Genetic diversity in the endangered dysploid larkspur Delphinium bolosii and its close diploid relatives in the series Fissa of the Western Mediterranean area

Allozyme diversity was evaluated in four closely-related taxa of the Delphinium series Fissa distributed throughout the Western Mediterranean area. All are considered threatened plants. Delphinium bolosii and Delphinium mansanetianum are narrowly endemic to the Eastern Iberian Peninsula, whereas Delphinium fissum ssp. sordidum is found in a few populations across the Peninsula. Delphinium fissum ssp. fissum is more widely distributed but often in small and isolated populations. In this group, Delphinium bolosii is dysploid (2 n  = 18) whereas the other taxa are diploid (2 n  = 16). A total of 12 populations were surveyed, including all known locations for D. bolosii , D. mansanetianum , and D. fissum ssp. sordidum . Eleven enzyme systems were assayed and 15 loci were resolved. Markedly depauperate values for genetic diversity were obtained for D. mansanetianum ( H _e = 0.013) and D. fissum ssp. sordidum ( H _e = 0.044). The estimates for D. fissum ssp. fissum ( H _e = 0.071) were below the values expected for widespread species. Small population size and marginal distribution have probably contributed to the low variability observed in this group. By contrast, D. bolosii exhibited comparatively larger populations and greater genetic diversity ( H _e = 0.138). We suggest that, apart from population size and local adaptation, genetic diversity during speciation may have been promoted by dysploidy through genomic recombination.  © 2007 The Linnean Society of London, Biological Journal of the Linnean Society , 2007, 92 , 773–784.     

Stability of genetic structure and effective population size inferred from temporal changes of microsatellite DNA polymorphisms in the land snail Helix aspersa (Gastropoda: Helicidae)

Temporal evolution of genetic variability may have far-reaching consequences for a diverse array of evolutionary processes. Within the polders of the Bay of Mont-Saint-Michel (France), populations of the land snail Helix aspersa are characterized by a metapopulation structure with occasional extinction processes resulting from farming practices. A temporal survey of genetic structure in H . aspersa was carried out using variability at four microsatellite loci, in ten populations sampled two years apart. Levels of within-population genetic variation, as measured by allelic richness, H _e or F _is , did not change over time and similar levels of population differentiation were demonstrated for both sampling years. The extent of genetic differentiation between temporal samples of the same population established (i) a stable structure for six populations, and (ii) substantial genetic changes for four populations. Using classical F -statistics and a maximum likelihood method, estimates of the effective population size ( N _e) illustrated a mixture of stable populations with high N _e, and unstable populations characterized by very small N _e estimates (of 5–11 individuals). Owing to human disturbances, intermittent gene flow and genetic drift are likely to be the predominant evolutionary processes shaping the observed genetic structure. However, the practice of multiple matings and sperm storage is likely to provide a reservoir of variability, minimizing the eroding genetic effects of population size reduction and increasing the effective population size.  © 2004 The Linnean Society of London, Biological Journal of the Linnean Society , 2004, 82 , 89–102.     

Low genetic diversity and allozymic evidence for autopolyploidy in the tetraploid Pyrenean endemic larkspur Delphinium montanum (Ranunculaceae)

Allozyme electrophoresis was conducted to survey the levels and distribution of genetic diversity in the tetraploid perennial larkspur Delphinium montanum (Ranunculaceae), which is endemic to the eastern Pyrenees of Spain and France and considered to be endangered. Seven populations were sampled, resolving 14 putative enzymatic loci belonging to eight enzymes. Banding patterns stained in gels revealed several enzymatic duplications attributable to autotetraploidy, such as the presence of both balanced and unbalanced heterozygotes and the lack of fixed heterozygosity. However, variability in D. montanum ( P  = 23.8%, A  = 1.48, and H _e = 0.082) was lower than that expected for an autotetraploid species. This, in addition to the scarcity of loci showing three or four different alleles, could indicate that this species has suffered genetic erosion through population bottlenecks, or, alternatively, that it is undergoing diploidization.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 155 , 211–222.     

Allozyme Diversity in Populations of Cymbidium goeringii (Orchidaceae)

Abstract: Using 14 allozyme loci, we investigated levels of genetic diversity within populations, and degree of genetic divergence among 24 populations of Cymbidium goeringii (Orchidaceae) in Korea and Japan. Cymbidium goeringii maintains high levels of genetic diversity both at population (mean expected heterozygosity, H _e = 0.238) and species levels (0.260). Means of H _e found in 24 populations were not significantly different from each other. About 90 % of the total variation in the species is common to all populations (mean G _ST = 0.108). No unique allele was found in any population. The indirect estimate of gene flow based on the mean G _ST was high ( Nm = 2.06). Nei's genetic identities for pairs of populations had high values (mean = 0.974 [SD = 0.013]). The Mantel-Z test showed a significant correlation between genetic distance and geographic distance. However, the mean G _ST value between 17 populations in Korea and seven Japanese populations was relatively low (0.029), even though the land connection between the southern Korean peninsula and southern Japanese archipelagos has not existed since the middle Pleistocene. Large numbers of small seeds of C. goeringii might travel long distances by wind from populations to populations both in Korea and Japan, increasing genetic diversity within populations and maintaining low genetic differentiation among populations.     

Population genetics of Dioon edule Lindl. (Zamiaceae, Cycadales): biogeographical and evolutionary implications

Dioon edule Lindl. (Zamiaceae) is a cycad endemic to Mexico, that occurs as one species D. edule and the geographical variety D. edule var. angustifolium (Miq.) Miq. Dioon edule has a north to south distribution in eastern Mexico. In this study, we analysed 14 allozymic loci in eight populations of D. edule from its total distribution range by sampling all known populations. Patterns of diversity and genetic variability, within and among populations, were obtained. The mean number of alleles per locus ( A ) was 1.44 and the percentage of polymorphic loci was relatively high ( P  = 54.78). The mean observed ( H _O ) and expected heterozygosity ( H _E ) were 0.27 and 0.24, respectively. F -statistics revealed an excess of heterozygous genotypes, locally and globally ( F  = −0.17 and f  = −0.27, respectively). The genetic variation explained by differences among populations was only 7.5%. We also detected a negative relationship between genetic diversity and latitude. On average, the gene flow between population pairs was relatively high ( Nm  = 2.98); furthermore, gene flow between population pairs was significantly correlated with geographical distances ( r  = −0.38, P  = 0.025). Therefore, patterns of genetic diversity in D. edule appear to be associated with the post-Pleistocene spread of the species, from its southerly (origin) to its northerly range (derived populations, including its central distribution). The biogeographical and evolutionary aspects of the results of this study are discussed. We recognize Dioon angustifolium Miq. for the northernmost disjunct populations.  © 2003 The Linnean Society of London, Biological Journal of the Linnean Society , 2003, 80 , 457–467     

Population divergence in the amphicarpic species Amphicarpaea edgeworthii Benth. (Fabaceae): microsatellite markers and leaf morphology

Comparative analyses of the genetic differentiation in microsatellite markers ( F _ST) and leaf morphology characters ( Q _ST) of Amphicarpaea edgeworthii Benth. were conducted to gain insight into the roles of random processes and natural selection in the population divergence. Simple sequence repeat analyses on 498 individuals of 19 natural populations demonstrate that a significant genetic differentiation occurs among populations (mean F _ST = 0.578), and A. edgeworthii is a highly self-fertilized species (mean selfing rate s  = 0.989). The distribution pattern of genetic diversity in this species shows that central populations possess high genetic diversity (e.g. population WL with H _E = 0.673 and population JG with H _E = 0.663), whereas peripheral ones have a low H _E as in population JD (0.011). The morphological divergence of leaf shape was estimated by the elliptical Fourier analysis on the data from 11 natural and four common garden populations. Leaf morphology analyses indicate the morphological divergence does not show strong correlation with the genetic differentiation ( R  = 0.260, P  = 0.069). By comparing the 95% confidence interval of Q _ST with that of F _ST, Q _ST values for five out of 12 quantitative traits are significantly higher than the average F _ST value over eight microsatellite loci. The comparison of F _ST and Q _ST suggests that two kinds of traits can be driven by different evolutionary forces, and the population divergence in leaf morphology is shaped by local selections.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 96 , 505–516.     

Allozymic variation and relationships of the endangered cyprinodontid genus Valencia and its implications for conservation

Allozyme analysis was used to determine patterns of genetic variation and relationships within the genus Valencia , a group with two allopatric species, V. letourneuxi inhabiting the Balkan Peninsula, and V. hispanica which is endemic to the Iberian Peninsula. Products of 25 gene loci were analysed with AK *, FH *, LDH -1* and LDH -2* monomorphic when Fundulus heteroclitus macrolepidotus was not taken into account. Nine loci were diagnostic for both species of the genus Valencia . Levels of genetic variation of V. hispanica ( P =0·416, H ^e=0·118) and V. letourneuxi ( P =0·160, H _e=0·040) were higher than values reported previously. High population subdivision ( F _st=0·321) among V. hispanica populations indicates a clear interruption of genetic exchange among populations. High genetic variation and differentiation exhibited by populations of V. hispanica suggest that a recovery program should be carried out with natural stocks from the same localities.     

Low population differentiation and high genetic diversity in the invasive species Carduus acanthoides L. (Asteraceae) within its native range in the Czech Republic

Colonizing species are predicted to suffer from reductions in genetic diversity during founding events. Although there is no unique mode of reproduction that is characteristic of successful plant colonizers, many of them are predominantly self-fertilizing or apomictic species, and almost all outcrossing colonizers are self-compatible. Carduus acanthoides comprises a species of disturbed habitats with wind-dispersed seeds that colonizes open spaces of various sizes. Population genetic diversity was expressed by assessing patterns of variation at nine putatively neutral allozyme loci within and among 20 natural populations in its native distribution range in the Czech Republic. Overall, C. acanthoides displayed high levels of genetic diversity compared to other herbaceous plants. The percentage of polymorphic loci was 84.5, with values of 2.37, 0.330, and 0.364 for the mean number of alleles per polymorphic locus ( A ), observed heterozygosity ( H _o), and expected heterozygosity ( H _e), respectively. There was only weak evidence of inbreeding within populations ( f  = 0.097) and very low genetic differentiation among populations ( θ  = 0.085). Analyses of the data provide strong evidence for isolation-by-distance for the whole study area. Even the colonizing species, C. acanthoides , currently supports a substantial amount of allozyme variation at both the species and population levels.  © 2009 The Linnean Society of London, Biological Journal of the Linnean Society , 2009, 98 , 596–607.     

Evolutionary relationships of the Lake Malawi Oreochromis species: evidence from allozymes

Three tilapiine species belonging to the endemic Lake Malawi species flock known as' chambo' Oreochromis ( Nyasalapia ) karongae , O. (N.) lidole and O. (N.) squamipinnis and the species Oreochromis (Oreochromis) shiranus were collected from the wild. These four species were analysed at 43 enzyme loci using starch gel electrophoresis. No significant deviations from Hardy-Weinberg equilibrium were observed after correcting for multiple simultaneous testing. The expected heterozygosity ( H _e) was lowest in O. (O.) shiranus ( H _e=0.082). The three chambo species had higher levels: H _e=0.103 to H _e=0.116. The three chambo species as a whole were polymorphic (99%) at 16 different loci sharing 10 of these in common. No fixed differences between the three chambo species were observed but highly significant allele frequency differences existed between all the chambo species. The F _ST calculated for the three chambo species was 0051, closer to intra-specific than to inter-specific levels. O. (O.) shiranus could be clearly separated from all chambo species at five fixed loci. Comparison of the allozyme data from these species with five species from the same sub genera support the hypothesis that the chambo form a monophyletic group and that O. (N.) macrochir or a related species represents the sister taxon.     

Allozyme differentiation and genetic structure of the Lady's slipper (Cypripedium calceolus) island populations in north-east Poland

The population genetic structure in the rare and endangered perennial plant, Cypripedium calceolus , from north-east Poland was investigated. A total of 11 loci were analysed, of which five were polymorphic for the species and in every population. The 11 loci resolved gave a total of 19 alleles with the mean number of 1.73 alleles per locus. The proportion of polymorphic loci (P) was the same for the species and in every population, 45.5%. The average observed and expected heterozygosities for the total Lady's slipper data set were 0.156 and 0.184, respectively. The average expected heterozygosity for the populations under study ranged from 0.176 to 0.188 and did not differ significantly among populations. Genetic diversity within C. calceolus populations from the Biebrza Valley is relatively high compared with rare taxa and taxa with similar life histories. The genetic diversity among populations of C. calceolus studied ( F _ST = 0.016) is much smaller than the genetic diversity within populations. In the species studied, there was no significant correlation between genetic and geographical distances ( r = −0.73, Mantel test). This may indicate the relatively recent origin of the populations studied, or a high level of gene flow among populations. © 2002 The Linnean Society of London, Botanical Journal of the Linnean Society , 2002, 138 , 433–440.     

High genetic variability in Neotropical myophilous orchids

The patterns of genetic variability of seven Bulbophyllum Thouars species were investigated using 14 enzymatic loci, and compared with those of other Neotropical myophilous orchid species. The genetic variability estimated was very high ( H _e = 0.39–0.61, P  = 86–100%, A  = 2.6–3.8), despite the small population sizes, which may be explained by the vegetative reproduction. Of the three species with multiple populations, only B. epiphytum showed moderate values of genetic structuring. Environmental characteristics and seed dispersal mechanisms are important in understanding the differences in the population structure observed. Almost all pairs of species showed low genetic similarity, indicating a long period of divergence. Our results suggest that B. adiamantinum and B. insectiferum , species currently classified in different sections of the genus, are more closely related than previously thought.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 33–40.     

Clinal genetic variation and isolation by distance in the European eel Anguilla anguilla (L.)

The genetic variability and structure of the European eel ( Anguilla anguilla L.) in populations throughout Europe was reassessed using 15 allozymic loci, seven of which were polymorphic. Seven sites were sampled on a latitudinal gradient across the natural continental range, extending from southern France to southern Norway. Heterozygosity ( H _e = 0.05) and level of polymorphism (P = 0.43) were comparable to other marine fish. Populations were poorly differentiated ( G _ST = 0.014, F _ST = 0.002), which is not surprising considering the high dispersal capability of the European eel. However, a significant geographical cline was detected at two alleles ( IDH-1 * 100 and GPI-1 * 110 ), and genetic distances ( D _CE) were concordant with geographical coastal distances. Mantel tests, pairwise F _ST's and multidimensional scaling analyses identify three distinct groups: Northern Europe, Western Europe and the Mediterranean Sea. We propose that the clinal genetic structure in the European eel may be due to (1) isolation by distance (as recently detected with microsatellites), (2) temporal reproductive separation, (3) post-larval selective forces, (4) contact between formerly separated groups or (5) some combination thereof.  © 2002 The Linnean Society of London, Biological Journal of the Linnean Society , 2002, 77 , 509–521     

Genetics of the peripheral populations of the alpine bullhead, Cottus poecilopus (Scorpaeniformes, Cottidae) in Poland

The alpine bullhead ( Cottus poecilopus ) in Poland is highly polymorphic ( P for examined populations in the range of 0.0–0.45, mean P  = 0.11, P for species 0.64) in seven out of 11 loci studied ( Pgd-1, Mdh-1, Ck-1, Gpd-1, Gpd-2, Gpi-1, Gpi-2 ). However, they reveal very low heterozygosity (mean H _o = 0.007, mean H _e = 0.019), which may result from population subdivisions, small genetically effective size ( N _e) of some populations and frequent inbreeding, the latter being probably mediated through polygyny. The index of genetic differentiation ( G _ST) for the Polish metapopulation equals 0.48, which is relatively high. By far, the strongest are genetic differences between the Sudeten-Carpathian group and the Hańcza population. Gene flow between the Sudeten-Carpathian group and the Hańcza population is very limited and their divergence may have started as early as the last (Eemian) interglacial period.     

Contrasting levels of genetic diversity between the common, self-compatible Liparis kumokiri and rare, self-incompatible Liparis makinoana (Orchidaceae) in South Korea

Levels of allozyme variation and intrapopulation spatial genetic structure of the two terrestrial clonal orchids Liparis kumokiri , a self-compatible relatively common species, and L. makinoana , a self-incompatible rare species, were examined for 17 ( N  = 1875) and four ( N  = 425) populations, respectively, in South Korea. Populations of L. makinoana harboured high levels of genetic variation ( H _e = 0.319) across 15 loci. In contrast, L. kumokiri exhibited a complete lack of allozyme variation ( H _e = 0.000). Considering the lack of genetic variability, it is suggested that current populations of L. kumokiri in South Korea originated from a genetically depauperate ancestral population. For L. makinoana , a significant deficit of heterozygosity (mean F _IS = 0.198) was found in population samples excluding clonal ramets, suggesting that pollen dispersal is localized, generating biparental inbreeding. The significant fine-scale genetic structuring (≤ 2 m) found in a previous study, in addition to the moderate levels of population differentiation ( F _ST = 0.107) and the significant relationship between genetic and geographical distances ( r  = 0.680) found here, suggests a leptokurtic distribution of seed dispersal for L. makinoana . Although populations of L. makinoana harbour high levels of genetic variation, they are affected by a recent genetic bottleneck. This information suggests that genetic drift and limited gene flow could be the main evolutionary forces for speciation of a species-rich genus such as Liparis .  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 41–48.     

Large variations in the ratio of effective breeding and census population sizes between two species of pond-breeding anurans

The viability of wild populations is frequently assessed by monitoring adult census sizes ( N _c). This approach is particularly useful for pond-breeding amphibians, because assemblages during the breeding season are relatively easy to detect and count. However, it is the genetic effective population size ( N _e) or surrogates such as effective breeding population size ( N _b) that are of primary importance for long-term viability. Although N _c estimates of one anuran amphibian ( Bufo bufo ) in Britain were much larger than those of another ( Rana temporaria ) at the same sites, the ratios of N _b to N _c were much smaller in B. bufo than in R. temporaria. These differences were sufficiently great as to reverse the effective size order at one site, such that N _b for R. temporaria was larger than that for B. bufo. Differences in adult sex ratios at breeding sites probably contributed to lower N _b values in B. bufo populations compared with those of R. temporaria . The relationship of N _b to N _c can therefore vary dramatically even between similar species, to the extent that just monitoring N _c can give misleading impressions of relative effective breeding sizes and thus of population viability. It will be increasingly important to estimate N _e or N _b in wildlife populations for assessment of conservation priorities.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 365–372.     

Genetic and morphological variability of the endangered Syngonanthus mucugensis Giul. (Eriocaulaceae) from the Chapada Diamantina, Brazil: implications for conservation and taxonomy

Allozymic and morphometric studies were carried out on ten populations of Syngonanthus mucugensis (Eriocaulaceae), a species from north-eastern Brazil threatened by extinction. Genetic and morphological variability was low or moderately low in all populations, being lower in populations from Rio de Contas/Catolés ( P _L = 14.3–21.4, A  = 1.1–1.2, H _e = 0.026–0.059, D2M = 26.893–33.157) than in those from Mucugê ( P _L = 28.6–35.7, A  = 1.3–1.5, H _e = 0.078–0.164, D2M = 28.999–45.077). A high coefficient of endogamy ( F _is = 0.257) was found, which can be explained by the reproductive characteristics and distribution of the species. The values for genetic and morphological structuring ( F _st = 0.512 and A _MRPP = 0.175, respectively) were high as a result of the differentiation between populations from the two areas. The mean genetic identity between populations from the two areas (0.812) was much lower than between populations from the same area (Mucugê, 0.980; Rio de Contas/Catolés, 0.997). These results indicate that we are dealing with two distinct taxa and, as a result of the nature of the morphological differences found, a new subspecies is described for the populations of the region of Rio de Contas and Catolés, Syngonanthus mucugensis ssp. riocontensis . Such conclusions raise important implications for the conservation of Syngonanthus mucugensis , and will be used in the drawing up of management plans for its conservation.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 153 , 401–416.     

Conservation Genetics of an Endangered Herb, Hanabusaya asiatica (Campanulaceae)

Abstract: Hanabusaya asiatica (Nakai) Nakai (Campanulaceae), a bee- pollinated, perennial herb, is restricted to the mountainous regions of the eastern-central Korean peninsula. Allozyme analyses for 348 individuals assessed the levels of genetic diversity for five populations. Spatial autocorrelation statistics were also used to examine the spatial distribution of allozyme polymorphisms. The species maintains high levels of allozyme diversity ( H _eS = 0.217) and it exhibits low allozyme differentiation among populations ( G _ST = 0.132) compared with other endemics (mean H _e = 0.096, G _ST = 0.248). There is an apparent pattern of isolation by distance among populations. These results suggest that H. asiatica is at a genetic equilibrium. A considerable deficit in numbers of heterozygotes suggests mating among relatives in populations. At least three populations of H. asiatica should be sampled or conserved to capture or maintain > 99 % of the genetic diversity in the species as a whole. Within local populations, individuals are distributed in a structured, isolation by distance, manner. Approximate genetic patch width in the populations of H. asiatica examined is 5 - 8 m. For conservation purposes, it is suggested that, in general, the sampling of H. asiatica should be conducted at intervals in order to efficiently sample the genetic diversity across an entire population.     

Population genetics and the conservation status of the threatened Iberian steppe grass Puccinellia pungens (Pau) Paunero (Poaceae)

Puccinellia pungens (Pau) Paunero is a narrowly endemic grass found in two continental saline lagoons of north-eastern Spain. This rare plant has been classified as 'at risk of extinction' in several national and European catalogues of endangered species. Recent demographic studies indicate that population sizes greatly exceed several million individuals, challenging that threat category. Our genetic analysis, based on allozymes, has shown that in spite of the large population sizes, very low levels of genetic variation were found in P. pungens . Genetic variation was similar in most populations, but the largest, Gallocanta lagoon as a whole, had less variation (35% polymorphic loci, 1.4 alleles/locus, H _T = 0.038) than the more restricted Royuela range (45% polymorphic loci, 1.5 alleles/locus, H _T = 0.056), suggesting a recent population expansion of the Gallocanta populations from few founder lines. The low genetic distances among populations also suggest a recent divergence. The low genetic variation observed cannot be explained fully by eventual clonal spread and rare seedling establishment in the hypersaline environment. This low variation seems to result from extreme recent population bottlenecks as a consequence of habitat conversion to agricultural fields. In the light of our data, it seems unlikely that reinforcement of populations could increase the genetic diversity of the populations. Hence, conservation efforts should focus on avoiding further habitat loss of this endangered steppe grass species.  © 2007 The Linnean Society of London, Botanical Journal of the Linnean Society , 2007, 154 , 269–281.     

Is the last glaciation the only relevant event for the present genetic population structure of the meadow brown butterfly Maniola jurtina (Lepidoptera: Nymphalidae)?

Phylogeographical studies are available for a considerable number of European species, but few analyses exist for temperate species with very large and fairly continuous populations that are also absent from Northern Europe. Therefore, we studied the butterfly Maniola jurtina as a model for this group. The species has two major genetic lineages (mean genetic distance between lineages: 0.033; F _CT: 0.052), most probably evolving in glacial differentiation centres in the western and eastern Mediterranean. The onset of this differentiation might have been the beginning of the last glacial stage maximum some 40 kyr bp . A hybrid zone between these two lineages exists in western Central Europe. No genetic substructures have been found within the two lineages ( F _SC: 0.017) and average genetic distances are very small. Therefore, it is highly probable that postglacial expansion was of the phalanx type. There is, at most, very limited differentiation at regional and local scales. However, the genetic diversity within populations is high (means: A : 2.68; H _E: 17.2%; P : 78%), as would be predicted for such a common species. Comparison of these results with a published allozyme analysis revealed a similar phylogeographical pattern, but lower genetic diversity in the latter. Morphological patterns of wings and genitalia show similar geographical patterns as allozyme data.   © 2005 The Linnean Society of London, Biological Journal of the Linnean Society , 2005, 85 , 419–431.     

Allozyme Variation and the Genetic Structure of Populations of the Rare Sedge Carex misera (Cyperaceae)

Abstract Carex misera is a rare sedge, endemic to rocky outcrops and mountain summits within the southern Appalachian Mountains from northern Georgia to northern North Carolina. We assessed allozyme diversity for 406 individuals from nine populations over most of the geographic range. Twenty-seven putative loci were assayed and eight (30%) were found to be polymorphic. Nei's gene diversity statistics ( H _T =0.043, H _S =0.019, G _ST =0.551) indicated low levels of variation but relatively highly differentiated populations, suggesting little gene flow. Significant deviations from genotypic expectations under Hardy-Weinberg equilibrium, high positive fixation indices, and the existence of small genetic neighborhoods within populations suggest that at least some inbreeding occurs. Cluster analysis of Nei's genetic identity statistics and principal component analysis of allele frequency data showed high similarity among the six southern populations with the two northern populations more differentiated from them and from each other. These results suggest that preservation of the northern populations is necessary to conserve the already low levels of genetic diversity within the species.