ENZYME ELECTROPHORESIS AND EVOLUTIONARY RELATIONSHIPS AMONG THREE SPECIES OF LASTHENIA (ASTERACEAE: HELIANTHEAE)

Enzyme electrophoresis was used to examine variation at 18 gene loci in Lasthenia burkei, L. conjugens, and L. femontii. The species consist of diploid annuals restricted to vernal pools in California; a variety of data has indicated that they are closely related. Populations of the three species are similar at isozyme loci; L. conjugens and L. fremontii are most similar with a genetic identity of 0.965. Lasthenia conjugens and L. burkei are next most similar (0.934), while L. burkei and L. fremontii have an identity of 0.909. Lasthenia burkei and L. femontii each contain a different subset of the alleles found in L. conjugens. Electrophoretic data are concordant with the view that these three species are closely related, but do not support the hypothesis that L. burkei is a stabilized hybrid derivative of L. conjugens and L. fremontii because alleles unique to each of the latter two species are not combined in the former. Rather, distribution of alleles among the three species suggests two possible alternative hypotheses of evolutionary relationships. One hypothesis considers Lasthenia conjugens a hybrid derivative of L. burkei and L. fremontii while the alternative explanation views the latter two species as independent derivatives of the former. Neither of these hypotheses appears to be concordant with morphological data.     

Restoration genetics of the vernal pool endemic Lasthenia conjugens (Asteraceae)

Restoration of habitat for endangered species often involves translocation of seeds or individuals from source populations to an area targeted for revegetation. Long-term persistence of a species is dependent on the maintenance of sufficient genetic variation within and among populations. Thus, knowledge and maintenance of genetic variability within rare or endangered species is essential for developing effective conservation and restoration strategies. Genetic monitoring of both natural and restored populations can provide an assessment of restoration protocol success in establishing populations that maintain levels of genetic diversity similar to those in natural populations. California’s vernal pools are home to many endangered plants, thus conservation and restoration are large components of their management. Lasthenia conjugens (Asteraceae) is a federally endangered self-incompatible vernal pool annual with gravity- dispersed seeds. Using the molecular technique of intersimple sequence repeats (ISSRs), this study assessed levels and patterns of genetic variability present within natural and restored populations of L. conjugens. At Travis Air Force Base near Fairfield, California, a vernal pool restoration project is underway. Genetic success of the ecologically based seeding protocol was examined through genetic monitoring of natural and restored populations over a three-year period. Genetic diversity remained constant across the three sampled generations. Diversity was also widely distributed across all populations. We conclude that the protocol used to establish restored populations was successful in capturing similar levels and patterns of genetic diversity to those seen within natural pools. This study also demonstrates how genetic markers can be used to inform conservation and restoration decisions.     

Genetic evidence for the origin of Californian wild beets (genus Beta)

The genus Beta L. is a morphologically and genetically variable group composed of wild, weedy, and domesticated forms that are used for sugar production or as vegetables. In this study, we have evaluated genetic variation in 64 germplasm accessions of wild and domesticated beets and examined the origin of wild beet accessions in California using allozyme analysis. UPGMA analysis showed overall that domesticated and wild beets form genetically coherent groups. Wild beets in California have two different origins, from European Beta vulgaris or from Beta macrocarpa. Population-level patterns of allozyme variation for wild California beets related to B. vulgaris suggest that those populations evolved from naturalized populations of the cultivated B. vulgaris ssp. vulgaris which had hybridized to varying degrees with the sea beets B. vulgaris ssp. maritima. Wild California beets related to B. macrocarpa are essentially genetically identical to European accessions. In addition, we found substantial evidence for hybridization and introgression of B. vulgaris alleles in one B. macrocarpa accession in California. The obligate outcrosser B. vulgaris exhibits more allelic diversity than the self-compatible B. macrocarpa. Beta vulgaris ssp. maritima exhibits more genetic diversity than domesticated B. vulgaris ssp. vulgaris. Received: 2 November 1998 / Accepted: 29 April 1999     

Genetic variation and population structure in Korean endemic species: III.Hosta minor (Liliaceae)

Hosta minor, an insect-pollinated and rhizomatous herbaceous perennial, occurs in eastern and southern Korean Peninsula. AlthoughH. minor is a Korean endemic species and most populations of the species are scattered, the species maintains considerably high levels of genetic variation within the species, with a moderate level of variation (14%) found among populations. Nineteen of the 29 putative loci resolved (66%) were polymorphic within the species, the mean number of alleles per locus was 2.10 across all populations and averaged 1.68 within populations. In addition, genetic diversity was a considerably higher (for species and population level, mean estimates of genetic diversity were 0.275 and 0.230, respectively) than the average for other long-lived herbaceous perennials. Indirect estimate of the number of migrants per generation (Nm=1.03, calculated from mean G_ST) was moderate. Factors contributing to the high levels of genetic diversity found within populations ofH. minor include population maintenance via sexual and asexual modes of reproduction, primarily outcrossing breeding system, long generation time, probable ancient polyploid origin of the species, and moderate levels of seed dispersal by wind. Human disturbance in South Korea such as road and apartment constructions appears to be the major threat to this genetically diverse species.     

A review of the subspecies status of the Icelandic Purple Sandpiper Calidris maritima littoralis

The Icelandic Purple Sandpiper Calidris maritima littoralis (C.L. Brehm, 1831) represents one member of a poorly understood subspecies complex. Currently, differences in size define two other subspecies: Calidris maritima belcheri Engelmoer & Roselaar, 1998, which breeds in north‐eastern Canada along the Hudson Bay and James Bay, and Calidris maritima maritima (Brunnich, 1764), which breeds along the Arctic coasts elsewhere in northern Canada, Greenland, Svalbard, Scotland, and Fennoscandia, to northern central Siberia. There are large size differences amongst populations of C. m. maritima, however. As an Arctic/Alpine breeding bird, C. m. littoralis could provide an interesting perspective on the evolutionary changes following a northwards expansion of a species after glacial retreat. Considering the extent of the ice sheet in the northern hemisphere during the last glaciation, and the short period of time since it ended, the correct attribution of subspecies status for C. m. maritima may reflect either rapid diversification from a single population or ancestral splits of distinct evolutionary lineages that survived in isolation at southern latitudes. We applied morphometric subspecies criteria, diagnosability by Amadon's rule, and genetic analysis of five nuclear introns, and the mitochondrial DNA markers cytochrome oxidase c subunit I (COI) and NADH dehydrogenase subunit 2 (ND2), to geographically separate breeding populations in order to examine the subspecies status of the Icelandic population. The results do not provide support for the subspecies status of the Icelandic population because the nominate and Icelandic subspecies fail to uphold Amadon's rule, and genetic analyses indicate that the study populations derive from a single shared refugium. © 2015 The Linnean Society of London     

ALLOZYMIC VARIATION IN LOCAL APOMICTIC POPULATIONS OF LEMNA MINOR (LEMNACEAE)

Flowering occurrence and allozymic variation were studied in eight local populations of Lemna minor in eastern Ontario, Canada. After 2 years of survey, not a single flower was observed. This absence of flowering suggests the possibility of loss of sexual reproduction. This may have had no net adverse effect on fitness given the simple life history and prolific vegetative propagation in L. minor. However, the absence of sexual reproduction may limit genotypic diversity. The allozymic analysis detected 18 loci from 13 enzyme systems. Large deviations from Hardy-Weinberg equilibrium were common because of an excess of homozygotes for several enzyme systems. The genotypic diversity within these eight populations had a mean D value of 0.973 with an average number of genotypes per population of 19.6. These results suggest that genotypic diversity within these populations is not severely limited by the rarity of sexual reproduction. The mean genotypic distance index (D₁₄ = 0.801) suggests a high degree of differentiation between populations. The mean number of populations per genotype was 1.78. Using a Mantel test, the genotypic distance matrix was not significantly related to the population-to-population distance matrix (t = -0.161, P = 0.413). Although rare events of sexual reproduction may help maintain genetic variation, somatic mutations and multiple origins of clones may be important factors maintaining genetic diversity both within and between populations of L. minor.     

The influence of sea currents, past disruption of gene flow and species biology on the phylogeographical structure of coastal flowering plants

Aim We investigate the geographical genetic structure of two coastal plant species, Cakile maritima Scop. (Brassicaceae) and Eryngium maritimum L. (Apiaceae), through three sea straits and along one continuous stretch of coast using amplified fragment length polymorphisms (AFLPs). The two species have a similar ecology in that they grow in sandy habitats, but differ in life‐form (annual vs. perennial) and dispersability of seeds by sea water as inferred from floating experiments. The sea straits differ in their geological history and their modern current systems. The primary goal of our study was to test the hypothesis that sea straits have an influence on the geographical patterns of genetic variation at the population level. Location The areas around the Strait of Gibraltar, the Dardanelles, the Bosporus and the Atlantic coast of western France. Methods For both species we investigated AFLP variation in several populations from each area. Bayesian clustering and diversity and differentiation measures were used to analyse the genetic data. Results In most areas the spatial genetic structure was similar between the two species. They share the presence of distinct genetic gaps along the coast through the Strait of Gibraltar and the Bosporus, and these genetic gaps coincide with the straits. Both species show genetic continuity along the coast of western France. A distinct genetic gap was found through the Dardanelles for C. maritima but not for E. maritimum. Main conclusions The study shows that sea straits have an influence on the geographical patterns of genetic variation. Sea currents are inferred to cause the genetic gap through the Strait of Gibraltar. In the Bosporus and, for C. maritima, through the Dardanelles, the genetic gaps found are explained by the past closure of these two straits as well as by present‐day factors. Simulations indicate that the lower differentiation of C. maritima through the Dardanelles than through the Bosporus cannot be explained by the difference in geological history of these two straits. The difference in seed dispersability between the two species is argued to be responsible for the observation that differentiation among genetic clusters is higher in E. maritimum than in C. maritima where a direct comparison is possible.     

THE ORIGIN AND RELATIONSHIPS OF LASTHENIA BURKEI (COMPOSITAE)

Lasthenia burkei (Compositae) is a narrowly restricted California endemic closely related to L. conjugens and L. fremontii. These three species differ from each other by pappus and phyllary characters and in geographical distribution. All are freely intercrossable, but L. fremontii forms rather sterile artificial hybrids with its two relatives which, in turn, form fairly fertile artificial hybrids with each other. Lasthenia burkei and L. conjugens have homologous chromosomes, four of which are homologous with four of those of L. fremontii. The remaining two chromosomes probably have reciprocal translocations which lead to multivalent formation during meiosis in interspecific hybrids. Pollen viability is restored in most F₂ generations, suggesting a close genetic relationship among the three species. The evolutionary relationship among these species may be a linear one with L. burkei occupying an intermediate position between L. fremontii and L. conjugens, although the direction of this linear phylogeny is not certain, or it may be one in which L. burkei has been derived from hybridization between its two relatives. Support for the latter hypothesis comes from the appearance of some individuals in F₁ progenies of L. conjugens × L. fremontii that are morphologically indistinguishable from L. burkei (although fairly sterile). The apparently rather simple genetic basis for the morphological characteristics of each of the species in this trio suggests that the morphologically heterogeneous genus Lasthenia may be considerably more homogeneous genetically than might be suspected. Because of the diverse kinds of relationships among these three Lasthenias, possible alternative taxonomies for the group are dependent upon those relationships that a taxonomist wishes to communicate. Nevertheless, the patterns of diversification in this group have led to reaffirmation of an earlier decision that three species should be recognized.     

Limited mitochondrial DNA variation within South Africa's black rhino (Diceros bicornis minor) population and implications for management

The taxonomy of African black rhinoceros (Diceros bicornis) remains unresolved. Maintaining levels of genetic diversity and species rescue by reintroduction and restocking requires its resolution. We compared the sequences of the mitochondrial DNA (mtDNA) control region for a total of 101 D. bicornis from three subspecies: D. b. minor, D. b. michaeli and D. b. bicornis. A single unique haplotype was found within the 65 D. b. minor samples from KwaZulu‐Natal (KZN) Province, South Africa, 55 of which came from Hluhluwe‐iMfolozi Game Park (HiP) and Mkuzi Game Reserve (MGR) source populations. However, six different haplotypes were represented in eleven D. b. minor samples from Zimbabwe. Similarly, published autosomal microsatellite data indicate low levels of diversity within the KZN D. b. minor populations. The low levels of mtDNA diversity within the KZN metapopulation point to the possible need for genetic supplementation. However, there is a need to determine whether the low levels of genetic variation within KZN D. b. minor are a result of the recent bottleneck or whether KZN historically always had low diversity.     

GENETIC DIVERSITY AND POPULATION STRUCTURE IN TRADESCANTIA HIRSUTICAULIS (COMMELINACEAE)

Tradescantia hirsuticaulis, the hairy-stemmed spiderwort, is an insect-pollinated perennial plant species found primarily on rock outcrops in Georgia, South Carolina, and Alabama. Although populations of T. hirsuticaulis are rare and scattered, local populations are frequently large. Levels of genetic variation were assessed for 13 populations representing the species' range in these three states. Despite the disjunct distribution of this habitat specialist and apparent lack of specialized seed and pollen dispersal mechanisms, exceptionally high levels of genetic variation are maintained within the species, with a moderate level of variation (18%) found among populations. Twenty-nine of the 33 loci resolved (88%) were polymorphic within the species; the mean number of loci polymorphic within populations was 54%. The mean number of alleles per polymorphic locus was 3.24 across all populations and averaged 2.37 within populations. Genetic diversity was 0.206 for the species, whereas mean population genetic diversity was 0.157, both much higher than the average for other short-lived herbaceous perennials. Estimated levels of gene flow were moderate (Nm = 0.95), and a significant association between geographic distance and genetic distance between populations was found (r = 0.68; P < 0.0001). Habitat destruction is the major threat to this genetically diverse species. Since gene flow among its highly dispersed populations is limited, diminution or extinction of local populations could jeopardize the long-term evolutionary potential of this species.     

Cryptic goldfields: a molecular phylogenetic reinvestigation of Lasthenia californica sensu lato and close relatives (Compositae: Heliantheae sensu lato)

Maximum parsimony analysis of DNA sequence data from the internal and external transcribed spacer (ITS and ETS) regions of 18S-26S nuclear ribosomal DNA and the 3' trnK intron of chloroplast DNA from over 60 populations of Lasthenia sect. Amphiachaenia yielded a well-supported tree showing that the most common species of Lasthenia, L. californica sensu lato (s.l.), is not monophyletic. Members of Lasthenia californica s.l. belong to two well-supported but morphologically cryptic clades. One clade includes members of L. macrantha; the other represents a basally divergent lineage in L. sect. Amphiachaenia. Members of each clade can be diagnosed by pappus morphology and by geographic distribution, except for epappose plants that occur in a broad region of sympatry in central California. Overall diversification in the clade corresponding to L. sect. Amphiachaenia has been accompanied by minimal morphological divergence, which has resulted in previously underappreciated cryptic diversity.     

A divergent lineage among Octopus minor (Sasaki, 1920) populations in the Northwest Pacific supported by DNA barcoding

Octopus minor (Sasaki, 1920) is an important economic fishery resource in China. In order to explore the stock information and the phylogeographic status of O. minor, mitochondrial DNA cytochrome c oxidase subunit I (COI, 565?bp) and 16S rRNA (493?bp) genes were amplified from 11 different sampling locations. Genetic diversity evaluated by haplotypic and nucleotidic diversity implied high diversity in Lianjiang, and relatively low diversity in Rongcheng, which suggests that effective measures to protect the O. minor resource in this area are urgently required. Private haplotypes and remarkable higher pairwise Φ_ST in Yilan are responsible for the deep genetic divergence between Yilan and the 10 other populations. Haplotypes networks and two clusters’ topological structure also support the distinct subgroups (lineages A and lineages B), which apparently possess smaller genetic variation than mean interspecies distance. Taiwan island and its strait may act as a natural barrier that restricts the gene flow from the mainland. Deep genetic divergence between mainland and Taiwanese east coasts suggests different genetic stock, indicating that different management strategies are required.     

Spatial genetic structure in Beta vulgaris subsp. maritima and Beta macrocarpa reveals the effect of contrasting mating system,influence of marine currents,and footprints of postglacial recolonization routes

Understanding the factors that contribute to population genetic divergence across a species' range is a long‐standing goal in evolutionary biology and ecological genetics. We examined the relative importance of historical and ecological features in shaping the present‐day spatial patterns of genetic structure in two related plant species, Beta vulgaris subsp. maritima and Beta macrocarpa. Using nuclear and mitochondrial markers, we surveyed 93 populations from Brittany (France) to Morocco – the southern limit of their species' range distribution. Whereas B. macrocarpa showed a genotypic structure and a high level of genetic differentiation indicative of selfing, the population genetic structure of B. vulgaris subsp. maritima was consistent with an outcrossing mating system. We further showed (1) a strong geographic clustering in coastal B. vulgaris subsp. maritima populations that highlighted the influence of marine currents in shaping different lineages and (2) a peculiar genetic structure of inland B. vulgaris subsp. maritima populations that could indicate the admixture of distinct evolutionary lineages and recent expansions associated with anthropogenic disturbances. Spatial patterns of nuclear diversity and differentiation also supported a stepwise recolonization of Europe from Atlantic‐Mediterranean refugia after the last glacial period, with leading‐edge expansions. However, cytoplasmic diversity was not impacted by postglacial recolonization: stochastic long‐distance seed dispersal mediated by major oceanic currents may mitigate the common patterns of reduced cytoplasmic diversity observed for edge populations. Overall, the patterns we documented here challenge the general view of reduced genetic diversity at the edge of a species' range distribution and provide clues for understanding how life‐history and major geographic features interact to shape the distribution of genetic diversity.     

Are bottlenecks associated with colonization? Genetic diversity and diapause variation of native and introduced Rhagoletis completa populations

The success of invasive species appears to be a paradox: despite experiencing strong population bottlenecks, invasive species are able to successfully establish in new environments. We studied how the walnut husk fly, Rhagoletis completa, was able to successfully colonize California from the Midwestern United States, by examining genetic diversity and diapause variation of native and introduced fly populations. Climate plays an important role in the successful establishment of introduced insects, because insect diapause is highly dependent upon external climatic conditions. We examined if: (1) fly populations show signs of a population bottleneck, (2) native and introduced flies differ in diapause length when exposed to California and Midwestern climatic conditions, and (3) population genetic diversity is related to variation in diapause length. We assessed if fly diapause conformed more to a model of establishment by local adaptation or to a model of a highly plastic “general-purpose genotype”. Our results indicate that only two populations close to the original introduced location showed signs of a population bottleneck, and native and introduced populations did not differ in genetic diversity. Genetic diversity increased in the northern introduced populations, suggesting that multiple introductions have occurred. Flies emerged about 2 weeks earlier under the Midwestern treatment than the California treatment, and introduced flies emerged about a week earlier than native flies. All flies emerged when walnuts are typically available in California. Although variance in diapause length differed between populations, it did not vary between populations or regions. Furthermore, genetic diversity was not associated with diapause variation. Therefore, multiple introductions and a “general-purpose genotype” appear to have facilitated the fly’s invasion into California.     

Population genetic diversity and species relationships in the genus Rhinanthus L. based on microsatellite markers

The genus Rhinanthus L. is complex, containing many taxonomically unresolved taxa. In this paper we studied genetic variation and species relationships in 15 populations of six Rhinanthus species from three sections. For this purpose, we developed new microsatellite primers for R. osiliensis and used them to investigate genetic variation in two narrow endemics (R. osiliensis, R. javorkae) and in four widespread species (R. rumelicus R. wagneri, R. angustifolius and R. minor). Species‐specific private alleles were found in all species except R. osiliensis and R. angustifolius. The Bulgarian endemic R. javorkae showed the lowest genetic variation, followed by widespread R. minor and Estonian endemic R. osiliensis. Rhinanthus javorkae and R. minor were genetically most differentiated. Section Cleistolemus is weakly structured genetically, indicating close affinity between R. osiliensis, R. rumelicus, R. wagneri and R. angustifolius.     

Low genetic diversity and recovery implications of the vulnerable Bankoualé Palm Livistona carinensis (Arecaceae), from North-eastern Africa and the Southern Arabian Peninsula

The Bankoualé Palm, Livistona carinensis is the only known species of Livistona occurring in Africa and is currently classified as vulnerable (IUCN 2004). This extreme outlier species of the genus is restricted to Yemen, Somalia and Djibouti, where all populations are in rapid decline. In Djibouti the palm is confined to three valley systems within the upland plateau of the Goda Massif. This study used microsatellite markers to investigate the genetic diversity and relationships within the species. At the species level L. carinensis contained very low genetic diversity. Most variation was due to the variation between the samples from Yemen and Somalia compared with those in Djibouti. The Djibouti populations were almost monomorphic across the nine loci tested. Interestingly, and despite the small sample sizes, the individuals from botanic gardens collections of the Yemen and Somalia populations were more genetically diverse than the Djibouti populations. This study indicates that the populations in Yemen and Somalia are highly significant for the conservation of the species genetic diversity. Given the lack of genetic diversity both within and among L. carinensis populations in Djibouti, plants could be cultivated for in-situ population enhancement from any seed that is available from within Djibouti with no significant genetic impacts of provenance mismatch. Clearly the populations from Djibouti, Somalia and Yemen are different genetic provenances raising some issues for the conservation and recovery of L. carinensis.     

Phylogeographic evidence for the postglacial colonization of the North and Baltic Sea coasts from inland glacial refugia by Triglochin maritima L.

We investigated the geographical distribution of genetic variation in 67 individuals of Triglochin maritima from 38 localities across Europe using AFLP markers. Analysis of genetic variation resulted in the recognition of two major genetic groups. Apart from few geographical outliers, these are distributed (1) along the Atlantic coasts of Portugal, Spain and France and (2) in the North Sea area, the Baltic Sea area, at central European inland localities, the northern Adriatic Sea coast and the Mediterranean coast of southwest France. Considering possible range shifts of T. maritima in reaction to Quaternary climatic changes as deduced from the present-day northern temperature limit of the species, Quaternary changes of coastline in the North Sea area and the very recent origin of the Baltic Sea, we conclude that the coastal populations of T. maritima in the North Sea and Baltic Sea areas originated from inland populations.     

Microsatellite and chloroplast DNA analyses reveal no genetic variation in a beach plant Surianana maritima on the Paracel Islands,China

Genetic variation within and among six natural populations of beach plant Suriana maritima on the Paracel Islands of China was assessed using SSR and chloroplast DNA sequence analysis. The results showed a complete absence of genetic variation between individuals either within or among populations. The cumulative consequences of a founder effect, selfing, and inbreeding may be responsible for the loss of genetic diversity in S. maritima.     

High genetic divergence characterizes populations of the endemic plant Lithophragma maximum (Saxifragaceae) on San Clemente Island

Narrowly-ranging species frequently harbor less genetic variability relative to widespread relatives and face graver extinction threats due to the heightened impacts of stochastic events on ecological and genetic diversity. In this study, we examined the impact of historical and current threats to the maintenance of genetic variation in Lithophragma maximum (Saxifragaceae), a perennial herb endemic to San Clemente Island, California. This species exists as small populations confined to canyons along 4 km of the southeast coastline of the island. In 15 populations analyzed with 10 microsatellite markers, we identified an average of 2.05 alleles per locus and 58.7% polymorphic loci. Significant departures from Hardy–Weinberg equilibrium existed in six populations; five of these exhibited heterozygote deficiency. Bayesian inference of genetic structure indicated a significant amount of structure among populations and canyons and infrequent gene flow even over very short distances. We also identified a significant and positive correlation between genetic and geographic distances, indicative of isolation by distance. There was no evidence of recent bottlenecks in any of the sampled populations, but older bottlenecks were detected in two populations. These results suggest that populations of L. maximum have historically been small and isolated, which is likely due to the rugged habitat in which this species occurs and limited pollen and seed dispersal. Given the high degree of structure observed across populations, we suggest that conservation efforts should focus on preserving populations in multiple canyons, maintaining large population sizes to preserve genetic variation, and controlling the spread of invasive species in areas where L. maximum occurs.     

Does habitat fragmentation reduce genetic diversity and subpopulation connectivity?

The estimation of levels of genetic variation has received considerable attention because it is generally thought to be indicative of overall species vitality and the potential for evolutionary responses to environmental changes. Here, we use allozymes markers and two distinct collections of Cakile maritima, an annual species from sandy coastal habitats (2000 generation and 2005 generation collected from 9 populations in their natural habitats), to assess the magnitude of expected genetic change. We compared genetic diversity between generations (all populations combined), and then between populations at each generation. Based on 13 loci scored from the eight enzymes examined, a high genetic diversity was detected at both the population and generation level as compared to other herbaceous species. However, allelic richness reduction in the 2005 generation suggested restricted gene flow and a high risk of future genetic bottlenecks, if larger tracts of coastal areas disappear. Most loci showed deviation from Hardy‐Weinberg equilibrium due to excess of heterozygotes in all populations suggesting that this species has an allogamic mode of reproduction. It appears most likely that this species has experienced a recent decrease in population size, and that genetic drift in small populations has resulted in a loss of alleles occurring at low frequency. Despite the deterioration process, maintenance of high genetic diversity suggests that there are some ecological factors determining population structure.