CO2 + H2O

photosynthesis

sugars

Saponins Cyanogenic
Cardiac glycosides
respiration
glycosides Glucosinolates

acetyl − CoA amino acids

Terpenoids
Sterols protein
malonyl − CoA Alkaloids
Phenols

Flavonoids Tannins Lignin

fatty acids
lipids

27
Secondary Metabolites

The sum of all of the chemical reactions that take place Some secondary metabolites are also involved in
in an organism is called metabolism. Some aspects defense against invading pathogens, a subject that is
of metabolism, such as the metabolism of carbon and also addressed in this chapter.
nitrogen assimilation and energy conversions, have been
addressed in earlier chapters. Most of that carbon, nitro-
gen, and energy ends up in molecules that are common 27.1 SECONDARY METABOLITES:
to all cells and are required for the proper functioning of
cells and organisms. These molecules, e.g., lipids, pro-
A.K.A NATURAL PRODUCTS
teins, nucleic acids, and carbohydrates, are called primary The distinction between primary and secondary
metabolites (see Appendix). Unlike animals, however, metabolites is not always easily made. At the biosyn-
most plants divert a significant proportion of assim- thetic level, primary and secondary metabolites share
ilated carbon and energy to the synthesis of organic many of the same intermediates and are derived from
molecules that may have no obvious role in normal the same core metabolic pathways (Figure 27.1). In
cell function. These molecules are known as secondary the strictest sense, however, secondary metabolites
metabolites. are not part of the essential molecular structure or
In this chapter, we examine some of the broader function of the cell. Secondary metabolites generally,
aspects of secondary metabolites. The focus will be the but not always, occur in relatively low quantities and
biosynthesis, physiology, and ecological roles of four their production may be widespread or restricted to
major classes of secondary metabolites: particular families, genera, or even species. Also known
• terpenes, including hormones, pigments, essential as natural products, these novel phytochemicals were
oils, steroids, and rubber, of little interest to biologists because of their apparent
lack of biological significance. They were known,
• phenolic compounds, including coumarins, flavo-
however, to have significant economic and medicinal
noids, lignin, and tannins,
value and were thus of more than a passing interest to
• glycosides, including saponins, cardiac glycosides, natural products chemists. Natural products have found
cyanogenic glycosides, and glucosinolates, and use in antiquity as folk remedies, soaps, and essences.
• alkaloids. They include drugs and other medicinal products,

459
460 Chapter 27 / Secondary Metabolites

FIGURE 27.1 A schematic to illustrate CO2 + H2O

biosynthetic relationships between prin-
cipal primary and secondary metabolites photosynthesis
(circled).

sugars

Saponins Cyanogenic
Cardiac glycosides
respiration
glycosides Glucosinolates

acetyl − CoA amino acids

Terpenoids
Sterols protein
malonyl − CoA Alkaloids
Phenols

Flavonoids Tannins Lignin

fatty acids
lipids

dyestuffs, feedstocks for chemical industries (gums, The terpene family includes hormones (gibberellins
resins, rubber), and a variety of substances used to flavor and abscisic acid): the carotenoid pigments (carotene and
food and drink. In recent years, however, it has become xanthophyll); sterols (e.g., ergosterol, sitosterol, choles-
increasingly evident that many natural products do terol) and sterol derivatives (e.g., cardiac glycosides);
have significant ecological functions, such as protection latex (the basis for natural rubber); and many of the
against microbial or insect attack. essential oils that give plants their distinctive odors and
flavors. Cytokinin hormones and chlorophyll, although
not terpenes per se, do contain terpenoid side chains. It
is apparent from this list that many terpenes have signifi-
27.2 TERPENES cant commercial value as well as important physiological
roles. Many terpenes and terpene derivatives may be
27.2.1 THE TERPENES ARE considered primary metabolites. The hormones abscisic
A CHEMICALLY AND acid and gibberellin, the carotenoid and chlorophyll
FUNCTIONALLY DIVERSE pigments, and sterols (steroid alcohols) all play signifi-
GROUP OF MOLECULES cant roles in plant growth and development. The vast
majority of terpenes, however, are secondary metabo-
With nearly 15,000 structures known, terpenoids are lites, many of which appear to act as toxins or feeding
probably the largest and most diverse class of organic deterrents to herbivorous insects.
compounds found in plants. As discussed earlier in
Chapter 19, the unifying feature of terpenes is that
they are generally lipophilic polymers based on the sim- 27.2.2 TERPENES ARE CONSTITUENTS
ple 5-carbon unit 2-methyl-1,3-butadiene, or isoprene, OF ESSENTIAL OILS
which is derived via either the mevalonic acid pathway
Many plants, such as citrus, mint, Eucalyptus, and vari-
or the MEP pathway (Chapter 19, Section 19.3).
ous herbs (sage, thyme, etc.), produce complex mixtures
Terpenes can be grouped into several classes, based
of alcohols, aldehydes, ketones, and terpenoids, known
on the number of carbon atoms (Figure 27.2). This
generally as essential oils (essence, as in perfume).
large chemical diversity arises from the number of basic
Essential oils are responsible for the characteristic odors
units in the chain and the various ways in which they
and flavors of these plants but they are also known
are assembled. Formation of cyclic structures, addi-
to have insect-repellant properties. The terpenes and
tion of oxygen-containing functions, and conjugation
terpene derivatives found in essential oils are predom-
with sugars or other molecules all add to the possible
inantly hemi-, mono-, and sesquiterpenes, which can
complexity. The name terpenoid derives from the fact
be moderately to highly volatile. Several examples are
that the first compounds in the group were isolated
shown in Figure 27.3. In most plants, the essential oils
from turpentine (Ger. terpentin), an essential oil (chiefly
are synthesized in special glandular trichomes (hairs) on
pinene) distilled from the resins of several coniferous
the leaf surface (Figure 27.4), although the essential oils
trees.
Number
of
Carbons Class Example
COOH
5 Hemiterpenoid CH 3 C CH CH 2 Tigilic acid
CH 3 CH 3
10 Monoterpenoid CH 3 C CH (CH2)2 C CH CH2OH Geraniol

OH
10 Cyclic
Menthol (peppermint oil)
monoterpenoid

15 Sesquiterpenoid HOCH 2 Farnesol (widespread)

20 Diterpenoid Phytol (chlorophyll)

CH 2OH

Squalene
30 Triterpenoid
(a steroid precursor)

Stigmasterol
30 Triterpenoid (a sterol)

HO

40 Tetraterpenoid -carotene
(a carotenoid)
FIGURE 27.2 Terpenoids are classified according to the number of carbon atoms in
the basic skeleton.

461
462 Chapter 27 / Secondary Metabolites

H3C H3C O
Extracellular
CHCH2CH2OH CHCH2C cavity

H3C H3C H Cuticle

iso-Amyl Alcohol iso-Valeraldehyde

(Mentha, Eucalyptus) (Eucalyptus)
Secretory cells

H3C Stalk cell

H3C CH3 CH3

Epidermis

O
CH3 Mesophyll

HC O CH3 FIGURE 27.4 A schematic diagram of an epidermal glan-

C dular hair in cross-section. Essential oils are produced in
the secretory cells and accumulated in a cavity that forms
H between the secretory cells and the overlying cuticle.
Glandular hairs are found on the leaf surface, where it is
Geranial 1:8 Cineole thought they might serve to deter feeding by herbivores.
(Ctenium aromaticum) (Artemesia)
(lemon grass)
trace quantities. The extremely low level of cholesterol
CH3 allows plant oils to be marketed as ‘‘cholesterol-free.’’
Sterols are constituents of plant membranes, which
is perhaps their most important known function in
HOCH2 plants. Because sterols are planar molecules, their pack-
ing properties are different from phospholipids that
make up the bulk of the membrane bilayer. Sterols pack
H3C CH3 more tightly than phospholipids and therefore tend to
increase the viscosity and enhance the stability of mem-
Farnesol branes. Otherwise, little is known about the function of
(widespread) sterols in plants. Unlike the steroid hormones in ani-
FIGURE 27.3 Representative terpenes that commonly mals, there is no known hormonal role for sterols in
occur in essential oils include: hemiterpenes (isoamyl plant development. Some sterols may have a protective
alcohol, isovaleraldehyde), monoterpenes (geranial, cine- function, such as the phytoecdysones, which have a
ole), and the sesquiterpene, farnesol. structure similar to the insect molting hormones. When
ingested by insect herbivores, phytoecdysones disrupt
of citrus are produced by glands in the peel. The the insect’s molting cycle. Other sterols are present as
resins of certain conifers, for example, also accumu- glycosides, which give rise to a number of interest-
late mixtures of terpenes, including the monoterpenes, ing and economically significant secondary products.
α- and β-pinene, and myrcene (Figure 27.5). Steroid glycosides are discussed below.

27.2.3 STEROIDS AND STEROLS ARE 27.2.4 POLYTERPENES INCLUDE

TETRACYCLIC TRITERPENOIDS THE CAROTENOID PIGMENTS
AND NATURAL RUBBER
Steroids and sterols are synthesized from the acyclic
triterpene squalene, although they generally are modi- Larger terpenes include the tetraterpenes (40-carbon)
fied and have fewer than 30 carbon atoms. Steroids with and the polyterpenes. The principal tetraterpenes are
an alcohol group, which is the case with practically all the carotenoid family of pigments (Chapter 6). The only
plant steroids, are known as sterols. The most abundant important isoprene derivatives with a greater molecu-
sterols in higher plants are stigmasterol and sitosterol lar mass than the tetraterpenes are rubber and gutta.
(Figure 27.6), which often make up more than 70 percent Rubber is a polymer consisting of up to 15,000 isopen-
of the total sterols. However, plants also contain a large tenyl units. The polymer may be linear, as shown in
number of the more than 150 other sterols known to Figure 27.7, or cross-linked into more complex config-
occur in nature. Plant sterols include cholesterol which, urations. The only difference between rubber and gutta
although widespread in occurrence, is present in only is the configuration of the double bonds. In rubber the
 27.3 Glycosides 463

H3C CH3 FIGURE 27.5 Pinene, myrcene, and men-

CH3 CH3
thol are monoterpenes. Pinene and
myrcene are found in the resins of some
CH2 conifers. Menthol is the principal con-
stituent of the essential oil of peppermint
(Mentha piperta). Pinene also has insecti-
OH cidal properties.

CH2 H3C CH3

α-Pinene Myrcene Menthol

double bonds are all cis configurations, while in gutta

the double bonds are all trans. CH3 H CH3 H CH3 H
In the plant, rubber occurs as small particles sus- C C C C C C
pended in a milky-white emulsion called latex. Latex CH3 CH2 CH2 CH2 CH2 CH3
production is widespread in plants, with estimates rang- n
ing from a few hundred to several thousand species that FIGURE 27.7 Rubber is a linear polymer of isoprene units
produce latex in some form. The principal commer- where the value of n may range from a few hundred to
cial source is Hevea brasiliensis, a rubber tree native to several thousand.
the Amazon rainforest. Others include the ornamen-
tal rubber tree (Ficus elastica), milkweed (Asclepias), and produced in the phloem, accumulating in a series of
the Russian dandelion (Taraxacum Kok-saghyz). Latex long, interconnected vessels called laticifers.
contains about 30 to 40 percent rubber and 50 per- The best-known source of gutta is a desert shrub,
cent water. The balance is a complex mixture of resins, Parthenium argentatum, which grows in northern Mexico
terpenes, proteins, and sugars. In most plants, latex is and southwestern United States. Parthenium (commonly
known as guayule) may contain as much as 20 percent
latex by weight, which is stored not in laticifers but in
the vacuoles of stem and root cells. Guayule was at one
time a significant commercial source of gutta for use in
H3 C CH3 rubber products. However, while a single rubber tree, if
properly tapped, can continue to produce for up to 30
CH3 years, guayule plants must be harvested (and, of course,
CH3 CH3 replanted) annually.
CH3 Finally, there is a connection between terpenes
and air pollution. Many of the essential oils, especially
hemiterpenes, monoterpenes, and sesquiterpenes, are
highly volatile and are given off in large quantities by
HO plants, particularly during warm weather. Known gen-
Stigmasterol erally as volatile organic carbon (VOC), these natural
emissions from plants contribute to the formation of
haze and cloud, and are involved in the formation of
H3 C CH3 toxic tropospheric ozone.
CH3
CH3 CH3 27.3 GLYCOSIDES
CH3
Some of the more interesting, if not important,
derivatives synthesized by plants are glycosides. Most
glycosides are thought to function as deterrents to
HO
herbivores. The term glycoside (Gr. glykys, sweet)
Sitosterol refers to the bond formed (called a glycosidic bond)
FIGURE 27.6 Stigmasterol and sitosterol differ only in when a sugar molecule condenses with another
the presence or absence of a double bond (highlighted). molecule containing a hydroxyl group. Sugars may form
These are the two most abundant plant sterols. glycosidic bonds with other sugars, such as when linked
464 Chapter 27 / Secondary Metabolites

together to form polysaccharides, or with hydroxyl sugar gives saponins the properties of a surfactant or
groups on noncarbohydrate molecules, such as steroids detergent. When agitated in water, saponins form a sta-
or amino acids. The sugar most commonly found in ble soapy foam. The name saponin is in fact derived from
glycosides is glucose, although specific glycosides often Saponaria (soapwort), which at one time was employed
contain rare sugars. as a soap substitute.
Three particularly interesting glycosides are the The principal role of saponins appears to be as
saponins, the cardiac glycosides (cardenolides), and the a preformed defense against attack by fungi. Evidence
cyanogenic glycosides. A fourth family, the glucosino- indicates that saponins form complexes with sterols con-
lates, although technically not glycosides, are a similar taining an unsubstituted 3-β-hydroxyl group. When the
structure and so are included here. saponins react with sterols in the membranes of invading
fungal hyphae, the result is a loss of membrane integrity.
In a classic example of one-upmanship, however, many
27.3.1 SAPONINS ARE TERPENE
pathogenic fungi have developed strategies, such as the
GLYCOSIDES WITH DETERGENT development of detoxifying enzymes, for circumvent-
PROPERTIES ing this defense mechanism. Oat (Avena), for example,
Saponins may take the form of (1) steroid glycosides, produces a triterpenoid saponin, avenacin A-1, which
(2) steroid-alkaloid glycosides, or (3) triterpene glyco- is localized in the root epidermal cells and effectively
sides (Figure 27.8). Saponins may also occur as aglycones protects against an invasion by a fungal pathogen (Gaeu-
(e.g., the terpene without the sugar), which are known mannomyces graminis var. tritici) that infects the roots of
as sapogenins. In much the same way as soap, which both wheat (Triticum) and barley (Hordeum). However,
is the sodium salt of a fatty acid, the combination of another strain of G. graminis (var. avenae) produces an
a relatively hydrophobic triterpene with a hydrophilic enzyme, avenacinase, that detoxifies avenacin A-1 and

FIGURE 27.8 Saponins are triterpenoids or steroids H3C CH3

containing one or more sugar units. Medigenic
acid glucoside is a triterpenoid saponin from alfalfa
(Medicago sativa). Disogenin glycoside is a steroidal
saponin isolated from clover (Meliotus spp.). The
addition of a hydrophilic sugar group to a normally COOH
hydrophobic terpenoid gives saponins surfactant CH3 CH3
HO
properties similar to soap.
CH3

O
Glucose CH3 COOH

Medigenic acid glucoside

O
CH3
H 3C

CH3
O

CH3

O
Glucose

Rhamnose

Rhamnose

Disogenin-glycoside
 27.3 Glycosides 465

allows the pathogen to invade oats as well as wheat Digitalis purpurea or Grecian foxglove, D. lanata. The
and rye. two principal cardenolides in digitalis are digitoxin and
The effect of saponins on eukaryotic membranes its close analog digoxin. Digitalis is also the source of a
is highly nonspecific and it is not clear how plants saponin, digitonin.
protect their own membranes against the deleterious Since the late eighteenth century, digitalis has been
effects of their own saponins. One possibility is that the used for its therapeutic value in treating heart conditions
saponins are stored in the form of a biologically inac- such as atherosclerosis. Because they disrupt the heart
tive molecule, called a bisdesmosidic saponin, which muscle Na+ /K + -ATPase pumps (hence the appellation
has two sugar chains rather than one. When under cardiac), cardenolides are highly toxic to vertebrates. The
attack, the inactive form may be converted to the active extreme toxicity of cardenolides has long been exploited
monodesmosidic form by hydrolytic removal of the by African hunters, who coated their arrows and spears
second sugar chain. Alternatively, biologically active, with cardenolide-rich extract from plants. In therapeutic
monodesmosidic saponins may be sequestered in vac- use, however, carefully regulated doses can both slow
uoles or organelles whose membranes contain a high and strengthen the heartbeat. Unfortunately, the lethal
proportion of sterols with a protected 3-β-hydroxyl and therapeutic doses are very close, so the therapy must
position. be carefully monitored.
The effect of saponins on animals is somewhat vari- Other common sources of cardenolides are the
able. While not significantly toxic to mammals, saponins milkweeds, Asclepias and Calotropis. These two species
do have a bitter, acrid taste and will cause severe gas- are known as ‘‘milkweeds’’ because they produce a
tric irritation if ingested. Saponins will hemolyze red milky-white, cardenolide-rich latex. The milkweeds are
blood cells, however, if injected into the bloodstream. particularly interesting because they are the principal
This action is presumably because of their detergent host for ovipositing monarch butterflies. The emerg-
properties and their ability to disrupt membranes gen- ing larvae feed on the milkweed leaves and sequester
erally. On the other hand, saponins are highly toxic to the cardenolides without ill effect. The cardenolides
fish and have been used as fish poisons. Saponins have are retained through metamorphosis and are present
also been implicated in reports of livestock poisoning. in the adult monarchs. When birds, such as blue jays,
Alfalfa saponins, for example, can cause digestive prob-
lems and bloating in cattle. At the same time, there are
reports that saponins contained in alfalfa sprouts will
lower serum cholesterol levels. Commercially, saponins
from the bark of Quillaja saponaria have been used as CH CO
C
surfactants in photographic film, in shampoos, liquid
CH3 CH2 O
detergents, toothpastes, and beverages (as emulsifiers).
The saponin glyscyrrhizin from licorice (Glyscyrrhiza
glabra) has been used in medicines and as a sweetener CH3
and flavor-enhancer in foods and cigarettes.
OH

O
27.3.2 CARDIAC GLYCOSIDES ARE Sugars
HIGHLY TOXIC STEROID
GLYCOSIDES
Digitoxin
The cardiac glycosides (or, cardenolides) are struc-
turally similar to the steroid saponins and have similar
detergent properties. They are distinguished from other H H H H
O
steroid glycosides by the presence of a lactone ring
(attached at C17) and the rare sugars (found almost H3C C C C C C
exclusively in this group of steroids) that form the gly- O O O H H
coside (Figure 27.9). Like the saponins, cardenolides H H H
occur naturally as either the glycoside or the aglycone
(or genic).
The cardenolides have a wide distribution; they have Digitoxose
been recorded in more than 200 species representing FIGURE 27.9 Digitoxin, a cardiac glycoside. The sugar
55 genera and 12 families and are a principal agent component of digitoxin consists of 1 molecule of glucose
in accidental poisonings of humans. Perhaps the best and 1 molecule of acetyl-digitoxose. The structure of
known is digitalis, a mixture of cardenolides extracted digitoxose, one of the rare sugars found in cardiac glyco-
from the seeds, leaves, and roots of purple foxglove, sides, is shown below.
466 Chapter 27 / Secondary Metabolites

attempt to feed on monarchs, the accumulated carede- and lyase enzymes are found in the mesophyll cells. Only
nolides induce an emetic reaction that forces the bird to when the tissue is crushed and the contents of the two
vomit. The bird then wisely avoids attempting to feed cells are mixed will cyanogenesis occur.
on monarch larvae for some time. There is some evidence that the presence of
cyanogenic glycosides deters feeding by insects and
other herbivores, although most animals have the
27.3.3 CYANOGENIC GLYCOSIDES
ability to detoxify small quantities of cyanide. Clearly
ARE A NATURAL SOURCE OF the effectiveness of cyanogenic glycosides as a deterrent
HYDROGEN CYANIDE depends on many factors, such as the amount present,
It might seem odd that plants synthesize chemicals capa- the rate of release of cyanide, and the ability of the
ble of releasing deadly hydrogen cyanide or prussic acid animal to detoxify. The level of cyanogenic glycosides
(HCN), but more than 60 different cyanogenic com- in plants is highly variable, influenced by both genetic
pounds of plant origin have been described from more control and environmental stress. The latter is a
than a dozen plant families. Predominant among these concern when using Sorghum for livestock forage.
are the cyanogenic glycosides. A common cyanogenic Dhurrin accumulates rapidly and can cause livestock
glycoside is amygdalin (Figure 27.10), which occurs poisoning when Sorghum plants are stressed by drought
in many representatives of the family Rosaceae. It is or frost.
found in the seeds of apples and pears and in the bark, Many common food plants naturally contain
leaves, and seed of the stone fruits (apricot, peaches, cyanogenic glycosides in concentration sufficiently low
plums, cherries). Most cyanogenic glycosides appear to that they are not normally a health hazard. These
be derived from one of four amino acids (phenylala- include soy and other beans (Fabaceae); apples, apricots,
nine, tyrosine, valine, and isoleucine) or from nicotinic peaches, plums, and other fruits in the family Rosaceae;
acid. Intact cyanogenic glycosides are not themselves and flax seed (Linum), which is a popular health
toxic, but when the plant is damaged by a herbivore, food. One food source that contains large amounts
the glycoside undergoes an enzymatic breakdown and of cyanogenic glycosides is cassava, a potato-like root
cyanide is released. Cyanide, a noncompetitive inhibitor of the tropical plant Manihot esculenta. Cassava, also
of cytochrome oxidase, is acutely toxic. known as manioc or, in North America, tapioca, is a
The enzymatic breakdown of cyanogenic glycosides major source of starch for millions of people in tropical
is a two-step process (Figure 27.10). First, the sugars are countries. However, poisoning is avoided by careful
released by the enzyme β-glycosidase. The resulting preparation of the plant. This includes grinding the
hydroxynitrile is moderately unstable and will slowly root and expressing the fluids, or boiling the root in
decompose, liberating HCN in the process. Normally, several changes of water.
however, decomposition is accelerated by a second
enzyme, hydroxynitrile lyase. Enzymatic release of 27.3.4 GLUCOSINOLATES ARE SULFUR-
cyanide does not normally occur in intact plants because CONTAINING PRECURSORS
the enzymes and the substrate are spatially separated.
TO MUSTARD OILS
In some cases, separation is maintained within the
cell, but in others, the enzymes are in one cell and Glucosinolates are found primarily in the mustard
the cyanogenic glycosides in another. In Sorghum, for family (Brassicaceae) and related families in the order
example, the cyanogenic glycoside dhurrin is synthe- Capparales. They are precursors to the mustard oils,
sized and stored in epidermal cells, while the glycosidase an economically important class of flavor constituents

FIGURE 27.10 Amygdalin is a cyanogenic glycoside O Glu Glu OH

found in large quantities in seeds of common fruits
in the family Rosaceae. Hydrolysis of amygdalin is a CH
C CH + 2 Glucose
C
two-step process, resulting in the release of highly β-Glycosidase
toxic hydrocyanic acid (HCN). CN CN
Amydalin Mandelonitrile

H
C O
+ HCN

Benzaldehyde Hydrocyanic acid

 27.4 Phenylpropanoids 467

that gives the pungent taste to condiments such as repelled by the presence of glucosinolates in a plant,
mustards and horseradish as well as the distinctive flavor while others have adapted to use the glucosinolates
of cabbages, broccoli, and cauliflower. or mustard oils as attractants to stimulate feeding and
All glucosinolates are thioglucosides (thio, sulfur) ovipositing.
with the general structure shown in Figure 27.11A. Glucosinolates, or rather their absence, have had
The sugar is always glucose. The diversity encoun- a significant impact on the oilseed industry. Rape seed
tered in structure and properties is due to the R (principally Brassica napus) is a good source of veg-
group, which may range from a simple methyl group etable oil, but its high content of glucosinolate together
to large linear or branched chains containing aromatic with high erucic acid (a 22-carbon fatty acid) gives the
or heterocyclic structures. The biological activity of oil undesirable taste and poor storage properties. New
glucosinolates depends primarily on their hydrolysis to strains have been bred with low glucosinolates and erucic
mustard oils (Figure 27.11B). Hydrolysis of glucosi- acid. These strains, called canola in order to distin-
nolates is catalyzed by an enzyme called myrosinase guish them from normal rape, are now an economically
(a thioglucosidase). The hydrolysis product is unstable important oil source.
and immediately undergoes a rearrangement to form a
thiocyanate or isothiocyanate. Like the cyanogenic gly-
cosides, glucosinolates are spatially separated from the 27.4 PHENYLPROPANOIDS
hydrolytic enzymes so that the mustard oils are normally
formed only when the cells are disrupted, allowing the Aromatic amino acids may be directed toward either
enzyme and substrate to come together. As with other primary or secondary metabolism. Also known as phe-
defense compounds, some herbivores are deterred or nolics, or polyphenols, phenylpropanoids are a large

A. FIGURE 27.11 (A) All glucosinolates are thioglu-

– cosides with the same basic structure. In a
N O SO3 thioglucoside the sugar is linked to the rest of
R C the molecule via a sulfur atom. Variation is intro-
duced by the composition of the R group. (B)
S glucose
Enzymatic removal of glucose from a glucosi-
nolate creates an unstable product that sponta-
neously rearranges to form the pungent mustard
B. oils in the thiocyanate, isothiocyanate, or nitrile
forms.

–
N O SO3
CH2 CHCH2C Sinigrin
S glucose

(thioglucosidase)

–
N O SO3
CH2 CHCH2C
S–

–
CH2 CHCH2 N C S + SO42
spontaneous
iso-thiocyanate
rearrangement
–
CH2 CHCH2 S C N + SO42
thiocyanate
–
CH2 CHCH2 C N + S + SO42
nitrile
468 Chapter 27 / Secondary Metabolites

OH three end-products: tyrosine, phenylalanine, and trypto-

phan. In a similar manner, the conversion of chorismate
to prephenate is inhibited by both phenylalanine and
tyrosine, while the conversion of chorismate to anthrani-
late is inhibited by tryptophan.
FIGURE 27.12 Phenylpropanoids are derivatives of the Another step of interest in the pathway is
simple hydroxylated aromatic ring, phenol. the second step in the conversion of shikimate to
3-enolpyruvylshikimate-5-P. The enzyme 3-enolpy-
ruvylshikimate-5-phosphate synthase (EPSPS) is
family of secondary metabolites derived from the aro- inhibited by the herbicide glyphosate (marketed
matic amino acids. Phenylpropanoids are a chemically commercially as RoundUp™). Plants treated with
diverse family of compounds ranging from simple phe- glyphosate will die because they are unable to synthesize
nolic acids to very large and complex polymers such the aromatic amino acids and their derivatives,
as tannins and lignin. Also included are the flavonoid especially protein. Thus glyphosate-treated plants
pigments that were described earlier in Chapter 6. effectively die of protein starvation.
The basic structure is phenol, a hydroxylated aromatic
ring (Figure 27.12). As with other secondary products,
many phenolics appear to be involved in plant/herbivore 27.4.2 THE SIMPLEST PHENOLIC
interactions. Some (e.g., lignin) are important structural MOLECULES ARE ESSENTIALLY
components, while others appear to be simply metabolic DEAMINATED VERSIONS OF THE
end-products with no obvious function. CORRESPONDING AMINO ACIDS
Synthesis of most secondary phenolic products begins
27.4.1 SHIKIMIC ACID IS A KEY with the deamination of phenylalanine to cinnamic acid
INTERMEDIATE IN THE (Figures 27.14, 27.15). The enzyme that catalyzes this
SYNTHESIS OF BOTH AROMATIC reaction, phenylalanine ammonia lyase (PAL), is a
AMINO ACIDS AND key enzyme because it effectively controls the diversion
PHENYLPROPANOIDS of carbon from primary metabolism, such as protein
synthesis, into the production of phenylpropanoids.
The biosynthesis of most phenylpropanoids begins An alternate route, the deamination of tyrosine to
with the aromatic amino acids phenylalanine, tyrosine, p-OH-cinnamic acid, appears to be limited largely, if
and tryptophan. These aromatic amino acids are, not exclusively, to grasses.
in turn, synthesized from phosphoenolpyruvate and Cinnamic acid is readily converted to
erythrose-4-phosphate by a sequence of reactions p-OH-cinnamic acid by the addition of a hydroxyl
known as the shikimic acid pathway (Figure 27.13). group (Figure 27.15). The sequential addition of
The shikimic acid pathway is common to bacteria, hydroxyl and methoxy groups gives rise to caffeic acid
fungi, and plants, but is not found in animals. and ferulic acid, respectively. None of these four simple
Phenylalanine and tryptophan are consequently among phenols appear to accumulate to any extent. Their
the 10 amino acids considered essential for animals principal function appears to be as precursors to more
(including humans) and represent the principal source complex derivatives such as coumarins, lignin, tannins,
of all aromatic molecules in animals. Tyrosine is not flavonoids, and isoflavonoids.
classified as essential because animals can synthesize it
by hydroxylation of phenylalanine.
Synthesis of the aromatic amino acids begins with 27.4.3 COUMARINS AND COUMARIN
the condensation of one molecule of erythrose-4-P, DERIVATIVES FUNCTION AS
from the pentose-phosphate respiratory pathway, with
ANTICOAGULANTS
one molecule of phosphoenolpyruvate (PEP) from gly-
colysis. The resulting 7-carbon sugar is then cyclized The coumarins (Figure 27.16) are a widespread fam-
and reduced to form shikimate. Shikimate is then con- ily of lactones called benzopyranones. More than 1,500
verted to chorismate, a critical branch point that leads examples are known, from more than 800 species of
either to phenylalanine and tyrosine or to tryptophan. plants. The biosynthesis of coumarins is not well under-
The shikimic acid pathway is an excellent example of stood because the putative enzymes and their genes
feedback regulation, where the concentrations of prod- have yet to be isolated. It is most likely, however, that
uct molecules regulate the flow of carbon through the two simplest forms, coumarin and 7-OH-coumarin
key enzymes. The first key enzyme in the pathway (umbelliferone), are formed by an ortho-hydroxylation
is the aldolase that catalyzes the initial condensation of cinnamic acid and p-coumaric acid, respectively, fol-
erythrose-4-P with PEP. Aldolase is inhibited by all lowed by ring closure (Figure 27.17).
 Erythrose-4-phospahate + Phosphoenolpyruvate
1

NADPH

+
NADP
_
COO

HO OH
OH
SHIKIMATE
ATP

ADP

EPSPS Glyphosphate

Pi
_
COO

O _
HO C COO

CH2
CHORISMATE 2
Arogenate
3 +NH3
+

Anthranilate
+ +
NH3 NH3
+
NH3 _ _
_
H2C CH COO H2C CH COO
CH2 CH COO

N
H
TRYPTOPHAN
HO
TYROSINE PHENYLALANINE
FIGURE 27.13 The shikimic acid pathway for biosynthesis of aromatic amino acids
in plants. Initial precursors are erythrose-4-P from the pentose-phosphate path-
way and phosphoenolpyruvate from glycolysis. Enzymes indicated as 1, 2, and 3 are
subject to feedback inhibition and thus are important regulatory enzymes in the path-
way. Those enzymes are (1) 3-deoxyarabino-heptulosonate-7-phosphate synthase;
(2) anthranilate synthase; (3) chorismate mutase. The enzyme EPSP synthase
(EPSPS), which catalyzes the second of three reactions in the conversion of shikimate
to chorismate, is inhibited by the herbicide glyphosate. Glyphosphate thus prevents
the synthesis of the amino acids tyrosine and phenylalanine.

469
470 Chapter 27 / Secondary Metabolites

Pentose- P
pathway Glycolysis

Erythrose-4- P P -enol Pyruvate

Protocatechuic acid
Shikimic acid Hydrolyzable
Gallic acid
pathway tannins

Alkaloids
Tyrosine Tryptophan
Indole-3-acetic acid
Phenylalanine (IAA)
4-OH- Cinnamic acid
(ρ-Coumaric acid) PAL
CS
Cinnamic acid Flavonoids
Coumarin Condensed
ρ-Coumaric acid tannins

Ferulic acid

Lignin
FIGURE 27.14 The shikimic acid pathway plays a central role in the synthesis of
numerous primary and secondary metabolites. PAL = phenylalanine ammonia lyase.
CHS = chalcone synthase.

As a family, coumarins are noted for their roles lowing damage by the European corn borer). The
as antimicrobial agents, feeding deterrents, and germi- invading Aspergillus produces a group of mycotoxins
nation inhibitors. The simplest example, coumarin, is called aflatoxins (from A. fla. + toxin), believed to be
the product that gives new-mown hay its characteristic the most potent and carcinogenic of natural toxins
pleasantly sweet odor. Coumarin is also a constituent of (Figure 27.16). Deaths have been recorded from inges-
Bergamot oil, an essential oil that is used to flavor pipe tion of maize contaminated with as little as 6 to 15 parts
tobacco, tea, and other products. While coumarin itself per billion (or, 6 to 15 μg kg− 1) of aflatoxin. Aflatox-
is only mildly toxic, many of its derivatives can be highly ins have multiple effects: they are mutagenic; they bind
toxic. One derivative, dicoumarol (Figure 27.16), is to DNA and prevent RNA transcription; they compete
typically found in moldy hay or silage containing sweet with hormones for binding sites, impair the immune sys-
clover (Meliotus spp.). Dicoumarol causes fatal hemor- tem, and damage the liver and kidney. Peanut products
rhaging in cattle by inhibiting vitamin K, an essential are a common source of aflatoxin poisoning in humans
cofactor in blood clotting. The discovery of dicoumarol and all raw, shelled peanuts in the United States must be
in the 1940s led to the development of a synthetic inspected for their aflatoxin content. Aflatoxins in cattle
coumarin derivative, Warfarin™, widely used as a rodent feed is another particular problem. When cattle ingest
poison. Scopoletin (Figure 27.16), the most prevalent the toxin, they convert it to another equally toxic form
coumarin in higher plants, is often present in seed coats. and secrete it into the milk.
It is suspected of being a germination inhibitor that
must be leached out of the seed coat before germination
can proceed. 27.4.4 LIGNIN IS A MAJOR STRUCTURAL
The most toxic coumarin derivatives are synthe- COMPONENT OF SECONDARY
sized not by plants, but by fungi. The fungus Aspergillus CELL WALLS
flavus commonly infects foodstuffs such as livestock Lignin is a highly branched polymer of three
feed, peanuts, and maize (the latter especially fol- simple phenolic alcohols known as monolignols
 27.4 Phenylpropanoids 471

+ +
NH3 NH3

H2C CH COO– H2C CH COO–

Phenylalanine OH Tyrosine

PAL

HC CH COO
–
HC CH COO
–
HC CH COO– HC CH COO –

HO H3CO
OH OH OH

Cinnamic ρ-Coumaric Caffeic Ferulic

acid acid acid acid
FIGURE 27.15 Phenolic building blocks. Deamination of phenylalanine followed by
hydroxylation to form p-coumaric acid are the first two steps in phenylpropanoid
biosynthesis. In grasses, p-coumaric acid may be formed directly by deamination
of tyrosine. Phenylalanine ammonia lyase (PAL) is a critically regulated enzyme that
controls the diversion of phenylalanine from protein biosynthesis to phenylpropanoid
biosynthesis.
(Figure 27.18). Gymnosperm lignin is comprised xyloglucans, its presence decreases the digestibility of
mainly of coniferyl alcohol subunits while angiosperm these polymers as well. Also, when fungal pathogens
lignin is a mixture of coniferyl and sinapyl alcohol enter host cells, they do so by enzymatically degrading
subunits. The biosynthesis of lignin is not well the host cell wall. Several studies have shown that lignins
understood, but it is believed that the alcohols are and other phenolic derivatives accumulate at the site of
oxidized to free radicals by the ubiquitous plant enzyme, fungal penetration, presumably slowing the rate of cell
peroxidase. The free radicals then react spontaneously wall degradation.
and randomly to form polymeric lignin.
Lignin is found in cell walls, especially the sec-
ondary walls of tracheary elements in the xylem. In spite
of its abundance (second only to cellulose), the struc- 27.4.5 FLAVONOIDS AND STILBENES
ture of lignin is not well understood. Lignin is a very HAVE PARALLEL BIOSYNTHETIC
large polymer; it is insoluble in water and most organic PATHWAYS
solvents, and impossible to extract without consider- As noted earlier in Chapter 6, flavonoids include the
able degradation. Moreover, the three basic monomers anthocyanin pigments that serve to attract insect polli-
may link together in a variety of ways to form a multi- nators and the isoflavonoids that function as phytoalex-
branched, three-dimensional structure. The complexity ins, or antibacterial and antifungal agents. Flavonoids
is so great that, like snowflakes, each lignin ‘‘molecule’’ have a role in symbiont recognition between rhizo-
may be unique. For the same reason that monolignols bia and host roots. It has also been demonstrated that
are able to form extensive linkages with each other, exposure of plants to UV radiation increases the con-
the lignin polymer is able to form numerous cross-links tent of flavonoids, suggesting that flavonoids such as
with other cell wall polymers. The result is a high degree kaempferol may offer a measure of protection by screen-
of mechanical strength and rigidity in lignified woody ing out harmful UV-B radiation.
stems. Lignin is in fact what makes wood, wood! Flavonoids represent a very large class of pheno-
Although the principal function of lignin is struc- lic derivatives (more than 4,500 representatives are
tural, it has also been implicated as a defensive chemical. known), with a variety of functions. Biochemically, how-
Lignin itself is not readily digested by herbivores and, ever, all flavonoids share a common structure consisting
because it is covalently linked to cellulose and cell wall of three rings, labeled A, B, and C (Figure 27.19).
472 Chapter 27 / Secondary Metabolites

COOH COOH

O O HO
Cinnamic acid -Coumaric acid
Coumarin

HO O O
O O HO O O
Umbelliferone Coumarin 7-Hydroxy coumarin
FIGURE 27.17 Biosynthesis of coumarins. Coumarin and
H3CO 7-hydroxycoumarin (umbelliferone) are formed from
cinnamic acid and p-coumaric acid, respectively. Biosyn-
thesis involves a trans/cis-isomerization, or rotation
HO O around the C-C double bond (arrows) in the side chain,
O
followed by ring closure.
Scopoletin

O OO O committed step in flavonoid biosynthesis. CHS cat-

alyzes the stepwise condensation of three molecules of
malonyl-CoA with one molecule of p-coumaryl-CoA
C to form 4,2 ,4 ,6 -tetrahydroxychalcone, or, naringenin
H2 chalcone. A chalcone is a C6 —C3 —C6 pattern wherein
OH OH ring C is not yet closed. The enzyme chalcone isomerase
Dicoumarol then catalyzes the closure of ring C to form narin-
genin. Naringenin is the precursor to flavonols such
O as kaempferol and quercitin as well as the anthocyanin
pigments.
O Stilbenes are a group of C6 —C2 —C6 compounds
that have a chemical defense role, primarily against
fungal invasions of heartwood (the distinctively colored
core of the tree). Stilbenes are one of several phenyl-
OCH3 propanoid derivatives that are continually infused into
O O
the heartwood of trees after the cells have been lignified.
Aflatoxin B1 A defensive role is suggested by the localized deposition
FIGURE 27.16 Coumarins. More than 300 coumarins of stilbenes in regions of sapwood as well, when the sap-
have been reported from more than 70 plant fami- wood is attacked by insects of pathogenic fungi. The role
lies. Coumarin and 7-hydroxycoumarin (umbelliferone) of stilbenes in this case appears to be one of preventing
are derived from cinnamic acid and p-coumaric acid, the spread of the invading insect or pathogen.
respectively. Scopoletin is probably the most widespread Stilbene biosynthesis, like the flavonoids, involves
coumarin in plants. Dicoumarol is a powerful anticoag- the sequential condensation of malonyl-CoA units with
ulant found in moldy hay. Dicoumarol derivatives and either cinnamoyl-CoA or p-coumaroyl-CoA, except
synthetic coumarins are commonly used to thin blood that the enzyme catalyzing the reaction is stilbene
in cardiac patients. Aflatoxin B1 , a mycotoxin, is among synthase.
the most carcinogenic and toxic of naturally occurring
compounds.
27.4.6 TANNINS DENATURE PROTEINS
AND ADD AN ASTRINGENT
Both ring B and the 3-carbon bridge that makes up TASTE TO FOODS
ring C are derived from the shikimic acid pathway via
phenylalanine and p-coumaric acid. The six carbons The name tannin is derived from the historic practice
that make up the A ring are derived from malonic of using plant extracts to ‘‘tan’’ animal hides, that is,
acid, in the form of a malonyl-coenzyme A complex, to convert hides to leather. Such extracts contain a
malonyl-CoA. (Malonyl-CoA is also a principal inter- mixture of chemically complex phenol derivatives that
mediate in fatty acid synthesis.) The key enzyme is bind to, and thus denature, proteins. For years, the most
chalcone synthase (CHS), which catalyzes the first common test for tannins was the capacity to precipitate
 OH OH OH
O CH3 CH3 O O CH

CH CH CH
CH CH CH
CH2OH CH2OH CH2OH
Coumaryl alcohol Coniferyl alcohol Sinapyl alcohol
FIGURE 27.18 Monolignols are the principal lignin monomers. Extensive
cross-linkages most commonly form between the ring alcohol group and the
double-bonded carbon atoms.

O O

(3) Malonyl - CoA C C

O-
CH2 S - Co A

+
Cinnamoyl CoA
(R1, R2 = H) R1

Coumaroyl CoA
R2
( R1 = OH, R2 = H)
CoAS

Chalcone synthase Stilbene synthase R2

R1

OH HO
B
HO OH
A OH
Naringenin Stilbenes
chalcone

OH O

OH
B
HO O
A C
Naringenin Flavonols
(a flavonone) Anthocyanins

OH O
FIGURE 27.19 The flavonoid ring structure and stilbenes are both synthesized by
sequential condensations of 3 molecules of malonyl-coenzyme A (CoAS) with either
cinnamoyl-coenzyme A or p-coumaroyl-coenzyme A. Malonyl-coenzyme A is in
turn formed by the carboxylation of acetyl-CoA, the first step in the formation of
long-chain fatty acids.

473
474 Chapter 27 / Secondary Metabolites

COOH H3 C
C CH
H3C
COO R
HO OH
OH H3C
Gallic acid CH3
FIGURE 27.20 Gallic acid is the basic structural unit of Pyrethrin I
hydrolyzable tannins.
FIGURE 27.21 Pyrethrin, a monoterpenoid ester, is a
commonly used organic insecticide. Pyrethrin is isolated
from thedried, unexpanded flowerheads of Chrysanthe-
gelatin (a protein). Two categories of tannins are now mum cineraiifolium (Asteraceae).
recognized: condensed tannins and hydrolyzable tan-
nins. Condensed tannins are polymers of flavonoid units
linked by strong carbon-carbon bonds. These bonds membrane. Pyrethrins are popular organic insecticides
are not subject to hydrolysis but can be oxidized by because they have a relatively low mammalian toxic-
strong acid to release anthocyanidins. The basic struc- ity. Natural pyrethrins are also readily inactivated by
tural unit of hydrolyzable tannins is a sugar, usually oxidation and so do not persist in the environment.
glucose, with its hydroxyl groups esterified to gallic acid Many other terpenes, while not insecticides per se,
(Figure 27.20). Gallic acid residues are in turn joined to do help plants to repel invasions by insects. Many plants
form an extensively cross-linked polymer. respond to insect attack by producing additional quan-
Like other phenolics, the biological role of tannins tities of toxic or repellant metabolites. Unfortunately,
is not clear. Tannins do appear to deter feeding by many in some cases chemical deterrents can turn against the
animals when tannin-free alternatives are available. plant. As insects evolve resistance, they are able to use
This effect could be related to the astringency—a sharp, the same chemical as a host-recognition cue to help
somewhat unpleasant sensation in the mouth—for them locate hosts they can feed on without ill effect.
which tannins are noted. The astringent property of At least one group of flavones, the isoflavones,
tannins is a component in the flavor of many fruits has become known for its anti-microbial activities
as well as drinks such as coffee, tea, and red wine. (Figure 27.22). Isoflavones are one of several classes
As well, tannins tend to suppress the efficiency of of chemicals of differing chemical structures, known
feed utilization, growth rate, and survivorship. The as phytoalexins, which help to limit the spread of
conventional interpretation has been that tannins bacterial and fungal infections in plants. Phytoalexins
reduce digestibility of dietary protein, presumably by are generally absent or present in very low concentra-
binding with protein in the gut. Other studies, however, tions, but are rapidly synthesized following invasion by
have cast doubt on this interpretation, suggesting that bacterial and fungal pathogens.
tannins may be toxic in other, yet unknown, ways. The details of phytoalexin metabolism are not yet
clear. Apparently a variety of small polysaccharides,
glycoproteins and proteins of fungal or bacterial origin,
27.5 SECONDARY METABOLITES serve as elicitors (L. elicere, to entice) that stimulate
the plant to begin synthesis of phytoalexins. Studies
ARE ACTIVE AGAINST with soybean cells infected with the fungus Phytophthora
INSECTS AND DISEASE indicate that the fungal elicitors trigger transcription
27.5.1 SOME TERPENES AND
ISOFLAVONES HAVE HO O
INSECTICIDAL AND 2
ANTI-MICROBIAL ACTIVITY A C
3
One of the better-known and commonly used natural
B
insect toxins is pyrethrin, a monoterpene ester that is H O
produced in the ovaries of flowers of Chrysanthemum O
(or, Pyrethrum) cineraiifolium (Figure 27.21). C. cinerai- FIGURE 27.22 Structure of the isoflavone formononetin,
ifolium is widely cultivated in Montenegro, Japan, and isolated from red clover (Trifolium pratense). In a flavone,
eastern Africa, where the unexpanded flower heads are the B ring is attached to the 2 position of the C ring. In
dried and powdered. Pyrethrin is a neurotoxin that an isoflavone, the B ring is attached to the 3 position of
interferes with sodium channels in the insect nerve the C ring.
 27.5 Secondary Metabolites are Active Against Insects and Disease 475

of mRNA for enzymes involved in the synthesis of function of their products is not known. One pos-
isoflavones. The production of phytoalexins appears to sibility is that avirulence genes encode enzymes for
be a common defense mechanism. Isoflavones are the the production of elicitors and resistance genes encode
predominant phytoalexin in the family Leguminoseae, receptors that recognize elicitors. A variety of elicitors
but other families such as Solanaceae, appear to use have been identified, most of them extracellular micro-
terpene derivatives. bial products commonly associated with cell walls of
bacteria and fungi. For example, fungal elicitors include
27.5.2 RECOGNIZING POTENTIAL β-glucans, chitosan (a chitin subunit),1 and arachi-
PATHOGENS donic acid (an unsaturated lipid). Other elicitors include
various polysaccharides, glycoproteins, and small pep-
As discussed earlier in Chapter 13, plants challenged tides. Even pectic fragments, resulting from initial
by insects or potentially pathogenic organisms also degradation of the plant cell wall pectins, or mechan-
respond with a hypersensitive reaction, characterized ical damage are capable of eliciting a hypersensitive
by changes in the composition and properties of the cell reaction.
wall and other factors, including necrotic lesions at the Recognition of elicitors by the plant cell likely
site of the invasion, that limit the spread of the invading takes place at the plasma membrane. It is expected that
pathogen. Because the hypersensitive reaction is a form some form of signal transduction pathway is required
of developmental response, we must assume that a signal to relay this information to the nucleus in order to
detection and transduction chain is involved. effect gene transcription. A variety of common signal-
Attempts to explain the susceptibility of plants to ing agents have been suggested, including changes in
infection have shown that disease has an underlying pH, and ion fluxes (especially potassium and calcium).
genetic basis. In fact both pathogens and potential host For example, a transient uptake of Ca2+ (and efflux
plants carry genes that determine whether or not disease of K+ ) was observed when cultured cells were chal-
will occur and disease will occur only when those genes lenged with a fungal elicitor. Moreover, expression of
are compatible. Compatibility can be explained by the defense response genes can be regulated by regulat-
gene-for-gene model, which predicts that resistance ing intracellular Ca2+ levels. Thus, defense responses
will occur only when the pathogenic microorganism can be activated by stimulating Ca2+ uptake with Ca2+
carries a dominant allele at the avirulence (Avr) locus ionophores or inhibited by blocking Ca2+ channels.
and host plants carry a complementary dominant allele Other early events in elicitor-treated cells include pro-
at the resistance (R) locus (Figure 27.23). Any other tein phosphorylation and the production of active oxy-
combination of dominant and recessive alleles leads to gen species (O− 2 and H2 O2 ), known as the oxidative
a successful infection. A matching pair of dominant burst. The precise role of these various signals and how
pathogen avirulence genes and dominant plant resis- they interact in the signal cascade is unknown. It is
tance genes initiates a hypersensitive reaction in the a topic that is under active investigation in numerous
plant. laboratories.
Although a number of avirulence genes have been
isolated from both bacteria and fungi, the specific
27.5.3 SALICYLIC ACID, A SHIKIMIC
ACID DERIVATIVE, TRIGGERS
Host plant genotype SYSTEMIC ACQUIRED
R r RESISTANCE
Avr/R Avr/r In many cases, the hypersensitive reaction leads to a
Avr incompatible compatible general immunity against infection known as systemic
Pathogen genotype

acquired resistance or SAR (Chapter 13). Most of the

no disease disease evidence suggests that the signal for SAR is salicylic
acid (SA) or its volatile methylated derivative methyl-
avr/R avr/r
salicylate. The role of SA in SAR is indicated by several
avr compatible compatible
lines of evidence. For example, SAR is characterized by
a rise in the levels of SA along with the activation of
disease disease defense-related genes and the appearance of their prod-
FIGURE 27.23 The gene-for-gene model predicts that ucts, pathogenesis-related (PR) proteins. In addition, SA
incompatibility between a pathogen and host requires
complementary dominant avirulence and resistance
1
genes in the pathogen and host, respectively. All other The principal carbohydrate in most fungal cell walls is chitin
genotypes are compatible and lead to invasion of the host rather than cellulose. Chitin is a polymer of N-acetylgluco-
by the pathogen or, i.e., disease. samine that forms microfibrils similar to cellulose.
476 Chapter 27 / Secondary Metabolites

is synthesized from chorismate via isochorismate, a reac- receptor in the plasma membrane. The elicitor-receptor
tion catalyzed by the enzyme, but Arabidopsis mutants complex activates a membrane-bound phospholipase
that failed to accumulate SA after SAR induction were that releases linolenic acid. The formation of jasmonic
shown to be deficient of this enzyme. Accumulation of acid then involves several steps in which the linolenic
SA in infected plants carrying mutations of the isocho- acid undergoes successive oxidizations and cyclization.
rismate synthase gene is no more than 5 to 10 percent Central to jasmonate signaling is a family of
of wildtype plants and systemic resistance is severely transcriptional repressors (JAZ) and an F-box protein
compromised. (COI1) that is structurally and functionally related to
The molecular genetics and biochemistry of SAR the TIR1 auxin receptor (Chapter 18). Because of this
is slowly yielding to investigation. A central player is similarity, the current model for jasmonate signaling
the regulatory protein NPR1 (NON EXPRESSOR bears a close resemblance to that of auxin-mediated
OF PATHOGENESIS-RELATED GENES1). NPR1 degradation of AUX/IAA transcriptional repressors
is expressed at low levels throughout the plant, but its previously described in Chapter 18. According to this
messenger RNA levels rise some two- to threefold fol- model, jasmonates promote binding of JAZ repressor
lowing pathogen infection or salicylic acid treatment. proteins to the E3 ubiquitin ligase (SCFCOI1 ) and
In addition, several Arabidopsis mutants that were not their subsequent degradation via the ubiquitin-26S
responsive to salicylic acid were found to have muta- proteasome system, thus allowing for the activation of
tions in the NPR1 gene. NPR1 protein interacts with a jasmonate-responsive genes.
group of transcription factors known as the TGA fam-
ily of transcription factors; an interaction that requires
salicylic acid and stimulates the transcription of PR 27.7 ALKALOIDS
genes.
Of particular interest is the recent discovery that 27.7.1 ALKALOIDS ARE A LARGE FAMILY
changes in the redox status of the cell play an impor- OF CHEMICALLY UNRELATED
tant role in the NPR1/TGA interaction. A change MOLECULES
in the cellular redox potential, from an initial burst
of reactive oxygen species to a more reducing envi- As a group, alkaloids share three principal characters:
ronment, is induced by salicylic acid and appears to they are soluble in water, they possess at least one
facilitate the transcription of PR genes by triggering the nitrogen atom, and they exhibit high biological activity.
translocation of NPR1 from the cytosol into the nucleus Often the nitrogen will accept a proton, which gives it
where it is free to interact with the TGA transcription a slightly basic, or alkaline, character in solution (hence
factors. the name alkaloid). Alkaloids are for the most part het-
erocyclic, although a few aliphatic (noncyclic) nitrogen
compounds, such as mescaline and colchicine, are con-
sidered alkaloids. Altogether, some 12,000 alkaloids have
27.6 JASMONATES ARE LINKED been found to occur in approximately 20 percent of the
TO UBIQUITIN-RELATED species of flowering plants, mostly herbaceous dicots.
PROTEIN DEGRADATION
As previously shown in Chapter 13, a plant’s resistance 27.7.2 ALKALOIDS ARE NOTED
to insects and disease is also mediated by the fatty acid PRIMARILY FOR THEIR
derivative jasmonic acid and its methyl ester (methyl- PHARMACOLOGICAL
jasmonate), collectively referred to as jasmonates (See PROPERTIES AND MEDICAL
Figure 13.16). Jasmonates were first recognized for their
APPLICATIONS
ability to promote senescence of detached barley leaf
segments, but a role in disease resistance was suggested The word ‘‘alkaloid’’ is virtually synonymous with the
when phytoalexin biosynthesis in cell cultures was linked word ‘‘drug’’; as recently as 1985, 10 of the 12 com-
to jasmonic acid content. It is now known that jasmonic mercially most important plant-derived drugs were
acid accumulates in wounded plants and in plants treated alkaloids. Alkaloids generate varying degrees of phys-
with many elicitors. Jasmonic acid has also been linked iological and psychological response in humans, most
to the activation of a number of genes encoding proteins often by interfering with neurotransmitters. In large
with antifungal properties. doses, most alkaloids are highly toxic, but in smaller
Jasmonic acid is synthesized by the oxidation and doses they may have therapeutic value.
rearrangement of linolenic acid, an unsaturated fatty acid From prehistory to the present, alkaloids or
(see Appendix). Plant membranes are a rich source of alkaloid-rich extracts have been used for a variety of
linolenic acid as a constituent fatty acid of phospholipids. pharmacological purposes, such as muscle relaxants,
It is thought that elicitors first bind with an unknown tranquilizers, antitussives, pain killers, poisons, and
 27.7 Alkaloids 477

mind-altering drugs. One of the oldest known is opium, Berberine, however, is found in several sources, includ-
an exudate obtained from the immature seed capsule ing seeds of barberry (Berberis vulgaris). Barberry was
of the opium poppy (Papaver somniferon). The use of at one time a common horticultural hedge but is now
opium mixed with wine to induce sleep and relieve pain out of fashion due to the high incidence of poisoning in
was noted on Sumerian tablets dating back to 2500 BC. young children attracted to its bright-red berries.
The species name somniferon was chosen by Linnaeus The quinolizidine alkaloids, such as lupinine, are
because of its sleep-inducing properties. Opium is frequently called lupine alkaloids because of their high
a latex gum containing a mixture of more than 20 abundance in the genus Lupinus. Although range ani-
different alkaloids, including morphine, codeine, and mals are deterred from eating lupines because of their
papaverine (Figure 27.24). The opium poppy has been bitter taste, grazing on lupines is still a common cause
traditionally cultivated in the Golden Crescent of the of poisoning of grazing cattle. The highest concen-
eastern Mediterranean (presently Iran, Afghanistan, tration of alkaloids occurs in the seed, so livestock
and Pakistan) and in parts of southeast Asia. The losses are generally highest in the fall. Other alkaloids
genus Papaver contains 9 other species, many of them that cause poisoning of livestock include senecionine
common garden ornamentals, but none of which (Senecio, groundsel), lycotonine (Delphinium, larkspur),
produce alkaloids of medical interest. In 399 BC, the scopolamine (Datura stramonium, jimson weed), and
Greek philosopher Socrates was executed by consuming atropine (also known as hyoscyamine) from black
an extract of hemlock (Conium spp.) containing the henbane (Hyoscyamus niger). Agricultural crops are not
alkaloid coniine. In the Western Hemisphere, the without some risk to human consumption. The family
Aztecs and other native cultures used the peyote cactus Solanaceae is noted for genera such as Datura, Hyoscya-
(Lophophora williamsii), containing mescaline and a mus, and Atropa belladonna (the deadly nightshades),
large number of other alkaloids, for its hallucinogenic all containing high amounts of toxic alkaloids. Edible
properties. members of the Solanaceae include potato (Solanum
Alkaloids are generally classified on the basis of tuberosum) and tomato (Lycopersicum esculentum), which
the predominant ring system present in the molecule produce the steroidal alkaloids α-solanine and tomatine,
(Figure 27.25). In spite of the extensive variation in respectively. The solanine alkaloids are cholinesterase
structure, however, alkaloids are generated from a lim- inhibitors that interfere with nerve transmission. For-
ited number of simple precursors. Most alkaloids are tunately, the alkaloid content of both potato tubers and
synthesized from a few common amino acids (tyrosine, ripe tomato fruit is well below toxic levels, although
tryptophan, ornithine or argenine, and lysine). The green vines have higher levels and are potentially toxic
tobacco alkaloid nicotine is synthesized from nicotinic if eaten. Potato tubers that have been exposed to strong
acid and caffeine is a purine derivative. sunlight and have begun to green may also synthesize
Although a few alkaloids are found in several genera toxic levels of the α-solanine and should not be eaten.
or even families, most species display their own unique, The indole alkaloids are often referred to as
genetically determined pattern. As with other secondary terpenoid-alkaloids because, although the basic indole
metabolites, individual alkaloids may be restricted to ring structure is derived from tryptophan, the rest
particular organs, such as roots, leaves, or young fruit. of the molecule is derived from the mevalonic acid

HO HO H3C CO O

NCH3 NCH3 NCH3

O O O

H3CO HO H3C CO O
Codeine Morphine Heroin
FIGURE 27.24 Codeine, morphine, and heroin are structurally related alkaloids.
Codeine and morphine are naturally occurring alkaloids isolated from the seed cap-
sules of the opium poppy (Papaver somniferon). Heroin is a semisynthetic alkaloid
produced by acetylation of morphine. Codeine is commonly used as a cough-
suppressant and local anesthetic. Morphine is used primarily as an analgesic or pain
killer. Heroin was originally synthesized by pharmaceutical chemists in the late nine-
teenth century as part of an effort to find a more effective alternative to morphine.
 Alkaloid Class Example Other Representatives
CH CH2
Quinoline
HO
H3CO N

N quinine
N
Isoquinoline H3CO
morphine
papaverine codeine
N
H3CO berberine
OCH3
N

OCH3
Indole
N
vinblastine
vindoline reserpine
strychnine
OH
N N OCOCH3

CH3 COOCH3

Pyrrolizidene HO CH3
O
H3C
CH3 O senecionine retrorsine
O O
N

N
Quinolizidine OH

lupinine cytisine
N N

O
Tropane
H scopolamine
N NCH3 O C C atropine cocaine
H2COH

Piperidine

N nicotine coniine

N CH3
H N

O CH3
Purine H H3C
N N
N N
caffeine

N N O N N

CH3
FIGURE 27.25 Alkaloids are a chemically diverse group of heterocyclic,
nitrogen-containing compounds that are classified according to the predominant
type of ring structure.

478
 Summary 479

pathway. Two well-known terpenoid-indole alkaloids, tree Cinchona officinalis and Rauwolfia alkaloids are con-
vinblastine and vincristine, are produced by the Mada- centrated primarily in the root bark. Although alkaloids
gascar periwinkle, Catharanthus roseus. In the 1950s it have been studied extensively for their pharmacological
was discovered that vinblastine and vincristine arrest and medicinal value, there is much yet to be learned
cell division in metaphase by inhibiting microtubule with respect to their physiology and chemical ecology.
formation. They have since been used in the treatment
of Hodgkin’s lymphoma, leukemia, and other forms
of cancer. Unfortunately, C. roseus produces a complex
array of indole alkaloids of which vinblastine and vin- SUMMARY
cristine represent only a very small proportion (about Although the products of plant metabolism are often
0.05% of leaf dry weight). This makes their extraction designated as either primary or secondary metabolites,
and purification difficult and costly. Recent efforts have the distinction between the two is not easily made.
led to the development of cell culture systems in order to Primary metabolites such as protein, lipid, carbohy-
produce higher yields at reasonable cost. Unfortunately, drate, and nucleic acids comprise the basic metabolic
because alkaloid production is so often tissue-specific, machinery of all cells. Others, such as chlorophyll and
production in cell culture has not been highly success- lignin, are more restricted in occurrence but are equally
ful. Other well-known alkaloids such as cocaine (from essential to the growth and development of the organ-
leaves of the coca plant, Erythroxylum coca), codeine ism. Secondary metabolites, on the other hand, may be
(from the opium poppy, Papaver somniferum), nicotine found only in specific tissues or at particular stages of
(tobacco), and caffeine (coffee beans and tea leaves) are development and have no obvious role in the develop-
widely used as stimulants or sedatives. ment or survival of the organism.
Three metabolic pathways that give rise to both
primary and secondary metabolites are the mevalonic
27.7.3 LIKE MANY OTHER SECONDARY acid and MEP pathways and the shikimic acid pathway.
METABOLITES, ALKALOIDS The mevalonic acid and MEP pathways give rise to
SERVE AS PREFORMED two 5-carbon compounds, isopentenyl pyrophosphate
CHEMICAL DEFENSE and dimethylallyl pyrophosphate, that form the basis
for the terpenoid family. Terpenoids include primary
MOLECULES
metabolites such as phytol (a portion of the chlorophyll
Whether alkaloids have any specific function in plants molecule), membrane sterols, carotenoid pigments, and
has been a subject of debate for many years. Some the hormones gibberellin and abscisic acid. Secondary
plants appear to invest a significant proportion of their products include a range of 5-carbon hemiterpenoids
resources, especially nitrogen, in a diverse array of alka- through 40-carbon tetraterpenes and the polyterpenes,
loids and yet they have no obvious function in the rubber and gutta.
physiology of the plants themselves. Like other sec- The shikimic acid pathway produces the
ondary products, however, there are at least two general aromatic amino acids phenylalanine, tyrosine, and
arguments supporting a defensive role. Firstly, most tryptophan—all primary metabolites required for
alkaloids have a bitter taste, which is considered a uni- protein synthesis. Deamination of phenylalanine to
versally repellant character for all animals, including cinnamic acid, catalyzed by the enzyme phenylalanine
insects. Secondly, all alkaloids are biologically active ammonia lyase (PAL), effectively diverts carbon from
and many are significantly toxic to insects and other primary metabolism into the synthesis of a wide range
animals. Nicotine, for example, is a potent insect poison of secondary metabolites—coumarins, lignin, tannins,
and one of the first insecticides used by humans. flavonoids, and isoflavonoids— based on simple
Many alkaloids have antibiotic properties and may phenolic acids.
have a role in defense against microbial infection. Inter- Salicylic acid is involved in establishing the hyper-
estingly, it has been known for a long time that alkaloid sensitive reaction to insect herbivory and pathogen
concentrations are very low when measured on a whole infection. The gene-for-gene model predicts that no
plant basis, but, as noted earlier, may be very high in infection will occur when the pathogen and host
selected organs or tissues. Often the tissues that accumu- plant carry dominant avirulence and response genes,
late alkaloids are those most vulnerable in terms of plant respectively. An early event in this sensing/signaling
fitness (young influorescences or seed pods, for example) pathway is the activation of defense-related genes and
or peripheral tissues that would be first attacked by her- synthesis of their products, pathogenesis-related (PR)
bivores. Thus the morphine content of whole young proteins. The plant immune response continues with
poppy capsules is less than 2 percent, but 25 percent the development of a general immune response known
or more in the latex that exudes when the capsule is as systemic acquired resistance (SAR). SAR signaling
wounded. Quinine accumulates in the outer bark of the includes the protein NPR1, which responds to a change
480 Chapter 27 / Secondary Metabolites

in cellular redox potential by migrating into the nucleus it often takes a week before any signs of
where it interacts with transcription factors to stimulate injury are observed. Explain this delay.
the transcription of PR genes. 9. In what ways do plants have some control over
Other interesting and useful secondary metabolites insect herbivory?
include the saponins (terpene glycosides), cardiac gly-
10. How do plants respond to invasion by microbial
cosides, cyanogenic glycosides, and glucosinolates. The
pathogens?
alkaloids are a heterogeneous group of nitrogenous
compounds with significant pharmacological prop-
erties. Although the physiological roles of secondary
metabolites are poorly understood, most are toxic to FURTHER READING
some degree and appear to serve primarily in defense
against microbial infection and attack by herbivores. Cordell, G. (ed.). 1997. The Alkaloids. San Diego: Academic
Press.
Durrant, W. E., X. Dong. 2004. Systemic acquired resistance.
Annual Review of Phytopathology 42:185–209.
CHAPTER REVIEW Fobert, P. R., C. Després. 2005. Redox control of systemic
acquired resistance. Current Opinion in Plant Biology
1. Distinguish between primary and secondary
8:373–382.
metabolism.
Grant, M., C. Lamb. 2006. Systemic immunity. Current Opin-
2. What are terpenes? How do they originate within ion in Plant Biology 9:414–420.
the plant and what functions do they serve? Harborne, J. B., F. A. Tomas-Barberan (eds.). 1991. Ecological
3. Explain why sterols may be considered Chemistry and Biochemistry of Plant Terpenoids. Oxford:
both primary and secondary metabolites. Clarendon Press.
4. What do saponins, cardenolides, and amygdalin Klein, R. M. 1987. The Green World: An Introduction to Plants
have in common? and People. New York: Harper & Row.
O’Kennedy, R., R. D. Thornes (eds.). 1997. Coumarins: Biol-
5. Why are amino acids such as phenylalanine
ogy, Application, and Mode of Action. Chichester: Wiley.
and tryptophan essential in the human diet?
Rosenthal, G. A., M. R. Berenbaum. 1991. Herbivores: Their
6. What are EPSPS and PAL? What key metabolic Interactions with Secondary Metabolites. Vol. 1: The Chemi-
roles do they play? cal Participants. 2nd ed. San Diego: Academic Press.
7. Alkaloids are, for the most part, chemically Staswick, P. E. 2008. JAZing up jasmonate signaling. Trends
unrelated. What is the basis for grouping them in Plant Sciences 13:66–71.
together?
8. How does the herbicide glyphosate kill plants?
When plants are treated with glyphosate,