206 fly, see flies above a-L-fucosidase, 562
flavan-3-ols, 159, 193,200-204,206,210,212 fly, cabbage root, 309 fucosidases, 544
2,3-cis-flavan-3-ols, 203 fly, carrot rust, 113 fucosterol, 444
2S-flavan-3-ols, 202 fly, onion, 227 fucoxanthin, 495
flavan-4-01s, 200 foam, 316, 456, 468 Fucus, 495
flavanone, 151, 156, 158, 164, 173, 175, 179, Foeniculum vulgare, 332 fuels, 12
188,201 foliage, 682 Fumaria capreolata, 9, 588
flavanone 3-hydroxylase, 59, 162 folic acid. 97 Fumaria parvijlora, 241
(2S)-flavanone 3-hydroxylase, 162 folicanthine, 665 Fumariaceae, 303, 581, 598, 600, 601, 603,
flavanonols, 156 Folin-Denis assay (phenols, tannins), 210 605,610
flavin mononucleotide, 303 food additives, see additives, food fumonisin, 70
flavone C-glycosides, 160, 162 foods, 319, 321, 346, 349, 376, 377, 393, 394, fumonisin B I , 70
flavone glycosides, 4, 151-153, 167, 168, 172, 447,456,460,466,499,517,564,663,689, fungal endopolygalacturonase, 402
173, 178, 188 692,708 fungal metabolites, 3, 8, 35, 64, 65, 70-72, 84,
flavone sulfate, 159 forage, 179,219,227,274,290,437,456,460, 85,99, 133-135, 138, 142, 148, 183, 184,
flavone synthase I, 158 470 186, 193,210,238,245,253,254,260-262,
 Index 731

270,273,288,319,339,342,343,367,368, galanthine, 620 geophagy, 684

372,374,376,378,397,402,408,411,414, Galbulimina, 542, 543 Geraniaceae, 68. 195, 197, 337
415,420,447,486,501,502 Galega, 569, 570 geraniaI, 341-344, 346
fungal mycelia, 561 Galerina, 237 geranicardic acid, 68
fungal parasites, 659 Galeruca tanace/i, 88, 92 geranii herba, 209
fungi, vii. viii, 2, 3, 8, 9, 12, 17,26.32,42, Galerucinae, 447 geraniin, 197,208-210
45,47-49,53,56,63-65,70-74,76-78, Galium, 85, 86 geraniol, 326-328, 333, 340, 342-344, 346,
80,84-87,91,97, 104, 106, 107, 117, 121, Galium molJugo, 87, 241 349, 353, 354, 383, 396, 611
122, 125, 134, 135, 137, 145, 148, 164, 167, gallic acid, 193-196, 198,208,209,212,213 geranium, 68, 74
173, 174, 183, 184, 186, 188,217,218,234, gallic acid glycosides, 123 Geranium, 195, 197, 208
235,237,240,242,249,252-254,261,262, ( - )-gallocatecbin, 172 Geranium maculatum, 209
269, 273, 286, 288, 290, 295, 306, 309, 312, ( + )-gallocatecbin, 202 Geranium pyrenaicURl. 195
315,318,319,324,329,342,343,367,368, gal1otannins, 193, 194, 196, 210, 212, 213 geranyl diphosphate, see geranyl
374,376-378,380,390,395-397,408,411, galloyl D-glucose, 207, 210 pyrophosphate
414, 415, 427, 433, 442, 444, 459, 486, 488, O-galloylation, 206 geranyl phosphate, 328
495,497,501,502,514,517,520,543,558, galls, 194,514; see also crown gall, crown-gall geranyl pyrophosphate (GPP), 82, 315, 324,
559, 561, 562, 566, 573, 574, 591, 594, 601, tumors 326-333, 335, 336, 342, 368-370, 431, 488,
616,655,658-660,666,682,684,697,705, gambier, 204 669
706; see also endophytic fungi, filamentous gametes, 376, 396, 444 geranyl pyrophosphate synthetase, 326, 399,
fungi, fungi imperfecti. mycorrhizal gametophytic compounds, 368 401,488
fungal, rust fungi ganglionic blocking agents, 511 geranyl transferase. 488
fungi imperfecti. 235 gangrenous condition, 658 geranylcitronellol. 419. 423
fungi, rust, 110 Gardenia jasmino/des, 353, 358 geranylfamesyl pyrophosphate, 422
fungicidal compounds, 35, 145,377,459 Gardenieae, 612, 638 gemnylgeranial, 419
fungistatic compounds, 175 gardenoside, 358 gemnylgeraniol, 399,420, 750
fungitoxic compounds, 173, 175, 179, 184, 186 Gardneria, 638 geranylgemnyl pyrophosphate (OOPP), 370,
fungus, black-rot, 377 garlic, 226 399,401,402,407,414,415,425,486,488
fungus, blue stain, 342 Garrya, 673 geranylgeranyl pyrophosphate-geranyl
fungus, brown rot, 183 Garryaceae, 321, 360, 361, 408, 668, 673 transferase, 488
fimgus, red rot. 145 garryine, 673 gemnylgemnyl pyrophosphate synthase, 399,
fungus, snow mold, 273 gas chromatography, see chromatography, gas 401, 402, 488
fungus, take-all, 459 Gascardia madagascariensis, 422 gemnylhydroquinone, 336, 337
Funtumia. 677. 689 gascardic acid, 422 geranyUinalyl pyrophosphate, 399, 405, 412
furanocoumarins, 7, 113, 130, 132-135, 137, gastroenteritis, 460, 561 Gerbera jameson;;, 287
138,316,484,571,575 gastrointestinal tract, 237 gennacrane A epoxide, 374
furanocoumarins. angular, 132-134, 136 gastrointestinal tract, tone and peristalsis, 664 gennacranes. 374
furanocoumarins, linear, 133, 134-137 Gastrolina depressa, 84 gennacranolide lactones, 386
furanoses, 247. 248 Gastrolobium, 29, 660 gennacranolides, 386-388
furoquinoline alkaloids, 568, 570, 571, 573, gaudichaudioside, 420 gennacrene A, 372, 383, 387
575, 576, 610 Gaultheria procumbens, 122 gennacrene D. 383
(25R)-5a-furostan-3p,26-diol, 678 gas chromatography/mass spectrometry (GC/ Gennany, 168, 553, 641
fusaric acid. 237. 543 MS), 30, 38, 40 gennination, vii, 6, 24, 31, 32, 54, 78, 83, 84,
Fusarium, 70, 74, 186,237,238,269,378 GDP-glucose, 259 92,93,99, 105, 109, lIO, 118, 125-128,
Fusarium amygdali. 415 gedunin, 476, 477 132, 135, 167, 175, 183,220,231,240,265,
Fusarium javanicum, 186 Geigera. 390, 573 287,288,309,311,317,340,344-346,350,
Fusarium moniliforme. 70, 414 geissoschizine, 2, 632, 634, 635, 642 376, 378, 379, 389-392, 394, 396, 414, 415,
Fusarium oxysporum, 237. 543 gel,26O 435,459
Fusarium sambucinum. 135 gelsemicine, 646 gennination, seed, 317, 390, 395, 415
Fusarium solani, 145, 186. 442 Gelsemieae, 638 Gesneriaeeae, 87, 88, 164, 173, 174, 361
Fusicladium effusum, 84 gelsemine. 646 Giardia intestinalis, 652
fusicoccin, 415, 750 Gelsemium, 640, 646 Gibberella fujikuroi, 408
Gelsemium eiegans. 646 Gibberella pulicaris, 135
GABA. see ,),-aminobutyric acid, Gelsemium sempervirens, 646 gibberellic acid, 3, 408, 411
Gabon, 647 generalist feeder, 10, 92, 124, 135, 136, 361 gibberellins, 6, 10, 132, 361, 378, 398,407,
Gabunia, 637 genes, 3, 4, 8, 16,41, 153, 189-191,239,245 408, 411, 414, 423, 425, 426
Gagea, 269 genes. ras, 321 ginger, 114
Gaillardia pulchella, 672 Genipa, 700 gingerol, 114
D-galactan, 259 geniposide, 361 Ginkgo biloba, 173,200,202,413,420
galactans, 260 geniposidic acid, 360, 361 Ginkgoaceae, 68
meso-galactitol, 262, 264 Genista canariensis, 560 ginkgolides, 413, 420
galactoglycerols, 30 Genisteae. 549, 557 ginseng, 48, 460
O-a-D-galactopyranosyl-(l-6)-O-a-D- genistein, 167, 178, 179, 184, 191 girinimbin,663
galactopymnosyl-(1-6)-O-a-D- genistein,7-0-glycoside, 167 Gisekia, 708
glucopyranosyl-(1-2)-/3-D-fructofuranoside, ginseng, Siberian, 462 glabrine, 231
254,256 gentian, 359-361, 364 glands, see adrenal, anal, Dufour's, mammary,
4-0-/3-D-galactopyranosyl-D-glucopyranose, Gentiana, 670 mandibular. Nasonov, oil, scent and thyroid
254 Gentiana lutea, 364 glands, glandular swelling, and glandular
galactose, 172, 244, 248, 249, 260, 456 Gentianaceae, 149, 359, 361, 364, 366 trichomes
D-galactose, 248, 259, 260, 456 Gentianales, 88, 360 glandular swelling, 662
a-galactosidase, 561 Gentiananae. 360 glaucarubalone, 482
/3-galactosidase, 561, 562 gentianine, 669, 670 glaucine, 588, 591
galactosides, 266 gentiobiose, 254, 266 glaucolide-A, 390
galacturonic acid, 248, 260, 261 gentiopicrin, 364 glaucoma, 603, 664, 692
D-galacturonic acid, 248, 260, 456 gentiopicroside, 359, 699 Gleditisia triacanthos, 215, 226
galanthamine, 622, 623 gentisic acid, 82, 106. 113 Globulariaceae, 358, 361
732 Index

Glochidion, 693 7-0-glucosyltransferase, 132 Giycyrrhiza echinata. 182

glomerin, 569 N[15(,8-glucosyl}oxy-8-hydroxypalmitoyl]- Glycyrrhiza giabra, 184, 460, 462, 463
GIomeris margilUlta, 569 taurine, 37 glycyrrhizae radix, 460
Glomus etunicatum, 167 p-glucosyloxymandelonitrile, 280 glycyrrhizic acid, 460, 462, 463
Glomus macrocarpum, 167 p-glucosyloxymandelonitrile, 4'-caffeoyl ester, glycyrrhizin. 460. 462, 463
Gloriosa, 617 280 glyoxylic acid. 222. 273. 274. 579
D-glucans, 254, 260 glucotropaeolin, 303, 304 glyoxysomes, 24
glucobarbarin, 303 D-glucuronic acid. 248. 252. 253. 260. 264. Glyphosate. 97. 195
glucobrassicin, 306, 309 456 Glyptopetaium, 452
glucocapparin, 308 ,B-glucuronidase. 294. 561- Gnetales, 173
,B-glucogallin, 197 glutamate receptors, 698 gnetins, 140
,B-O-glucogallin, 196 glutamic acid. 80. 215. 217. 218. 220, 226. Gnetum, 140, 149
glucoiberin, 308 697 Gnidia giaucus, 404
gloconasturtiin, 303, 304 y-glutamic acid-(cysteine)n-g1ycine, 241 Gnidia iatifolia, 404
D--d-gluconolactone, 629 glutamine. 98. 220. 226, 228. 253 gnidiglaucin, 404
6-D-glucopyranosyl esters, 30 ,B-glutamylaminopropionitriIe. 226 gnidilatidin, 404
3'·O-,8-D-glucopyranoside of 4-(3',4'- y-glutamylcysteine,241 Gnophaela ialipennis, 552, 565
dihydroxyphenyl)butan-2-one, 110 )"glutamyl-cy.teine-glycine ()"glu-<:ys-gly). goat•• 390, 411. 676. 705
[4'-O-(,8-D-glucopyranosyl-3'-O-(,B-D- 241 goitrigenic derivatives, 300, 307
galuctopyranosyl)-,B-D-glucopyranosyl]olean- )"glutamyl-3-cyuooalunine.231 goitrin,307
12-ene-28-oic acid, 460 ()"glutamylcysteine).-,B-alunine, 241 goldenseal, 603
3-0-,B-D-glucopyranosyl-D-glucopyranose, 254 y-glutamyl-S-methyl-L-cysteine.241 Golgi bodies, 338
4-0-a-D-glucopyranosyl-D-glucopyranose, 254 4-glutamyIsemialdehyde. 226 gouada, 111
4-0-,8-D-glucopyranosyl-D-glucopyranose, 254 glutathione. 234. 240. 241 goniothalamin, 139
6-0-{J-D-glucopyruoosyl-D-glucopyranose, 254 glutathione reductase, 240 Goniothalamus giganteus, 67, 139
a.D-glucopyranosyl-a-D-glucopyranoside, 254 glyceollin. 182. 184. 261 goniothalenone, 139
O-,B-D-glucupyranosyl-(l-6)-,B-glucose, 266 glyceollin I. 182. 184 Goodeniaceae, 257, 361
4-,B-glucopyrano.yloxy-3- glyceollin II. 182. 184 Goodeniales, 360, 361
hydroxymethylbutyronitril-2-ene, 285 glyceollinm. 182. 184 gorlic acid. 29
4'-O-,B-D-glucopyranoside of 4-(4'- glyceraldehyde. 247. 249 Gossypium. 13.35.77.78.369,378
hydroxyphenyl)butan-2-one, 110 glyceridea. vii. 16. 25. 26. 30. 32. 35. 38-40 Gossypium hirsulum, 35, 369
O-<>-D-glucopyranosyl-(l-3)-,B-D- glycerokinase, 23 gossypol. 8. 376. 378. 380
fructofuranosyl a.D-glucopyranoside, 257 glycerol. 16. 18, 19. 23. 35, 40 (- )-gossypol. 376
3,B-D-glucopyruoosyloxy-4-methyl-2-(5H)- glycerol glucosides, 267 gout. 619, 673
furanone, 285 glycerol-3-phosphate. 23 goyazensolide, 391
D-glucosamine, 248, 250 sn-glyceroI3-phosphate. 21. 23. 24 grain, see cereal grain
glucose, 3, 13. 19. 52. 132. 162. 172. 174. glycerophospholipids, 17 gramicidin S. 234, 236
194-196. 249. 253, 256, 257, 260. 264. 269, glycine. 273. 591 gramiue. 513. 514. 523. 525
290. 291, 328,456, 588 Glycine max. 158. 182. 184. 221. 238. 241. Gramineae, see Poaceae
D-glucose. 207, 210. 247. 248. 256, 265. 267. 261.293 grandifoliolenone.473
456.465 glycine neurochemical activity, 646, 670 grape. 142. 145. 171
glucose mimic, 254 glycine receptor, 642 grapefrui4 175. 340
a-glucosidase, 561 glycinol, 182 grass. 30. 78. 101. 107. 115. 179. 230. 238.
a-glucosidase inhibitors, 254, 561, 562 glycoalkaloida, 678. 680. 682, 690 284. 290, 294. 346, 352, 391, 514, 564, 659.
,8-glucosidase inhibitors 207, 254 glycogeo, 254. 259. 270 666,667; see also bluegrass, ryegrass
a.glucosidases, 253, 254 glycolipids. 16. 17, 30 grass. "drunk." 659
{J-glucosidases. 78. 85. 115, 117. 131. 132. glycolysis. 3. 248 grass, orchard. 659
138.207.254.280.287-290.294-296.298. glycolytic pathway, 19 gras•• pereunial rye. 179.659
300, 561. 562. 629. 630. 682 glycoproteins. 72. 206. 237. 244. 259. 561. 562 gras•• rye. 376
D-glocose 6-phosphate. 252 ,B-glycosidase, 273, 280, 286-289, 294, 296 grass, "sleepy," 659
7-0-glucoside-3-0-rutinoside,8 glycosidase inhibitors, 535 grasshopper. 124, 125.317.384.468.522
glucosides, 266; see also betalain glucoside, glycosidases, 561 grasshopper, migratory. 316, 479
cucurbitacin E glucoside, cyanidin 3-0- glycoside sulfates, 458 grasshopper. striped. 559
glucoside, 7,8- glycosides. 151-153. 158-160. 162. 166. 167, grasshopper, two-striped, 682
dehydroiridotrial glucoside, delpbinidin 171-174. 177, 178. 189. 190. 192, 201. 244. grassluod. 125
3-glucoside.2.4-dihydroxy-1,4(2H)- 247.249.250.252-254.256.266.267.269. Gratiola, 444
benzoxazin-3-one (DmOA) glucoside, 2.4- 273. 274. 280. 282, 285, 286, 288-290. 296, Gratiola officinalis, 444, 445
dihydroxy-7-methoxy-I.4(2H)-benzoaxazin- 300.312.441; see also uothroquinone. gravid females, 309, 468
3-one (DIMBOA) glucoside, 7-0-glucoside- cardiac, chalcone, coumaryl, cyanogenic, grayuoin. 277
3-0-rutinoside, glycerol dehydrodiconiferyl alcohol, digitalis. flavone grayanotoxin I. 675
glucosides, o-hydroxybenzyl-,8-D-glucoside, C~, flavone, flavonol, gallic acid, iridoid, grayuootoxins. 398. 408. 411-413
3-hydroxyindole ,8-D-glucoside, hypolaetin kaempferol,lactone-fonning, monoterpene, grazing. 8. 126. 179. 296
8-0-glucoside, iridodial glucoside, iridotrial nitrile, phenylethyl alcohol, phenolic, Greeks. 658
glucoside, isoxazolinone glucosides, prinasin phenylpropanoid, pseudocyanogenic, green leaf volatile complex, 31, 309
2'-glucoside, quercetin 3-g1ucoside, quercetin, secoiridoid, sesquiterpene, steroid, greeobugs. 363, 365. 562
secoiridoid glucoside, .13-isoxazolin-5-one stilbene, strophanthidin, and triterpene greenbugs. biotype B. 113
glucoside glycosides Griffonia simplicifolia, 228
glucosinolates. 6. 8. 209. 266. 287, 290. 295. C-glycosidea. 152. 160. 162. 192 Griselinaceae, 361
300.302-308.310.311,468 glycosidic alkaloids, 249 griseofulvin, 64
D-glucosone, 265 Glycosmis, 568, 574, 663 griseophenone B. 64
6-0-glucosylaucubin, 363 Glycosmis arborea, 568 Qros.ulariaceae. 359
6-C-glucosylchrysin, 162 C-glycosyl flavones, 151, 162, 172, 173 groundsel. 550
8-C-glucosylchrysin. 162 glycosyl transferase, 321 growth inhibiting compounds, 33, 118, 510
,B-glucosyltransferases, 276 glyco.ylation, 151. 160. 162. 170, 190.206. growth regulators. 164. 367.437.445.486.
C-glucosyltransferase, 162, 190 276 500. 501, 510
 Index 733

guaiacol, 124 hardwood, 115 hemagglutinins, 234, 240, 242, 244

guaianes, 374 hares, snowshoe, 208, 451 hemiacetals, 247, 630
a-guaiene, 373, 376 harmalan. 660 hemicellulose, 259, 262
guaianolides, 387, 394 harmaline, 660 hemigossypol, 378
Guam, 226, 294 harmalol, 660 hemiparasite, 78, 364, 552, 559
L-guanidinohomoarginine, 226 harman, 568, 570, 655, 660 Hemiptera, 289, 658
guangdougen. 168 harman alkaloids, 8, 568, 576, 655, 660 hemiterpene alkaloids, 692
guanosine diphosphate-D-glucose, 259 harmine, 10, 570, 576, 660 hemiterpenes, 312, 315~318, 335, 655, 692
guar gum, 260 harmoI,660 hemoglobin, 234, 242, 246
guarana, 700 Harmonia axyridis, 363 hemolymph, 460
Guarea, 477 harringtonine, 625, 626, 627 hemolysis, 459
Guarea guidona, 481 Harrisonia, 475, 477 hemolytic, 458, 459
guayule. 109, 127,312,319,320 Harrisonia abyssinica, 481 Hemsleya carnosiflora, 444
Guettarda heterosepala, 649 harrisonin, 481 Hepatica, 267
guettardine, 649 hashish, 336 Hepaticae, 412
Guettardoideae, 612, 637 hasubanan alkaloids, 592, 615 hepatitis. 460
Guignard test (HCN), 273, 294, 296 hasubanonine, 592, 613 hepatotoxic compounds, 168, 209, 227, 232,
guinea pig, 543, 564, 600 haustorium, 78, 92,452, 659 378, 552, 553
gulose, 249 Haworth perspective fonnulas, 246 heptaketide, 64
gum, 260; also see carob gum, chewing gum, hay, 40, 132, 137 heptanal, 113
guar gum, gum acacia, gum arabic, gum Hazunta, 637, 647 n-heptane, 52, 337
mastic headache, 570, 640 Heracleum, 136
gum acacia, 260 headspace trapping, 326 Heracleum sphondylium, 515
gum arabic, 260, 271 heart, 35, 244, 462, 581, 697 he,bicide, 93, 97, 99, 105, 127,352,496
gum mastic, 447 heart conditions, 651 herbivores, vii, 7, 9~11, 13, 15, 32, 34, 121,
Gunneraceae, 197,337 heart failure, 674 128, 135-138, 207-209, 212, 213, 289, 290,
gut, 207, 208, 213, 245, 253, 340, 343, 460 heartwood, 78, 85, 88, 121, 136, 145, 148, 310,339,340,365,510,511,515,546,558,
gutta, 320, 321 149, 187,324 566,659
gutta percha, 312, 318, 320, 321 Hebenstretia dentata, 358 he,bivO'y, 9,10,14,30,53,135-137,209,
Guttiferae, see Clusiaceae Hectorella, 708 288,289,339,340,345,380,420,441,452,
Gymnema sylvestre, 463 Hedeoma drummondii, 338 696
gymnemagenin, 463 Hedeoma reverchonii, 338 he,bs, 36, 47, 110, 125, 126, 130,346
gymnemic acids, 462 Hedera, 610 Hemandiaceae, 581, 609
gymnomitr-8(12)-en-9a-ol, 368 Hedera helix, 459, 612 hemanduIcin, 385
gymnomitr-8(12)-en-9-one, 368 hederagenin, 459 heroin, 511, 597
gymnosperms, 37, 115, 140, 164, 173, 191, hederasaponin C, 458 herpes simplex virus, see virus, herpes simplex
194,226,264,286,290,324,337,338,382, a-hederin, 458 hesperetin, 175. 176
401,402,405,412,413,420,442,510,538, Hedyotideae, 612, 638 heteratisine, 673
543,624,626,677,709 Heimia, 694 heterodendrin, 284
gynocardin, 285, 289, 298 Heimia salicifolia, 694 (S)-heterodendrin, 282
Gyrinops, 444 helenalin, 390, 392 Heterodera g/ycines, 125, 128
Gyrostemonaceae, 304, 708, 709 Helenieae, 387 Heterophyllum, 287
He/enium amarum, 390 Heteroptera, 285
habitat, 4, 7, 31,125,136 He/enium microcephaium, 390 Heteropteris, 290
Hackelia cali/ornica, 552, 565 heliangin, 389 Heterotheca subaxillaris, 125
Haemadoraceae, 257 Heliantheae, 36, 46, 387 heteroyohimbine types, 631
haemanthamine, 621 ~623 Helianthemum scoparium, 126 Hewa brasiliensis, 5, 287, 288, 296, 297, 312,
haemanthidine, 621 Helianthus, 389, 390, 415 318-320,322,327
hair loss, 220 Helianthus annuus, 47, 415 hexadecanal, 30
hairpencils. 383, 552 Helianthus occidentalis, 415 (7Z)-hexadecenal,,31
hairy root cultures, see cultures, hairy root Helianthus tuberosus, 257, 389, 520 (9Z)-hexadecenal, 31
Haiti,537 Helichrysum, 45 (11Z)-hexadecenal,31
Halictus, 37 Heliconius, 289, 297 (E)-3-hexadecenoic acid, 17, 26
Halimeda, 419 Helicoverpa zea, 1,30,92,169,175,265,316, hexadecanol, 52
halimedatrial,419 381,390,437,682 hexadecatrienoic acid. 17
hallucinations, 523, 658 Heiicteres, 444 hexahydroxydiphenic acid, 194, 213
hallucinogens, 12, 111, 148,361,514,523, Heliothis, 316, 380 hexahydroxydiphenoyl group, 207, 212
525,535,546,559,581,647,655,658-660, Heliothis virescens, 169,220,231,415,481 hexaketide, 63, 64
694, 559, 697 Heliothis zea, see Helicoverpa zea hexanal, 113
halogenated secondary metabolites, 340 heliotrine, 549, 553 hexanoic acid, 86
Ha[ophytum, 708 Heliotropium, 547, 552 I-hexanol, 31
Haloragaceae, 197 Heliotropillm indicum, 551, 553 (2E)-hexenal, 31
Haloragidaceae, 337 Heliotropillm popovii, 553 (3E)-hexenal, 31
HamameIidaceae, 195, 197,252,361 Helix pomatia, 559 E-2-hexenal, 113
Hamamelis virginiana, 252 Heileborus, 267 E-2-hexenol, 31, 32
hamamelitol, 264 Helleborus joetidus, 515 E-3-hexadecenoate, 26
D-hamamelitol, 252 helminthosporal, 378 hexapeptides, 700
hamamelose, 251, 252, 264 helminthosporin, 64, 65, 85 hexen-I-ol, 31, 32
D-hamamelose, 252 He/minrhosporium, 378, 396, 423 (3Z)-hexenol, 31, 32
Hamelieae, 612, 638 Helminthosporium carbonum, 179, 184, 186 Z-3-hexen-l-01, 31, 32
Hannoa klaineana, 662, 667 Helminthosporium gramineum, 64 hexenyl acetate, 309
Hannoa undulata, 482 He/minthosporium maydis, 72 (3Z)-hexenyl acetate, 30
Haplopappus heterophyllus, 317 Helminthosporium sacchari, 378, 396 Z-hex-3-enyl acetate, 309
Hap!ophyton cimicidum, 641 Helminthosporium sativum, 378 2-hexuloses, 248
Haplophyton crooksii, 641, 653 Helopeltis, 650 D-arabino-hexulose, 250
734 Index

D-arabino-3-hexulose.250 honey, 177,256,257,343,385,402,553 hydrogen peroxide, 79, 117, 118, 592

Hibiscus, 77 honey locust, 215, 226 hydrogen sulfide, 227
Hibiscus eiatus, 77 honeybee, 35, 78, 177, 343, 445 hydrogenation, 507
Hibiscus sabdariffa, 171 honeydew, 99, 105,257,385,468,550,559, hydrolase, 162, 188,229,231,261
hide-powder method (tannins), 210 561 hydrolysis, 23, 24, 52, 83, 87, 89, 130, 136,
hides, 193,210 hops, 316, 363 193, 194,206,208,212,213,221,226,227,
higenamine.591 hordenine, 513, 514, 522, 523 253,256,259,267,269,288,290,294,300,
Hillia, 611, 612 Hordeum distichum, 522 302,305-307,309
Hillioideae, 612, 637 Hordeum vulgare, 288, 298, 522, 523 hydrolyzable tannins, 12, 193-195, 197,
Himantandraceae, 543, 610 honnona! activity, 408, 414, 500, 501 207-210, 212
Hintonia, 444 honnone, 3, 6, 12, 32, 33, 165,231,297,317, 13{S)-hydroperoxylinolenic acid, 32
Hippocastanaceae, 217, 218, 222, 223, 231, 318,322,367,383,384,393,395,396,398, Hydrophyllaceae, 177, 337
273,458 419,425,437,439,444,452-455,484,514, hydroquinone terpenoids, 336, 351
Hippocratea excelsa, 671, 690 516,518,520,659; see also antijuvenile hydroquinones, 76-79, 83, 94,104,124-126
Hippocrateaceae, 668, 670, 671 honnones, ecdysones, honnone binding sites, w-hydroxy acids, 54
hippocrateine I, 671 juvenile honnones, juvenile honnone hydroxamic acid, 98, 99, 105
Hippomane, 403 mimics, molting or moulting honnones, a-hydroxyacetosyringone, 123
Hippuridaceae, 337, 361 phytohonnones, plant honnones 3-hydroxyacyl-ACP dehydratase, 20
Hippuridales, 360, 361 honnone binding sites, 520 12-hydroxyamoorastatin, 481, 482
Hiptage, 290 hormones, plant, 6, 12 3-hydroxyanthranilic acid, 568
hiptagin, 291 homwonn, tobacco, 30, 49, 221, 526 y-hydroxy-L-arginine, 223
hhcinol, 146, 148 horse chestnut, see Aesculus hippocastanum, 4-hydroxy-L-argbrine, 223
hirsutin,310 232 m-hydroxybenzoic acid, 148
histamine, 168,517,692 horseradish, 168, 300, 303, 306 p-hydroxybenzoic acid, 77, 82, 104, 125, 126,
histidine, 168, 215, 575, 692, 693 horseradish peroxidase, 117, 645 165,277
histidine decarboxylase, 168 horses, 70, 676 hydroxybenzoic acids, 104, 106, 108, 121, 122,
histtionicotoxins, 708 Hortia, 569, 570 128,148
HIV, 209, 210, 213 host plants, vii, 70, 98, 363, 364, 366, 441, o-hydroxybenzyl-~D-glucoside, 122
HIV-I, 91, 254, 521, 562, 616, 698, 711 468 8-hydroxybergapten, 133, 134
HIV-I cell binding, 462 hosts, vii, 9, 10, 13,37,39,41,53,54,67,71, (2R,3R)-3-hydroxy-2-'H-butenoate, 21
lHV-l reverse transcriptase, 521, 600, 601. 613 74,78,84,92,93,98, !lO, 128, 177. 183, 2-hydroxy-3-butenylglucosinolate, 305
HIV-2,698 209,210,217,221,235,237,238,244,261, (2R)-hydroxy-3-butenylglucosinolate, 307, 308
HMO-CoA, see (3R)-3-hydroxy-3- 270,285,288,289,294,297,308,309,311, D-3-hydroxybutytyl-S-ACP, 20
methylglutaryl coenzyme A 545, 552, 559, 564, 565, 566, 574, 582, 591, IO-hydroxycamptothecine,648
HMG-CoA reductase, see 659, 669, 689, 690, 703 9-hydroxycanthin-6-one, 662
hydroxymethylglutaryl-CoA reductase Hottentots, 623 lO-hydroxycanthin-6-one, 662
HMO-CoA reductase, see ~hydroxy-~ houseflies, 35 {N-[3-(3-hydroxy-3-carboxypropylantino)-2-
methylglutaryl coenzyme A reductase human immunodeficiency virus, see virus, hydroxy-3-carboxypropyll-acetidine-2-
HMO-CoA synthase, see 3-hydroxy-3- human immunodeficiency (1llV), 210 carboxylic acid}, 230
methylg1utaryl-CoA synthetase humans, 31, 35, 36,47,67,99, 110, 123, 132, 16~hydroxy-5a-cholestanol, 680
Hodgkin's disease, 645 137, 138, 165, 167, 168, 174, 177, 191,213, 2O-hydroxycholesterol, 680
hodgkinsine, 665 226, 227, 244, 245, 253, 265, 267, 290, 294, 22~hydroxycholesterol, 439
Hodgkinsoniafrutescens, 665 299,306,313,317,321,324,346,361,378, (25R)-26-hydroxycholesterol, 678
HODMAT, see 4-dimethylallyltryptophane, 380,382,389,402,403,417,462,466,478 26-hydroxycholesterol, 678
hydroxy derivative hummingbirds, 177 4-hydroxycinnamic acid, 155
Hokkaido, 673 humulanes, 375 hydroxycitronellol, 353-355
holaphylline, 686 humulene, 371, 375, 393 IO-hydroxycitronellol, 353, 354, 358
Holarrhena, 677, 686, 689 humulene cyclase, 375 (S)-( - )-IO-hydroxycitronellol, 358
Holarrhena antidysenterica, 686 humulone, 316 7-hydroxycoumarin, 132
Holarrhena mitis, 700 Humulus lupulus, 316, 363 hydroxydanaidal, 550, 551, 552
holocalin, 287 humus, 207 ( - )-(R)-hydroxydanaidal, 550
(R)-holocalin, 280 Hunteria alkaloids, 634, 635, 637 ( + )-(S)-hydroxydanaidal, 550
HolocaJyx balansae, 280, 297 Huperezia serrata, 708 D-3-hydroxydecanoic acid, 35
holoparasite, 78 huperzine A, 708, 709, 711 7 a-hydroxy-3-desoxyzaluzanin C, 390
holothurins, 458 Hura, 403 8-hydroxydihydrobiochanin A, 186
Holygarna lactone, 269 Hyadaphis, 383 a-hydroxydihydrochalcone, 174, 189
Homaliaceae. 522 Hyalophora cecropia, 384, 444 2-hydroxy-2,3-dihydrogenistein, 179
Homalium pronyense, 521 hybrids, 8, 164, 177, 188,287,337,338,344, 7-hydroxydihydromatatadiether, 362
Homeosoma electellum, 415 351,376,437,535,552,558,559 4-hydroxy-3,5-dimethoxyphenylethylamine,
homoarginine, 223, 517 hydnocarpic acid, 36 523
homocysteine, 513 Hydnocarpus anthelminthica, 28 5-hydroxy-3,4-dimethoxyphenylethylamine,
homoerytbrina alkaloids, 617, 624-626, 677 Hydrangea, 142, 569 523
Homoesoma electellum, 390 Hydrangea macrophylla, 171 20-hydroxyecdysone, 440, 442, 444, 454
homeostasis, 460 Hydrangeaceae, 142, 143, 359, 361, 569 (3S,5R,6S)-3-hydroxy-5,6-epoxy-5,6-dihydro-~
homogentisic acid, 82, 106 ( - )-~hydrastine, 603 ionol,5oo
homohaningtonine, 625, 626 Hydrastis, 269, 603 18-hydroxy-9,10-epoxystearic acid, 53
homology, 9, 142 Hydrastis canadensis, 598, 603 3-hydroxyflavanone reductases, 200
homolycorine, 620 1,3-hydride shifts, 370 2-hydroxyflavanones, 162
homomethionine, 304 hydrocarbons, 16,44,51-54,324,332,339, 3-hydroxyflavanones, 175,201,202
homomonoterpene, 341, 342 367, 407, 425 3-hydroxyflavones, 200
Homoptera, 89 hydrocinnamic acid, 175 9-hydroxygeraniol, 354
homoserine. 217, 220, 222 Hydrocotyloideae, 134 IO-hydroxygenutiol, 354, 355, 358
homosesquiterpene, 341, 383 hydrogen cyanide, 231, 273, 274, 287-290, 5-hydroxy-6-C-glycosylflavones, 173
homospennidine, 548, 549 294, 296, 345 3-hydroxyheterodendrin, 282, 295
sym.-homospennidine, 520 hydrogen ion concentration, 28 E-4-hydroxyhex-2-ena!, 553
 Index 735

y-hydroxyhomoarginine, 223 15-hydroxyprostaglandin dehydrogenases, 167 Icacinaceae, 360, 361, 639, 648
erythro-4-hydroxyhomoarginine, 223 6-hydroxyprotopine, 600 ice cream, 708
4-hydroxyhomoarginine, 223 p-hydroxypyruvic acid, 106 ichneumonid parasitoids, 31
I-hYdroxy-2-(hydroxymethyl)anthraquinone,87 4-hydroxy-2-quinoline, 571 Ichneumonidae, 227
6,B-hydroxyhyoscyamine epoxidase, 535 14-hydroxy-4,14-retro-retinol, 498, 503 ichthyotoxic compounds, 85
3-hydroxyindole {3-D-glllcoside, 267 hydroxyrutacridone epoxide, 574 idose, 249
hydroxyindolizidine alkaloids, 561 12-hydroxysolanidine, 684 /lex guayusa, 700
a-hydroxyisopropyldihydrofuranocoumarins, 16-hydroxytabersonine, 634 /lex paraguariensis, 700
133 12,B-hydroxyteinemine, 684 llliciaceae, 387
15a-hydroxy-( - )-kallr-16-en-19-oic acid, 418 22-hydroxytingenone, 452 Illicium religiosum, 94
17-hydroxy-( - )-kaur-15-en-19-oic acid, 418 6-hydroxy-2,4,7-trimethoxyphenanthrene, 148 illudane, 376
hydroxylase, 26, 102 5-hydroxytryptophan, 228, 229 Ilybius, 440
6a-hydroxylase, 182 5-hydroxytryptamine, 514, 640 imagos,591
hydroxylation, 87, 107, 121, 142, 151, 152, 5-hydroxy-L-tryptophan, 215 imidazole alkaloids, 575, 610, 692, 693, 709
158,159,173,201,206,220,226,514,571, N-hydroxytyrosine, 275 imino-aldol type condensations, 507
573,587,615,620-622,624,656-658 3-hydroxyuridine, 703, 711 immune response, 6, 261, 562
13-hydroxylupanine, 559 hydroxywilfordic acid, 528 Impatiens balsamina, 81
13a-hydroxylupanine, 554, 555 5-hydroxyxanthoxin, 133, 134 imperatorin, 135. 136
(+ )-13a-hydroxylllpanine, 554 Hyenanche, 385 import, 19
17-hydroxylupanine, 559 Hyenanche globosa, 385 inandenine A, 521
8-hydroxylysergic acid, 659 hygrine, 531, 532, 539 inandenine B, 521
6a-hydroxymaackiain, 186 (R)-hygrine, 532, 534 incense, 413
( + )-6a-hydroxymaackiain, 182 Hygrocybe, 705 indenobenzazene, 610
p-hydroxy-(R,s)-mandelonitrile, 276, 277 Hygrophorus, 705 India, 265, 288, 389, 424, 478, 539, 553, 599,
5-hydroxymatatabiether, 362 Hylotrupes baju/us, 340 603, 609, 613, 640, 663, 686
6a-hydroxymedicarpin, 186 Hy/urgops, 340 Indian tobacco, 540
N-(4-hydroxy-3-methoxybenzyl)alkyl amides, Hymenaea,413 Indians, 523, 535, 559, 610, 658, 660, 677
517 Hymenocardiaceae, 700 Indians. American, 290, 523, 649, 694
N-(4-hydroxy-3-methoxybenzyl) amides, 517 Hymenoptera, 289, 340, 397, 426 Indians, Andean, 535
2-hydroxy-5-methoxy-3-[(8'Z,II'Z)-8',II', 14'- hymenovin, 390 Indians, Aymara, 684
pentadecatriene]-p-hydroquinone, 78 Hymenoxys odorata, 390 Indians, Aztec, 385
l-hydroxy-2-methylanthraquinone,87 hyoscyamine, 511, 535, 537 Indians, Hoti, 645
2-hydroxymethylanthraquinones, 88 (R)-hyoscyamine, 537 Indians, Jivaro, 591, 660, 700
3'-hydroxy-N-methy1coclaurine, 587 (S)-hyoscyamine, 535, 537 Indians, Juris, 588
3-hydroxymethyl-2(5H)-fllranone, 269 Hyoscyamus, 532, 535, 537 Indians, Tarahumara, 148
Hyoscyamus albus, 532, 535 indican, 267
(2S,4S)-4-hydroxy-4-methylglutamic acid, 226
Hyocyamus niger, 535 indicaxanthin, 705, 708, 711
(3R)-3-hydroxy-3-methylglutaryl coenzyme A
hyperglycemia, 591 indicine N-oxide, 553
(HMG-CoA),313
indigo, 267
,B-hydroxy-,B-methylglutaric acid, 580 Hypericaceae, see Clusiaceae
Indigo/era, 227, 232, 267, 290
3-hydroxy-3-methylglutaryl-CoA,313 hypericin, 76, 91, 92
Indigo/era spicata. 227, 232
{3-hydroxy-,B-methylglutaryl coenzyme A, 313 Hypericum androsaemum, 92
indole, 3, 6, 98, 113, 344, 514, 516
hydroxymethylglutaryl-CoA reductase (HMO- Hypericum drummondii, 69, 74
indole 3-acetaldehyde, 98
CoA reductase), 313, 314, 319, 326 Hypericum perforatum, 91, 93
indole acetaldehyde oxidase, 98
,B-hydroxy-,B-methylglutaryl coenzyme A hypersensitive response, 342
indole acetic acid, 3, 6, 98, 105, 165,514,516
reductase (HMOR), 313, 314 hypertension, 517, 640, 673, 686
indole 3-acetic acid, 94, 98, 165,437,514
,B-hydroxy-,B-methylglutaryl-CoA synthase hypertensive effects, 640
indole alkaloids, 353, 354, 358, 359, 365, 366,
(HMG-CoA synthase), 313 hypervitaminosis D, 437
560,563,575,610,615,628-632,634,636,
3-hydroxy-3-methylglutaryl-CoA synthetase, hyphae, 444
637,639,640,645,649,652-655,663,664,
313 hyphal elongation, 167
666-668, 686
2-hydroxynaringenin, 162 Hyphantria, 591, 603
indole, alkaloids, class 1, 630, 632, 634-636,
lO-hydroxynerol, 354 hypnotic, 537
641,645
hydroxynitrile lyase, 273, 282, 287, 289, 297, hypocotyl, 186,229,310,317,346
indole alkaloids, class 2, 632, 634
299 hypoglycemia, 222, 261, 647 indole alkaloids, class 3, 632, 634, 635, 645
a-hydroxynitriles, 273, 275, 285, 297 hypoglycin A, 222 indole alkaloids, class 5, 632, 634, 635, 645
18-hydroxyoleic acid, 53 hypoglycin B, 222 indole pyruvate decarboxylase, 98
16,B-hydroxy-22-oxo-5a-cholestanol, 680 hypolaetio 8-0-gl11coside, 167 indole 3-pyruvic acid, 98
16-hydroxypalmitic acid, 53 hypotension, 647, 697 indole-3-acetic acid, see indole 3-acetic acid
(IS,2S,3S)-3-hydroxy-2-(2'-cis-pentenyl) hypotensive effects, 591, 600, 606, 609, 640, indoleacetamide, 98
cyclopentane-l-acetic acid, 33 660, 670, 684, 686 indoleacetamide hydrolase, 98
I-hydroxyphaseollone, 186 Hyptis pectinata, 269 3-indoleacetonitrile, 306
4-hydroxyphenylacetaldehyde, 578, 586 hyptolide, 269 indoleamines, 517
(E)-p-hydroxyphenylacetaldehyde oxime, 275, hyrax, 468 indoleglycerol 3-phosphate, 98
276 indolizidine alkaloids, 533, 535, 543, 546,
1-(4'-hydroxyphenyl)-2-nitroethane, 290, 296 IAA, see indole 3-acetic acid 560-563, 565
m-hydroxyphenylpropionyl-CoA, 147 IAA oxidase, 132, 165 indolizidine system, 531, 533, 543, 546, 560,
p-hydroxyphenylpyruvic acid, 101, 102, 106 Iberis, 444 565,571
trans-4-hydroxypipecolic acid, 218 Iberis amara, 444, 445 indoloquinazoline alkaloids, 575, 610
trans-5-hydroxypipecolic acid, 218 iboga, 35, 634, 647, 648 indolyl glucosinolates, 302
trans-4-hydroxypipecolic acid-4-sulfate, 218 iboga alkaloids, 632, 634-637, 645, 647 3-indolylmethyl glucosinolate, 306, 309
12-hydroxy-4,6-pregnadien-3,20-dione, 440 ibogaine, 647 indospicine, 215, 227, 232
3-,B-hydroxy-5a-pregnane-20-one, 465 ibogaline, 647 inflammation, 392, 402, 517, 603
3-,B-hydroxypregn-5-en-20-one, 465 (- )-ibogamine, 635, 636 inflexin,415
4-hydroxyproline, 226 (+ )-ibogamine, 635, 636 inflorescence, 135, 136, 546, 659
trans-4-hydroxyproline, 226 ibogan alkaloids, 636, 637 infrared spectrophotometry (IR), 1; see also
trans-4-hydroxy-L-proline, 215 ibotenic acid, 697, 698 light
736 Index

ingenane, 402. 403, 424 invertase, 256 isobutyiamides, 36, 39

ingenol esters, 403 invertase inhibitor, 253 isobutylamine.513
inhibition, 26, 59, 78, 83, 95, 99, 123, 145, iodine, 307 3-isobutyl-2-methoxypyrazine, 703
146, 166, 188,209,311,510,517,520,521, iodoplatinate, 547 isochiorogenic acid, 104
523,526,535,537,540,544,549,550,558, ion channels, 651 isochondrodendrine. 606, 607
559,561,562,574,588,591,597,599-601, ion pair, 327, 329, 331, 351, 369-371, 396 isochorismate synthase, 97
603,606,607,612,613,616,619,623,625, ionization, 327, 329. 330, 369-371 isochorismic acid, 65, 80. 81, 88, 93, 97, 104,
627,640,641,645,647,648,650,651,654, ionones, 452 105, 122
656, 659, 660, 664, 673, 675, 676, 681, 682, a-ionone. 492 isocodonocarpine, 521
685, 696, 697, 700, 703, 708, 711 ,8-ionone, 384, 492 Isocoma wrightii, 317
inhibitors, 9, 13, 31, 33, 39, 40, 83, 88, 89, 93, ionophores, 65, 698 22-isodemissine, 684
99, 104, 105, 109, 110, 118, 125, 126, 128, ipecac alkaloids, 610, 611. 652 isodihydronepetalactone, 362
132, 135, 145-147, 164, 166, 187, 191, 195, ipecac root, 610 isodomedin, 415
231,234,242,246, see also ipecacuanha alkaloids, 610, 628 Isodan, 408, 415, 419
adenyltransferase inhibitors, cholinesterase ipecoside, 610, 611 Isodon inflexus, 415
inhibitors, a-glucosidase inhibitors, f3- Iphigenia.617 /sodon japonicus, 447
glucosidase inhibitors, glycosidase inhibitors, ipolamiide, 358 Isodon kameba, 415
invertase inhibitors, mixed function oxidase ipomeamarone, 378 Isodon shikokianus, 415
inhibitors, monoamine oxidase inhibitors. Ipomoea, 658 isodonal, 419
proteinase inhibitors, trypsin inhibitors Ipomoea halalas, 72, 377 isoelaeocarpine, 563
inhibitors of ,B-glucosidases, 254 Ipomoea purpurea. 7 ( + )-isoepoxydon, 71
inhibitors of glycosidase activity, 544 Ipomoea violacea, 655, 658 isoeugenol, 113
injury, 8, 30, 31, 122, 131,591,613 Ipomopsis, 444 isoflavanone, 179, 181, 186
Inocybe.697 Ipomopsis aggregala, 444, 452 isoflavans, 151, 179, 184
inositol,591 Ips, 340, 342 isoflavone,O-methyltransferase, 179
chiro-inositol. 264 Ips confusus, 343 isoflavone synthase, 178
D-chiro-inositol, 264 Ips paraconfusus, 343 isoflavones, 151, 178, 179, 181, 182, 184,
myo-inositol. 253, 264, 271 Ips pini, 343 186-188, 190,261,439
myo-inositol, hexakis-o-phosphate, 264 ipsdieno1, 343, 344, 352 isoqavonoids, 174, 178, 179. 186, 192. 316
scyllo-inositol,264 (+ )-ipsdieno1, 343 isogramine, 525
inotropic effects, 581, 591, 603, 692 ipseno1, 343 isohaningtonine, 625, 626
insect-host plant relationships, 441 lridaceae, 82, 87, 179,257,452,499 isoiridomynnecin, 362
insecticidal effects, 36, 40, 49. 111, 114. 186. iridal, 452 isoleucine, 217, 222, 232, 273, 274, 276, 282,
187,221,316,318,322,348,351,361,393, iridodial, 353-355, 361, 363, 366 286,287,300,302,513,547,549
398, 412, 418, 419, 424, 472, 478, 480, 483, epi~iridodial, 354
(2S)-isoleucine, 549
484,514,528,533,538,545,550,609,619, 8-epi-iridodial, 358 isoliquiritigenin, 158, 181, 182
isolysergic acid amide, 659
623, 642, 697 iridodial glucoside, 355
E-Z isomerization, 132
insecticides, viii. 36, 41, Ill, 118, 128, 186. iridodiol, 362
isoneomatatabiol, 362
348, 363, 412, 418, 419, 423, 478, 510, 525, iridoid glycosides, 353, 355, 358, 359,
526, 662, 686 isopavine alkaloids, 605
361-366
isopelletierine, 540
insecticides, carbamate, 110 iridoid monoterpene biosynthesis, 353
insecticides, organophosphate, 537 isopentenyl diphosphate, see isopentenyl
iridoid monoterpenes, 46, 87, 88, 124,324,
pyrophosphate
insectivorous, 52, 240 337, 353-355, 358-364, 366, 552, 579, 610,
isopentenyl diphosphate A-isomerase, 315
insects, viii, 8-10, 13, 14,35-37,39-42,49, 611, 648, 668-670; see also monoterpene isopenteny1 pyrophosphate, 312, 315, 319, 321,
50,52-54,83,84,89,92,93,99,111,118, iridoid alkaloids, monoterpene derived 326-329, 335, 348, 368-370, 401, 488, 494
122, 124, 127, 129, 130, 132, 135-138, 169, indole alkaloids isopentenyl pyrophosphate isomerase, 488
173, 174, 176, 177, 179, 184, 187, 189, 191, iridomynnecin, 353, 361. 362 isopentenyl pyrophosphate A3-A 2-isomerase,
207-209,212,213,217,220,221,227,228, Iridomyrmex, 353, 361, 362 315
231, 232, 244, 254, 260, 262, 269, 273, 285, Iridomyrmex humilis, 362, 703 isopentenyl pyrophosphate:dimethylallyl
288-290,293-295,297,298,300,305, Iridomyrmex nitidiceps, 669 pyrophosphate isomerase, 315
307-311,313,317,318,322,324,329,338, Iridomyrmex nilidis, 362 isophorone, 124
340-343,346,348,353,361,363,365-367, iridotrial,354 isophyllocladene, 412
379, 380, 382-384, 390, 395, 396, 398, epi-iridotrial, 354, 358 isopimaric acid, 415
416-419, 422, 437, 439, 440-442, 444, 445, 8-epi-iridotrial, 358 isopimpinellin, 133-136
447,452,454-456,460,466,468,472,478, iridotrial glucoside, 355 isopiperitenone, 333
479,486, 498, 504, 510, 511, 514, 515, 526, iris, 452 ( - )-trans-isopiperitenol. 333, 335
528, 537, 538, 540, 543, 545, 550, 559, 563, Iris germanica, 452 ( - )-isopiperetinone, 333
567, 570, 582, 583, 588, 591, 600, 603, 615, Iris pallida, 452 isoprene, 312, 315, 319, 321, 322, 326, 346
641, 652, 653, 682, 690, 696, 703; see also Iris pseudacorus, 452 isoprene rule, 312, 346, 371; also see
insect vision iris, yellow flag, 452 biogenetic isoprene rule
insects, scale, 89 iron, 171,230,510,521,545 isoprene units. 312, 369
insects, social, 343 irone, 452 isoprenoid compounds, 313, 425
interactions, vii,S, 9, 10, 14, 15,32,37,71, irritant effect, 346, 378, 382, 402, 403, 423, isopropyldihydrofuroquinoJines, 571
76, 78, 79, 121, 122, 125; see also plant- 424,460 (+ )-cis-isopulegone, 333
animal, pIant~funga1, plant-herbivore, plant~ Isalis, 267 /sopyrum, 269
insect, plant~microbe, and plant~plant Isalis linctoria, 267 isoquinoline alkaloids, 441, 578-582, 584,
interactions iso-a-acids,316 612, 638, 652, 668
intennediates, Schiff~base, 508 isoalantolactone. 392, 396 isorubijervine, 684
intennediates, synunetrical, 526, 531, 532, 534, isoamyl acetate, 343 isosafro1e. 110
537, 540, 548, 554, 608, 694 isoasarone, 113 isosalipurposide, 158, 173
intennediates, unsymmetrical. 535, 548 isobalfourodine, 573 isosiphonodin. 269
introgression, 337, 338 isobergapten, 135 isotabtoxin, 238
Inula royaleana, 673 isobicyclogennacrenal, 376 isothebaine. 591, 592, 594
Inuleae, 387 isoboldine. 590, 591 isothlocysnates, 266, 302, 305, 306, 308-311,
inulin, 257 isobruceine A, 482 383
 Index 737

[( - )·I·isothiocyanoato·(4R)· kamebanin, 415 lacrymation, 226, 227, 561, 697

(methylsulfmyl)]butane, 306 kangaroos, rat, 29 p..lactam ring, 234
( - )M3M isothujone, 340 KAS I, see ketoacyl-ACP synthase (KAS I) lactation, 317, 659
isotopicallY-labelled precursors, 326 KAS II, see ketoacyl-ACP synthase (KAS II) lactic acid bacteria, 520
isotrlglochinin, 280 KAS ill, see ketoacyl-ACP synthase (KAS ill) lactobacillic acid, 27
isovaleric acid diamide, 533 katemfe, 245 lactone forming glycosides, 247, 267, 269
isovestitol, 184 (- )-kauran-16a-ol, 418 lactones, 37, 59, 117, 130
isovincoside, 359, 628, 629, 654 kaurene, 673 lactose, 254
isovitexin, 162, 166 ent-kaurene, 398, 407, 408, 411, 420 Lactuca, 537, 599, 650
isoxazolinone glucosides, 293 I-kaurene, 408 lactucarium, 389
LJ3M isoxazolin-5--one, 293, 298 kaurenoic acid, 415 Iactucin, 389
LJ3M isoxazolin-5M one glucoside 1,2,293 kaur-l6--en-19-oic acid, 418, 420 Ladenbergia, 640
,B--(isoxazolin-5-on-2-yl)-~alanine, 290 kava, 139, 140 ladybugs, 542
isozymes, 5, 97, 102,300 kavakava, 139 Laetisaria arvalis, 26
Ithomiinae, 550, 551 kavalactones, 140 laetisaric acid (9Z,12Z,8-
Iva axillaris, 390, 396 kawa, 74, 139 hydroxyoctadecadienoic acid), 26, 27
ivy, 459 kawain, 140 Lagerstroemia subcostata, 694
kawakawa, 139 Lagodon, 420
jacarubin, 148 K.edde's reagent, 267 Lagodon rhomboides, 420
Jacquinia pungens, 452 kerosene, 535 lambs,685
jacquinonic acid, 452 ,B-ketoacyl-acyl carrier protein, 156 Lamiaceae, 49, 50, 77, 108, 168, 257, 269,
jam, 708 3-ketoacyl-ACP reductase, 19,20 280, 326, 328, 330-332, 337, 338, 346, 358,
Jamaica, 222 3-ketoacyl-ACP synthase (KAS I), 19, 21 361,371,387,405,406,408,412,413,415,
Japan, 362, 383, 460, 472,600,649,689 3-ketoacyl-ACP synthase (KAS II), 19-22 416, 418-420, 426, 442, 447, 451, 459, 672
japonicin A, 69 3-ketoacyl-ACP synthase (KAS ill), 19-22 Lamiales, 88, 360, 361
Jasminum, 32 ketocarotenoids, 498 Lamianae, 360, 361
Jasminum officinale, 326 l1-ketocycloartenol, 444 Lamiananae, 360
jasmonate·induced proteins, 33 a-keto acids, 508 Iamiide, 358
jasmonic acid, 16, 32-34, 39 ,B--keto esters, 58 laminarabiose, 254
jasmonic acid methyl ester, 32, 33, 39 a-ketoglutararic acid, 65, 80, 81, 158, 159 laminarin, 260
Jatropha curcas, 244, 402, 424 ketones, 48, 51, 52, 76, 288, 317, 339, 507, lantadene B, 451
Jatropha gosSYPifo/ia, 402 579,689 Lando/phia, 319
Jatropha multifida, 244 ketoses, 247, 248, 250, 253, 262 lanosterol, 431, 433, 436
jatronilizine, 598-600 khat, 210, 213, 522, 530, 668, 671 lantadene A, 447, 451
Java, 227, 649 Khaya nyasica, 480 Lantana camara, 447
jays, see blue jays Khaya senegalensis, 480 !apachol,85
Jerusalem artichoke, 257 khellin, 137 a-lapachone, 82
jerveratrum alkaloids, 684 khivorin, 476 larch,257
jervine, 684-686 kidneys, 174, 380 Lardizabalaceae, 610
jhanic acid, 500 kievitone, 182, 186 Larix iaricina, 415
jojoba, 51, 52, 55; see also wax, jojoba Kiggelaria africana, 289 Larrea divaricata, 120, 210
Juglandaceae, 82-84, 197, 337, 598 kinase C, 402 larvae, 9, 10, 35, 40, 53, 67, 84, 88, 92, 93,
Jug/ans, 81, 82, 84, 363 kinetin, 3, 6 111, 123, 124, 128, 132, 135, 136, 138, 175,
Juglans nigra, 83 ldaineanone, 482 179, 184, 190,208,217,220,221,227,228,
Juglans regia, 81, 83, 265 Klamath weed, 91, 92 232, 245, 253, 265, 269, 285, 288, 289, 294,
jugione, 81, 83-85, 92 knockdown effect, 348 296,297,307,308,310,311,316,317,
Julianaceae, 67 Know/tonia, 269 340-342, 363-366, 380, 381, 390, 394, 402,
julisporine, 562 Koenoline, 663, 666 412,415,416,418,424,437,441,444,445,
Jungermannia, 412 kohlrabi,306 447,459,460,466,468,470,471,479,480,
Jungermannia subulata, 143 kokusagin, 136 522, 526, 529, 535, 537, 544, 550-552, 558,
Juniperus, 179,337 kola, 700 559,562,565,591,597,603,609,613,615,
Juniperus ashei, 338 kolavenol, 380 619, 627, 640-642, 650, 653, 658, 660, 664,
Juniperus pinchotii, 338 kolavenyl pyrophosphate, 408 682, 683, 686, 696, 703, 709
Juniperus scopulorum, 338 Korea, 460 lasalocid A, 67
Juniperus virginiana, 338 K-PLMF I, 123 Lasiantlulea fruticosa, 380
Junonia coenia, 364, 365 Krameria, 35, 210 lasidiol angelate, 380
juribidine, 677, 681 Kramericeae, 35 Lasiocampidae, 437
Justicia hayatai, 120 Krebs cycle, 3, 6, 226 lasiokaurin, 420
justicidin A, 120 Kreysigia, 617 Lasius carniolicus, 419
justicidin B, 120 Laspeyresia pomone/la, 381, 468
juvabione, 383 laballenic acid, 49 latex, 318-322, 402-404, 466, 587, 593, 596,
juvenile honnone, 317, 318, 367, 383, 384, labdadienols, bicyclic, 405 597
395,398,419,440,442,444,479 labdadienyl pyrophosphate, 407 lathyrine, 700
juvenile honnone ill, 384 9,,BH-labdadienyl pyrophosphate, 414 L-Iathyrine, 215, 223, 226
juvenile honnone mimics, 367, 383, 384 labda-8(17),13-dien-15-yl (copalyl) lathyrism, 226
pyrophosphate, 407 Lathyrus, 174, 189,215,223,226
kaempferol, 152, 158, 166, 188 labda-8(l7),13-dien-15-yl pyrophosphate, 406, Lathyrus cicera, 223
kaempferol glycosides, 172 407 Lathyrus clymenum, 223
kaempferoJ 3-0-xylosyIgaIactoside, 168 ent-/-Iabda-8(17),13-dien-15-yl pyrophosphate, Lathyrus hirsutus, 223
kaffir beer, 210 408 Lathyrus iatifo/ius, 223
kairomones,9, 10, 1I0, 168,300,307,324, labdane, 404-406, 420 Lathyrus odoratus, 174, 189, 223, 293
339,341,352,363,367,380,381,447,454, Labiatae, see Lamiaceae Lathyrus pusillus, 223
498,511 Laburnum anagyroides, 142 Lathyrus sativus, 223, 226, 290, 518
kale, 306 laccase, 140 Lathyrus sylvestris, 223
Kalmia, 411 lacewings, 362, 363 Lathyrus tingitanus, 226, 700
738 Index

Iaticifers, 318, 319, 320, 465 (- )-lepidozenaI, 376 limonin D-ring lactone hydrolase. 483
Latin America, 319, 499 Lepidozia vitrea, 376 Limonium, 571
Latlfa, 537 Lepioto, 237 limonoate dehydrogenase, 483
laudanosine, 587, 588, 591 Lepipolys, 364 limonoate D-ring hydrolase, 483
laudanosoline, 605 leprosy, 36, 85 limonoic acid A-ring lactone, 483
Lauraceae,46, 1l0, 121, 139,340,380,387, leptioe I, 682 limonoids, 11,473, 475-485, 576
419, 510, 528, 563, 585, 592, 598, 605, 610, Leptinotarsa, 537, 597, 619, 650 B-seco-limonoids, 480
614,660 Leptinotarsa decemlineata, 31, 231. 412. 682 C-seco-limonoids. 476, 480
Iaurate, 35 Leptochloa dubia, 125 D-seco-limonoids, 476, 477
laureline, 591, 592 Leptocoris isolata, 285. 289, 294 Linaceae, 282, 549
Laurencia, 381 Leptopyrum. 269 linalool, 31, 337, 339-341, 344, 346
Laurencia pinnata, 442 Leptosphaeria macuians, 309 linaIyl acetate, 341
lauric acid, 17, 25, 35 Lepus americanus, 208,451 linalyl alcohol, 346
Laurus nobilis, 340 Lespedeza sericea, 78, 452 linalyl diphosphate, see linaIyl pyrophosphate
Lavandula officinalis, 130 lethal dose, 526, 677, 685 linaIyl pyrophosphate, 328, 329, 331, 332
lavender, 130 lethal substance, 522, 526, 537, 558, 562, 582, (3R)-linalyl pyrophosphate, 332
lawsone, 81, 87 609,619,640,642,673,677,685,703 (3S)-linaIyl pyrophosphate, 332
laxative, 35, 91, 517 lettuce, 83, 118, 346, 389, 459, 558, 703 linamarin, 9, 276, 277, 282, 287-290, 295,
leachate, IS, 83, 109, 126 Leucaena leucocephala, 219, 220 297, 299, 517
leaf hopper, 385 Leucathoe grayana, 411 Linaria, 569, 570
leather, 193, 204, 210 leucine, 226, 282, 285, 300, 302, 513, 578, 580 Lindero, 380
leaves, 7, 14, 16, 17, 18,22,24,29,30,31, D-Ieucine, 234 liniment, 122,346
35, 39, 40, 41, 51, 54, 55, 72, 78, 83, 84, L-leucine, 215, 217, 274, 625 linoleic acid, 17, 18,22,24,26,27,30,32,34,
88,91,92,99, 105, lll, 123-132, 135-138, (2S)-leucine, 549 35, 42, 43, 53
142, 148, 152, 171-174, 184, 193, 194, 196, leucoantbocyanidins, 162. 163, 200. 206 9Z,l2Z-linoleic acid, 26
197,206-210,212,220,226,227,238,240, leucoanthocyanins, 201 linolenic acid, 17, 18, 22, 24, 26, 30, 32, 34,
242,256,269,277,280,282,287-290,294, leucoapigeninidin, 164 35
296, 298, 300, 303, 307, 308, 320, 326, 330, leucoblasts, 326 a-linolenic acids. 17
338, 340, 341, 345, 351, 362, 371, 373, 381, leucocyanidin, 163 2-linoleoyl-I,3-dipahnitio,35
384,386,411-414,420,423,437,454, leucocytes, 591 a-linoleyl phosphatidylcholine, 22
457-459, 463, 466, 468, 470, 473, 476, 480, leucodelpbinidin, 163 linseed oil, 308
486,496,497,499,514,517,518,521,522, Leucojum aestivum, 623 Linum, 537
528,535,537,552-554,556,557,561, leucoluteolinidin, 164 Linumflavum.117
566-568,608, 611, 617, 647, 652, 658, 659, leucoplasts, 326 Linum usitatissimum. 274, 288
671, 673, 676, 689, 698, 700; see also hay leukemia, 145, 511, 627, 645, 648, 654 linustatin, 282, 287, 288, 299
leaf, cedar leaf oil, elm leaf chrysomelid, liana, 33, 671 Lipophis erysimi, 383
green leaf volatile complex, leaf beetles, leaf Libocedrus vateensis. 118 lipase, 24, 39
abscission, leaf cutter ants, leafhoppers, leaf lice,641 lipid, vii, 4, 6, 8, 14, 16-19,22-25,30,32,
lipids, leaf movement, South American leaf lichen, 56, 70, 77, 85-87 35, 39-41, 45, 49-55, 74, 92, 93, 135, 138,
blight, velvetleaf licorice, 462 152, 166, 191, 207, 208, 298; see also
lecanoric acid, 70 licorice, European, 184 cyanolipids. glycerophospholipids,
Lecithydaceae, 197 Liebennann-Burchard test. 456 glycoJipids, phospholipids
lectins, 234, 240, 242, 244, 245, 246, 261, 271, light, vii, 3, 4, 7, 8, 13, 26, 30, 40, 42, 50, 78, lipid metabolism, 4
272 91,92, lll, 120, 126, 128, 129, 131-138, lipid peroxidation, 209
Lecythis ollaria, 229 145, 153, 163, 164, 177, 206, 209, 237, 289, lipids, leaf, 17, 18
leek, 51, 226 320,328,414,477,486,491,494-497,554, lipids. seed. 32
legumes, 3, 6, 78, 145, 179, 181,218,220, 634 lipoquinones, 9
221, 226, 228, 229, 232, 241, 242, 244-246, light gap specialists, 7 lipoxygenase, 24, 26
256, 261, 280, 290, 391, 398,460, 514, 543, light screens, 165 5-lipoxygenase, 209. 267
549, 552, 554, 558, 559, 566, 567 light-harvesting complexes, 496 Lippia dulcis, 385
Leguminosae, see Fabaceae lignanolides, 117 Lippia rehmanni, 447. 451
Leishmania, 606 lignans, ll, 54, 102, 106, 117, 118, 120, Liquidambar formosana, 196
Leishmania mexicana amazonensis. 660 127-129, 190 liquidambin, 196
leishmaniasis, 599, 612 lignification. 11,259 liquids. supercritical, 326
leishmanicidal, 599 lignin, 3, 11,54, 102, 106, 108, 114, 115, 117, liquiritigenin, 158.201
Lemaireocereus thurberi, 582 123, 127, 128,208,213,259,288 (2S)-liquiritigenin, 178
lemmatoxin, 460 Ligusticum, 136 Liriodendron tulipi/era. 591, 613
Lemna, 29, 40, 537, 540, 543, 591, 613, 640, Liliaceae, 3, 35, 51, 85-87, 173,222,226, liriodenine,591
641,650 231,257,267,269,280,444,457,459,466, D-(-)-liriodennitol,264
Lemna minor. 145, 439 468,510,543,617,619,625,677,684 lirioferine.591
Lemna paucicostata. 289 Lilium longifIorum, 267. 271 liriotulipiferine. 591
Lemnaceae, 439 Lilium pardarinum, 458, 472 Litchi chinensis, 223
lemon. 346 limes, 137,257,528,535 lithiumsalts,16
Lens culinaris, 48 Limnanthaceae, 52, 304, 308 Lithospermum, 196
Lentibulariaceae, 361 Limnanthes douglasH, 52, 55 Lithospermum erythrorhizon. 85
lentil,48 limonene, 331, 332, 337, 339, 340, 342, 343, little bluestem, 125, 346
Leonotis nepetaejolia, 49 345, 346, 351 livOI, 65, 168,209,237,313,380,552,553
I..eontice, 558 (- )-limonene, 331, 335, 340, 351 liver, cirrhosis, 460. 619
Leontinus edodes, 700 (+)-limonene, 331, 340 liver, rat, 361, 431
Lepidium, 526, 537, 540, 558, 599, 601, 641, (- )-(4S)-limonene, 335 liverwort, 130, 136, 142, 145, 174,368,369,
650,673 d-limonene. 331 376, 386, 389, 412
Lepidoptera, 136, 365, 366, 390,437, 551, I-limonene, 331, 333, 335 livestock, viii, 29, 70, 91, 132, 136, 290, 306,
564,566,591,623,640,641,650,658,660, limonene cyclase, 331 307,317,378,380,383,390,402,403,447,
682, 703, 711 E-limonene oxide, 344 466, 517, 546, 552, 559, 561, 659, 684, 685,
lepidopteran caterpillars. 591 Z-limonene oxide, 344 689
 Index 739

Loasaceae, 361 (- )-lupinine, 554, 555 maize (Zea mays), 70, 98, 105,415,436,501,
Loasales, 360, 361 Lupinus, 179,229,510,553,554,558,559 517
Loasanae, 360 Lupinus angustifolius. 554, 700 Malacosoma americana, 288, 437
Lobelia, 531, 540 Lupinus argenteus, 552, 559 Malacosoma neustria, 437, 455
Lobelia berlandieri, 540 Lupinus fulcratus, 559 malaria, 392, 395, 482, 569, 599, 612, 640,
Lobelia inflata, 540 Lupinus luteus, 554, 556, 557 648, 649, 654, 662, 698
lobeline, 538, 540 Lupinus poiyphyllus, 3, 8, 9, 554, 557 malaria, tertian, 662
lobsters, 498 lutein. 486, 494-496, 505 malaria, vivax, 651
locoweed, 561 luteoforol, 164 male scent organ, 552
locust, 353, 379, 444, 460, 484; see also black luteolin, 152, 160, 164, 189, 191 Malesherbiaceae, 285, 287
locust, honey locust luteone, 179 malic acid, 160
locust, desert, 444, 478-480 lutoid particles, 318 Mallotus japonicus, 69
locust, migratory, 221 lyase, 158, 261 Maloideae, 264, 287
Locusta, 221, 228, 460, 618, 650 lycoctonine, 674 malonamide starter, 65, 67
Locusta migratoria, 53, 253, 288 lycomarasmin, 237 malonamido-CoA, 65
Locusta migratoria migratorioides, 221 lycopene, 486, 490-492, 499 malonate, 152, 155, 156, 463, 465, 471
Loganiaceae, 149,361,510,511,615,636, E-lycopene, 492 malonic acid, 148
638,640,641,646,652,653,660 lycopersene, 490 malonyl-ACP, 19, 20
loganic acid, 354, 359, 669 Lycopersicon, 30, 39-41, 146,230,269,381, malonyl-CoA, 19, 21, 23, 51, 58, 139, 142,
loganin, 353-355, 358, 365, 610, 611, 501,677,682 147, 155, 156, 531, 535, 539
628-630, 649, 669 Lycopersicon esculentum, 30, 230, 677 Malouetia, 677
Lolium perenne, 271, 659 Lycopersicon hirsutum, 30, 39-41, 381 Maipighia glabra, 265
Lotium tementulum, 659 Lycopersicon pimpinellifolium, 680 Malpighiaceae, 265, 290, 361, 514, 660, 700
Lomalia, 82 Iycopodine, 540 maltose, 254
Lonchocarpus, 186, 253 Lycopodium, 510, 531, 540, 544. 545 Maluoetia, 686
Lonchocarpus costaricensis, 226, 543 lycorenine, 620, 621 Malva sylvestris, 241
Lonchocarpus sericeus, 543 lycoricidine, 510, 623 Malvaceae, 18,28,35,77,309,378,569,570,
long chain alcohol, see alcohols, long chain lycoricidinoi, 510, 623 662,697
longicyclene, 375 lycorine, 620, 622, 623 malvidin, 167
longifoiene, 375 Lycoris radiata, 510. 623 Mamestra configurata, 308
Lonicera japonica, 459 lygaeid bugs, 468 mammalian eyes, 171
Lonicera morrowii, 355 Lymantria, 591, 603, 689, 703 mammalian systems, 19,35,37,65,72
lophenol, 582 Lymantria dispar, 361, 380, 411 mammals, 8, 9, 35-37, 101, 104, 106, 125,
lophocereine. 581, 582, 662 lymphoma, non-Hodgkin's, 235 130, 132, 136, 137, 207, 208, 228, 253, 288,
Lophocereus, 580 lysergic acid amide, 655, 658, 659 290,306,307,312,313,340,361, 378,402,
Lophocereus schottii, 441, 580, 582 lysergic acid. 316, 511, 655-657, 659, 660 411,444,468,478,498,507,511,517,553,
Lophoco/ea heterophylla, 143 Lysergsaurediethylamid (LSD), 659 559,581,591,594,698
Lophopetalum toxicum, 470 lysergic acid, diethylamide (LSD), 316, 511, mammary gland, 659
Lophophora williamsii, 523, 580, 581 525,659 Mammea siamensis, 209, 212
lophophorine, 581 lysergylalanine, 657 Mancinella, 403
loquat, 148 lysergyltripeptide, 658 (R)-mandelonitrile, 289
Loranthaceae, 45, 197,244,246,466 Lysichiton,610 (R)-( + )-mandelonitrile lyase, 280, 295
lotaustralin, 9, 276, 277, 287, 288-290, 296, lysine, 215, 217-219, 507, 517, 531, 537-540, mandibular glands, 343
297,299 542, 543, 554, 561, 694, 703 Mandragora, 537
(R)-lotaustralin, 282 D-lysine, 217, 218, 538 Mandragora oificinarum, 537
Lotoideae. 220 L-lysine, 217, 218, 239 mandrake, 537
Lotononis, 549 lysine decarboxylase, 554 Manduca sexta, 30, 40, 208, 221, 245, 526
Lotus, 179 Lythraceae, 88, 149, 197, 554, 692-694, 709 Manettia, 612, 638
Lotus corniculatus, 184, 289 Iyxose, 249 manganese ion (Mn 2 +>, 315, 319, 428, 488
Lotus pedunculatus, 179, 291 Mangifera indica, 67
Lotus scoparius, 126 rna huang, 522 mangiferin, 148
Lotus uliginosus, 184 maackiain. 179, 181, 182, 186 mango,67
Lou Gehrig's disease, 226, 294 (- )-maaackiain, 179, 182 mangostin, 148
LPP, see linalyl pyrophosphate ( + )-maackiain, 182 mangrove, 204
LSD, see lysergic acid, diethyl amide macarpine, 601 Manihot esculenta, 276, 290, 297
lubimin, 377 Machaerium, 187 Manihot glaziovii, 319
lubricant, 16, 35, 52 Machaerocereus gummosus, 582 Manilkara sapota, 321
(R)-lucumin, 277, 279 Mackin/aya, 570 manna, 257
lunacrine, 571 Madura, 140 D-marman, 259
Lunaria biennis, 521 Maclura pomijera, 145 mannans, 259, 260
lunarine, 521 macIurin,148 Mannich condensation, 507, 531
Lunasia amara, 571 Macrorungia, 693 mannitol, 260, 262. 264
lunasine, 571 Macrosiphon a/bifrons, 559 D-mannitol, 262, 264
lung edema, 378 Macrotermes subhyalinus, 419 mannose, 248, 249, 253
Lunularia, 146 madder, 85, 87. 89 D-mannose, 248, 249, 259, 260
lunularic acid, 142, 143, 145, 146 Maesa lanceolala, 77 mannosidase, 544
lupanine, 9, 510, 554-556, 558, 559, 567 maesanin,77 ,B-D-mannosidase, 294
( + )-lupanine, 554, 555 maesaquinone, 77 mannosides, 266
lupeol,447 magnesium ion (Mg 2 +), 81,155,171,264, mannosidosis, 561
Luperini,447 315,319,326,371,375,401,428,488,656 manooi, 405, 412
lupine alkaloid biosynthesis, 554 Magnolia kobus, 118, 128 mantid,468
lupine alkaloids, 9, 514 Magnoiiaceae, 118, 121, 123,387,591,610 maple. 223, 231, 523
lupines, 2 Magnoliales, 109,286,610 Mappia.648
lupinic acid, 700 Magnoliidae, 280, 575 Marah macrocarpus, 411
lupinine, 554, 559 maitenine, 452 Marantaceae, 245
740 Index

Marchantia polymorpha. 143 Melanchra persicariae, 550 Merrillia, 575

margarine. 35, 499 Melanophryniscus.560 mesaconine, 675
marigold. Mrican, 48 Melanoplus, 528 mesaconitine. 674, 675
marijuana, 324, 336 Melanoplus bivittatus, 559 mescaline, 511, 513, 522, 523, 579, 580, 581
marine animals, 223 MeJanoplus difjerenlialis, 460 mesembrenol, 624
marine borers, 78, 85, 88 Melanoplus sanguinipes, 316. 318, 384, 460, mesembrine. 624
marine compounds, viii, 14, 32, 48. 340, 380. 479 mesembrine alkaloids. 624
381, 394, 396, 398, 419, 420, 423, 425, 442, Melastomataceae, 197 Mesembryanthemaceae, 617, 623
495, 506, 696, 700, 703 Melia, 484 Mesembryanthemum, 617, 623, 624
marine organisms, viii Melia azadirachta, 479, 480 Mesembryanthemum alkaloids, 624
mannesin, 133 Melia toosendanan, 480 mesophyll cells, 9, 282, 299, 557
(S)-mannesin, 133, 135 Meliaceae, 337, 361, 399, 473, 475, 477-480, Mespillus germanica, 269
(+ )-(S)-mannesin, 133 483-485,533,571,573,575,576,601,608, messages. see chemical messages
Martyniaceae, 361 663 metabolic cost, vii, 1.6,7, 13,46, 193,208,
3-epi-maslinic acid, 447 meliacin A2, 480 209, 245, 510
mass spectrometry (MS), I, 54, 55, 132, 138, melianone, 473, 480 metabolism, see amino acid metabolism, fatty
142, 170, 179, 191,212,234,247,300,302, meliantriol. 480 acid metabolism, lipid metabolism. terpeDoid
326,353,367,379,396,427,455,457,470, meliatoxin AI> 480 metabolism
487,506,554,645,655,695 meliatoxin A2, 480 metabolism, primary, vii. 1,3-6, 10, 16,94,
mass spectrometry. chemical ionization (CI), meliatoxin B 2, 480 96, 104, 166, 214, 248
457 meliatoxin Bb 480 metabolites, primary, vii. 3-6, 16.25,247.510
mass spectrometry. fast atom bombardment meliatoxins Ah 480 metabolites, shunt, 6
(FAB), 170, 212, 223, 457 Melica, 659 metal, 169, 170, 177,208,209,237,240,260,
mass spectrometry. tandem, 655 Melicope, 574 276
massoia lactone. 269 Melilo/us alba, 130, 132, 137, 138 metals, heavy, 240, 241, 242
masticatory, 522, 528, 538 Melilotus, 130 metaphase, 120
Mastixiaceae. 361 Melioideae, 477 Metarrhizium. 253
( - )-matairesino1, 117, 118 melizitose, 257 methadone. 597
mare, see yerba mare mellitoxin, 385 exo-cis-3,4-methanoproline, 218
matricaria acid, 49 Melochia, 571, 700 methiooine, 27, 45, 63, 186,215,217,226,
matricaria ester, 43 Melochia tomentosa, 666, 700 230,232,238,513,568,570,580,598,700,
matrine, 556, 559 Melodinus, 637, 647 702
Matthiala. 159 Melodorumjruticosum, 59, 74 12-methoxyabietic acid, 415
Matthiola incana, 163 Meloidae, 382, 393 lO-methoxyajmalicine, 640
Maurandya antirrhiniflora, 364 Meloidogyne incognita, 459 4-methoxy-l-allylbenzene, 113
maytanbutacine, 695 Meloidogyne javanica, 479, 482 4-methoxybenzaldehyde, 124
maytanprine. 695 melosatin A, 666 6-methoxybenzoxazolinone (MBOA), 94, 99,
maytansine, 676, 694-696 melosatin B, 666 100, 101, 104
maytansinoids, 692, 694-696, 711 membrane, 3, 9, 14, 16-19,22,23,32,35,41, a-methoxy-A<>:-butenolide. 269
maytanvaline. 695 49, 65, 115, 135, 153, 166, 179, 182, 191, 9~methoxycanthin-6-one, 662
Maytenus. 452 208,231,237,261,270,314,319,321,326, 4-methoxycinnamaldehyde, 113
Maytenus buchananii, 695 331, 376,401, 415, 427, 428, 430, 433, 440, 4~methoxydalbergione, 78. 79
May tenus ilicifolia, 695, 696 444,452,459,468,490,492,497,517,518, 5~methoxy-N,N-dimethyltryptamine, 514
Maylenus ova/us, 695 520, 528, 550, 556, 574, 598, 603, 606, 660, 9-methoxyellipticine, 645
Maytenus serrata, 695 675, 682, 685; see also chloroplast lO-methoxycamptothecine, 648
MBOA, see 6-methoxybenzoxazolinone membrane, nerve-cell membrane, plasma I-methoxycanthin-6-one. 662
meadowfoam, 52, 55 membrane, thylakoid membrane, tonoplast IO-methoxycanthin-6-one, 662
meadowsweet, 209 membrane 2-methoxycarbonyl-3-prenyl-l,4-
meal, see cottonseed meal, oahneal membrane penneabilility, 35, 338, 520, 682 naphthoquinone. 87
mealybugs, 561 memory impainnent, 709 16-methoxy-2,3-dihydro-3-dihydroxy-N(I)-
measles. 120 menaquinone, 80, 81, 92 methyltahersonine. 634
mechanism, cationic, 327 Mendonciaceae. 361 a-methoxy-,..hydroxy-Ll<>:-butenolide, 269
meconic acid, 596 Menispennaceae, 179,245,290,385,387,442, 6-methoxY-7-hydroxycoumarin, 133
medicagenic acid. 459 510,511,581,588,592,598,606,608-610, I-methoxy-3-hydroxymethylcarbazole, 663
Medicago sativa, 179. 183, 261, 456 700 3-methoxy-4-hydroxyphenylethylamine, 523
medicarpin, 181, 183, 186 Menispermum dauricum, 606 lO-methoxyibogamme,647
(- )-medicarpin, 181, 183 Menispermum, 385 methoxylation, 580
medicinal uses, viii, 10,48. 69, 91, 92, 114, menses inducer, 696 methoxymedicarpin, 183
122, 123, 128, 139, 168, 188, 189, 192, 193, Mentha piperita. 328, 332, 333, 335, 351 6-methoxymellein, 73. 129
209,213,229,244,313,324,346,353,367, Mentha spicata, 335, 351 6-methoxY-N-methy1-1,2,3,4-tetrahydro-,8-
376, 392, 395, 397, 398, 420, 424, 426, 452, p-menthane-3,8-cis-diol, 346 carboline, 660
455,456,460,462,463,468,470,471-473, meotho1, 335, 346 4'-methoxynOIprotosinomenine, 608
481-483,507,530,535,544,546,553,559, I-menthol, 333 2'-methoxyphaseollin-flavan, 186
564,566,576,593,603,615-617,623,627, menthol esters, 333 5-methoxypodophyllotoxin, 117
628, 634, 640, 648, 649, 653, 655, 659, 662, menthone, 333 4-methoxy-l-propylbenzene, 113
667,671,673,684--686,692,694,710,708 ( - )-menthone, 335 ( - )-5'-methoxysativan, 179
medicine, 12,91,350,362,452,455,460, 1-3,4-menthone lactone, 335 16-methoxytabersonine.634
506,522,535,537,570,593,601,609,613, Menyanthaceae, 361 4-methoxy-l-vinyl-,B-carboline, 660
640, 642, 645, 648, 669, 670, 671, 673, mercaptoethanol, 107, 155 N-methylanthranilate, 568
684-686, 690, 698, 700, 708 Mercurialis perennis, 513 N-methy1ation, 513, 587
medicine. Ayurvedic, 539 Merendera, 617 methyl (2E,6E)-famesoate, 384
medicocarpin, 183 Merilliodendron megacarpum, 648 methy13,B-isobutyryloxy-l-oxomeliac-8(30)-
megastigrna-5,7(E).9-trien-4-one. 500 Meris alticola, 364 enate, 480
Megoura viciae, 363 Meris paradoxa, 364 methyl angolensate, 476
melacacidin. 200 merogedunin. 477 methyl azoxymethanol. 290, 294
 Index 741

methyl chavicol, 341 7-methyl-~-nucleoside hydrolase, 702 Microplitis croceipes, 32

methyl cinnamate, 344 methylodoratol, 174, 189 microsomal fractions, 22. 23. 26, 107, 128,
methyl ester of triglochinin, 280 4-metbyl-2-oxopentanoic acid, 580 155. 158.209.275.276.277.290.302.335.
methyl esterification, 260 methylphloracetophenone, 58, 59 368. 428. 430-432. 436. 444. 447. 452
2-methyl famesyl pyrophosphate. 430 cis-3-metbyproline, 218 microtubule assembly, 618, 676
N-methyl groups, 508, 531 cis-4-methylproline, 218 microtubule protein polymerization, 696
methyl jasmonate, 16, 33, 34, 39 metbylpseudolycorine, 620 Microtus montanus, 101, 104, 105
methyl salicylate, 122,344 N-methylpseudopelletierine, 540 mildew, powdery, 309
methyl thujate, 340 N-methylputrescine, 532, 535 milk. 70. 210. 390. 553. 659. 667
methylamine, 513 O-methylpsychotrine, 613 milk sickness, 317
NP-methylamino-L-alanine, 226 N-methyl-..1 1-pyrrolideine, 532 milk thistle, 168
,8-N-methylamino-L-alanine, 294 N-methylpyrrolideine, 532, 533 milkweed bug. 49. 317. 468. 470. 479
4-N-methyIaminobutanal, 532 N-methylpyrrolidine. 531. 532 Milletia, 186
N'l-methyl-L-amino-2,3-diaminopropionic acid, N-methyl-..1 1-pyrrolinium cation, 532, 539 millipedes. 273. 289. 569
226 I-methyl-Al_pyrrolinium cation, 534 mimicry. 552. 564
O-methylandrocymbine, 617, 618 N-methyl-..1 1-pyrrolinium salt, 533 Mimosa, 218, 227
O-methylanhalonidine, 581 I-methyl-A'-pyrrolioium chloride, 535 mimosa extract, 206
methylation. 151. 152. 173.265.513.571. l-methyl-J2-pyrroline. 533 Mimosa pudica, 123, 218
573. 587. 588. 590. 595. 597. 623 6-methylsalicyIate synthetase, 56, 58 mimosine, 219, 220, 230, 232
C-methylation, 69 6-methylsalicylic acid. 56. 58. 59. 121. 122 L-mimosine, 219
O-methylation, 206 Se-methylselenocysteine, 229 L-mimosine synthase, 219, 220
C-methyl-branched inositols, 265 14a-methylsterol 14a-demethylase, 436 mimosoid legumes, 227, 514
3-methylcarbazole, 663 (S)-Z-N-methylstylopine. 600 Mimosoidae. 179.221.227.514.521.543
24R-methylcholest-5,7-dien-3,8-ol, 441 3-(metbylsulfinyl)propyl glucosinolate, 304 Mimusops balala, 321
24S-methylcholest-5.7-dien-3,B-ol. 441 E-3'-metbylsulfonylallyl E-cinnamate, 110 mint. 5. 49. 77. 257. 338
4a-methyl-7-cholestenol,582 4-methylsulfmyl-3-butenyl isothiocyanate, 310 miracle fruit, 245
N-methylcoclaurine. 586. 591 3-methyl-l,4-thiazane-S-carboxylic acid, Z27 miserotoxin, 290
N-methylconiine, 543 methylthioethers, 45 mistletoe, 244, 246
22-methyl-')'-cyc1oiridal, 452 methyltransferase. 85. 108. 133.531.587-599. mite, two spotted spider, 67, 68, 341
N-methylcytisine, 558, 559 601.702 mites. 9. 34. 445. 478
NI-methyl-L-2,3-diaminopropionic acid, 226 4'-O-methyltransferase, 179 mitigative effect, 346
3,3'-rnethylenebis(4-hydroxycoumarin), 132 7-O-methyltransferase, 179 mitochondria. 8. 18. 19.41.99. 107. 166. 187.
methylenebisphloroglucinol, 56, 69 N-methyltransferases, 587 191.209.313.318.338.361.419.420.601.
24-methylenecycloartenol, 436 N-methyltynunine, 522, 624 645
2-(methylenecyclopropyl)-3-methylalanine. 222 7-methylxanthine,7fYl mitogenic, 244
methylenedioxy bridge, 110, 118, 571, 598, methysticin, 140 mitosis, 618, 623
600.601 metronidazole, 392 mitotic poison, 682
4-metbyleneglutamic acid, 217, 218, 226 mevalonate. 11. 65. 81. 82. 85. 86. 132. 133. mitragynine. 630
a-methylene-,),-hydroxybutyric acid, 269 312.315.316.326.395.401.430.447.465. mixed function oxidases, 107, 118,333,348,
4-methyleneproline, 218 488.490.492. 655. 656. 669. 670. 678. 680. 656
Z,z-methylene interrupted system, 24 689.692 mixed function oxidases, P450 dependent, 276,
4a-methylergosta-8.24(28)-dien-3,B-ol. 436 mevalonate 5-diphophosphate, 315 333. 526. 600
4a-methylergosta-8,24(28)-dien-3,8-01 to 24- mevalonate kinase, 315 mixed function oxidases (pSMO or
methylenelophenol (J8-..::F)-sterol mevalonate pathways, II polysubsb'ate mixed function oxidases), 348
isomerase, 436 mevalonate 5-pyrophosphate decarboxylase, mixed function oxidase inhibitors (PSMO
4a-methylergosta-8.14.24(28)-trien-3,B-ol. 436 315 inhibitors), 110
methylethylketone, 276 mevalonic acid. 8. 87. 312-315. 319. 321. 326. mogroside V, 463
methyleugenol, 111, 113 328. 329. 348. 354. 355. 359. 368. 370. 383. (+ )-mollisacacidin. 206
E-methyleugenol, 136 384.401.442.447.486.490.573.581.656. 2,3-trans-3,4-( + )-mollisacacidin, 206
N-methylflindersine, 571, 573 689 mollugin,87
4-methylglutamic acid. 215. 217. 218. 226 (R)-mevalonic acid, 313 Molluginaceae, 171,708.709
erythro-4-methylglutamic acid, 215 mevalonic acid metabolism, 132 molluscicidal. 209. 390. 394. 459. 460. 470.
4-methyl-3-heptanol. 31 mevaloside. 269 471.559
N-methyl-trans-4-hydroxy-L-proline, 226 mexicanolide,476 molluscs, 559
D-I-O-methyl-muco-inositol, 264 Mexico. 52. 91. 148. 219. 298. 320. 388. 466. molt. 317. 318. 384. 440. 444. 468
ID-3-0-methyl-chiro-inositol, 264 540. 559. 658. 671. 673. 700 molting hormones, 479; see also moulting
ID-I-O-metbyl-muco-inositol,264 mezerein, 403 honnones
ID-I-0-methyl-myo-inositol, 264, 265 MFO, see mixed function oxidases momilactones, 414
ID-4-0-methyl-myo-inositol, 264, 265 minimum inhibitory concentration (MIC) monellin,245
lL-I-O-metbyl-myo-inositol, 264, 265 values. 683. 686 Moniliniafructicola, 183, 377
IL-2-0-methyl-chiro-inositol, 264 mice. 230.403.460.560.564.599.627.645. Monimiaceae, 581, 592, 610
lL-3-0-methyl-chiro-inositol, 264 677 monkey, 226
O-methyl-scyllo-inositol,264 mice, immunodeficient, 648 monoacylglycerols, 24
5-0-methyl-myo-inositol, 264 Michael addition, 389, 415 monoamine oxidase activity, 148
methylisoeugenol, 113 micha11amine B. 698 monoamine oxidase inhibitors, 514
E-metbylisoeugenol, 113 Michigan. 362 monoamine oxidases. 509, 514, 517, 660
2-methyljuglone, 80 microbes. 4. 17. 191.207.253 monoamines, 509, 513-515, 517
7-metbyljuglone, 63, 80 Microcyclus ulei, 288, 296, 319 monocotyledonous plants, 19,87, lIS, 164,
metbyllycaconitine, 675 Microcystis aeruginosa, 239, 245 172. 192. 206. 252. 280. 286. 324. 457. 510.
methylmalonate, 57 microfibrils, 259 517.562
methybnalonyl-CoA. 65 microfilament assembly, 72 monocrotaline, 549, 551, 552, 553
2-methyl-6-methoxyfuranobenzoquinone, 79 Micromelum, 575, 663 monogalactosyldiacylglycerol, 19,23
N-methylnicotinic acid, 708 microorganisms, 4, 42, 80. 88, 92, 94, 95, 106, monoIaurin, 35
O-methylnorbelladine, 622, 623 126. 149. 234. 259. 296. 508. 515. 520. 568. Monolinia fruticola, 377
6-0-methylnorlaudanosoHne, 587 667.711 monomeric G-actin, 237
742 Index

monomorine I, 563 moth, silk, 36 mydesmone, 380

monomorine VI, 563 moth, sunflower, 390, 415 mydriatic activity, 537
Monomorium, 533, 542 moth, urania, 253 myocardial fibrosis, 35
Monomorium pharaonis, 563 moths, 177,269,270,288,289,344,361,364, myocarditis, 35
monooxygenases, 59, 98, 108, 121, 158, 159, 517,535,545,550-552,565-567,669,71l myopathy, 623
179,181,182,202,601; see also moths, arctiid, 551, 552, 565 Myoporaceae, 46, 361, 380
cytochrome P-450~dependent moths, sphingid, 535 Myoporum deserti, 380
monooxygenases, polysubstrate, 348 motion sickness, 537 Myosurus, 269
monooxygenases, tryptophan 2~monooxygenase moulting honnones, 440, 441, 454; see also myotic effects, 692
monosaccharides, 244. 247-250, 256, 264, molting hormones myrabolans, 195
270,272 mountain, Andes 535 myrcene, 331, 337, 342-344
monoterpene biosynthesis, 322, 326-329, 331, mouse, 528, 552, 559, 588, 591, 597, 600, 601, ,B-myrcene, 343
333, 350, 394 607,612,619,623,640,645,658,660,664, Myricaceae, 197, 244
monoterpene cyclases, 329, 342, 354 673-676, 682, 685, 695, 702, 703 myricetin, 152, 166, 195
monoterpene-derived indole alkaloids, 628, movement, leaf, 193 Myrioge1losporae, 659
630, 632, 634, 636, 638, 640, 655 mucilage, 582 myristic acid, 17,25,35, 101, 127, 129
monoterpene glycosides, 346 Mucor, 501, 502 Myristica jragrans, 25
monoterpene iridoid alkaloids, 364 Mucor muceda, 502 Myristicaceae, 179,514,610
monoterpene synthesis, 326, 326 Mucorales, 50 l, 502 myristicin, Ill, 128
monoterpenes, 5, 31, 47, 78,133,312,322, Mucuna, 228, 229, 232, 706 Myrithecium, 378
324,326-333,335,337-346,348-354, Mucuna pruriens, 228, 517 Myrmeca ruginodus, 88
367-370,381,383,412,430,451,452,455, mugineic acid, 230 Myrmecaphodius excavatacollis, 53
575; homomonoterpenes, iridoid mulberry, 35, 145,253 Myrmica rubra. 124
monoterpenes, monoterpene alkaloids, mulberry, paper, 145 myrosinases, 300, 302
monoterpene iridoid alkaloids, monoterpene mule deer, 207 Myrsinaceae, 68, 77
derived indole alkaloids MU1ldulea, 186 Mynaceae, 59, 69, 136, 140, 142, 197,337
monoterpenes, acyclic, 324, 329 ( - )-munitagine, 605 Myrtales, 197
monoterpenes, bicyclic, 329 muramine, 603 myrtan,204
monoterpenes, cyclic, 324, 329. 331 Murraya, 575, 660, 663 myrtenal,451
monoterpenes, irregular, 346 Murraya koenigii, 663 Myrtiflorae, 197
monoterpenes, methylcyc1opentanoid, 363 Musa, 514 Myzus persicae. 173, 363, 550
monoterpenes, monocyclic, 329 Musa sapentium, 517
monoterpenoid alkaloids, 525, 668-670 Musaceae, 581
NAD-dependent dehydrogenation, 335
Monotropaceae, 361 Musca domestica, 35
NADH, 19,20,22,26,51, 117, 166, 167,226,
muscaaurin I, 705
Montanoa tomentosa, 420 252
muscaaurin II, 705
Montiniaceae, 361 NADH dehydrogenase, 166, 191
muscaflavin, 705, 706
mood changes, 596 NADPH, 20-22, 51, 56, 58, 97,107,108, 1l7,
muscarine, 511, 537, 692, 697, 711
moose, 207 121,155,158,163,164,167,178,179,181,
muscarine alkaloids, 697
mopanols, 200 200, 202, 204, 275, 276, 277, 333, 335, 355,
muscarinergic acetyl choline receptors, 537
Moquinea, 390 430,431,490,491,600,630,631
muscarinic action, 528, 697
Moraceae, 134, 136, 137, 138, 142, 145. 149. NADPH-cytochrome c reductase, 431
muscimol, 697, 698
179,253,319,466,533,543,563,571 nagilactone C, 420
muscle, see cardiac muscle, skeletal muscle,
moracins, 145 nagilactone P, 420
smooth muscle, uterine muscle
morin, 166 Na1ldina domestica, 280, 297
muscle lactate dehydrogenase, 642
Morinda, 9, 82, 85, 229 muscle weakness, 612 nandinin, 280
Morinda amplexicaulis, 229 naphthoate synthase, 81
muscone,37
Morinda lucida, 9, 82, 86 muscular relaxant actions, 670 naphthoquinone, 56, 63, 64, 76. 80-87, 94,
Morinda reticulata, 229 104
mushrooms, 35, 235, 237, 246, 496, 517, 666,
Maringa aleifera, 306, 310 697,709 1,4-naphthoquinone, 81
Moringa stenopetala, 306, 310 musk deer, 37 naphthylisoquinoline alkaloids, 698, 709
Moringaceae, 303, 361 Mussatia, 114, 128 naphthylphthaIamic acid, 166
morphinandienone alkaloids, 586, 592 mussatioside I, 114 narciclasine, 510, 623, 623
morphinandienones, 592 mustard, 300, 308-311 narciprimine, 510
morphine, 51l, 585, 592-597, 613, 615, 623 mustard greens, 306 narcissidine, 620
morphine alkaloids, 585, 592-594, 605 mustard oils, 302 Narcissus poeticus, 620
morphogen, 71, 552 mutagenic, 137.510,549,615,645 Narcissus pseudonarcissus, 620
Marus. 253, 543 mutants, 4, 41, 94-96, 156, 230, 303, 313, narcotic compounds, 588, 596, 603
Morus alba, 142, 145,341 411,496 narcotine, 595-597, 603
Morus nigra. 341 mutations, 3-5, 7, 9, 10, 12 a-narcotine, 596, 603
mosquito repellent, 346 Mutisieae, 387 ( - )-a-narcotine, 603
mosquitoes, 67, 344, 641 mutualistic relationships, 30, 35 naringenin, 156, 158-160, 164, 167, 168, 175,
mosses, 40, 46,130,142,172,173,179,497; T-muurolol,381 176,201
see also club moss muzigadial, 380 (2S)-naringenin, 159, 162, 164, 178
moth, burnet, 9, 517 MV A, see mevalonic acid naringenin 7-rutinoside, 175
moth, catalpa sphinx, 361 myasthenia gravis, 623, 709 naringenin-chalcone synthase, 156
moth, cinnabar, 550 mycelium, 501, 502 naringin, 175, 176
moth, codling, 381, 397, 468, 472 Mycobacterium smegmatis, 69 Narthecium ossifragum, 269
moth, diamondback, 308, 309, 311, 468, 472 Mycobacterium tuberculosis, 262 nartheside, 269
moth, giant silkwonn, 384 mycology, 12 Nasonov glands, 343
moth, gypsy, 361, 380, 394, 41l, 424 mycomycin, 48 nasturtium, 303, 309
moth, leek, 227, 459, 471 mycorrhiza, vii, 9, 149 Nasturtium, 363
moth, magpie, 551 mycorrhizae. vesicular arbuscular (V AM), 167 Nasturtium ojJici1lalis, 304
moth, oriental fruit, 34 mycorrhizal fungi, vii, 261, 561 Nasutitermes,419
moth, oriental tussock, 417 mycotoxins, 5, 13, 15, 56, 70, 73, 74, 378, natural products, viii, 6, 13, 14, 15,40,56,65,
moth, plume, 552, 566, 669, 690 379, 395, 396, 655 92,93, 114
 Index 743

natural selection, vii, 4-6 Nicotiana, 230, 232, 269, 525, 526, 528, 532. (R,S)-norcoclaurine, 595
Naucleeae, 638 677 (S)-norcoclaurine, 586
nausea, 612, 697 Nicotiana debneyi. 377 (S)-norcoclaurine synthase. 586
naval stores, viii, 398, 412 Nicotiana glauca, 528 nordihydroguaiuretic acid (NDGA), 120,210
Near East, 478 Nicotiana repanda. 526 norepinephrine, 640
necic acid, 549, 551, 552 Nicotiana rustica, 526, 528 norhannan, 660
necrosis, 378, 423 Nicotiana sylvestris, 526 norlaudanosoline, 587, 595
Nectandra colo, 139 Nicotiana tabacum, 526, 629 (S)-norlaudanosoline, 2, 3, 587
Nectandra megapotamica, 660 nicotinamide, 528 (S)-norlaudanosoline synthase, 2, 587
nectar, 176, 177, 362, 366, 550, 565 ( - )-nicotianamine, 230 nomicotine, 526
nectar guides, 177 ( + )-nicotianamine, 230 nororientaline, 588
nectar thieves, 176, 362, 366 nicotine, 510, 511, 513, 525. 526, 528. 531, norpluviine, 620, 621
nectaries. extrafloral, 362 534, 538, 540 nonuspolinone, 533
nectaries. foliar, 289 (S)-nicotine. 525 nors doring, 402
Nectria, 253 nicotinic acetylcholine receptor channel. 708 North America, 337, 338, 352, 365, 388, 411,
neem tree, 478-480 nicotinic acid, 274, 277, 286, 507. 525, 526, 441,466,550.558,563
Nelumbonaceae, 592, 610 528,529,531,538,545,708 norvaline, 513
Nelumbonales, 337 nicotinic receptors, 675 noscapine, 596, 597
nematicidal, 49, 50, 398, 403. 419, 559 Nigel/a, 269 Nothapodytes joetida, 648
nematode, root-rot, 340, 479, 482 Nigella damascena. 568 Notholaena, 174
nematode, soybean cyst. 125 Nigeria, 663 Notholaena standleyi, 423
nematodes, 8, 9, 42, 49, 67, 184.340,403, NIH shift, 522 notholaenic acid, 423
419,437,442,452,459,479,514 nimbalide, 479 NPP, see neryl pyrophosphate
neoajmaline, 640 nimbin, 476, 479 nuclear magnetic resonance (NMR), 1, 14,58,
Neodiprion setifer, 340 nimbolide, 473, 479, 482, 484 63,74,76,92,93,142,170,179,186-188,
neoflavonoids, 151, 187 nimbolidin A, 480 190,203,204,212,230,269,294,302,303,
neoherculin, 36 nimbolidin B, 480 311,318,326,365,367,423,427,431,454,
neohesperidin, 175, 176 ninhydrin, 217, 220, 226, 560 457,458,484-487,506,579,645
neolignan, 54, 106, 121. 127, 129, 140 nitidine, 601 DC-nuclear magnetic resonance (NMR), 1, 58,
neolinustatin, 282, 299 Nitidulidae. 37 117, 190,203,212,311,353,365,447,465,
neomatatabiol. 362 nitric oxide, 293 487,531,535,546,568,579,584,615,617,
(+)-neomenthol, 335 nitrile glycosides, 293 625, 628, 632, 655, 668, 692, 695
d-neomenthol, 333 nitriles. 275, 302, 306 DC-I3C-nuclear magnetic resonance (NMR)
( + )-neomenthyl glycoside, 335 nitro acids. 290 couplings, 58
( + )-neomenthyl glucosyl transferase, 335 nitro compounds, 276, 290, 295. 298, 299. IH-nuclear magnetic resonance (NMR), 1,58,
neonepetalactone, 362 303,591 63,76,92,203,212,311,365,423,435
neopulchellidine. 672 a-nitrocarboxylic acid. 303 2H-nuclear magnetic rsonance (NMR), 548,
Neoterpes graefiaria, 364 nitrogen. vii, 6-8, 208, 209, 219, 220, 229, 554
neoxanthin, 486, 494, 496 266,275,287,290,294,303,506-511,523, nuclear magnetic resonance, Fourier transform
9'-Z-neoxanthin, 501 531,544,546,554,565,567,580,581,584, methods, 1
Nepenthes, 52 617,621,622,649,668,669,675,676,678, nuclei, 318, 444, 502
Nepeta cataria, 358, 361 680, 689, 705 nucleic acid synthesis, 648
nepetalactol, 363 nitrogen ftxation. 83. 219 nucleic acids, 3,167,207,249,520,615,700,
nepetalactone, 358. 361-363 nitrogen fixing bacteria, 244, 261 709,710
Nephila clavipes. 551 nitrogen oxides, 293 5' -nucleotide phosphodiesterase, 208
neral, 341-343. 346 l-nitro-2-(p-hydroxyphenyl)ethane, 276, 296 nucleotide sugars, see sugars, nucleotide
neriifolin, 468, 472 l-aci-nitro-2-(p-hydroxyphenyl)ethane, 275, nucleus. 337, 378, 442, 465. 475, 488
Nerium oleander. 98, 468, 470 276 nudibranch, 380
nero1, 328, 349, 354, 365, 563 2-nitro-3-(p-hydroxyphenyl)propionic acid, 275 nudiflorine, 528
nerolidol. 370, 380 aci-l-nitro-2-methylpropane.276 Nuphar, 525, 554, 668, 670
nerolidyl pyrophosphate, 371, 430 l-nitro-2-phenylethane, 303 Nuphar alkaloids, 525
nerve transmission. 517 2-nitrophenylglucosinolate, 303 Nuphar japonicum, 670
nerve-cell membrane, 674 3-nitropropanol, 290 Nuphar [uteum, 670
nervous system. 510, 517, 623 3-nitropropionic acid, 290, 291, 294 nutmeg, 25, 111
neryl phosphate, 328 nitrous oxide, 290 nutrition, 12, 126
neryl pyrophosphate, 324, 326-332, 348 Nocardia acidophilus, 48 nuts, 229; see also betel nut, chestnut, coconut,
Netelia heroica, 32 noctuid caterpillars, 316 horse chestnut, nutmeg, peanut, tungnut,
neuralgia, 673 nod factors, 261 walnut
neurochemical activity, 591, 646 nodosin, 419, 420 nuts, macadamia, 290
neurolathyrism, 223, 226 nomiiin, 476. 479, 481, 483, 484 Nyctaginaceae, 708
neuromuscular action, 607 nona-2,6-dienal, 346 Nyctemera annuiata, 551, 564
neuromuscular blocking agents, 643 non-apparency, 209 Nymania capensis, 477
neuromuscular transmission. 675 nondepolarizing neuromuscular poison, 675 Nymphalidae. 364-366
neuron, 510 (2E)-nonenal, 37 Nympheaceae, 197, 516, 581, 670
neuropteran. 53 (2E,4E)-nonadienal, 37 NympheaIes, 337
Neurospora. 96 non-protein amino acids, vii, 6, 28, 215, 217, nymphs, 317.444,479
neurosporene, 490 218,220-223,226,228-232,234,240,241, Nyssaceae, 197, 360, 361, 648
neurotoxic syndrome, 70 274,277,285,289,294,303
neurotoxicity, 231, 296, 675 noradrenaline, 510, 591, 640, 659 oak, 194, 195,208,209,213,264
neurotoxin, 221. 223. 226. 299, 675 noraporphines, 588 oak, cork, 447
neurotransmitters, 510 norbelladine, 619-622, 624 oak, red, 252
New World, 447 O-norbelladine, 622 oatmeal, 451
New Zealand, 179, 385, 403, 563 (6S)-O-norbellidine, 622 oats, 37, 40, 70, 94, 98, 105, 230, 376, 459,
Nicandra physaloides. 532 norcimiphytine, 641 517
Nicandreae, 537 norcoc1aurine, 586, 591, 605, 615 obacunone, 481
744 Index

obtusaquinone, 78, 79 olive, 98 Orobanche, 391, 392

obtusifoliol. 435, 436 olivetol, 336 Orobanche rapum-genistae, 557
Oceania, 226, 294 ololiuqui, 655, 658 orotate, 226
Ochnaceae, 173 Omphalea, 253 orris root oil, 452
Ochromonas ma/hamensis, 432 Onagraceae, 188, 197 orsellenic acid, 57, 58
Ochrosia, 645, 652 Oncidium ceboletta, 148 orthoacetate, 453, 468
Ochrosia elliptica, 645 oncinotine, 521 Orthoptera, 559
ochtodene, 340 Oncinotis inandensis, 521 oryzalexin A, 414
Ochtodes secundiramea. 340, 351 Oncinotis nitida, 521 oryzalexin B, 414
ocimene, 341, 344 oncogenes, 321 oryzalexin C, 414
,8-ocimene, 340 Oncopeltus, 658 oryzalexins, 414
(E)-p-ocimene, 341 Oncopeltusfasciatus, 49, 317, 468, 479 Orzyaephilus surinamensis, 37
Ocotea pseudocoto. 139 Ondontomachus, 703 OSB, see o·succinylbenzoic acid
Oeotea rodiaei, 605 one eye or "cyclopism," 685 osladin, 463
Oeateo veraguensis. 121, 127 onion, 34, 226, 227, 376 Osmanthus absolute, 500
ocoteine, 591 Onoclea neriifolia, 431 Osmunda japonica, 173
oeotine,605 D.( + )·ononitol, 264 osteolathyrism, 223, 226
oct-l-ene-3-o1, 31 Ontario, 338 osthol, 113, 136
octacosanol, 52 oogoniol I, 444 Ostrinia furnacalis, 480
octadecanol, 52, 54 oogoniol 2, 444 Ostrinia nubilalis, 99, 104,417,696,711
18-octadecanolide. 37 oogoniol, 444 Osyris alba, 557
octodene, 340 Oomycetes, 260 ouabain, 468
(9Z,11E,13E)-octadecatrienoic acid. 35 Operculina, 658 ovaries, 111,419
7,8,11,12,7',8',11',12'-octahydro-.p,.p.carotene, ophiobolanes, 415, 422 "over-production" of metabolites, 6
490 ophiobolin A, 422, 423 ovicidal,49
Odocoileus hemionus, 207 opbiobolin B, 422 oviposition, 13,37, 113, 129, 136, 173, 190,
Odocoileus virginianus, 207, 390 ophiobolins, 422 191,300,308-311,340,350,381,394,397,
odor, 324, 339, 340, 344, 346, 349, 351, 353, Ophiobolus graminis, 459 437,441,468,472,566,591
381,382,385,452 Ophiorrhiza mungos, 648 ovules. 596
odoracin. 403, 419 Ophrys, 344, 381, 382, 393, 395 oxalic acid, 160, 265
odoratol, 174 Ophrys bombyliflora, 381 Oxalidaceae. 173
odoratrin,419 Ophrys insectifera, 381 NJ-oxalyl-2,3-diaminopropionic acid, 223
( - )-odorinol, 533 Ophrys lutea, 344, 381 NJ-oxalyl-L-2,3·diaminopropionic acid, 226
Oecophoridae, 136 Ophrys sph. litigiosa, 381
oxazolidine-2·thiones, 305
Oenanthe crocata, 47 Ophrys tenthredinifera, 381
oxidation, 506, 509, 549, 580, 598, 600, 613,
oenothein B, 210 ophthahnology,664
622, 656, 663
Oenathera hooken', 173 opiate receptors, 597
a--oxidation, 24, 30, 44, 106
Oenothera microntho. 126 Opiliaceae, 45
P-oxidation, 24, 30, 43, 44, 121, 195
oestrone, see estrone opines, 239, 245, 246
ohchinolid A, 480 opium, 585, 593, 595-597 oxidation. Baeyer-Villiger type, 45
oxidative coupling, 173, 196, 197,210,588,
ohchinolid B, 480 opium poppy, 585, 593, 596
oil cells, 324 590-594
Opossum, 698
oil glands, 335, 353, 367, 368 opsin,497 oxidative decarboxylation, 77, 104
oil of citronella, 346 Opuntia ficus-indica, 582 oxidative demethylation, 663
oil seeds, 19, 336, 603 Oraesia, 591 oxidative desaturase, 22
oils, 16, 18, 19,25-28,35,42,43,45,55, orange, 313, 315, 340, 381, 473, 482-484, 496 oxidative pentose cycle, 248
109, 122, 137; also see castor oil, cedar leaf Orcbidaceae, 147, 148, 337, 343, 385, 393, oxidative phosphorylation, 187, 231,419,601,
oil, coconut oil, cottonseed oil, croton oil, 510, 537, 549, 560, 581, 670, 693 640
edible oils, essential oils, linseed oil, orcbids, 147, 149, 343, 344, 350, 352, 381, oxidoreductase, 109, 115
mustard oils, oil cells, oil glands, oil of 395,670 2,3.oxidosqualene:cycIoartenol cyclase, 433
citonella, oil seeds, orris root oil, rosewood orchinol, 147 2,3·oxidosqualene:lanosterol cyclase, 433
oil, seed oils, tung oil Orchis militaris, 147, 148 3-oxoactinidiol, 500
Olacaceae, 45 orcinol, 73, 147-149 oxoaporphine alkaloids, 588, 591
old house borer, 340 ordeal ceremonies, 463; see also trial by ordeal 10-oxogeranial, 354, 355
Oldenlandia, 700 Oreina cacaliae, 552 2-oxoglutarate, 158, 159, 162,202,226
Olea europaea, 98 organeUes, 19, 40 10-0xoneral, 355
Oleaceae, 32, 98, 361, 458 organic acids, 3 12-oxophytodienoic acid, 32, 39
OleaIes, 360, 361 organisms. see eukaryotic organisms, marine 100oxosparteine, 556
oleander, 98 organisms, microorganisms, parasitic 17 ·oxosparteine, 556, 559
oleandrin, 468 organisms, prokaryotic organisms, soil oxyanthin, 277
L-oleandrose, 251 organisms oxyanthin 5"-benzoate, 277
oleanolic acid, 447. 459, 460 Oricia, 573 oxygen, 13, 19, 21, 22, 24, 26, 27, 42, 44, 58,
olefin, 4, 20, 23, 35, 332, 333, 370, 373, 374, oridonin, 419, 420 63,67,74,94, 126, 152, 155, 156, 158, 159,
402, 435, 486 orientalidine, 594 162,206,226,231,266,276,290,294,315,
oleic acid, 17, 18,21-24,26,27,30,35, orientaline, 587 327,332,333,370,387,398,430,431,442,
42-44, 52, 53 ( - )·orientaline, 592 463,473,476,490,492,587,600,601,676
oleoresin, 338, 343, 398 orientalinone, 592 oxygen, molecular, 22, 26, 27, 42, 43, 107,
oleyl-ACP, 21, 23 ( + )·orientalinone, 592 108, 155, 158, 159, 178, 290, 303
oley1-CoA, 22, 23, 26 orienrin, 162 oxygen, singlet, 49, 640, 642
1·oleyl-2-palmitylphosphatidic acid, 23 Ormosia, 557 oxygenation, 46, 56, 131, 133
oligomeric proanthocyanidin, 201 ornithine, 220, 221, 226, 507, 517, 521, 525, Oxylobium, 29
oligophagous, 340, 380, 460 529,531,532,534,537,545-548,564,708 oxyresveratrol, 142, 145
oligosaccharides, 208, 244, 247, 249, 254, 256, ornithine carbamoyltransferase, 238 oxytocic, 600, 650
257,261,262,270,271 ornithine decarboxylase, 517, 531, 548 Oxytropis, 253, 561
olivacine, 628, 645 Orobanchaceae, 78, 361, 362, 391, 392, 557 (- )·ozic acid, 415
 Index 745

Pachlioptera aristolochiae, 591 parasympatholytic drngs, 510, 692 Peganum harmala, 570, 660
Pachycereus, 580 parasympathomimetic drugs, 697 pelargonidin, 152, 163, 177
Pachycereus pringlei, 441 parenchyma, ISS, 280, 526 Pelargonium, 68
pachydictyol A, 420 parisin.376 Pelargonium X hortorum, 68
Pachysandra, 677, 686 parkeol, 444 pellotine, 579-581, 660
Pacific islands, 139 Parkinsonia aculeata, 222 "'-peltatin, 117, 120
Paederia scandens, 363 Parkinson's disease. 229 ,B-peltatin, 117, 120
paederoside, 363 Paropsis atomaris, 289 peltogynols, 200
paeonia lactiflora, 209, 346 parsley, 4, 14,48, 134, 137, 138, 153, 156, Peltophorum africanum, 218, 222
Paeoniaceae, 195 158, 188,251,261,270 Peltophorum inerme, 222
Paeoniae radix, 346 parsnip, 128, 135-138 penicillin G, 234, 235
paeoniflorin, 346 parsnip root, 137 penicillins, 234, 235, 568
paeony, 209 parsnip, wild, 7, 136-138 Penicillium, 56, 58, 59, 64, 74, 234, 658
Pagiantha, 637, 648 Parsonia, 549 Penicillium atrovenetum, 290
paints, 16 parthenin, 388, 390, 392 Penicillium citrinum, 63
PAL, see phenylalanine ammonia lyase Porthenium, 109, 313, 319, 320, 390, 395 Penicillium digitatum. 520
Palaquium gutta, 321 Parthenium argentatum, 109,313,319,320, Penicillium griseofulvum. 56, 64
Palicourea, 664 517 Penicillium potulum, 56, 58, 59
Palicourea fendleri, 665 Parthenium hysterophorus, 389, 392 Penicillium urtieae, 59
palmatine, 600 parturition, 660 Penstemon, 177
palmitic acid, 17, 22, 23, 26, 30, 35, 53 passibiflorin. 285, 286 Penstemon barbatus, 364
palmitoleic acid, 17, 18, 67 passicapsin, 285, 286 Penstemon virgatus, 364
palmilOyl-ACP, 19, 21, 23 passicoriacin, 285, 298 Penstemon whippleanus. 669, 690
pabnitoyl-CoA,21 Passiflora edulis, 287, 500, 660 penstemonoside, 669
I-palmitoyl-2,3-diolein,35 Passiflora species, 10, 282, 285, 286, 289, 660 Pentaceras, 662
I-palmitoyl-2-linoleoyl-3-olein, 35 Passiflora warmingii, 282 1,2,3,4,6-pentagalloylglucose, 196, 197
palmityl-ACP, 23 Passifloraceae, 187,285-287,296-298,660 ,B-pentagalloylglucose, 193, 194, 196
patins, 25, 52, 115 passion fruit, 500 ,B-penta-O~galloyl-D-glucose, 196, 197,207,
palustrine, 521 passisuberosin, 285 210
Panama, 209, 344 passitrifasciatin, 285, 286 pentaketide, 59, 63
Panax ginseng, 460 Pastinaca sativa, 7, 136, 137 pentanortriterpenes, 473, 477
Panax quinquefolium, 48, 460 a-patchoulene, 373 peonidin, 177
Pandaca, 637, 647, 648 ,B-patchoulene, 373 pepper, 110,328,351,377,415,494
Pandaceae, 700 y-patchoulene, 373 pepper, ashanti. 538
Pangium edule, 285 patchouli, 373, 376, 384, 393 pepper, black, 110,517,538
Papaver, 593, 605 patchouli alcohol, 384 pepper, long, 539
Papaver bracteatum, 593, 597, 603 P-patchoulin, 376 pepper, red, 486, 494, 504
Papaver orientale, 592 patchoulol, 373, 374, 393 peppennint, 328, 332, 335, 351
Papaver rhoeas, 603 ( - )-patchoulol, 373 peppery taste, 290
Papaver somniferum, 585, 587, 588, 590, patchoulol synthase, 374 peptide a1ka1oids, 657, 692, 698, 700, 711
592-596, 601, 603 pathogen, 9, 15,26,27,48,67,71,72,84,98, peptide bonds, 215
Papaveraceae, 286, 303, 458, 510, 557, 576, 128, 137, 164, 186, 188, 209, 237, 238, 245, peptides, 215, 234, 235, 237-242, 244-246,
581,587,592,598,600,601,603,605,610 261,270-272,288,319,340,342,368,376, 261, 597; see also dipeptides, ergot
Papaverales, 576 377,408,414,415,442,458, SIS, 543, 558, alkaloids, hexapeptides, lysergyltripeptides,
papavetine, 588, 596 561,682 peptide alkaloids, peptide bonds, tripeptides
paper, American, 383 pathway, see acetate-malonate pathway, peptides, chloroplast transit, 19
paper manufacture, 136, 145 biosynthetic pathways, eukaryotic pathway, peptides, cyclic, 235, 237
Popi/io, 123, 124, 136, 173, 179, 190, 191, glycolytic pathway, mevalonate peptides, y-glutamyl, 241
207,208,220,221,231,307,341 pathways, phenylalanine-cinnamic acid Peregrinis maidis, 288
Papilio brevicauda, 136 pathways, prokaryotic pathway, skildmic perfumery, viti, 12, 32, 109, 122, 346, 385,
Papilio glaucus canadensis, 123 acid pathway patulin, 5, 59, 70, 72 500,670
Papilio glaucus glaucus, 123, 124 Paullinia cupana, 700 Pergularia, 563
Papilio glaucus, 208 pavine alkaloids, 586, 605, 606 Peridroma saucia, 316. 479
Papilio polyxenes, 136,207,208,307 pawpaw, see Asimina triloba perilla, 335, 351
PapUio xuthus, 173, 191 pea. ISS, 165, 181, 182,220,447 Perillafrutescens, 280, 294. 326, 335, 351,
Papilionoideae, 179,220,221,231,549,553, peach, 175,277,288,290,346,415 447
557,558 peanut, 65, 145, 226 ( - )-perillyl alcohol, 335
papyriferic acid, 451 pear, 30, 381 periplanone B, 383
Parabenzoin trilobum, 380 pear, Japanese, 238 Peritassa, 452
Parachartergus aztecus, 53 peas. 559 periwinkle, 511
paracoto, 139 paean, 84 perloline, 510
Paraguay, 420, 696 Pecten maximus, 498 Peronospora tabacina, 377. 414
paralysis, 607, 643 pectenoxanthin, 498 Persea, 419
paralytic effects, 645 pectinases, 261 Persian lilac, 479
Paramecium caudatum. 564 pectins, 170,247,259,260,262,270,402 perspiration, 664
parasites, 40, 53, 92, 183, 227, 391, 392, 394, Pedaliaceae, 118,361 Peru, 114, 535
424,452,533,545,551,559,564,566,614, Pedicularis, 552, 559 Peschiera, 637
650,652 Pedicularis bracteosa, 552, 559 pesticides, 12, 73, 111, 671
parasitic organisms, 10,41,67,73,78,92, Pedicularis crenulata, 552, 559 Petalostylis, 660
364,367,391-393,452,454,466,479,546, Pedicularis groenlandica, 552, 559 petals, 29, 91, 173, 177,231,328,494,498,
552, 554, 564, 566, 576, 652, 659, 660, 666, Pedicularis racemosa, 552, 559 499
689,690 Pedieularis semibarbata, 559, 566 Petasites, 553
parasilOids, 31, 32, 39, 41, 343; see also pedunculagin, 196, 197 petiole, 153, 276
ichneumonid parasitoids peduncularine, 563 Petiveria, 708
parasorboside, 269 Pegonum, 569, 570, 576, 660 petroleum, 16, 51, 53
746 Index

petroselenic acid, 26 L-phenylalanine, 178, 303, 274, 277, 304 photophosphorylation, 525
Petroselinum, 14, 133, 136 phenylalanine ammonia lyase, 4, 107, 130, photorespiration, 6, 7
Petroselinum crispum. 133, 155. 156, 261, 262 133, 155, 156, 184, 186, 195,509 5-phosphoribosyl-pyrophosphate transferase, 98
Petroselinum hortense, 14, 133, 153, 155,251 L-phenylalanine ammonia lyase, 107 photosensitizers, 49, 50
Petteria ramentaeea, 559 phenylalanine decarboxylyase, 515 photosynthesis, 3, 6, 8, 76, 96, 145. 166, 248,
Petunia, 159, 162, 163,202,269,437,453 phenylalanine hydroxylase, 102 266,338,414,486,488,496,497
Petunia hybrida, 162, 163 phenylalanine tRNA, 558 photosynthetic photophosphorylation, 601
petuniasterone A, 437 phenylalanine-cinnamic acid pathway, 209 photosystem I, 496
petunidin, 167 ,B-phenethylamine, 244, 581 photosystem n, 167, 496
Peucedaneae, 134 phenylethylamines, 514, SIS, 517, 581 photosystem n reaction center P680, 496
peyophorine, 581 ,B-phenylethylamines, 58 I phototaxis, 497
peyoruvic acid, 579, 580 2-phenylethylglucosinolate, 303 phototoxic activity, 49, 92, 93,136,317,573,
peyote cactus, see cactus, peyote l-phenylhepta-I,3,5-triyne,49 660
pH, 8, 9, 99,170,171,207,208,217,276, ,B-phenylnitroethane, 290 phototoxins, 134
315,374,483,507,518 phenylpropanoid glycosides, liS Phratora, 124
Phaeelia crenulata, 336 phenylpropanoids,4, 11, 13,54,78, 102, 106, phthalideisoquinoline alkaloids, 600, 603, 610,
Phaedon cochleariae, 309, 311, 445 109,110,111,125,127,136,190,193,324, 615
phaenthine, 606 339, 538, 570 phycology, 12
Phaeophyceae, 260 phenylpropionic acid, 165 Phycomyces, 501
phagocytosis, 591 phenylpyruvic acid, 101, 102, 106 Phycomyces blakesleeanus, 502
phagorepellent, 597, 682, 689 2-phenyl-5-(I'-propynyl)-thiophene,49 phycomycete, 376, 444
phagostimulant, 35, 207, 288, 551 pheromones, 16,32,34,36,37,39,41,111, phyletic problems, 337
Phalarts arundinacea, 514 122, 124, 125, 128, 324, 339, 342-344, 352, Phylica, 610
Phalaris tuberosa, 514, 515 363,365,367,376,383,394,486,501,502, Phyliea rogersj;, 585
phalloidin, 237 542, 551, 552 PhylIanthoideae, 286
phalloin, 237 pheromones, alarm, 324, 339, 343, 383 Phyllanthus, 537
phallotoxins, 237 pheromones, sex, 343, 363, 365, 505 Phyllobates, 677
Phalfus impudicus, 174 Phidippus, 49 phyllocladene,412
pharmaceuticals, 12, 535, 653, 655 phloem, 257, 262, 320, 465, 468, 472, 547, Phyllodectina, 124
phaseic acid, 501 554, 559, 567 phylloquinones, 80, 82, 86, 488
phaseollin, 181-183, 186 phloridzin, 174 Phyllosticta, 72
phaseollinidin, 182 phloroglucinol, 57, 68, 69, 73, 149, 174, 187, Phyllotreta annoraciae, 168
phaseollinisoflavan, 186 200,210 Phyllotreta cruciferae, 308
phaseolotoxin, 238 phlorotannins, 210, 213 Phyllotreta nemorum, 445
Phaseolus, 146,313,341,488,501 Phoebe brenesii, 110 Phyllotreta strio/ala, 308
Phaseolus aureus, 222, 241 Phoma, 378 Phyllotreta tetrastigma, 445
Phaseolus lunatus, 287, 296, 341 Phoradendron flavescens, 244 Phyllotreta undulata, 445
Phaseolus multij1orus, 241 Phoradendron vi/losum, 244 phylogenetic studies, 337, 350, 367, 395, 405,
Phaseo/us radiatus, 313 phorbol, 402, 425 413,485
Phaseolus vulgaris, 156, 167, 183, 184, 190, phorbol esters, 402, 403 phylogeny, viii, I, 11, 12, 164,209
241,242,244,245,261,288,289,488 Phormia, 597, 640, 703 phylogeny, plant, viii, 164
phasin,244 Phormium, 444 physiological activity, 506, 647
phellandrene, 346 Phorodon humuli, 363 physiological effects, 510, 537, 581
a~phellandrene, 344, 345 phosphatases, 330, 488 physiology, plant, 12
,B-phellandrene, 342 phosphatidic acid, 22-24 Physostigma venenosum, 663, 664
Phellinaceae, 610, 625 phosphatidic acid phosphatase, 23 physostigmine, SID, 511, 537, 655, 663, 664
Phelline, 610, 625 phosphatidylcholine, 18, 22-24, 26, 39, 208, phytic acid, 264
Phellodendron, 575, 610 430 phytin, 264
phenacylpyrrolidines, 564 lyso-phosphatidylcholine, 26 phytoaiexins, 8,42,48,72,74, 130, 135, 137,
phenanthrenes, 146, 147, 149,588 phosphatidylethanolamine, 26 138,145,146,148,165,174,178,179,
phenanthroindolizidine alkaloids, 546, 563, 564 phosphatidylglycerol, 17, 19,23 182-184, 186, 188, 261, 262, 309, 367,
phenanthroquinolizidine alkaloid, 546, 564 3'-phosphoadenosine-5'~phosphosulfate 376-378,393,395,397,398,401,408,414,
phenet~yl alcohol, 113 (PAPS):desulfoglucosinolate 415,423,426,510
phenethyi alcohol glycosides, 267 sulfotranserases, 303 phytochelatins, 234, 240, 241
phenethylisoquinoline precursor, 626 phosphodiesterase, 588 phytochrome, 171
phenol, 124 phosphoenolpyruvate, 6, 94, 96, 97 phytodienoic acid, 32, 39
phenolic carboxylic acids, 194 6-phosphogluconate dehydrogenase, 132 phytoecdysteroids, 442, 444
phenolic compounds, 8, 13, 54, 56, 58, 72, 78, 6-phosphogluconic acid, 253 phytoene, 431, 486, 488, 490, 492, 494, 496,
79,108,113,114,120-122,124,125-129, phosphoinositides, 46 499,504
142,170-173,178,187,188,193,194,203, phospholipase, 24 Z-phytoene, 488, 490
206,209,210,212,213,232 phospholipids, 16,17,320 15~Z-phytoene, 490
phenolic glycosides, 124 (5R)-phosphomevalonate, 315 15,15'~E-phytoene, 488, 490
phenols, 170, 171, 190, 193,210,212,213 phosphomevalonate kinase, 315 15,15'-Z-phytoene, 488, 490
phenotypes, 9 N-(phosphonomethyl)glycine, 195 phytoene dehydrogenase, 501
phenyl acetate, 113 phosphopantotheine site of ACP, 19,21 phytoene synthase, 488
phenylacetaldehyde, 113 4'~phosphopantetheine prosthetic group, 19 E-phytofluene, 490
phenylacetic acid, 106, 165 phosphorus, 6 Z~phytofluene, 490
phenylacetothiohydroximate, 303 phosphorylases, 259 phytogeographic distribution, 209
phenylalanine, 3,4,94,96,97, 101, 102, phosphorylation, 315, 518, 520; see also phytohonnone, 33. 98
105-107,118,121,128,130,142,147, 155, dephosphorylation, oxidative phytol, 398, 399, 414, 423
156, 187, 195,203,208,209, 229, 273, 277, phosphorylation, photophosphorylation, Phylolacea, 459, 460, 708
280,286,287,297,300,302,507,509,513, photosynthetic oxidative phosphorylation Phytolacea americana, 244, 460
522,535,540,564,568,617,619,624-626, photocarcinogenic substances, 137 Phytolaeca dodecandra, 459
657,694 photodennatitis, 137,336 Phytolaccaceae, 244, 458-460, 521, 575, 708
D-phenylalanine, 234 photooxidation, 496, 499 phytophagous, 10, 12, 13, 129
 Index 747

Phytophthora drechsleri, 48 pine, Virginia, 257 planlains, 514

Phytophthora infestans, 377, 684 pinidine, 543 plantamajoside, 108
Phytophthora megasperma, 178,261,270 ( - )-pinidine, 543 plant-animal interactions, 110, 379
Phytophyta, 242 ( - )-pinidinol, 543 plant-fungal interactions, 376
Phytoseiulus persimilis, 341 D-pinilol, 264, 265 plant-herbivore interactions, 546
phytoslerols, 427, 433, 435-437, 440, 441, D-( + )-pinitol, 264 planthoppers, 173
447,458,582 L-( - )-pinilol, 264 plant-insect interactions, 49. 88,111.118,122,
phytotoxic compound, 175,237 ( - )-pinoresinol. 117 591,597
phytoloxins, 56, 71, 72, 84, 126-128,237, ( + )-pinoresinol, 117 plant-microbe interactions, 261, 272
238,351,367,377,378,398,415,422,423, pinosylvin, 140, 142, 145 plant-plant interactions, 76, 78, 122
459,531,543,562 (+ )-pinpollitol, 264, 265 plants, see aphid host plant compatibility, cell
phytotoxins, sesquiterpenoid, 378, 379 Pinus, 52, 136, 140, 337, 340, 370, 413 culture. cell walls, defense, dicotyledonous
phytOluberin, 377 Pinus banksiana, 338 plants, eggplant, host plants, insect-host
phytyl side chain, 81 Pinus contorta, 338 plant relationships. interplant
Picea, 140, 337 Pinus jeffreyi, 52, 337, 543 communication, monocotyledonous plants,
Picea breweriana, 337 Pinus pinaster, 368 honnones, phylogeny, physiology. plant
Picea engelmanU, 337, 543 Pinus ponderosa, 337, 343 animal interactions, plant fungal interactions,
Picea glauca, 337 Pinus resinosa, 142 plant herbivore interactions, plant insect
Picea mariana, 337 Pinus sabiniana, 543 interactions, plant microbe interactions, plant
Picea pungens, 337 Pinus sylvestris, 142. 340 plant interactions, sterols, sympetalous
Picea rubens, 337 pioneer species, 7 plants, systematics, thermogenic plants,
Picea sitchensis, 337 pipecolic acid, 217-219, 538, 561 tissue culture, tumorigenic plants
piceatannol, 142, 145 L-pipecolic acid, 217, 232, 538 plants, Chinese, 135, 168, 522, 528, 537, 544,
a-picolinic acid, 543 Piper, 36, 104, !lO, !l3, !l4, 538, 539 570, 593, 606, 648, 658, 670, 673, 686, 708
Picralima,640,641 Piper betle. 528, 538 plants, dioecious, 176
Picralima alkaloids, 628, 640, 641 Piper jutokadzura, 113, 118 plants, flowering. 85, 123
Picralima nitida, 641 Piper guineense, 538 plants, herbaceous, 152
picrasin A, 476 Piper hookeri, 104 plants, sympetalous, 108
Picrasma, 660 Piper iongum, 539 plants, thennogenic, 123
Picrasma javanica, 660 Piper methysticum, 136, 139, 140, 538 plants, tumorigenic, 666
Picrasma quassinoides, 473 Piper nigrum, 110, 114,517,538 Planynota flavedana, 92
picric acid, 273, 293, 294 Piperaceae, 36, 104, !lO, !l3, 121, 139, 514, plasma membranes, 574
picropodopbyllotoxin, 120 517,538 plasmid, Ri, 239
Picrorhiza, 444 Piperales, 610 plasmid, Ti, 47, 239
picrotoxinin, 385 pipercide. 114 Plasmodium berghei, 69
picroloxins, 367, 385, 387, 670 piperenone, 113 Plasmodiumfalciparum. 482, 599, 606, 607,
Pieris, 307, 308, 597, 641, 689 .JI-piperideine, 508, 556 614, 625, 640, 649, 651, 652, 660, 666
Pieris brassicae, 307-309, 650 .JI-piperideine carboxylic acid, 538 Plasmodium yoelii, 574. 625
Pieris japonica, 412 piperidine, 340 plastid., 8, 19,94, 106, 107,313,326,401,
Pieris rapae, 307-309, 468, 470, 472 piperidine alkaloids. 525, 529-531, 533, 411,488, 490, 494, 528, 708; see also
pig, 35 537-540, 543-545, 554, 561 proplastids, sieve-tube
pigmen~ 78, 80, 82, 83, 85, 91, 135, 152, 164, .J1-piperidine-2-carboxylic acid, 217 plastids
171, 174, 177, 189, 192,326,380,413,486, .J1-piperidine-6-carboxylic acid, 217 plastoquinones. 76, 86, 488
491,494-499,502,504,506,568,705,708, piperine, !lO, 253, 539, 543 plalydesmine, 573
709 piperlongumine, 539 Platyneris, 420
pilocarpine, 5!l, 692, 693 piperlongwninine, 539 Platyneris dumerilii, 420
Pilocarpus, 692 piperonal, !lO, 344 plalyphylloside, 121, 129
pilocereine, 582 piperonyl butoxide, 110 Platyptilia, 669
Pilocereus, 580 pipoxide, 104 Platyptilia pica, 364, 366, 552, 566
pimara-8(9),15-diene, 407 Piptadenia, 514 Plebejus icarioides, 559
9-,BII-pimara-7,15-diene, 414 Piqueria trinervia, 345 Plenckia, 452
pimaranes, 405, 406, 412 piquerol A, 345 plenolin, 393
pimaric acid, 406 piqnerol B, 345 PleurostyUa africana, 522
Pimelia prostrata, 403 pisatin, 181-183 plumhagio, 63, 64, 80, 84
Pimpinella, 109 (-)-pisatin, 181-183 Piumhaginaceae, 63, 80, 82, 197,571,708
Pimpinella anisum, 122 piscicide., 12, 120, 186, 209, 212, 390, 456, Plumbago, 63, 80
pimpinellin, 133, 135 460 plumeran, 636, 637
Pinaceae, !l7, 136, 173,338,405,413,543 Pissodes strobi, 340 Plumeria obtusifoUa, 361
pine, 315, 375, 393, 543 Pistacia ientiscus, 447 Plumerieae, 637
pinene, 346 pistil,269 Plumerioideae, 636
a-pinene, 331, 340-345 Pisum sativum, 182, 183,220,231,293,369. plumieride, 361
(- )-a-pinene, 331, 342, 343 708 plum., 290
(+ )-a-pinene, 331, 342, 343 Pithecellobium, '127 Plutella "1lostella, 308, 309, 311, 468, 472
d-a-pinene, 331 Pithecellobium lobatum, 227 pluviine, 620
l-a-pinene, 331 Pithecellobium saman, 521 Poaceae, 68, 98, 99, 105, 107, 118, 125,230,
,B-pinene, 331, 340-345 pithecolobine, 521 257,282,285,286,321,346,459,510,522,
(- )-,B-pioene, 331 Pittosporaceae, 45, 46, 134, 337 549, 659, 700
I-,B-pinene, 331 Pituranthos triadiatus, 136 Poanes hobomok, 362
( - )-pinene cyclase I, 331 Pityrogramma, 174 Podalyrieae, 557
(+ )-pinene cyclase I. 331 Plagiochila ovalifolia, 376 Podocarpaceae, 173, 179,405,420,442
d-pinene cyclase, 331 plagiochitin, 376 Podocarpus, 179,405, 420
i-pinene cyclase, 331 plant sterols, see phytosterols Podocarpus elata, 442
pine, jack, 338 Piantaginaceae. 304, 361, 363 Podocarpus !errugineus, 405
pine, lodgepole, 338 Plantago lanceolata, 363 Podocarpus macrophyllus, 405
pine, ponderosa, 343, 420 Plantago major, 358 Podocarpus nubigena, 174
748 Index

podogenin, 463 Polypodium vulgare. 442, 453. 462 primitive angiospenn families, 19
podophyllotoxin. 117. 118. 120. 128. 618. 676 polypodogenin, 463 Primula, 264
Podophyllum. 117. 118. 120. 128 polypodoside A, 462 Primulaceae, 264, 444
Podophyllum hexandrum, 118, 128 Polyporaies. 46 Pristiphora erichsonii, 415, 425
Podophyllum peltatum, 117 polysaccharides. 3. 244. 247. 252. 256. 257. Pristiphora geniculata, 289
pods. 174. 182. 193. 194.203.206.324.340. 259-262. 270-272 proacaciberin, 282
349.420. 439. 468. 554. 608 polysaccharides. algal, 247, 260, 271 (S)-proacaciberin. 282
Poekilocerus bujonius, 468 Polystictus versicolor, 91 proanthocyanidin biosynthesis, 164
Pogonopus, 612, 638, 652 polyvinylpyrrolidone, 208 proanthocyanidin dimers, 206
Pogostemon cablin, 373, 393 pomegranate, 539 proanthocyanidins. 164. 191. 193. 198. 200.
Pogostemon heyneanus, 373 ponasterone A. 444 20\.203. 204. 206-210. 212. 213
Pogostemon patchouli, 373, 671, 672 Poncirus, 337 (2R.3S)-proanthocyanidins. 201
poison. 506. 507. 511. 514. 517. 537. 543. poplars. 257 proaporpbine alkaloid. 591. 592
544. 546. 552. 553. 559. 561. 563. 564. 583. poppies. 593, see also opium poppies Procavia capensis syriaca, 136
584.588.596.603.607.617.641.646.658. Populus. 122-124. 128 procevine, 684
659,666; see also arrow poison, fish poison, Populus deltoides, 124 procyanidin dimers, 203, 204, 206
mitotic poison, nondepolarizing Populus grandidentata, 124 procyanidin oligomers, 203
neuromuscular poison, poison hemlock, Populus tremuloides, 124 procyanidins. 193.200.203.204.206
poison ivy, rat poison Poranthella, 542 2,3-cis-(2R,3R)-procyanidins, 206
poison hemlock, 543 Poranthera corymbosa, 554 prodelphbtidins. 200. 206. 2\0
poison ivy, 67 pore, see sieve-tube pores Prodenia, 591
poison-dart frog, 677 Poronia punctata, 71 Prodenia eridania, 289
poisoning. 10. 12. 32. 229. 237. 239. 244. 254. Porphyra, 260 profisetinidins, 206
290.294.306.317.390.403.412.447.466. Portulacaceae, 708 (2R)-profisetinidins. 206
468. 514. 540. 546. 552. 553. 559. 561. 596. postpartum bleeding, 420 (2S)-profisetbtidins. 206
603.607.646.658.659.676.684.697 postpartum contractions, 660 progestagens. 427. 433. 437. 439. 463
poisonous compounds, 417, 223. 232, 235, postpartum hemorrhage, 570 progesterone, 439, 462, 465
237.244. 246. 288. 290. 402. 403. 439. 441. potassium. 16.260. 300. 307-309. 348. 517 progoitrin. 307. 308
452.466.506-507.517.537.544.552.559. potassium chloride (KCn. 9 epi-progoitrin, 307
583.584.617.641.673.674.676.689.705 potassium pennanganate, 535 proguibourtinidin, 206
Polemoniaceae, 177. 189,444 potato. 14. 31. 33. 39. 41. 99. 149. 166. 245. prokaryotic organisms. 4, 5, 18, 19,234
Polistes metricus, 53 313.314.345.352.372.377.383.397.651. prokaryotic organisms, thennophilic. 26
pollen. 30. 39. 176. 352. 389.437.498.515 677.682-684 prokaryotic pathway, 16, 19, 22, 23
pollen tube. 132. 389. 439 potato blight. 269 prolactin release, 640. 659
pollen tube growth. 389 potato leaves, 682 proline. 207. 208. 213. 215. 217. 219. 222.
pollination. vii. 9, 26. 30. 151, 165. 171, 176, potato peels, 684 226. 232. 657. 705. 708
177, 189.343.344.352.362.385.393.515. potato peels, fried. 684 prolycopene. 491. 494
516. 530. 550 potato sprouts. 684 prolyl t-RNA synthetase, 222
pollinators, 152, 176, 177 potato vines, 684 promelacacidins, 206
polyamine alkaloids. 520. 521. 529 potatoes, cull, 684 promOlphinanes, 592
polyamine biosynthesis, 518 potatoes, "greened," 684 promoter, CaMV 35S, 629
polyamines. 517. 518. 520. 521. 564 potentillin. 197 promoters. 514
polychaetes. 381. 420 potions, 506 propanaJ. 513
PolygaJaceae. 149. 458 PR transferase, 98 syn-propanethiai S-oxide, 227
polygodiaJ. 379 preakuammicine, 632, 634 S-(prop-l-enyl)cysteine, 227
Po1ygonacese. 76. 85. 86. 88. 162. 187. 197. precipitation reactions, 507 propionaldehyde. 227
267. 379. 387. 543. 708 precocenes. 312, 316-318 propionyl-CoA. 27. 65
polygonaquinone, 76 predators. 343. 361. 382. 447. 466. 470. 498. proplastids. 18. 19.22.26.401
Poiygonatum ofjicinalis, 222 551.552 S-n-propy1cysteine. 227
Polygonum. 76. 206. 267. 379 pregnenolone. 439. 465. 686 prorobinetinidin, 206
Poiygonum stagninum, 209 prehelminthosporai, 378 Prosopis, 218, 543, 562
Polygonum tinctorium. 267 prelnnularic acid. 143 prostaglandins. 29. 32. 34.40. 123. 167.420
polyhydroxylated tropane alkaloids. 535 prenyl pyrophosphate hydrolases. 488 Proteaceae. 68. 82. 286. 537
polyhydroxypiperidine alkaloids, 531, 544 4'-prenyloxyresvemtrol, 145 proteacin, 280
polyisoprene, 318-322 prenylated proteins 321 protease. 242
Z-l.4-polyisoprene, 319 prenylation. 82. 86. 87. 132-134. 173. 182. protein. 156. 158. 167. 184. 193.206-210.
polyisoprene E-allylic diphosphate initiator. 186 212.213.215.217.220-222.231.232.234.
319 3-prenylindoles, 663 240. 242. 244-246. 259. 261. 272. 276. 280.
polyketide biosynthesis, 58, 139, 142 prenyltransferase. 316. 319. 322. 326. 329. 300.312.320-322.371.373.444.490.492.
polyketide derived alkaloids, 543 368-370 498. 503. 532. 549. 557. 561. 709. 710; see
polyketide synthetase, 57 prephenate aminottansferase, 106 acyl camer proteins, ADP/ATP carrier
polyketides. 41. 56-72. 74. 76. 77. 85-87. 91. prephenate dehydrataae. 101. 177 protein, chromoproteins, flavoproteins,
92. 121. 139. 148. 155.237.507.538.543. prephenate dehydrogenase, 173 glycoproteins, jasmonate-induced proteins,
698 prephenic acid, 101, 102. 104, 106 fJ-.k.etoacyl acyl carrier proteins, microtubule
polymeric F-actin, 237 prephytoene. 348. 488 protein polymerization, non-protein amino
polymerizatioo. 5.115.117.206.237.262. prephytoene pyrophosphate, 488, 490 acids, prenylated proteins, proteinase
319.327,370 presquaIene, 350, 430 inhibitors, proline-rich proteins, protein
polymorphism. 7. 296. 298 presqualene alcohol, 348. 350, 430, 455 amino acids, toxic proteins
polymyxin B. 234. 235 presqualene pyrophosphate, 428, 430, 431, 488 protein amino acids, 215, 216, 226, 231, 232,
polymyxin D, 234 press cake. 244, 307 303
polyneuridine aldehyde. 640 pre-tazettine. 621, 623 protein biosynthesis, 613, 623
polyols. 247. 262. 264 prickly pear. 582 protein synthesis. 323. 452. 482. 520. 623.
polypeptides, 19 primeverose. 254, 266, 280 625. 627. 645. 676. 696. 700
polyploidy. 5. 588. 597. 619 2-fJ-.primeverosyloxy-2-phenyl-2S-acetonitrile. proteinase inhibitors, 33, 39, 40, 245
Po/ypodium glycyrrhiza. 463, 471 277 proteins, prenylated, 321
 Index 749

proteins, proline-rich, 207 Psychotria beccarioides, 665 pyruvate dehydrogenase, 19

prothoracic glands, 439 psychotridine, 665 Pythium aphanidermatum, 47
protoalkaloids, 513 Psychotrieae, 612, 638 Pythium ultimum, 26
protoanemonin, 269 psychotrine, 613, 616
protoberberine, 586-601, 610 Ptaeroxylaceae, 478, 575, 576 quadrigeminine A, 665
proIoberberine alkaloids, 586-601 Ptaeroxylon, 478 quail, 179,517
protoberberine alkaloids, quaternary, S99 ptaquiloside, 375 Quassia amara, 482
prolOcatechualdehyde, 619-621 Ptectinophora gossypiella, 169 quassimarin, 482
protocatechuic acid, 96 Pte/ea, 571, 573, 574 quassin, 473, 477
protohyperlcin, 91 Pteiea trifoliata, 571 quassinoids 473, 476-478, 482, 484, 485
protolimonoids, 473 Pteridium aquilinum, 414, 442 Quebec, 378
proton extrusion, 415 pterocarpans, 179, 181, 182, 184, 186 ( - )-qoebrnchamine, 635
proton pumps, 520 Pterodon pubescens, 420 ( + )-quebrachamine, 635
protopine, 594, 600, 601, 603, 610, 616 Pterophoridae, 364 L-( - )~quebrachitol, 264
protopine alkaloids, 600, 601 Puccinia coronata, 98 quebrncho, 204, 208, 213
protoplasts, 2, 9, 15, 518 pulchellidine, 672 quercetin, 152, 159, 160, 166, 167, 195
prolOSlepbenine, 592, 613 pulchellin, 672 quercetin 3-glucoside, 169
protoveratrine A, 684-686 pulegone, 333, 345 quercetin 3-0-galactoside, 167
protoveratrine B, 684-686 (+ )-pulegone, 333, 345 quercetin 3-rhanmoside, 169
protoverine, 684, 686 Pulsatilla, 269 quercetin glycosides, 172
protoverine, polyesters, 684, 686 pumiliotoxin A, 560 D-quercitol, 264
protozoans, 392, 431 pumiliotoxin B, 560 proto-quercitol, 195
prnnasin, 272, 217, 279, 280, 286-289, 294, pumpkin, 33, 315, 370, 399 scyllo-quercitol, 195
297 pungeot principle, llO, ll4, 300, 302, 452, Quercus, 195, 264
(R)-pronasin, 277, 280 517,530,538 Quercus robur, 195, 196
prunasin 2'-glucoside, 277, 280 Punica granatum, 539 Quercus rubra, 252
pronasin hydrolase, 280, 296 Punicaceae, 539 Quercus suber, 447
prunasin-6-malonate, 277 pupa, 99, 440, 552, 591 quinazoline allraloids, 568-570, 575, 576, 610
Prunus, 269, 272, 274, 277. 280, 286, 288, 363 pupil dilation, 537 quinazolinocarboline alkaloids, 568, 571, 576
Prunus domestica, 280 purgative, 69,444, 445, 452 quinic acid, 94, 95, 99, 102, 104, 108, 121,
Prunus laurocerasus, 274 purine alkaloids 700 171, 195,210
Prunus maximowiczii, 269 pnrine bases, 507, 700, 702, 703 ( - )-quinic acid, 94
Prunus persica, 288 purine nucleoside phosphorylase, 702 quinic acid esters, 108
Prunus serotina, 277, 280, 287, 288, 295, 299 puromycin, 234 quioidine, 506, 5ll, 649-651
Prussian blue assay (tannins), 210 Purshia, 444 quinine, 10, 175, 361, 506, 5ll, 569, 571,
pseudoaconitine, 673 Purshia tridentata, 208 649-651, 653
Putranjiva roxburghii, 304
pseudoalkaloids, 668 quinine sulfate, 650
pntrescine, 510, 517, 518, 521, 526, 529, 531,
pseudocereals, 460 quinoline alkaloids, 568, 570, 571, 573, 575,
534,547-549, 565
Pseudococcus iongispinus, 561 610, 612, 615, 638, 649
putrescine-N-methyltransferase, 531
pseudocopulation, 344, 381 quinolizidine alkaloid biosynthesis, 554, 561.
Putterlickia verrucosa, 695
pseudocyanogenic glycosides, 273, 294 567
Pyracantha, 499
pseudoguaianes, 374 quinolizidine alkaloids, 2, 3, 364, 544, 546,
Pyralidae, 390
pseudohypericin, 91 pyranocoumarin, 137
552-554, 556-560, 564-567, 690, 709
pseudojervine, 686 pyranoquinoline alkaloids, 570, 571, 573 quinolizidine ring systems. 693, 694
pseudolycorine, 620, 623 quinolone alkaloids, 568, 570, 571, 573-575
pyranoquinoline alkaloids, linear, 570
Pseudomonas, 83 pynmoses, 247, 248 4-quinolone alkaloids, 568, 570, 571, 574
Pseudomonas syringae, 98, 237, 238 pyrazine alkaloids, 692, 703 quinonemethide structures, 63, 117, 212
Pseudomyrmex, 289 pyrazines, 703, 711 quinonemethides, pentacyclic nortriterpene,
Pseudomyrmex !errugineus, 53 pyrelhrins, llO, 346, 348, 430 452
d-pseudonorephedrlne, 522 Pyrethrum, 348 quinones, 8, 14,56,65,76-80,92,93,97, 104
pseudopelletierine, 539 Pyricularia oryzae. 414, 423, 543 ortho-quinones, 76
pseudopurpurio, 87 pyridine alkaloids, 513, 525 para-quinone, 77. 78
Pseudotsuga, 337 pyridoxamine-5~phosphate, 508
quinovose, 456
Pseudotsuga menziesii, 200, 338 D~quinovose, 251
pyrimidine bases, 135, 700
D-psicose, 250 Pyrolaceae, 361
Psila rosae, 113, 136 pyrolysis, 312 rabbit, 380, 451, 600, 646, 658, 674, 685
psilocin, 525 pyrooes, 155 Rabdosia, 408, 415, 419
Psilocybe, 525 2-pyrones, 139, 140 Rabdosia japonica, 432, 447
psilocybin, 513, 525 pyrophosphatases, 9, 330 races, 8
psilostachyin, 387 pyrophosphomevalonate decarboxylase, 315 radial diffusion method (tannins), 212
psilostachyin B, 387, 388 pyranoquinoline alkaloids, angular, 570 radical coupling, 64 , 82, 117
psilostachyin C, 388 Pyrrhallta luteola, 88 radicle, 78, 175, 317, 346, 390, 523, 526, 537,
Psilotales, 173 Pyrrhocoridae, 383 540, 558, 599, 601, 641, 650, 673, 697
Psi/otum, 173 Pyrrhocoris apteris, 383 radish, 300, 303, 306, 309, 314
PSMO inhibitors, see mixed function oxidase pyrrole esters, 552, 553 Radula variabilis, 174
inhibitors pyrrolidine alkaloids, 531-534, 538-540, 545 raffinose, 256, 261
PSMOs, see mixed function oxidases A1-pyrroline-5-carboxylic acid, 226 ragweed, 388
Psophocarpus tetragonolobus, 293 pyrrolizidine alkaloids, 321, 364, 466, ragwort, 550
psoralen, 133-135, 137, 138 546-554,559,564-567,683,703 Ranales, 576
psoralen synthase, 133 8a-pyrrolizidioe, 549 Rangifer tarandus, 207
psorospennin, 148 pyrrolizidine N-oxides, 546, 547 Rannnculaceae, 267, 269, 272, 277, 286, 290,
Psorospermum!ebrifugum, 148 pyrroloquinazoline alkaloids, 568, 569, 570 408,466,510,549,557,568,581,588,592,
psychoactive substances, 336 Pyrus serofina, 238 598, 600, 603, 605, 606, 610, 660, 673
psychotomimetic activity, 660, 694 pyruvate, 6, 19,97, 101,273,274,578-580, Ranunculales, 576, 610
Psychotria, 611, 612, 638 660,697 ranunculin, 269, 272
750 Index

ranunculoside, 269 resistance, disease, 174, 184,300,309 Rhizophora man~/e, 204

Ranuncu{us, 269, 363 resistant effects, 179, 183, 184, 193,207,210, Rhizophoraceae, 88, 197, 537, 549
rapanone, 77 221,242,269,288,308,309,526,535,607, Rhizopus, 658
rape, 30, 35, 306 614,652,659,660,676 Rhizopus stoionifer, 261, 401, 414
rapeseed, 307 resorcinol, 200 rhodanese, 290
Raphanus sativus, 306, 309, 345, respiration, 3, 4, lO, 16,59,92,99, 120, 166, rhodanine, 212. 213
raspberry, 110, 128,209 338,361,515,517,601,645, '103 Rhodnius prolixus, 479
raspberry ketone, 110 respiratory arrest, 597, 640 Rhododendron japonicum, 411
rat, see bush rats, rat liver. rat poison respiratory failure, 607, 674 Rhododendron mol/e, 412
rat poison, 641 respiratory paralysis, 675 rhodojaponin III, 412
Rathbunia alamosensis, 582 respiratory stimulation, 702 Rhodophyceae, 260, 260, 513, 515
rats, 29, 35, 132,209,460,553,591,641,651, resveratrol. 140, 142, 145 rhodopsin, 497
682, 685, 698, 702 E-resveratrol, 145 Rhodymenia palmata, 260
rattle box, 705 Z-resveratrol, 145 rhoeadane alkaloids, 603
rattle bush, 705 Retanilla, 610 rhoeadine, 603
Rauvolfia, 640 reticuline,9 rhombifoline, 556
Rauvolfia canescens, 33
(- )-reticuline, 561, 585, 595 Rhopalosiphum padi, 99
(+ )-reticuline, 561, 585, 592, 598, 600
Rauvoifia serpentina, 242, 354, 355, 366, 629, rhubarb, 76, 91
( - )-(R)-reticuline, 586, 592, 594, 595, 598,
640,653 Rhus, [95, 196,206
600
Rauvolfia tetraphylla, 640 Rhus typhina, 195, 196
(+ )-(S)-reticuline, 586, 588, 590, 595, 599,
Rauvolfia vomitoria, 640 ribalinine, 573
603
Rauvolfieae, 637 (R)-reticuline, 586, 592 ribose, 99, 250
reaction, see Diels Alder reaction, elimination, (S)·reticuline, 586-588, 590, 598, 600 D-ribose, 250
phytosystem IT reaction center, preciptiation Reticulothermes flavipes, 340 ribosomal subunit, 60-S, 623
reaction, transamination reaction retina, 497 ribosomal subunit, 80-S, 700
rearrangement, ally lie, 43 retinal, 497 ribosomes, 19, 318, 482, 488, 558
rearrangement, Claisen, 101, 106 E-retinal. 497 ribulose 5-phosphate, 253
rearrangement, dienol-benzene, 591, 592 l1-Z-retinal,497 ribulose-l,S-bisphosphate carboxylase, 208,
rearrangement, dienone-phenol, 591, 592, 608 E-retinoic acid, 497 213,242
rearrangement, E-Z (trans-cis), 131 retinol, 497 rice, 6, 14,33,35,52,53,70, 173,408,414,
rearrangement, merolimonol, 477 retronecine, 548-550, 552, 565 423,437
rearrangements, Wagner-Meerwein, 329 retrorsine, 548, 549 Richardella dulcijera, 245
Reboulia hemisphaerica, 368 (9S)-retrorsme, 548 Richardsonia,612
receptor, 510, 525, 591, 609, 649, 659, 660, retroviruses, 561 ricin, 244
664, 675, 698, 708; see also acetylcholine Retziaceae, 361 ricinine 513, 528
receptors, antennal reverse transcriptase, 521, 599-601, 616, 623 Ricinocarpoideae,412
receptors, dopamine receptors, glutamate reward,550 ricinoleic acid, 24, 26, 35
receptors, glycine receptors, muscarinergic reynosin, 392 ricinoleyl phosphatidylcholine, 24. 26
acetylcholine receptors, nicotinic Rhabdodendraceae, 197 ricinoleyl-CoA, 24
acetylcholine receptors, nicotinic receptors, Rhabdodendroideae, 575 Ricinus communis, 24, 26, 244, 261, 401, 407,
opiate receptors, postsynaptic receptors, Rhabdodendron, 197 408, 414, 424, 425, 528, 530
tarsal receptors Rhabdosia, 420 rimuene, 412
receptor, postsynaptic, 510 Rhagoletis cerasi, 37, 39 rinds, 337
red blood cells, 244 Rhamnaceae, 85, 86, 88, 149, 173,240,244, Rio Japuras, 588
reductant, 21, 22, 51 575, 581, 585, 610, 695, 700 rishitin, 376
reductive pentose cycle, 248 6'-0-( a-L-rhamnopyranosides of epivolkenin Rivea, 658
regulators, see growth regulator and taraktophyllin, 285 Rivea corymbosa, 655, 658
regulatory processes, 7 26-0-a-L-rhamnopyranosylpolypodogenin·3-0- RNA, 4, 237, 520, 618
Rehmannia glutinosa, 108 a-L-rhamnopyranosyl-(1-2)-/3-D- mRNA, 134, 156, 184,237
Reichsteinia cardinalis, 164 glucopyranoside, 462, 463 tRNA, 520, 558, 601
reindeer, 207 rhamnose, 172, 456 RNA polymerase, 521
Reinecke salts, 507 L-rhamnose, 247, 250, 251, 260, 456 RNA synthesis, 135, 452, 520, 599, 648, 676,
Rejoua,637 4(a-L-rhamnosyloxy)benzylisothiocyanate. 306 696
relaxant, see also muscular relaxant 7-a-L-rhamnosyl-6-methoxyluteolin, 168 rRNA transcription, 45 S, 648
Rhamnus, 85, 88, 610
relaxant, skeletal muscle, 136, 607 RNA-dependent DNA polymerase, 623
Rhamnusjrangula,91
relaxant, smooth muscle, 600, 702 RNA-polymerase IT, 237
Rhamnus purshianus, 91
religious practice, 139,647,697 roasted materials, 700, 703
Rhapheolepis, 206
Remija, 611, 649 Robillia pseudoacacia, 244
Rhazya stricta, 242, 629
repellents, 10,35, 169,212,309,324,339,
Rheum, 76, 85
robustaol A, 69
340,342,346,350,361,365,395,484,511, Rheum officinale, 91 rock hyrax, 136
522, 542, 582, 591, 682, 683; see mosquito rheumatism, 564, 673 rodenticide, J32
repellent, phagorepellent rhexifoIine, 364, 552. 669 rodents, 132, 136, 345, 380
repolarization, 685 rhipocephaJio, 381, 420 Rollinia,67
reproduction, 340, 395, 398, 419, 444, 455, rhipocephanal, 381, 420 Romans, 658
479,498,501,502 rhizobitoxin, 238 Rondeletieae, 612. 638
reproductive failure, 420 Rhizobium, 167, 191,244 root, 2, 5, 7, 8, 18,26,30,42,46-48,50,78,
reptiles, 511 Rhizobium japonicum, 238 83,85,92,99,104,109,111,117,125,127,
Resedaceae, 304 Rhizobium leguminosarum, 167 129, 130, 136, 139, 140, 152, 155, 165, 167,
reserpiline, 630 Rhizobium meUloti, 189, 190,261 183, 187, 190, 191, 193,209,231,238,242,
reserpine, 511, 628, 640, 647 Rhizoctonia, 253 256,257,261,270,290,291,310,311,313,
reserve carbohydrate, 257 Rhizoctonia leguminicola, 561, 565 320,335,345,346,351,353,364,391,396,
resins, 30, 40, [20, 136,3[8,324,339,340, Rhizoctonia repens, 148 411,413,419,423,439,447,452,457,460,
342,343,350,375,398,401,405,412,413, rhizoma iris, 452 462,468,479,482,490,492,494,526,528,
426 rhizome, 117, 147,364,444,452,462,463, 529,531,532-535,537,544-547,553,554,
resistance, 517, 607, 682, 689 471,600,670,681 559,561,562,566,588,597,601,608,610,
 Index 751

611,614,640,645,647,653,658,663,666, rutin, 164-167, 169 Santolina chamaecyparissus, 348

667, 673, 678, 700, 708, 709; see also rutinose, 266 santolinyl skeleton, 346
bloodroot. celery root, root cultures, ipecac Rutoideae. 571, 573. 575, 576 santonin, 392
root, root beer, root nodulation, root-rot Rwanda, 652 a-santonin, 393
nematode, rootwonn, white snakeroot Ryania speciosa, 418 sap, 514, 591
root beer, 110 ryanodine, 418 Sapindaceae, 46, 217, 222, 223, 231, 273,
root nodulation, 165 rye, 98, 99, 658-660 284-287,458
rootwonn, northern corn, 113 ryegrass, annual, 659 Sapium, 403
rootwonn, southern com, 113 Sapium japonicum, 403
rootwonn, western corn 113 sabinene, 332, 333, 338 Sapium sebiferum. 45, 50
raridins, 379 d-sabinene, 332, 333 sapogenins, 456-458, 470, 472, 678
Rorippa indica, 310, 311 sabinol, 333, 346 (25R)-sapogenins. 678
Rorippa nasturtium-aquaticum, 304 cis-sabinol, 333 (25S)-sapogenins, 678
Rosa, 265, 499 d-sabinyl acetate, 332 saponic alkaloids, 457, 460
Rosa canina, 242 saccharides, 244, 247, 261 saponic materials, 582
Rosa damascena, 326 saccharin, 462 saponification, 16
Rosa dilecta, 328 Saccharomyces cerevisiae, 245 saponins, 441, 447, 456-460, 462, 463, 468,
Rosaceae, 125, 134, 148, 175, 179, 197,208, Saccharum officinarum, 145 470-472
244,264,269,279,284,286,287,289,337, safflower, 48 saponins, bidesmosidic, 458
444,499,514,515,673 saffron, 499 saponins, ginseng. 460. 462
Rosales, 197 safrole,ll0 Sapotaceae, 245, 279, 318, 321,458,549,639,
rosane, 406, 412 safynol,48 703
rose hips, 265 sage, 319, 326, 330, 346, 371, 393 sapwood, 324, 591. 613
roselle, 171 sagebrush, 552, 559 Sarcocca, 686
rosenonolactone.412 sago, viii Sarcococca, 677
rosewood oil, 528 saguaro cactus, see cactus. saguaro Sarcopharyngia, 637
Rosidae, 277, 576 saikosaponin a, 460 D-sannentose, 250, 251, 465
Rosiflorae, 197 saikosaponin d. 460 sannentosin, 282
rosin, 398, 413 Salada, 452 sannentosin epoxide, 282
rosmarinic acid, 108 salannin, 479 Sarothamnus scoparius, 554, 559
rotenoids, 110, 151, 186-188,316 Salicaceae, 121-124 sarpagine alkaloid, 640
rotenone, 120, 186. 187 salicin, 121, 124 Sarracenia, 240, 543
roundwonns, 392, 539 salicortin,124 Sarraceniaceae, 360, 361
rubber, viii, 288, 296, 312, 318-323 salicylaldehyde. 124 sativan. 179, 186
rubber biosynthesis, 318, 319 salicylic acid, 104, IDS, 122, 123. 126, 129, sawfly, 340
rubber, natural, 318-321 650 sawfly, larch, 415, 425
rubber transferase enzyme, 319 saligenin, 122, 124 sawfly, mountain ash, 289
ruberytbric acid, 87, 89 saliva, 207, 208, 212, 561, 664, 697 sawfly, turnip, 418
Rubia akane, 700 salivation, 561, 697 Saxifragaceae, 88, 355, 359
Rubia cordl/olia, 700 Salix, 122, 124, 363 saxitoxin, 703
Rubia tinctoria, 85, 87, 89 Salpiglossideae, 537 scallop, giant, 498
Rubiaceae, 10, 76, 85-89, 108, 187, 229, 353, salts, see N-methyl-J'-pyrrolinium salt, Scarabaeidae. 179
358,360,361,363,405,412,444,458,481, Reinecke salts, sodium salts, lithium salts sceletenone, 624
510, 533, 579, 598, 610, 611, 615, 636-638, salutaradine. 592, 595 Scelelium, 623
640, 648, 649, 652, 653, 655, 660, 662, 664, salutaradinol I, 595 Sceletium alkaloids, 624
665, 700, 708 Salvadoraceae, 304 Scenedesmus, 494
rubijervine, 681, 684-686 Sa/via, 77, 345, 346, 371, 375, 394, 407, 413 scent glands, 670
Rubioideae, 612, 637 Salvia leucophylla, 345 Schelhammera, 625
RuBPC, 208 Salvia miltiorrhiza, 413 schelhammeridine, 626
Rubus idaeus, 209 Salvia offidnafis, 326. 330-332, 346, 371, Schiff base, 98, 497, 507, 509, 518, 531
Rudbeckia hirta, 125, 177 375,393 Schiff base formation, 507, 531, 620, 628
rugosin D, 208 Salvia sciarea, 407 Schiff-base intermediates, see intermediates,
rugosin E, 210 SAM, see S-adenosylmethionine Schiff-base
Rumex, 76, 85, 91, 363 samandarine, 677 Schinopsis, 204, 206
Rumex crispus, 91 Samanea saman, 521 Schinopsis balansae, 204
Rumex tenuifolius, 439 Sambucaceae, 361 Schinopsis lorentzii, 204
ruminants, 120, 340. 460 Sambucus, 499 Schistocerca, 686
RuspoJia hypercrateriformis, 533, 545 sambunigrin, 277, 279, 287 Schistocerca gregaria, 444, 460. 478-480
ruspolinone, 533 (S)-sambunigrin. 277 Schistosoma, 393
rust. see carrot rust fly. crown rust, rust sandalwood, 384 Schistosoma japonicum. 230
diseases. rust fungi, rusHed flour beetle sandaracopimaradiene, 407, 414 Schistosoma mansoni, 209, 420
rust diseases, 414 d-sandaracopimara-8(14)-diene, 408 schistosomiasis, 209. 380, 393, 420, 459, 460,
Ruta,574 d-sandaracopimara-8(14),16-diene, 408 612
Ruta graveolens, 133, 134, 138, 510, 571, 574 ent-sandaracopimara-8(14),15-diene.407 schistosomula, 230
rutabaga. 306 sandaracopimaridiene, 414 Schizachyrium scoparium, 125, 346
Rutaceae, 11, 12,36,117.133-135.138,158, sandaracopimaric acid, 407, 415 Schizaeaceae, 414
175,337,458,473,475-478,510,514,515, Sanguinaria canadensis, 601 Schizaphis graminum, 113, 129, 173, 363, 365
568-571,573-577,598,601,610,660,662, sanguinarine. 26, 961, 594, 601, 603 Schizolobium parahybum. 222
663, 692, 693 Saniculoideae, 134 schizophrenia, 514
rutacridone alkaloids, 8 Santalaceae, 45,197,521.549,557 Schotia brachypetaia, 145
rutacridone epoxide, 510, 574 Santalales. 45 schottenol, 441, 582
rutaecarpine, 570 santalenes, 384 Sdrpophaga incertu/as, 480
Ruta1es, 197,337,473,483-485,568,577, (+ )-(E)-a-santalen-12-oic acid, 381 Scirpus maritimus, 145, 149
616,667 Santa/um album, 521 scIareol. 405, 407. 412, 414
Ruteae,575 santamarin, 392 13-epi-scIareol. 406, 407, 414
752 Index

sclerotia, 658 seeds, vii, 5-7, 9, 13, 14, 16-19,22-33,35, 501,517,575,670,671; see also
Sclerotinia sclerotiorum, 137 39-43,47,49,51-53,55,73,78,83,84, homosesquiterpenes, sesquiterpene
Scolypopa australis, 385 99, 105, 109, 118, 121, 122, 125-127, 130, phytotoxins
Scolytus, 40, 72, 342 132,136,137,152,167,171,175,179,183, sesquiterpenes, acyclic, 370, 382
Scolytus multistriatus, 83, 168 189,190,203,206,208,215,217,219,220, sesquiterpenes, eremophilane, 371
Scolytus quadrispinosus, 84 222,223,226-229,231,232,241,242,244, sesterterpenes, 370, 398, 415, 420, 422-424,
Scoparia dulcis, 101, 104 245,249,251,256,260,264,265,277,279, 488
scopolamine, 511, 534, 535, 537 280,282,287,288,294-300,305-307, Sevin, 110
scopoletin, 130-133,288 309-311,345,376,379,390,391,396,402, sex cells, 376
( - )-scoulerine, 600 403,411,414,435,445,459,460,468,472, sex detennination, 439
( - )-(S)-scoulerine, 603 473,478-481,483,499,528,529,537,544, sex expression, 437
(S)-scoulerine, 9, 588, 598, 600 552-554,558,559,561,565,567,568,597, shade, 7, 177,219
(13R)-scoulerine, 603 609,615,617,627,634,635,641,645,653, sheep, 136,212,220,390,411,451,466,515,
(13S)-scoulerine, 603 655,658-660,663,667,669,671,700,705, 559, 676, 685, 705
14-S-scoulerine, 603 710,711 shikimate-3-phosphate, 94, 97
Scrophularia raamosa, 358 Selaginaceae, 361 (- )-shikimate-3-phosphate. 97
Scrophulariaceae, 78, 85, 88, 92, 101, 108, Selaginallales, 173 shikimic acid, 3, 8, 11,64,65,76.77,80,82,
158,173,174,177,179,358,361,363,364, Seiaginella, 173 85-87,94-97, 102, 104, 121, 126, 139, 142,
366,387, 391,444,452,458,466,468,500, selection pressure, 9, 11, 12 148, 193, 195, 215, 324, 507, 568
522,549,552,557,559,566,569,570,669 selenium, 229 (- )-shikimic acid, 94, 195
Scrophulariales, 88 selenocystathlonine, 229 shikimic acid pathway, 11, 64-66, 76, 77, 94,
scrub, Florida, 125, 127 self-inhibition, vii, 125 95,97, 125
scutellum, 240 Semiaqui/egia, 269 shikimi-no-ki, 94
Scutia buxifolia, 700 semiochemical, 339, 365, 367 shikonin, 85
scutianine A, 700 sendanin, 480 shiromodiol diacetate, 380
scutianine B, 700 senecic acid, 549 shoots, 136, 138, 145, 175,231,238,345,408,
scutianine C, 700 Senecio, 510, 547, 549-552, 559 411,415,423
scutianine D, 700 Senecio jacobaea, 553 Siberia, 673
scutianine F, 700 Senecio longilobus, 553 Sibine stimulea, 390
scutianine G, 700 Senecio spathulatus, 552 sickness, see milk sickness, motion sickness,
sea cucumbers, 458 Senecio triangularis, 552, 559 vomiting sickness
sea urchin, 340, 381, 420, 696 Senecio vulgaris, 546, 547, 549. 550, 553, 565, Sida, 569, 570
seasonal variations, 543
566 Sideritis, 420
Secale, 99, 101
Senecioneae, 46, 387, 510, 549. 552 Sideritis mugronensis, 168
Secale cerale, 99
senecionine, 548. 549, 550, 552, 553, 559. 566 siderophores, 230, 240, 521
Sechium edule, 31
senecionine N-oxide, 550, 553, 566 Sierra Leone, 29
secodine, 632, 635
senepoxide, 104 sieve plates, 259
secoiridoid glucosides, 353, 358
senescence, 518, 554 sieve tubes, 320
secoiridoid glycosides, 359
seniciphylline N-oxide, 552 sieve-tube plastids, 708
secoiridoids, 354, 358-361, 365
Senna. 88, 91 sieve-tube pores, 259
(- )-secoisolariresinol, 117, 118
sepals, 91 signal transducer, 33, 39
secologanin, 2, 353, 358, 359, 366, 579, 610,
sequestration, 9, 124, 208, 220, 324, 340, 353, signals, 8, 33, 41, 53, 165, 167, 191,261
611,628,630,636-638,641,648,652,668,
361,363,364,418,442,466,550-552,564, Silene cucubalus, 242
669
secondary compounds, viii, 1,3-12, 14.40, 683,703 silk, 136, 341, 361
76,85,94,97,98, 101, 138, 139,232,249, sequoyitol, 264 silkwonn, 35, 128, 169, 187, 341, 440, 454
287, 296, 311 serendipity berries, 245 silkworm, cecropia, 444
secondary metabolites, vii, 1-16,32,34,53, serine, 215, 217, 220, 222, 226, 290 silybin, 168
58,76,77,80,92,94, 101, 102, 104, 106, D-serine, 215, 217 Silybum marianum, 168
136, 179,247,252,254 L-serine. 215, 217 silychristin, 168
secondary phloem, 320 (2-serine )tabtoxin, 238 silydianin, 168
secophthalideisoquinoline alkaloids, 603, 610 serotonergic, 510 silymarin, 168
secret societies, 647 serotonin, 228, 229. 510, 514, 516, 517, 525. Simaba multiflora, 481, 482
secretory canals, 324 640, 659, 660 simalikalactone D, 482
secretory cells. 326 serotonin antagonist, 640 simarolide, 476
Securinega alkaloids, 703 serpentine, 630-632, 645, 653, 654 Simaroubaceae, 46. 88,197,337,473,
Securinega suffruticosa, 703 sesamin, 118, 120 475-478,481,482,485,575,576,660,662,
securinine, 703 Sesamum indicum, 118 666
sedamine, 540 Sesbania, 705 Simmondsia chinensis, 51, 52
sedative. 460, 640, 670, 673 Sesbania alkaloids, 705 Simmondsiaceae, 51, 293
sedges, 384, 659 Sesbania drummondii, 705, 711 sinalbin, 300, 308, 310
Sedum, 531, 539 sesbanimide, 705, 711 sinapic acid, 108. 113. 115
Sedum cepaea, 282 sesbanimide B, 705 sinapine, 300, 302, 310
Sedum sarmentosum, 282 sesbanimide C, 705 Sinapis alba, 308, 310
seed coats, 275, 277 sesbanine, 705 sinapyl alcohol, 115
seed dispersal, see dispersal, seed seselin, 130 Sindris albimaculatus, 29
seed dormancy, see dormancy, seed sesquiterpene alkaloids, 375, 525. 670, 671 ,B-sinesal, 370
seed germination, see germination, seed sesquiterpene biosynthesis. 370, 371, 401 sinigrin, 266, 303, 307-309
seed oil. 35, 36, 45, 118, 285 sesquiterpene cyclase, 368 Sinningia cardinalis, 164
seedlings, 34, 47, 78, 92, 99, 109, 125, 127, sesquiterpene glycosides, 378 sinoacutine, 592
132, 135, 145, 158, 178, 183,220,229,230, sesquiterpene lactones, 316, 351, 365, 367, sinomenine, 592
238,275,276,280,282,287,288,290,296, 376,386-397,415,425,454 Sinomenium, 592, 593
310,314,315,317,344,345,369,376,390, sesquiterpene olefin cyclase, 374 Sinomenium acutum, 592
401,407,408,413-415,424,425,436,437, sesquiterpenes, 13,30,46,78.312,335, siphonidin, 269
444,452,484,501,504,634,645,652,673, 337-339,342,344,346,350,367-387, Siphonodon australe, 269, 272
702, 703, 708 391-394,396-398,412,415,419,420,500, siphonoside, 269
 Index 753

sirenin, 376 Solanum X ajanhuiri, 684, 690 spindle-tree, 269, 270

I-sirenin, 376 solasodine, 677, 678, 680, 683, 684 Spiraea, 673
Sitophilus zeamais, 437 (25R)-solasidine, 680 Spiraea japonica, 673
sitosterol, 436, 459, 582 solasonine, 683 Spiraeoideae, 264, 284
,B-sitosterol, 35, 320, 427, 435, 436, 439, 441, Solenopsis, 533, 542 spirobenzylisoquinoline, 610
458 Soienopsis saevissima, 35 Spirodela po!yrhiza, 162
skeleton, see artemisyl skeleton, cadalane Soienopsis xenovenenum, 552 spirosolanes, 677, 678. 680, 681
skeleton, cevane skeleton, chrysanthemyl Solidago, 418, 423 spirostanols, 681
skeleton, santolinyl skeleton Solidago altissima, 49 Spodoptera, 591, 603
Skimmia japonica, 573 solon, 168 Spodoptera eridania, 135,361,390
skimmianine, 573 solorinic acid, 86 Spodoptera exempta, 361, 380, 415, 418, 481
skin, 381, 498 solvent, 324, 326, 339, 348, 439, 496 Spodoptera exigua, 390
,B-skytanthine, 669 solvent extraction, 324, 326 Spodopterajrugiperda .. 145,390,412,480,
slaframine, 561 Sonneratiaceae, 88 481,658,666
slime mold, 71 Sonora, 582, 615 Spodoptera littoralis, 379,418,550
slobbering, 561 Sophora secundiflora, 559 Spodoptera fitura, 113, 136, 138,396,417,
smallpox, 237 Sophora subprostrata, 168 425,480
smartweed,76 sophoradin, 168 Spodoptera ornithogalli, 390
smooth muscle tonus, 588 Sophoreae, 557 Spondias mombin, 210, 212
snail gut acetone powder, 294 sophorose, 266 sponges, 380, 700
snails, 209, 289, 380, 393, 459, 460 soporific drug, 136, 538 spores, 110, 167,377,414,503,520
snake bites, 591 Sorbaria arborea, 284 Spreckeliajormosissima, 621
snakes, 179 sorbitol, 262, 264 spruce, 337, 543
snapdragon, 158, 189 D-sorbitol, 264 spruce, black, 337
snuff, 462, 514 D-sorbose, 250 spruce, blue, 337
soap, 16 L-sorbosone, 265 spruce, Engelmann, 337
sodium, 207, 260, 348, 468 Sorbus, 269, 289 spruce, red, 337
sodium carbonate, 294 Sordaria fimicola, 71 spruce, Sitka, 337
sodium channels, 348, 560, 674, 675, 677, 685, sorghum, 101,208,210,213,274-276,280, spruce, white, 337
703 282, 288, 290, 299 squalene, 3, 427, 428, 430-432, 444, 447, 452,
sodium channels, voltage-dependent, 560 Sorghum bie%r, 78, 274, 275 455, 490, 689
sodium metabisulfite, 207 Soulamea soulameoides, 481, 482 squalene biosynthesis, 428
sodium picrate paper (HCN test), 294 soulameolide, 477 squalene epoxidase, 431
sodium potassium-ATPase (Na+ +K+- squalene 2,3-epoxide, 427, 431, 432, 444, 447,
sour oranges, 340
ATPase), 468, 601, 603, 615, 660, 676 473
South America, 120, 139, 179, 229, 296, 388,
sodium salt, 16 437,460,525,535,550,561,605,6\0,643, (3S)-squalene epoxide, 447
sodium transport, 660 (3S)-2,3-squa1ene epoxide, 431, 432
649, 660, 684, 696, 700
softwood, 37, 115 squalene mono-oxygenase, 431
South American leaf blight, 288
soil, 4, 8, 26, 53, 83, 125-127, 191,207,229, squalene-2,3-oxide-cycloartenol cyclase, 432
South Pacific, 226. 294, 538
242,262,340,345,378,391,441,459,700 squalene synthase, 372, 428, 430, 452, 455
southeastern Asia, 210, 319
soil nutrients. 8 St. Anthony's fIre. 658
South-West Cape Province, 623
soil organisms, 4 S1. John's wort, 91
Soviet Asia, 210
solacauline, 682 Stachelameisen (ants), 703
solacongestidine, 681, 683 Soviet Union, 378, 623
stachyose, 256, 261
soladulcidine, 678, 683 soyasapogenol B, 78, 452
Stachyuraceae, 442
soiadulcine, 682 soybean, 178, 181, 182, 184, 188, 191, 192, Stachyurus, 442
solafloridine, 683 238,245,256,261,265,270,290,472 stalking game, 647
solamargine, 682, 683 spadix, 515 stalks, 608
Solanaceae, 3, 30, 31, 88, \08, 110, 134, 163, Spanish fly, 382 Stanhopea pulla, 344
183,242,377,395,453,458,494,506,5\0, Sparganothis sulfureana, 291 Stanhopea tricornis, 344
517,525,533,534,537,539,544,550,557, sparteine, 9, 554-556, 558, 559 Stanleya, 305
560,660,671,677 ( - )-sparteine, 554, 555 Staphyleaceae, 514
Solaneae, 537 sparteine sulfate, 558 Staphylococcus aureus, 69
solanidanes, 677 Spartium junceum, 559' starch, 249, 254, 257, 259, 260, 270-272, 290;
5a,22,BH,25,BH-solanidan-3,B-ol, 684 spasmolytic. 136, 591 also see floridean starch
22,BH,25aH-solanid-5-en-3,B-ol, 684 spathe, 515 starch synthesis, 249
solanidine, 458, 677-683 Spathelia, 477 starch synthetase, 259
solanidine alkaloids, 680, 684 Spathelioideae, 571, 575 starch, floridean, 260
Solaniflorae, 360 speannint, 335, 351 starfish, 458
solanine, 682 sperm tube growth, 340 steam distillation, 324, 452
a-solanine, 458, 682-684 sperm whale, 52 stearic acid, 17,22,26,35,51,53
soianocapsine, 678, 683 Spermacoce,612 stearoyl desaturase, 21
Solanum, 535, 677, 678, 680, 681, 683, 684 Spennacoceae, 612, 638 stearoyl-ACP, 19,21, 22
Solanum alkaloids, 249, 668, 678, 680, 681, spennicide, 376, 460 stearyl-ACP,51
683,684 spermidine, 517, 518, 520, 521, 529, 547 stearyl-CoA, 51
Solanum berthaultii, 269, 383 spermidine alkaloids. 521 Stegnospermaceae, 708
Solanum eleagnifolium, 683 spermine, 517, 518, 529 stemborer, yellow, 480
Solanum lycocarpum, 557 spermine-DNA complexes, 520 Stemmadenia pubescens, 632
Solanum malacoxylon, 437 sphondin, 133. 135, 136 stemmadenine, 632, 634, 635
Solanum mammosum, 460 spices, 12,47, 110, 130,340,346,379 stemofoline, 533
Solanum marginatum, 242 spider, 49, 80, 551, 564 Stemonaceae, 179
Solanum melongena, 677 spider, giant tropical orb, 551 Stemonaceae, 531, 533
Solanum nodiflorum, 460 spiders, jumping, 49 stemonine, 533
Solanum rostraturn, 677, 683 Spilanthes oiearacea, 36 stemospironine, 533
Solanum tuberosum, 31, 41, 377, 535, 677, spinach, 40, 94, 99, 442, 520 stems, 7, 30,117,152,193.207.269.514,
683,684 Spinacia o/eracea, 3, 557 522, 608, 658, 700
754 Index

Stenocarpus, 82 strawberries, 346 sulfuretin, 173

Stenocereus alamosensis, 582 Streptanthella, 305 sulfuric acid, 227, 535
Stenocereus gummosus, 441, 582 Streptanthus, 305 Sumatra, 187
Stenocereus thurberi, 441, 582 streptimidone, 705 sunflower, 390, 415
Stenosolen, 637 Streptocarpus dunnii, 87 surface active agents, 459, 460
Stephania, 592, 593 Streptomyces, 65, 97, 105,234,253,568 sutherIandin, 284
Stephania tetrandra, 606 Streptomyces erythreus, 65 Swainsona, 254, 561
stephanine, 591 Streptomyces griseus, 262 Swainsona canescens, 561
steptomycin, 253 Streptomyces verticillus, 235 swainsonine, 254, 270, 561, 562, 565
Sterculia foetida, 35 streptomycin, 262 swainsonine N-oxide, 561
Sterculiaceae, 28, 46, 304, 444, 466, 571, 581, streptovitacin A, 705 Swartzia madagascariensis, 460
655, 666, 700 stress, 8,13,134,174,184,189,190,274, sweet clover, 130, 132, 137
sterculic acid, 27 290,296,337,339,495,517; also see sweet compounds, 99,175,209,245,253,257,
stereochemistry, 21, 37, 40, 67, 97, 101, 117 abiotic stress, water stress resistance 261,264,367, 385, 398, 408, 420, 424, 444,
sterigmatocystin, 65 Strictocarpia, 658 456,462,463,471
Sternechus tuberculatus, 220 strictosamide, 648 sweet potato, 7, 72, 313, 377
steroid glycosides, 441 strictosidlne, 2, 359, 628-631, 635, 645, 648, Sweetenioideae, 488
steroid saponins, 456 649, 652-654 sweroside, 359
steroidal alkaloids, 510, 560, 668, 675, 677, strictosidine synthase, 2, 629 Swertia caroliniensis, 359
682, 686, 689, 690 Striga, 78, 92, 391, 392 swertiamarin, 359
steroidal sapogenins, 456 Striga asiatica, 78, 92, 392 swine, 460, 480
steroids, 3, 10,37,312,322,348,397,427, strigol, 78, 391, 396 Sylvifagus floridana, 390
428,432,433,439,440,447,453,455,471, stroma, 8, 19,23 sympathetic nervous system, 510
501,504; see also, corticosteroid, strophanthidin glycosides, 468 Symphonia globubfera, 148
ecdysteroid, steroid alkaloids, steroidal Strophanthus, 463, 465, 468 Symphytum, 552, 553
Strophanthus boivinii, 463 Symphytum officinale, 3, 557
glycosides, phytoecdysteroids
Symplocaceae, 361, 575, 610
sterol esters, 18 Strophanthus gratis, 469
sterol-J22-desaturase, 436 Symplocos, 610
Strophanthus kombe, 465, 469
synapse, 510, 662
J14_sterol reductase, 436 strychnan, 636
synergists, 13,35,50,110,111,113,128,348,
sterols, 312-314, 349, 370, 401, 427, 431, 433, Strychneae, 638
435-437,439-442,452-455,458,471,582; strychnine, 511, 607, 628, 641-643, 703
383
synomones, 9, 30, 39, 110, 176, 324, 339, 343,
see also C2rsterols, C29-sterols, campesterol, Strychnos, 511, 638, 640, 643, 652, 660
Strychnos alkaloids, 631-635
344,367,380,381,498,516
cholesterol, cholesterol binding, 20,22-
Synsepalum dulcificum, 245
dihydroxycholesterol, ergosterol, fucosterol, Strychnos dinklagei, 645
Syntermes na, 372
(25R)-26-hydroxycholesteroI, 20- Strychnos nux-vomica, 641
synthetase complexes, 19
hydroxycholesterol, 22,8-hydroxycholesterol, Strychnos usambarensis, 652
Syntomis, 591, 597, 603, 613, 619, 640, 641,
26-hydroxycholesterol, lanosterol, 411'- Stylidiaceae, 257, 360, 361
650, 658, 662, 664, 686, 703
methylergosta-8,24(28)-dien-3,8-01 to 24- stylopine, 600 syringic acid, 78, 126
methylenelophenol (J8_.J7)-sterol (S)-stylopine, 600
syringin.115,117
isomerase, 1411'-methylsterol 1411'- Stylopyga rhombifolia, 136 systematics, plant, 12
demethylase, 2,3-oxidosqualene:lanosterol Styraceae, 412 Syzygium aromaticllm, 59
cyclase. phytosterols, ,8-sitosterol, styrylpyrones, 139
stigmasterol, taraxasteroI, UDP:sterol suberin, 51, 53-55, 127
T -2 toxin, 378
transglucosylase subterranean clover, 179
Tabebuia guayacan, 85, 92
sterol-S-adenosylmethionine methyl transferase, succinic acid, 160
Tabemaemontana, 635, 637, 640, 645, 647,
436 succinic semi aldehyde, 81
652,654
steryl ferulates, 437 o-succinylbenzoic acid (OSB), 80-82, 87, 93
Tabemaemontaneae, 637
Stevia, 407 o-succinylbenzoic acid (OSB) CoA synthetase.
Tabernanthe iboga, 628, 647
Stevia rebaudiana, 420 81 Tabernanthe subsesselis, 647
stevio!, 408, 420, 425 o-succinylbenzoic acid (OSB) synthase, 81 tabersonine, 632, 634, 635, 645, 652
stevioside, 420 N'-succinyl-2,6-diaminopimelic acid, 217 tabtoxin, 237
7 -stigmastenol, 582 sucrose, 175,245,247,253-257,261,270, tachycardia, 674
stigmasterol, 427, 435, 436 271, 385, 420, 460, 462, 463 tadpoles, 460
Stilbaceae, 361 sucrose esters, 269 taeniacide, 539
stilbene biosynthesis, 142, 155, 156 sucrose synthetase, 256 D-tagatose, 250
stilbene glycosides, 140 sugar, 3, 15, 24, 30, 52, 96, 108, 132, 160, Tageteae, 387
stilbene synthases, 142, 156 164,170-172,175,194,197,208,244, Tagetes, 47
stilbenes, 8, 136, 139, 140, 142, 143, 145, 147, 247-253,256,257,259-262,264,266,270, Tagetes erecta, 48
149,156 273, 279, 280, 287, 290, 544, 583; see also tagetone, 346
stimulants, 12, 78, 128; see also feeding blood sugar, 2-deoxy sugars Talguenea, 610
stimulants, phagostimulants sugar beets, 3, 222, 256, 260, 460; also see tall fescue. 510, 659, 667
stimulants, central nervous system, 703 sugar beet wire worm tall morning glory, 7
stimulants, ganglionic, 540 sugar cane, 52, 78, 145,256,290,414 L-ta!omethylose, 251
stimulants, growth, 53 sugar carboxylic acids, 247, 463 talose, 250
stimulants, heart, 392 sugar mimics, 253 tamarack, 415
stimulants, skeletal muscle, 702 sugars, amino, 247, 253 Tanacetum. 328, 331, 332, 348
stimulants, uterine, 640 sugars, branched-chain, 247, 251 Tanacetllm vulgare, 88, 326, 328, 330-332
stinging hairs, 517 sugars, methoxy, 247, 251 tannase, 212
Stipa,659 sugars, nucleotide, sugars, phosphorylated, 248 tannic acid, 208
Stipa robusta, 659, 667 sulfatase, 160, 188, 294 tannin assays, see acid butanol assay, Folin-
stipitatonic acid, 59 sulfation of desulfoglucosinolates, 303 Denis assay, hide powder method
stizolobic acid, 706 sulfoquinovosyl diglyceride, 18 tanning, 200, 206, 207, 210
stomatal closure, 500, 501 sulforaphane, 306 tannins, 8, 12, 102, 152, 159, 163, 170, 171,
stomatal opening, 415 sulforaphene, 310 191, 193-197,204,206-210,212,213,289;
storage, 220, 240, 241, 247, 257, 259, 264, sulfur, 42, 45, 173, 226, 227, 229, 231, 266, see also condensed tannins. ellagitannins,
287,295 303,309 gallotannins, phlorotannins
 Index 755

tansy ragwort, 553 a-terthienyl, 48 Thelypodieae, 305

Tanzania, 652 Tessaria dodoneifolia, 175 Thelypodium, 304. 305, 310
tapewonn, 539 testosterone, 53, 440 Thelypodium paniculatum, 305
tapping, 320 tetanus, 607 Thelypodium eucosmum, 305
taraktophyllin, 285, 286 tetrabase, 294 Theobroma, 700
Taraxacum kok-saghyz, 319 tetracycline, 56, 65, 67, 74, 234 Theobroma cacao, 700
Taraxacum officinaie, 159 12-0-tetradecanoylphorbol-13-acetate, 403 theobromine, 700, 702, 703
taraxasterol, 447 Tetradium, 569, 570, 574, 575, 610 Theophrastaceae, 452
taraxerol, 447 Tetradium rutaecarpa, 570 theophylline, 511, 700, 702, 703
tarennoside, 358 l,2,3,6-tetragalloylglucose, 196 therapeutic function, 511, 612, 613, 642, 645.
taro, 26 Tetragono[obus, 184 648, 698, 708
tarsal receptors, 309 tetrahydroalstonine, 630, 631 thennopsine, 558
tartaric acid, 265 tetrahydroanabasine, 507, 508 Thermopsis, 552, 558
taspine, 591 tetrahydrobenzylisoquinoline alkaloids (TBIQ), Thermopsis divaricarpa, 552, 559
taste, 171, 175,210,237,245,253,261,264, 578,584,585,587,588,598,610 Thevetia thevetioides, 468, 472
267,269,300,302,316,339,340,346,361, I-tetrahydrobenzylisoquinoline alkaloids, 575, D-thevetose, 251, 465
L-thevetose, 251, 465
380,386,389,390,417,444,452,456,460, 610
thiamine pyrophosphate, 81
462,463,466,478,483,511,525,539,552, tetrahydroberberine, 598, 599
thiarubrine A, 48
641,673,684; also see bittersweet taste, (- )-L1-1-tetrahydrocannabinol, 336
thioctic acid, 237
bitter taste, distasteful properties, peppery L1 1(6ttetrahydrocannibinol, 336
thiocyanates, 266, 290, 302, 305, 306, 310
taste, sweet compounds tetrahydrocolumbamine, 598, 599
thioesterases, 21, 23
Taxaceae, 401, 402, 413, 626, 676 (S)-tetrahydrocolumbamine, 598 thioglucosidases, 300
taxane alkaloids, 402 tetrahydrohannan, 660 f3-thioglucosidases, 302
taxanes, 398, 401, 402, 413 tetrahydroisoquinoline alkaloids, 575, 581, 605 thiohydroximate O-sulfonates, 306
taxifolin, 159 tetrahydrolathyrine, 226, 232 thiohydroximates, 303-304
taxine, 626 tetrahydropalmatine, 599 thiohydroximates, S-glucosylation, 303
taxine I, 676 tetrahydroprotoberberine alkaloids, 598 thiohydroximic acids, 303
taxine II, 676 (S)-tetrahydroprotoberberine oxidase, 598 thiokinases, 23
taxiphyllin, 276, 277, 280, 286 tetrahydropteridine cofactor, 132 thiophene, 45, 48, 49
(R)-taxiphyllin, 276, 277, 280 4,2' ,4'.6'-tetrahydroxychalcone, 158 thioredoxin, 8
Taxodium distichum, 420 2a,3a,22a,23a-tetrahydroxy-24a-methyl-f3- thiosulfate sulfur transferase, 290
taxo1, 676, 677 homo-7-oxa-5a-cholestan-6-one, 437 Thladiantha grosvenorii, 463
taxonomists, 12 1,6,7 ,8-tetrahydroxyoctahydroindolizine, 562 threonine, 217, 220, 234
Taxus, 288, 402, 499, 626, 676, 677 1,3,5,6-tetrahydroxyxanthone, 148 threose, 249
Taxus alkaloids, 668, 676, 689 tetrahymanol, 431 Thuja, 337
Taxus brevifolia, 677, 691 Tetrahymena pyriformis, 431 Thuja occidentalis, 346
Taxus wallichiana, 677, 690 tetraketides, 58, 59 Thuja p!icata, 340
tazettine, 622, 623 4,4'-tetramethyldiaminodiphenylmethane, 294 a-thujaplicin, 340
TBIQ, see tetrahydrobenzylisoquinoline tetrandrine, 606 ,B-thujaplicin, 340
alkaloids ( + )-tetrandrine, 606, 607 thujaplicinol, 339
TCA cycle, see tricarboxylic acid cycle tetranortriterpenes, 473, 475, 477, 484 a-thujene, 333
tea, 165, 171, 172, 191,210,213,219,506, Tetranychus urticae, 67, 341, 445 l-a-thujene, 332
553, 700, 702 tetraphyllin A, 285, 286 thujic acid, 340
tea, black, 172, 210 tetraphyllin B. 285, 286, 298 thujone, 340, 346
teak. 88 tetraphyllin B sulfate, 285 3-thujone, 332, 333
Teclea,574 tetraploid. 337 (+ )-3-thujone, 340
Tecoma slans, 568, 669 tetraterpene biosynthesis, 431 d-3-thujone, 332, 333
Tectona grandis, 88 tetraterpenes, 46, 348, 367, 384, 431, 486-488, Thunbergiaceae, 361
telemagrandin I, 197 490 thylakoid membranes. 17, 18, 326, 518
telemagrandin II, 197 tetrodotoxin, 674 ThymeJaeaceae, 134,398,401-404,413,419,
teleology, 4 teucrein, 358 423, 424, 444
tendril,33 Teucrium, 358, 406 thymol,346
Tenodera ardifolia sinensis, 468 Teucrium marum, 358 thyroid glands, 220
Ti-plasmid, see plasmid, Ti
Tenthredinidae, 289 Texas, 338, 380, 387, 388, 472, 540
tigliane, 402, 403, 424
tenuiin, 390, 392, 394 Thailand, 698, 711
tiglic acid, 321
Tephrosia, 186 L1-1-THC, see L1-1-tetrahydrocannabinol
13-tigloyloxylupanine, 558
Tephrosia elala, 187 thalicarpine, 606
tigogenin, 680. 681
tephrosin, 187 thalicmine, 591
Tiliaceae, 466
teratogenic, 538, 543, 558, 559, 668, 684 Thalictrum, 242, 269, 588, 592, 605, 667 tingenone, 452, 671
teratological. 685 Thalictrum aquilegifolium, 277 tirucalloi, 473, 473
teratomas, testicular, 235 Thalictrum dipterocarpum, 242 apo-tirucaUol, 473
Terminalia chebula, 194 Thalictrum faheri, 606 L11 -tirucallol. 473, 476
tennites, 53, 85, 89, 340, 343, 372, 419, 500 Thalictrum foetidum, 660 tissue culture, 33, 85, 153, 326, 350, 365, 370,
Ternstroemia japonica, 460 thalidasine, 606 394,447,457,465,470,506,526,529,534,
Temstroemiaceae, 460 thaumatin, 245 544,546,554,576,610,645,652,666
terpenes, 177,312,327,345,384,396,424, Thaumatococcus danie/li, 245 tissue culture, plant, 2. 3, 13, 50, 80, 85, 86,
425,427, 428, 452, 483, 485, 504 THBIQ, see tetrahydrobenzylisoquinoline 92, 127, 137, 149,526,593,594,629,645,
terpenoid biosynthesis, 8, 313, 327, 370 alkaloids 649
terpenoid metabolism, 312-314, 507 L11(6tTHC, see L1'(6)-tetrahydrocannibinol 1LC, see chromatography, thin layer
terpenoids, 8, 11, 104, 507; see also Thea sinensis, 171, 210, 460, 700, 702 toads, 466, 560, 708
hydroquinone terpenoids Theaceae, 171, 197,460,700 tobacco, 30, 49, 50, 105, 110, 121, 128, 129,
terpinen-4-oI. 333 theaflavins, 172, 210 131,132,171,221,237,372,401,414,415,
a-terpineol, 331, 333 thearubigins, 172, 210 462,500,525,526,529,531,533,538,545,
terpinolene, 331, 342, 343 thebaine, 593, 595-597, 615 648, 653; see also tobacco smoke, tobacco
terramycin, 65 Theligonaceae, 361, 708, 711 homwonn, Indian tobacco
756 Index

tobacco smoke, 526 triacetic acid lactone, 58 tnsporic acid C, 502

tocopherols, 488 I-triacontanol,53 trisporic acid E, 502
a-tocopherol, 86 triacylglycerides, 16,23,24,26,27,42,43,45 trisporic acids. 501, 502, 504
Tocoyena, 611, 612, 638 triacylglycerols, 16-18,24-26,38,43,49 trisporin. 502
Toddalia, 610 trial by ordeal, 663, 673, 676; also see ordeal trisporol, 502
Toddalioideae, 571, 573, 575 ceremonies triterpene biosynthesis, chair-hoat-chair-boat
tomatidenol, 683 Tribolium castaneum, 35, 460 transition state, 427, 432
tomatidine, 458, 678, 680, 682 Tribolium confusum, 562 triterpene biosynthesis, chair-chair-chair-boat
(25S)-tomatidine, 680 Tribulus terrestris, 457 transition state, 427, 447, 473
tomatilladine, 683 tricarboxylic acid (TCA) cycle, 3, 24, 81 triterpene glycosides, pentacyclic, 459
tomatine, 458, 510, 690 tricetinidin, 172 triterpene glycosides, 441, 453, 456, 459, 460,
a-tomatine, 681, 682, 684, 689, 690 Trichilia, 477 463, 470, 582, 583
tomato, 30, 33, 39-41, 83, 109, 126,237,381, Trichilia emetica, 480 triterpenes, 78, 39, 322, 348, 369, 370, 380,
415,479,482,486,488,490,491,494,499, Trichilia roka, 480 422,427,428,431,432,444,447,451-456,
501,510,677,681,682,689,690 trichilin A, 480 458, 470, 473, 477, 483, 504, 575
tonic, 364, 670 trichirokanin, 480 triterpenes, B,D-seco-limonoids, 476
tonoplast membrane, 9, 598 Trichocereus pachanoi. 523 tnterpenes, B,D-seco structures, 477
toonacilin, 480, 484 Trichoderma viride, 459 tnterpenes, metabolically altered, 379, 473,
toosendanin, 480 Tricholomataceae, 35. 700 478
toothpastes, 603 trichomes, 30, 269, 324, 351, 383 triterpenes, pentacyclic, 431, 447
topoisomerase I, 645, 648 trichomes, glandular, 324, 386, 394, 395, 414, triterpenoid sapogenins, 458
topoisomerase II, 645 423 triterpenoid saponins, 456, 458, 460, 470
Torreya nucifera, 382 Trichomycetes, 260 Triticale, 101
Torricelliaceae, 361 Trichophyton rubrum, 683 Triticum, 101
Tovariaceae, 304 Trichoplusia, 660 Triticum monococcum, 282
toxic proteins, 244 Trichopsenius, 53 Triton X-loo (detergent), 432
toxic substances, 168, 169, 173, 227, 228, 242, trichothecenes, 378, 379, 395 trixagol, 500
244,340,591,640,650,676,683,685 Trichothecium, 378 Trogoderma granarium, 35
toxicity, vii, viii, 12,72,111,117,120, trichotomine, 666 Trollius, 269
135-137,316,317,346,348,380,411,415, Triclisia patens, 606 Tropaeolaceae, 303, 304, 308
441,456,459,460,479,480,482,510,529, tricoccin 5 22 , 476 Tropaeo/um majus. 303, 308, 309
552,553,559,564,583,591,612,613,619, Tricop/usius ni, 390 tropane alkaloids, 528, 531-535, 537, 539,
640, 646, 673, 683, 708 2-tridecanone, 30, 40, 41, 682 544, 545, 678
toxicity, mammalian, 187 trifarin, 380, 419 tropic acid, 535, 545
Toxicodendron radicans, 67 Trifoliumpratense, 179, 186,460 (S)-tropic acid, 535
toxiferine, 643 Trifolium repens, 179,289,296,297 tropine, 511, 535
C-toxiferine, 643 Trifolium subterraneum, 179 tropinone, 535
C-toxiferine I, 643 1 ,2,6~trigalloylglucose, 196 tropoloisoquinoline alkaloids, 588
toxin, 29, 30, 72, 78-80, 83, 125, 126, 138, Triglochin maritima, 275, 277 trout, 35
342,345,351,377,378,396,439,441,466 triglochinin, 277, 280, 290 trypanocidal, 599
TPP, see thiamine pyrophosphate triglycerides, vii, 16,26,27,35,41,262,320 Trypanosoma cruz;, 452, 479, 599, 645, 650,
ent-trachylobane, 408 Trigonella, 184 660
I-trachylobane, 408 Trigonellafoenum~graecum, 457, 528 trypanosomes, 479
trachyloban-19-oic acid, 415 trigonelline, 231. 528, 708 trypsin, 208, 242, 245
trachylobanoic acids, 415 3,5,3'~trihydroxybibenzyl, 147 trypsin inhibitors, 245
trail marking pheromone, 324, 343, 419, 437, 4,2',4'-trihydroxychalcone, 179 tryptamine, 2, 10, 359, 514-517, 525, 563,
542 2,5,7-trihydroxyflavanone, 162 628,629,630,648,649,652,654,655,660,
tranquilization, 139, 364, 640 7,2',4'-trihydroxyisoflavan, 184 663, 665, 667, 668
transamination, 101,508,513,581,681 5,7,4'-trihydroxyisoflavone, 178 tryptophan, 3, 94, 97-99, 209, 228, 267, 300,
transamination reaction, 217 6,7,4'-trihydroxyisoflavone, 179 302,507,515,516,525,568,570,575,656,
transcription, 568 1,4,5-trihydroxynaphthalene,83 660,665
transduction, 8, 16 9,12, 13-trihydroxy-(E)-1 O-octadecenoic acid, L-tryptophan, 3, 656
transglycosylation, 303 27,40 tryptophan amino transferase, 98
translation, 8, 226 9,1O,18-trihydroxystearic acid, 53 tryptophan dimethylallyltransferase (DMAT), 3
translocated, 256 Trimeniaceae, 610 tryptophan dimethylallyltransferase synthetase
transport, 3, 6, 8,9,18,19,65,114,115,117, 2,4,5~trimethoxy-l-propenylbenzene, 113 (DMAT) syntetase, 3, 656
166, 190,230,231,240,241,257,266,271, 2,6,6~trimethylcyclohex-2-ene-l,4-dione, 124 tryptophan 2-monooxygenase, 98
287,288,328,345,483,488,521,528,534, (2E,4E,6E,8E)-3,5,7-trimethyl-2,4,6,8- L-tryptophan synthase, 98
546,547,554,562,567,574,588,618,640, decatetraene, 37 Tsuga, 337
645, 660; see also cardiomuscular calcium 1-(2,3,6-trimethylphenyl)-but-2-ene-l-one, 500 T-toxins,72
transport, electron transport chain, sodium 4,5',8-trimethylpsoralen, 137 tuberculostatic effects, 149, 599, 600, 603
transport (3E,7E)-4,8,12-trimethyl-I,3,7,11- tuberculostearic acid, 27
transposable elements, 9 tridecatetraene, 341 tubers, 14, 31, 33, 147, 148, 166,257,314,
traumatin, 32 1,3,7-trimethylxanthine, 700 345,397
Trautvetteria, 269 tripdiolide, 420 Tubiflorae, 359
tree, see neem tree, spindle-tree, tulip tree tripeptides, 657 tubocurarine, 511, 605, 607, 675, 676
a,a-trehalose, 254 Triphyophyllum, 698 (+ )-tubocurarine, 676
trematode, 393 Triplostegiaceae, 361 d-tubocurarine, 643
tremetone, 317 Tripsacum, 101 tubulin, 120,617,618,645
tremulacin, 124 Tripterygium wilfordii, 420, 528, 671 tubulosine, 613, 652
tremuloidin, 124 TriptoIide, 420 tulip tree, 591
Trewia, 528 Trirhabda canadensis, 418 Tulipa, 269
Trewia nudijlora, 695-697 triricinoleoylglycerol, 26 tuliposide A, 269
trewiasine, 696, 697, 711 trisaccharide esters, 108 tuliposide B, 269
triacanthine, 700 trisporic acid B, 502 tumor, Ehrlich ascites, 210
 Index 757

tumor, human non-small cell lung, 420 Umbellularia cail/ornica, 340 veatchine, 673, 674
tumor-promoting substances, 402, 403 umbels, 135, 136 vegetables, 171, 306
(umon;, 209, 321, 402, 403, 511, 512, 546, Uncaria, 204, 206 velvetleaf, 309, 310, 697, 711
553,559,564,566,568,574,577,617,660, Uncaria gambia, 204, 481 venecurine, 645
671,695,696,711; see also antitumor 2-undecanone, 30 venereal warts, 120
substances, cancer, carcinomas, cell cultures, undecylamine, 513 Venezuela, 645
tumorigenic plants, tumor-promoting Ungnadia speciosa, 6, 14, 287, 298 venom, 179,343
substances United States, 35, 39, 52, 78, 84, 91, 92, 127, veno-occlusive disease, 552, 553
tumors, colon, 420 130, 136, 165,210,213,235,237,244,246, Vepris, 570, 573, 574
tumors, colo-rectal, 306 294,296,297,311,317,320,322,362,378, veratramine, 684, 686
tumors, crown gall, 239 383,390,396,460,549,559,561,582,615, veratric acid, 674
tumors, nasopharyngial, 591 646, 648, 673, 684, 705 veratridine, 675, 685, 686
tung, 35, 39, 74 unsaturation, 3, 16, 26 veratrine, 684, 686
tung nut, 244 uracil, 226 veratrole, 113, 124
tung oil, 244, 402 Uragoga, 611 Veratrum, 677, 684-686
turgorins, 123, 193 urea, 220, 221, 305 Veratrum alkaloids, 668, 677, 684-686, 690
turkey red, 89 uredospores, 110 Veratrum grandiflorum, 681, 684
Tumeraceae, 285, 286, 287 urinary antiseptics, 392 verazine, 677, 680, 681, 686
turnip, 306, 309 urine, 174,288 Verbena hybrida, 159
turnover, 324, 351, 510, 526, 574, 594 uronic acids, 456, 456 Verbena o/ficinalis, 355
turpentine, 312, 398, 412 ursolic acid, 346, 447, 452 Verbenaceae, 86, 88, 159, 337, 361, 364, 385,
turreanthin, 473 Urtica dioica, 517 405,406,413,415-418,442,447,458,655,
Turreanthus africanus, 473 Urticaceae, 88, 517, 563, 693, 700 666
tutin, 385, 670 urushiol, 68 verbenalin, 355
tyiocrebrine, 564 usambarensine, 652 verbenol, 343
Tylophora, 533, 563, 564, 576 uscharidin, 466 cis-verbenol, 343
Tylophora asthmatica, 571, 573 usnic acid, 58 verbenone, 343
tylophorine, 563, 564 usumbarensine, 652 verbenyl acetate, 343
tylophorinine, 564 uterine bleeding, 600 vernalization, 411
tyramine, 244, 515, 518, 522, 523, 579, 587, uterine contracting agents, 420, 600, 660 vemolepin, 389, 393
592, 620, 624 uterine muscle, 600 vernolic acid, 26, 27
Tyria jacobaea, 550 Utetheisa ornatrix, 551 vemolide, 392
tyrosine, 3, 64, 80, 94, 97, 101, 102, 104, 106, UV, see ultraviolet radiation Vernonia, 387, 389, 390
107,220,229,273,280,286,300,302,507, Uvaria, 67 Vernonia colorata, 392
509,522,523,564,579,587,591,595,608, Uvaria catocarpa, 175 Vernonia jlaccidifolia, 390
617,619,620,624-626,703,705 Uvaria zeylanica, 105 Vernonia gigantea, 390
L-tyrosine, 274, 275, 277, 290 Vernonia glauca, 390
tyrosine ammonia lyase (TAL), 107,509 vaccenate, 35 Vernonia hymenolepis, 389
L-tyrosine ammonia lyase (TAL), 107 Vaccinium, 171 Vernonieae, 387
tyrosine decarboxylase, 515, 599 Vaccinium myrtillus, 554 Veronica, 387
tyrosine hydrolase, 229 vacuoles, 9,13-15,131,151,152,155,177, versicolorin A, 65
tyrosine kinase, 145 186, 193,319,465,526,546,557,588,598, vertebrates, 510
629,631,652,705 verticillatine, 694
ubiquinone, 76, 86, 104, 125 vaginidiol, 135 vertigo, 640
Udoteaceae, 381, 420 valepotriates, 364, 365 vertine, 694
UDP-L-arabinose, 253 valerian, 353, 361, 364 vesicate, 137
UDP-D-galactose, 249 Valeriana, 353, 365 vesication, 307
UDP-galactose:diacylglycerol Valeriana edulis, 364 vesicles, 9, 596, 598, 618
galactosyltransferase, 23 Valeriana ojficinalis, 364, 670 Vespa, 343
UDP-D-galacturonic acid, 253, 260 Valeriana wallichii, 364 Vestia, 660
UDP-glucose, 162, 190, 196,249,256,266, Valerianaceae, 46, 353, 361, 364 vestitol, 184
267,276,277,293,296,299,303,335 Valerianella discoidea, 353 (- )-vestitol, 179
UDP-D-glucose, 249 Valerianella locusta, 353 ( - )-(3R)-vestitol, 179
UDP-glucose:aldehyde cyanohydrin ,B-glucosyl valerie acid, 35, 36 vestitone, 181
transferase, 282 valine, 234, 273, 274, 276, 282, 286, 287, 300, Vibumaceae, 361
UDP-glucose o-dihydroxycoumarin 7-0- 302,513 vicenin-2, 173
glucosyltransferase, 132 L-valine, 234 Vicia, 223, 226, 229, 280, 700
UDP-D-glucose-4-epimerase, 249 Vallesia,640 Vicia angustifolia, 287
UDP-glucosyl transferase, 276, 277 vallesiachotaman, 636, 638 Vicia Jaba, 48, 223, 229, 244, 309, 363, 623
UDP-D-glucuronic acid, 253 valonea, 195 Vida sativa, 223, 226, 232
UDP-D-xylose, 253 valpotriates, 353 vicianin, 254, 287
UDPG:sterol transglucosylase, 456 vanilla, 122 (R)-vicianin, 280, 287
UDPG:thiohydroximate glucosyltransferases, vanillic acid, 95, 113, 125, 126 vicianose, 266
303 vaniJIin, 122, 344 vicine, 700
ulcer, 420, 460, 462 vanillin assay, 212 Victoria, 516
ulceration, 420 Vanillosmopsis, 390 Vietnam. 379
Ulmaceae, 244 variation, 1,3-5,7,8, 10, 14,35,47,52, 121, Vigna unguiculata, 215, 217
ultraviolet light (UV), 1,4,42, 129, 131, 135, 136, 177, 198,206,212,213,286,287,294, vinblastine, 511, 645. 653, 654
137,138,145,159,177,179,188,188,212 305,311,337,338, 342, 395, 457, 480 Vinca, 637, 645
ultraviolet spectrophotometry, 1 varnishes, 413 Vinca minor, 640
ultraviolet-A radiation, 49, 135 vasicine, 570 vine amine, 640
ultraviolet -B radiation, 166 vasicinol, 570 ( + )-vincamine, 640
Ulva, 495 vasicinone, 570 Vincetoxicum, 563
Umbelliferae, see Apiaceae vasoconstriction, 517, 660 vincosan, 636, 638
umbelliferone, 130-135 vasodilation, 137,346,517,528,640,697 vincoside, 629
758 Index

vincristine, 506, 511, 645, 653, 676, 696 water cress, 304 xanthone, 65, 139, 148, 149
vindoline, 355, 630, 632, 634, 635, 645 water hemlock, 47 xanthone aglycones, 148
a-viniferin, 145 water stress resistance, 500 xanthophyll cycle. 497
e-viniferin, 145 water-beetles, 439 xanthophylls, 367, 494, 497, 501
(S)-5-vinyloxazolidine-2-thione, 307 watermelon, 499 X8nthorrheaceae, 88
Viola, 290 wattle, 204, 206 xanthotoxin, 113, 127, 128, 133-138
Violaceae, 290 wattle, black, 206 xanthotoxol, 133, 134
violaxanthin, 486, 494, 496, 497 wax ester, 51, 52, 54, 55 xanthoxin,501
E-violaxanthin, 501 wax, camauba, 52 xanthyletin, 130
9'-Z-violaxanthin, 501 wax, leaf. 53 Xeranthemum annuum, 26, 27
violaxanthin de-epoxidase. 497 wax, jojoba, 51, 52, 55 Xeranthemum cylindraceum, 280, 297
violaxanthin pool, 497 wax, sugar cane, 52 xeranthin, 280
viral activity 184, 191, 192 waxes, 16, 17, 51-55, 152, 191,422 x-ray crystallography, I, 610
viral infectivity, 562 webwonn, parsnip, 136, 137 D-xylan, 259
Virola, 514 Wedelia asperrima, 420 xylans, 260
virotoxins, 440 wedeloside, 420 Xyleborus ferrugineus, 442
virulence (Vir) gene, 123 weeds, 92, 99, 300, 310, 317, 351, 390, 391, meso-xylitol, 264
virus, 49, 105, 120, 121, 128, 129, 134, 520, 394, 396, 418, 466, 471, 472; also see Xylocarpus, 361, 481, 571
535,616,648; see also adenovirus, alligator weed, duckweed, Klamath weed, Xylocarpus granatum, 361,481
cytomegalovirus, retrovirus, RNA viruses locoweed, ragweed, smartweed Xylocarpus moluccensis, 361, 481
virus, avian myeloblastosis, 623 weevil, bean, 245 xylomollin,361
virus, herpes simplex, 210, 623, 648 weevil, bean, adult, 340 xylose, 172,248,249,456
virus, herpes simplex type I, 120 weevil, rice grain, 437 D-xylose, 248, 259, 260, 456
virus, human immunodeficiency (mv), 210 weevil, southern cow pea, 215, 217 6-0-P-D-xylosyl-D-glucose,254
virus, Moloney murine leukemia. 254 weevil, white pine, 340
virus, potato X, 651 Welwitschia, 140, 149 yage,660
virus, Rauscher leukemia, 623 West Indian cherry, 265 yarn, 33, 105, 147
virus, Rauscher NIH/3T3, 623 West Indies, 177 Yang, 460
virus, vaccinia, 648 wheat, 35, 41, 70, 98, 105, 115, 173,238,245, yangonin, 140
viruses, RNA, 648 361,415,501,517,553 yeasts, 261, 264, 270, 313, 428, 430, 441, 452,
viscotoxin A, 244 wheat germ, 245 517,520,582,583,601,613
viscotoxin B, 244 white snakeroot, 317, 321 yeasts, cactophilic, 583
viscotoxin, 244, 246 Wieland-Gumiich aldehyde, 643 yellow jessamine, 646
Viscum album, 244, 246 wighteone, 179 yellow rain, 379
vision, 344, 365, 382, 486, 497, 498 wildfire toxin, 237 yellow rice disease, 63
vision, insect, 498 wilfordic acid, 528 yellow starthist1e, 317, 322
visnadine, 137 wilIardiine, 700 yellow wooly bear, 390
Vitaceae, 140, 171 willow, 121, 122, 124 Yemen, 522
vitamin A, 312,497 wilt, verticillium, 378, 395 yerba mate, 213, 700
vitamin A aldehyde, 497 wine, 165, 171,210,708 yew, 676
vitamin B6, 508 Winteraceae, 387, 610 yingabaosu A, 392
vitamin C, 265 wintergreen, 122 ylangenes, 375
vitamin D, 437 witch hazel, 252 yobimban system, 640
vitamin K, 80 witchcraft, 537 yohirubine, 511, 628, 630, 640, 654
vitamin KJ. 80, 81 Withania somnifera, 539 yolks,498
vitamin K2, 81 woad,267 Yponomeuta cagnagellus, 269
vitamins, 167 wood, 11, 13,37,39,40,50,53-55,77-79, Yponomeuta evonymellus, 288
vitamins, group B, 264 85, 88, 92, 93, 105, 114, 115, 136, 143, 145, yuan-hoa, 404
vitellogenin, 221 149, 194, 340,473 yuanhuacine, 404
Vitex, 442 workers, grocery, 137 yuanhuarline, 404
vitexin, 162, 173 World War II, 649 yuanhuafme, 404
Vitis vinifera, 145, 171 wonn, see also bollwonn, budwonn, cutwonn, yuanhuatine, 404
(+ )-vitrenal, 376 giant silkwonn moth, homwonn, rootwonn, yuechukene, 575
vittae. 130, 136 roundwonn, silkwonn, tapewonn, webwonn, Zabrotes subfasciatus, 245
Voacanga africana, 635 wonnwood
Vochysiaceae, 533 wonn, sugar-beet wire, 36 Zaire,662
volatile fractions, 324, 326 wonn, tomato, 682 Zamia floridana, 294
vole, montaine meadow, 101 wonnwood, 346 Zanlhorhiza, 269
volkenin, 285, 286, 296 wort, 315 Zanthoxylum, 569, 570, 575, 610, 662
vomiting, 452, 612, 697 wot tip glycoside, 459 Zanthoxylum chalybaea, 573
vomiting sickness, 222 wound healing, 591 Zanthoxylum coco, 573
Vonones sayi, 80 wound infections, 591 Zanthoxylum holstii, 573
Vouacapoua, 145 wounds, vii, 8, 32, 39, 40, 104, 319, 324, 339, Zea, 65, 70, 101
342, 398, 420, 445 Ze. mays, 65, !O5, 173,435,504,700,703
walking stick, 341 Wrightia tomentosa, 686 zearalenol, 70
wallflower, 309 Wunnbaeiodeae, 617 zearalenone, 70
walnut, 83, 265 wyerone,48 zeatin,7oo
Waltheria, 700 wyerone epoxide, 48 trans-zeatin, 700
warburgana1, 379, 380, 419 zeaxanthin, 494, 496-498
Warburgia stuhlmannii, 379 xanthine, 700, 702 zeaxanthin epoxidase, 497
Warburgia ugandensis, 379 xanthine alkaloids, 700 zierin, 280, 287, 297
warlarin, 132, 138 xanthine base, 692 (S)-zierin, 280
Washington, 338 xanthinin, 389 zierinxyloside, 280
wasps, 41, 53, 54, 177, 343, 514 Xanthium, 389 zinc, 242, 543
waste product hypothesis, 1,5,7 Xanthocercis, 253 Zingiber officinale, 114
 Index 759

Zingiberaceae, 87, 88, 114, 179,337 zoapatie, 420 Zygaena, 289

zingiberene, 30 zombies, 537 Zygaenajilipendulae, '517,529
ziziphin, 700 Zonocerus variegatus, 290, 551, 564 Zygaena lonicerae, 703
zizyphi fructus, 700 zooparasites, 533 Zygaena trifolii, 9
Zizyphus, 610, 700 Zosteraceae, 533 Zygomycetes, 502
Zizyphus jujuba, 460, 700 zwitterions, 217, 705 Zygophyllaceae, 120, 458, 459, 569, 570, 576,
zoapatanoi, 420 Zygadenus, 677 660