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Defining Evolutionarily Significant Units for conservation

inferences about the evolutionary past. Thus, the term significantin ESUshould CraigMoritz is at the Dept of Zoology and be seen as a recognition that the set of Centre for Conservation Biology, populations has been historically isoThe University of Queensland, lated and, accordingly, is likely to have a Qld 4072,Australia. distinct potential. According to this view, the emphasis is on historical population structure rather than current riting in the first issue of TREE, adaptation. This departs from the more Ryder brought the term Evolution- usual concern that we should seek to w maintain the full array of differently arily Significant Unit(ESU) to the attention of a broad audience of ecologists and adapted geographic variants within a species3J. I suggest that to focus on evolutionary biologists. The ESU concept was developed to provide a rational basis maintaining the full array of locally adapted variants is not only difficult in for prioritizing taxa for conservation practice, but also negates the evoleffort (e.g. captive breeding), given that utionary process that we seek to mainresources are limited and that existing tain, insofar as preservation of variants taxonomy may not adequately reflect adapted to previous conditions may reunderlying genetic diversity*. With the explicit recognition of the genetic com- tard the response to natural selection. There may, of course, be other nonponent of biodiversity in conservation evolutionary reasons (e.g. ecological, legislation of many countries and in the economic, aesthetic) for ascribing Convention on Biological Diversity, the conservation value to a particular popuESU concept is set to become increasingly significant for conservation of natural lation. The recognition of ESUs is primarily as well as captive populations. However, the ESU remains poorly de- relevant to long-term management issues, that is, defining conservation priorities fined, both conceptually and operationally. Most definitions suggest than an ESU and setting strategy, although in the short should be geographically discrete, but term it may be prudent to avoid trans,~. genetic criteria range from significant di- locating individuals between ESUS vergence of allele frequencies3 through some level of genetic distance to congru- Criteria for recognizing an ESU Defining an ESU as a historically isoently structured phylogenies among lated set of populations leads to a qualigene+. Several authors have argued tative criterion based on the distribution that an ESU should display concordant divergence for both molecular and non- of alleles in relation to their phylogeny molecular traitG6. Although all are try- (Fig. 1). Simulation studies suggest that it takes about 4N generations from the ing to achieve the same end, it seems time that two populations separate for that the operational definitions vary according to the biological and legislat- there to be a high probability of their ive context. The purpose of this essay is having reciprocally monophyletic allelesg. to revisit the ESU concept in relation to Because of its relatively low effective recent developments in molecular popu- population size and high substitution rate, lation genetics. The suggested defi- animal mitochondrial DNA (mtDNA) is nitions and criteria are not supposed to expected to achieve this condition more be proscriptive - rather, the intention is rapidly than nuclear alleles. Indeed, welldifferentiated sister species may have to promote debate on the purpose and practice of using genetic information to reciprocally monophyletic mtDNA but phylogenetically unsorted alleles at nudefine conservation units. clear loci (e.g. northern versus southern elephant sealslo). To require reciprocal Conservation goals: what do we monophyly for both nuclear and mtDNA mean by significant ? The overriding purpose of defining genes (as required for genealogical ESUs is to ensure that evolutionary heri- concordance4) seems overly restrictive. Nonetheless, significant divergence in tage is recognized and protected and nuclear allele frequencies should be rethat the evolutionary potential inherent across the set of ESUs is maintained. For quired to avoid misclassifying populations a given set of populations we cannot pre- linked by nuclear, but not organellar, gene diet future outcomes, but we can make flow.

100

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Craig Moritz

0-

200

400

600

800

Generations

1wh Wy

II paraphyly

ab cd A-E-

abc __
A

d BA

a -

bed 0

Ill reciprocal monophyly

ab T Fig. 1. Development
mg of alleles

cd B

of phylogenetlc structurbetween populations. The graph (modified from Ref. 8) shows the results of 100 simulations with two daughter populations founded with 20 and 30 lndlviduals and allowed to grow rapidly to N = 200. P indicates the probability of alleles (a-d) within the two populations (A, 6) being (I) polyphyletic. (II) paraphyletic or (Ill) reciprocally monophyletlc. After the division of one populatton Into two, the phylogenetic relations of the alleles in the two daughter populations typlcally proceed from polyphyly, through various paraphyletlc conditions to reclprocal monophyly as ancestral polymorphisms are sorted and replaced by derived statesg. The rate depends on effective population size, usually taking at least 4N generations to achieve reciprocal monophyly, and is also influenced by mutation rate. population demography and the phylogeographlc dlstributlon of alleles before the separation of the two populations.

The above theory suggests a genetic criterion for recognizing an ESU: ESC s
should be reciprocally monophyletic for mtDNA alleles and show significant divergence of allele frequencies at nuclear locr.

Although such a definition may seem to be overly restrictive in some cases (see below), it has the advantages of being theoretically sound and of avoiding the issue of how much divergence is enough? that plagues quantitative criteria such as allele frequency divergence and genetic distance. It considers the pattern rather than the extent of sequence divergence, as it is not the intention to ascribe conservation value to an ESU in relation to mtDNA distance.

Contrast with management units and stocks

In practice, genetic analyses often
differences between sampled populations ranging from reciprocal monreveal

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combines the two types of unit. Dizon et al.5 attempted to clarify the definition of stocks using a hierarchy of phylogeographic pattern in conjunction with other evidence. Although their scheme was explicit, it remained unwieldy and did not recognize the different conservation goals. I suggest that the term stock be restricted to short-term management issues (e.g. monitoring harvests, etc.) and, in relation to genetics, be treated as synonymous with MUs as defined above. The identification of ESUs as defined above requires information on the distribution and phylogeny of mtDNA alleles and on the distribution of nuclear alleles. In contrast, only information on allele frequency is directly pertinent to the delineation of MUs, although for small samples and loci with high substitution rates, sequence information may provide more power for detecting population subdivisioni6. These types of data are accumulating rapidly for threatened or For green turtles exploited species 11J5,17. (Chelonia mydas), the genetic data suggest two ESUs - one in the Atlantic Ocean and the other in the lndoPacific - each consisting of numerous MUs (Fig. 2). Here, the black turtle (C m. agassizi) represents just one MU within the larger lndo-Pacific ESU. The humpback whale (Megaptera novaengliae), another intensively studied species, appears to represent a single ESU with numerous MUs, many of which correspond to major stocks recognized from migration routes18. As with any evolutionary property of populations or species, the definition of the ESU needs to be applied with common sense. In some circumstances it may seem overly restrictive. For example, where there has been rapid speciation or recent hybridization, mtDNA alleles may not yet be sorted between otherwise discrete taxa. However, the failure to define these as separate ESUs should

ophyly, through substantial but incomplete phylogenetic separation, to minor but statistically significant differences in allele frequency. Populations that do not show reciprocal monophyly for mtDNA alleles, yet have diverged in allele frequency, are significant for conservation in that they represent populations connected by such low levels of gene flow that they are functionally independent. The recognition of such Management Units (MUs) is fundamental to proper short-term management of the more inclusive ESUs, in that MUs are the logical unit for population monitoring and demographic study. MlJs are therefore recognized as populations with significant dive%ence of allele frequencies at nuclear or mitochondrial loci, regardless of the phylogenetic distinctiveness of the alleles. The distinction between ESUs and MUs is important, as it affects ways in which genetic evidence is obtained and interpreted. To use genetic information effectively, we should therefore distinguish between two types of conservation units, both important for management: ESUs,concerned with historical population structure, mtDNA phylogeny and long-term conservation needs; and MUs, addressing current population structure, allele frequencies and short-term management issues. The concept of discrete stocks as used for marine speciessJ2 sometimes

Application and limitations

The foregoing treatment was written with animal populations undergoing predominantly divergent evolution in mind. In practice, ESUswill usually camp lement rather than replace species defined under traditional, predominantly morphological criteria (although ESUs and species would be synonymous under some species conceptslJ4). Given the shortage of resources for managing major ecosystems, let alone previously described species, it is logical to focus genetic studies on species of greatest concern. However, an exciting extension is to apply these principles to whole communities - using comparative phylogeography to define geographic areas where component species have evolutionary histories separate from their conspecific+. This could have considerable significance for planning of regional reserve systems.

Atlantic-Mediterranean . Indo-Pacific , \
Aves (1) Costa Rica (15) Florida (,21) Florida ,(3) Cyprus (10) Aves (7) Suriname

@I
Florida Costa Rica Venezuela Suriname Ascension Brazil

FlA

C6R

VEN SUR ASC BRA AFR CYP OMA AUS JAP HAW MEX-B GAL

Hawaii (16)

hi-

(15)

Africa Cyprus Oman Australia Japan

Ascension (1) Ascension Brazil (15) (34)

Hawaii (6) Galapagos (8) Mexico (7) Oman(15) I

Hawaii
Brazil (1) Guinea Biaaau (1 2)

Mexico-B ~~apacJ~

0.8

0.6

% Sequence

divergence

P>O.O50

P<O.O60

P-ZO.005

Fig. 2. Definrng conservation units for green turtles (Chelonia mydas). (a) UPGMA dendrogram of mtDNAs analysed from 15 rookeries of green turtle spannrng most of the global distribution (redrawn from Ref. 20). (b) Tests of heterogeneity of allele frequencies at mtDNA (below) and five nuclear loci (above) among the same 15 rookeries. The first eight rows or columns are the Atlantic-Mediterranean rookeries (redrawn from Ref. 21). Rookerres throughout the global distribution of the green turtle have been screened for restriction fragment length polymorphisms of mtDNA>o and anonymous single copy nuclear sequence+. A major phylogeographic break is evident between mtDNAs from Atlantic-Mediterranean and Indo-Pacific rookeries (a), supported by slight, but significant variation in nuclear genes. Structuring of allele frequencies among rookeries within either area was substantial for mtDNA and less marked, but still significant for the nuclear loci (b). Accordingly, the species should be managed as two ESUs (Atlantic-Mediterranean and Indo-Pacific) each consisting of multiple MUs. Reanalysis of mtDNAs by sequencing of control region sequences can greatly increase the resolution of MUs (e.g. from 3 to 9 in the Australasian region22) but has not altered the perception of ESUs.

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100~~ not affect conservation priorities because the taxa in question are probably already recognized as species on broader biological criteria. A group of populations, such as North Atlantic humpback whales, which shows substantial but incomplete phylogenetic sorting of mtDNA allelesr8, would not be classified as a separate ESU, but still warrant conservation attention as separate management units. Conversely, the criteria may be oversensitive in some cases in species of very low vagility where most local populations are strongly differentiated for mtDNA and nuclear genes (e.g. Ensatir~G). In this circumstance, the genetic differences need to be interpreted in the context of the total variation within the species. An additional caveat is that the identification of ESUs and MUs is susceptible to error because of insufficient sampling: the analysis of too few individuals or populations could lead to the false recognition of ES&.; sampling too few nucleotides or too few nuclear loci could lead to failure to recognize important genetic patterns. Future directions The concepts and criteria for ESUsand MUs expounded above seem logical and theoretically valid, but it remains to be seen whether they are practical. Pertinent data are expanding rapidly. but there is

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a need for further theoretical study of the dynamics of allele distribution and phylogeny in demographic contexts relevant to threatened and exploited species. This is certainly a field where close interaction between experimental and theoretical biologists would pay off. Acknowledgements Thanks to John Avise, Brian Bowen,
Peter Dwyer, Peter Hale, Shane Lavery, Nancy Fitzsimmons and Steve Palumbi for critical reviews of the manuscript, to Anita Heideman and Lyn Pryor for preparation of figures and to the inmates of the Conservation Genetics lab for inspiration. Supported by grants from the Australian Research Council.

7 Ryman. N. C 1991) J F&h Em/ 39 (SuppI. A

8

21l-224 Avise. J.C. (1994)

Xatural Histoy

Molecular

Murken.

Chapman & Hall Neigel. J. and Avise. J.C. (1986) in

and Eoolutron. Processes and Theory (Nevo, 1..

Euolutionuy

and Karlin, S.. rds). pp. 515-534. Academic Press 10 Slade. R.W..Moritz. C. and Heideman. A. (1994) hlol. Brol. Ecol 11,341-X6 11 Moritz. C. (1994) ,bfo/ Era/. 3. 403-413 12 Gauldie, R.W. (1991) Gun..! F&7 Aquat. Sri. 48,
722-731

13 Cracraft, J. (1983) Cinr

Omitho/. 1. 159-187 14 Frost, D.R.and Hillis, D.M. (1990) Herpetologrca 46. 87-104

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(1992) syst Biol 41. 73-291

Dizon, A.E.. Lockyer, C., Perrin, W.F., Demaster, D.P. and Sisson. J. (1992) Cons. BioL
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Mol. Ecol 3. 3Kb-37:i

The early evolution of life: solutionto Darwin s dilemma

J. William Schopf
William Schopf is at the Dept of Earth & Space Sciences, Center for the Study of Evolution and the Origin of Life, institute of Geophysics and Planetary Physics, and Molecular Biology Institute, University of California, Los Angeles, CA 90024, USA.

What has been learned? As in any emerging area of science, numerous new generalizations have been drawn, four of which stand out as being particularly significant. (1) Life originated very early in Earth history, much earlier than had been assumed.

n 1859, Charles Darwin stated

I problem:

the

If the theory [of evolution] be true it is indisputable that before the lowest Cambrian stratum was deposited long periods elapsed . . and that during these vast periods the world swarmed with living creatures . . . . [However], to the question why we do not find rich fossiliferous deposits belonging to these earliest periods . I can give no satisfactory answer. The case at present must remain
inexplicable . ..* (Ref. 1, Ch. X)

Surprisingly, it was not until more than a century later, with publication of three pivotal papers in 19652-4and of a major monograph in 19685, that search for the missing Precambrian fossil record was demonstrated to be a fruitful area of scientific inquiry. Since that time in a scant three decades - more than 3000 taxonomic occurrences of microscopic fossils have been discovered in nearly 400 Precambrian geological formations6.7; the new field of Precambrian paleobiology has emerged, matured and become established worldwide as a viable subdiscipline of the natural science+; and most recently, two mammoth compendia, prepared by international groups of respected experts, have summarized the status of this interdisciplinary area of science, . To a major extent, Darwin s dilemma has been resolved - much of the missing fossil record has been uncovered. What has been learned? Where does this young field go from here?

Before the discovery of the Precambrian fossil record, few imagined that the well documented history of Phanerozoic life the familiar progression from seaweeds to flowering plants, from trilobites to humans -was merely the tip of the evolutionary iceberg. Indeed, the recent discovery of diverse cellularly preserved microorganisms in the 3465 t 5-millionyear-old Apex chert of Western Australia indicates that the Phanerozoic temporally encompassed less than 15%of all of evolution, and that living systems have existed for more than three-quarters of the history of the planet (Fig. 1). Moreover, because most of the 11 species described from this earliest known fossiliferous deposit are comparable to extant (oscillatoriacean) cyanobacteria - oxygen-producing photoautotrophs that are among the most highly evolved of all eubacteria - it seems certain that life must have originated substantially (and probably hundreds of millions of years) earlier.