Using dense locality sampling resolves the subtle genetic population structure of the dispersive fish species Plecoglossus altivelis

In dispersive species with continuous distributions, genetic differentiation between local populations is often absent or subtle and thus difficult to detect. To incorporate such subtle differentiation into management plans, it may be essential to analyse many samples from many localities using adequate numbers of high‐resolution genetic markers. Here, we evaluated the usefulness of dense locality sampling in resolving genetic population structure in the ayu (Plecoglossus altivelis), a dispersive fish important in Japanese inland fisheries. Genetic variability in, and differentiation between, ayu populations around the Japan–Ryukyu Archipelago were investigated in 4746 individuals collected from 120 localities by genotyping 12 microsatellite markers. These individuals represented the two subspecies of ayu, namely the Ryukyuan subspecies (Plecoglossus altivelis ryukyuensis) and both amphidromous and landlocked forms of the nominotypical subspecies (P. a. altivelis) along the archipelago. We successfully detected an absence of genetic differentiation within the landlocked form and subtle but significant differentiation and clear geographic patterns of genetic variation among populations of the amphidromous form, which had been considered genetically homogeneous. This suggests that dense locality sampling effectively resolves subtle differences in genetic population structure, reducing stochastic deviation in the detection of genetic differentiation and geographic patterns in local populations of this dispersive species. Resampling analyses based on empirical data sets clearly demonstrate the effectiveness of increasing the number of locality samples for stable and reliable estimations of genetic fixation indices. The genetic population structure observed within the amphidromous form provides useful information for identifying management or conservation units in ayu.     

Phylogeography of a vanishing North American songbird: the Painted Bunting (<Emphasis Type="Italic">Passerina ciris</Emphasis>)

The breeding distribution of Painted Buntings (Passerina ciris) is comprised of two allopatric populations separated by a 550-km distributional gap in the southeastern United States. Curiously, the boundary between the two recognized P. ciris subspecies does not separate the two allopatric breeding populations but instead runs roughly through the center of the interior population. Genetic relationships among these subspecies, and the allopatric breeding populations of Painted Bunting, have not been assessed. Given the recent decline in overall abundance of this species, such an assessment is warranted. We sampled birds from 15 localities (138 individuals) and identified 35 distinct haplotypes, six belonging to the Atlantic Coast population and 26 to the interior population, with three shared by both populations. AMOVA results showed that a significantly greater portion of the total genetic variance is explained when grouping birds by the interior and Atlantic Coast populations rather than by subspecies. Furthermore, our data indicate that the Atlantic Coast and interior populations represent independently evolving taxa, with no measureable gene flow between them. Although recently diverged (26,000–115,000 years ago), these isolated bunting populations represent incipient species. For development of conservation strategies, we suggest that the Atlantic Coast and interior populations be recognized as separate management units.     

Population structure and genetic diversity of red deer (<Emphasis Type="Italic">Cervus elaphus</Emphasis>) in forest fragments in north-western France

Red deer have been subjected to anthropogenic interference for many centuries. Most populations are managed according to hunting schedules, some have been kept long-term in enclosures and other populations have been restocked with foreign deer. The red deer in the Brittany region of north-western France only occupy the largest forests in the region, reaching quite high densities in restricted areas. Here, we aimed to assess the extent of the genetic variability of the populations in four forest fragments and investigate their population genetic structure. We show that, despite relatively large expected heterozygosity values, these geographically isolated populations are genetically impoverished relative to individuals from large continuous forests in other parts of Western Europe. We provide evidence for population genetic structure with large genetic differentiation between geographically close populations, suggesting the absence of effective exchange between the forests. Using samples from the most likely source population, we show that at least two populations were non-indigenous. In order to limit further loss of genetic diversity, it should be a management objective to reduce isolation of the different forests, rather than further increase it by fences and hunting practices that could limit free movement of red deer.     

Consistency of population genetics parameters estimated from isozyme and RAPDs dataset in species of genus <Emphasis Type="Italic">Prosopis</Emphasis> (Leguminosae,Mimosoideae)

Genetic variability, population structure and differentiation among 17 populations of 5 species and 2 natural interspecific hybrids of section Algarobia of genus Prosopis were analyzed from data of 23 isozyme and 28 RAPD loci. Both markers indicated that the studied populations are highly variable. P. alba populations in average showed lower values of genetic variability estimates from isozyme data, but this trend was not observed for RAPD markers. The hierarchical analyses of the distribution of genetic variability showed that the highest proportion of variation occurred within populations, the differentiation among species was intermediate and the lowest component was observed among populations within species. The consistency between results from both dataset implies that they are not biased and reflect the actual genetic structure of the populations analyzed. The matrices of Euclidean distances obtained from the two sets of markers were highly correlated according to Mantel test. In both cases the corresponding phenogram and MDS plot tended to cluster conspecific populations while hybrid populations were not intermediate between putative parents. Some disagreements between isozyme and RAPD phenograms were observed mainly in the affinities of hybrid populations. Such inconsistencies might result from reticular rather than dichotomic evolutionary relationships. The phenetic associations retrieved gave no support to the division of the section Algarobia into series.     

Genetic structure of kestrel populations and colonization of the Cape Verde archipelago

Genetic diversity and population structure were studied in eight populations of the kestrel Falco tinnunculus to identify the genetic consequences of spatial distribution and to infer the colonization patterns of the Cape Verde archipelago. We studied genetic differentiation and gene flow among seven island populations and one mainland population using nine microsatellite loci. Within the archipelago, differentiation was strong and genetic diversity and heterozygosity were low but variable among populations. Two subspecies F. tinnunculus neglectus on the northwestern islands and F. tinnunculus alexandri on all the other islands were identified as genetically distinct units. F. t. alexandri could be further separated into two groups on eastern and southern islands. Populations are probably founded by birds originating from the mainland. Immigration is more likely to the eastern and southern populations, whereas the northwestern islands with the lowest genetic diversity and highest differentiation are likely to exhibit fewer founding events by immigrants. The number of founding events on each island may depend not only on geographical distance to neighbouring populations, but also on directional immigration due to the northeastern trade winds. This may explain differences in genetic differentiation and diversity between populations and subspecies and may enable allopatric speciation.     

Population genetic structure of the European kestrel Falco tinnunculus in Central Poland

We analysed the genetic structure of the European kestrel population of Central Poland using nine highly polymorphic microsatellite loci. Samples were collected in two urban locations (Warsaw and ?ód?) and two rural areas. Sampling locations were at nearly equal distances from each other along an east to west line. We performed genotyping in a total of 99 birds. The results revealed the presence of a genetic structure in the population investigated. Bayesian clustering indicated that samples originated from more than one population. Genetic differentiation was less pronounced among the birds nesting in Warsaw and in the two rural sites, whereas all pairwise comparisons with the ?ód? population indicated moderate and significant genetic differentiation. The observed pattern of differentiation might have been caused by two factors: changes in allele frequency between seasons and/or the founding of the urban population of ?ód? from a different source population than the urban population from Warsaw. Additionally, we found a rather high gene flow among kestrels from the Warsaw urban area and the two investigated rural areas.     

Genetic variation of chloroplast and nuclear markers in natural populations of hazelnut (Corylus avellana L.) in Germany

Corylus avellana L. (hazel) is a long-lived, monoecious and wind-pollinated shrub species, widespread all over Europe. In Germany, hazel is intensively traded and planted, and thus is of central interest from a nature conservancy point of view. To assess the within- and between-population differentiation of hazel, 20 natural populations (18 from Germany, one from Italy and one from Hungary) were investigated genetically. Seven isozyme systems comprising 11 gene loci were analysed in up to 100 samples (average 92.6) per population, amplified fragment length polymorphisms (AFLP) were analysed in up to 50 samples (average 47.4) and nine cpDNA-SSR markers were assessed in 20 samples per population. Results for overall isozyme variability with Na 2.46 alleles per locus, allelic diversity (Ne) 1.39, expected heterozygosity He 21 % and 79 % polymorphic loci were in accordance with the findings of previous studies. The respective values for AFLPs were lower, but both marker systems revealed the same level of about 3.5 % differentiation between populations. For cpSSR only the Italian sample showed within-population variation and the two haplotypes were completely differentiated from all other populations expressing a unique genetic structure with one single haplotype. Among the three marker systems AFLPs showed the best ability to differentiate between populations. While only one isozyme locus revealed significant differentiation, 41 AFLP loci showed highly significant differentiation between all populations, but 26 loci when only German populations were considered. Consequently geographic differentiation analyses focused mainly on molecular markers. Mantel tests showed significant correlations between genetic and geographic distance, but in the unweighted pair-group method with arithmetic mean analyses, adjacent populations did not always form clusters. While chloroplast markers were able to clearly distinguish only the Hungarian population, the nuclear markers revealed clear spatial genetic structures. The correlations between geographic and genetic distance was high for AFLPs. The correlograms illustrate this effect for all populations as well as for the German populations.     

Genetic differentiation of southeast Baltic populations of sea trout inferred from single nucleotide polymorphisms

Sea trout (Salmo trutta m. trutta) is a migratory form of brown trout common in the Baltic Sea. Nine populations from the southeast Baltic (Poland; Lithuania; Denmark, Bornholm; Estonia and Russia) were genotyped using iPLEX Gold technology (Sequenom) with 62 informative SNPs. A diagnostic panel of 23 SNPs was applied to estimate genetic differentiation and assess the population structure of Baltic sea trout. The highest level of pairwise F_ST differences was observed between the Russian (East Gulf of Finland) and Polish (Baltic main basin) populations. The lowest differences were between the two Polish and the Polish and Lithuanian populations. A genetic similarity was noted between the Estonian Riguldi River and Danish Bornholm populations, and this finding was supported by a Bayesian and factorial correspondence analysis. Diversity within populations was highest for populations from Estonia and lowest for the Lithuanian population. Genetic structure analysis indicated that individuals from the nine populations were clustered into four groups.     

Genetic structure of the Eurasian lynx population in north-eastern Poland and the Baltic states

We analyzed the genotypes of Eurasian lynx (Lynx lynx) from three populations in the westernmost part of the species main range. One population was situated at the distribution edge (NE Poland) and the two other (Latvia and Estonia) were located within the main, contiguous range of the species. The aim was to determine if the genetic composition varied among these populations and if there was evidence of isolation among them. Based on microsatellite allele frequencies, we found the allelic richness in Polish lynx to be lower than that in lynx from Latvia and Estonia. We also found significant differentiation among the lynx populations, with the NE Poland population forming a distinct genetic group relative to the two other populations (R _ST = 0.15 and 0.22, P < 0.0001). We suggest that genetic differentiation among lynx populations is the result of habitat insularisation that limits gene flow. This finding emphasizes the necessity to consider the lynx genetic differentiation in conservation planning of this species in Poland.     

Population differentiation in the redshank (<Emphasis Type="Italic">Tringa totanus</Emphasis>) as revealed by mitochondrial DNA and amplified fragment length polymorphism markers

The redshank (Tringa totanus) is declining throughout Europe and to implement efficient conservation measures, it is important to obtain information about the population genetic structure. The aim of the present study was two-fold. First, we analysed the genetic variation within and between populations in the Baltic region in southern Scandinavia. Evidence of genetic structure would suggest that different populations might require separate management strategies. Second, in an attempt to study large-scale genetic structure we compared the Baltic populations with redshanks from northern Scandinavia and Iceland. This analysis could reveal insights into phylogeography and long-term population history. DNA samples were collected from six breeding sites in Scandinavia presumed to include two subspecies (totanus and britannica) and a further sample from Iceland (subspecies robusta). Two methods were used to study the population genetic structure. Domain II and III of the mitochondrial control region was analysed by DNA sequencing and nuclear DNA was analysed by screening amplified fragment length polymorphism (AFLP) markers. Mitochondrial DNA showed no variation between individuals in domain II. When analysing an 481 bp fragment of domain III seven haplotypes were found among birds. On the basis of mtDNA sequences, redshanks showed some evidence of a recent expansion from a bottlenecked refugial population. Bayesian analyses of AFLP data revealed a significant genetic differentiation between suggested subspecies but not between populations within the Baltic region. Our results indicate that populations of redshanks in Europe constitute at least three separate management units corresponding to the recognised subspecies.     

Significant genetic differentiation between Poland and Germany follows present-day political borders, as revealed by Y-chromosome analysis

To test for human population substructure and to investigate human population history we have analysed Y-chromosome diversity using seven microsatellites (Y-STRs) and ten binary markers (Y-SNPs) in samples from eight regionally distributed populations from Poland (n=913) and 11 from Germany (n=1,215). Based on data from both Y-chromosome marker systems, which we found to be highly correlated (r=0.96), and using spatial analysis of the molecular variance (SAMOVA), we revealed statistically significant support for two groups of populations: (1) all Polish populations and (2) all German populations. By means of analysis of the molecular variance (AMOVA) we observed a large and statistically significant proportion of 14% (for Y-SNPs) and 15% (for Y-STRs) of the respective total genetic variation being explained between both countries. The same population differentiation was detected using Monmoniers algorithm, with a resulting genetic border between Poland and Germany that closely resembles the course of the political border between both countries. The observed genetic differentiation was mainly, but not exclusively, due to the frequency distribution of two Y-SNP haplogroups and their associated Y-STR haplotypes: R1a1*, most frequent in Poland, and R1*(xR1a1), most frequent in Germany. We suggest here that the pronounced population differentiation between the two geographically neighbouring countries, Poland and Germany, is the consequence of very recent events in human population history, namely the forced human resettlement of many millions of Germans and Poles during and, especially, shortly after World War II. In addition, our findings have consequences for the forensic application of Y-chromosome markers, strongly supporting the implementation of population substructure into forensic Y chromosome databases, and also for genetic association studies.     

Genetic differentiation of the Thorn‐tailed Rayadito Aphrastura spinicauda (Furnariidae: Passeriformes) revealed by ISSR profiles suggests multiple palaeorefugia and high recurrent gene flow

Nucleotide sequence data (cytochrome b) and ISSR genomic fingerprints were used to analyse the genetic variation and population differentiation in Thorn‐tailed Rayadito, a widespread Patagonian forest bird. We included samples from eight populations of Thorn‐tailed Rayadito covering most of the distribution range of the species: from fragmented patches of Olivillo forest in northern Chile to Isla Navarino forests in the extreme south of South America. Low levels of genetic diversity were found among populations, with a large within‐population molecular variance indicating high levels of gene flow. The multivariate and cluster analyses based on ISSR markers show that the subspecies bullocki (from Mocha Island) differs significantly from all other populations. The subspecies fulva (Chiloé Island) shows less differentiation than bullocki, sharing several alleles with continental populations. Bayesian analyses suggest that the Mocha Island population contributes most to the total genetic diversity observed in the species. Mantel tests revealed no significant correlation between geographical distance and pairwise genetic distance and cytochrome b sequence analyses failed to detect differentiation among subspecies. Mocha Island might have been a palaeorefuge and this population may have diversified by genetic drift after the last glacial maximum. There is also the possibility of a postglacial colonization of the Thorn‐tailed Rayadito from an austral palaeorefugium, supporting a multiple refugia hypothesis. This study illustrates the usefulness of the rarely used ISSR genomic fingerprint method in avian phylogeography.     

Genetic Analysis of the <Emphasis Type="Italic">Indri</Emphasis> Reveals No Evidence of Distinct Subspecies

Subspecies were traditionally defined by identifying gaps between phenotypes across the geographic range of a species, and may represent important units in the development of conservation strategies focused on preserving genetic diversity. Previous taxonomic research proposed that phenotypic variation between scattered Indri indri populations warranted the naming of two distinct subspecies, I. i. indri and I. i. variegatus. We tested these subspecific designations using mitochondrial sequence data generated from the control region or D-loop (569 bp) and a large section (2362 bp) of multiple genes and tRNAs known as Pastorini’s fragment and nuclear microsatellite markers. This study used 114 samples of I. indri from 12 rainforest sites in eastern Madagascar, encompassing the entire range of the species. These genetic samples represent multiple populations from low- and high-elevation forests from both putative subspecies. Molecular analyses of the mitochondrial sequence data did not support the two proposed subspecies. Furthermore, the microsatellite analyses showed no significant differences across the range beyond population level differentiation. This study demonstrates the utility of incorporating multiple lines of evidence in addition to phenotypic traits to define species or subspecies.     

Genetic variation of teak (Tectona grandis Linn. f.) in Myanmar revealed by microsatellites

Genetic variation of teak (Tectona grandis Linn. f.) in 16 populations in Myanmar was investigated using ten nuclear microsatellite markers. Eight population pairs from two main regions in the north and the south of Myanmar were sampled. Each population pair consisted of an unlogged and a recently logged forest, each represented by 50 adult trees and 50 seedlings from the natural regeneration. For comparison, two land races from Benin (West Africa) were included. The major objectives of the study are to characterize the patterns of genetic variation of teak in natural populations, to examine genetic differentiation between adult trees and natural regeneration, and to investigate the impact of selective logging on genetic structures of teak. Genetic variation was high in all investigated populations. Slightly elevated levels of inbreeding were observed in the regeneration in comparison to the adults. Populations from the northern and the southern regions were strongly differentiated, but the differentiation between adults and natural regeneration and between unlogged and logged forests was low and not significant. Mantel tests indicated an isolation by distance (IBD) within the northern and the southern regions. High genetic diversity was also observed within the land races from Benin, which grouped to the southern populations. We failed to detect effects of logging on genetic diversity patterns or inbreeding in adults and regeneration, suggesting that high genetic diversity can even be sampled and maintained in disturbed forests. The observation of significant IBD and high differentiation between the populations of the north and the south of Myanmar suggests to include populations from widely separated forests in conservation programs, and to delineate provenance regions for the harvest and transfer of teak seeds and seedlings.     

Tigers of Sundarbans in India: Is the Population a Separate Conservation Unit?

The Sundarbans tiger inhabits a unique mangrove habitat and are morphologically distinct from the recognized tiger subspecies in terms of skull morphometrics and body size. Thus, there is an urgent need to assess their ecological and genetic distinctiveness and determine if Sundarbans tigers should be defined and managed as separate conservation unit. We utilized nine microsatellites and 3 kb from four mitochondrial DNA (mtDNA) genes to estimate genetic variability, population structure, demographic parameters and visualize historic and contemporary connectivity among tiger populations from Sundarbans and mainland India. We also evaluated the traits that determine exchangeability or adaptive differences among tiger populations. Data from both markers suggest that Sundarbans tiger is not a separate tiger subspecies and should be regarded as Bengal tiger (P. t. tigris) subspecies. Maximum likelihood phylogenetic analyses of the mtDNA data revealed reciprocal monophyly. Genetic differentiation was found stronger for mtDNA than nuclear DNA. Microsatellite markers indicated low genetic variation in Sundarbans tigers (He= 0.58) as compared to other mainland populations, such as northern and Peninsular (Hebetween 0.67- 0.70). Molecular data supports migration between mainland and Sundarbans populations until very recent times. We attribute this reduction in gene flow to accelerated fragmentation and habitat alteration in the landscape over the past few centuries. Demographic analyses suggest that Sundarbans tigers have diverged recently from peninsular tiger population within last 2000 years. Sundarbans tigers are the most divergent group of Bengal tigers, and ecologically non-exchangeable with other tiger populations, and thus should be managed as a separate “evolutionarily significant unit” (ESU) following the adaptive evolutionary conservation (AEC) concept.     

Genetic insights into the historical distribution of <Emphasis Type="Italic">Polylepis pauta</Emphasis> (Rosaceae) in the northeastern Cordillera Oriental of Ecuador

Scattered patches of Polylepis forest that occur within the 3,000–4,500 m altitudinal belt of the Andean Cordillera from Venezuela to Argentina have been hypothesized to be remnants of once continuous forests whose range became fragmented through anthropogenic activities that probably preceded the Spanish conquest. Allozyme variation of Polylepis pauta from 12 forest populations in three adjacent watersheds in Northeastern Ecuador was investigated to assess whether observed patterns of gene diversity were consistent with a more continuous historical range of the species and to evaluate the populations’ potential for restoration. Genetic diversity and polymorphism in P. pauta populations were higher than mean values for most wind pollinated and dispersed temperate and tropical tree species with regional distributions. Genetic differentiation among watersheds was lower than among populations within each watershed. Isolation by distance was not evident and several populations from different watersheds were more genetically similar than populations from the same watershed. Larger forest patches with broader altitudinal ranges had more alleles. Forest patches on steeper slopes and at higher elevations supported populations with less genetic diversity; this might have resulted from the predominance of vegetative reproduction in these landslide prone areas. The amount of genetic diversity maintained by P. pauta, coupled with low genetic differentiation among populations within and among watersheds, is consistent with a more continuous historical range of the species in Northeastern Ecuador and point to the Oyacachi basin as having the highest levels of genetic diversity.     

Population isolation rather than ecological variation explains the genetic structure of endangered myrmecophilous butterfly Phengaris (=Maculinea) arion

Genetic variation of the globally threatened obligatorily myrmecophilous Large Blue butterfly Phengaris (Maculinea) arion (Lepidoptera) was studied, using six microsatellite markers, in a country where its decline is dramatic (Poland). Material was collected on 13 sites showing considerable ecological variation as far as biotope, larval food plant and host ants of the butterfly were concerned. Genetic variability, estimated in terms of number of alleles and heterozygosity, was the lowest in the most isolated populations. However on sites localized in areas where suitable biotopes were extensive and interconnected, P. arion still held relatively high genetic diversity. Pairwise F _ST values indicated small and moderate differentiation among samples (F _ST = 0.01–0.15), with the exceptions of two isolated localities (0.20). We did not find clear evidence of isolation by distance. The presence of four or five genetic clusters was indicated. Analysis of the membership of each individual to each cluster showed that the vast majority of individuals from three isolated populations were clustered in three separate genetic groups. The most distinct population was the one, which had been found to be specialized towards Myrmica lobicornis in previous studies. Individuals from the remaining populations could not be clustered in separate genetic groups, however some dominance of different clusters in geographical regions was observed. Some portion of the population’s genetic variability could be explained by geographical distribution, however the percentage of variation, explaining the differences between two main regions (S and NE Poland), was very low. We conclude that the main factor shaping the current genetic structure of P. arion in Poland is the recent isolation of populations related to habitat fragmentation but local ecological specializations may be also a potential factor. Therefore the necessity of activities aiming to halt the further reduction of genetic variability, as well as the monitoring of priority populations (e.g. those belonging to unique host races), should be emphasized in future action plans in Central Europe.     

中国红原鸡和泰国红原鸡遗传多样性分析

利用29个微卫星DNA标记对来自中国的红原鸡Gallus gallus spadiceus亚种和来自泰国的红原鸡Gallus gallus gallus亚种进行遗传多样性分析, 评估亚种内的遗传变异和亚种间的遗传分化, 结果表明: 共检测到168个等位基因, 每个位点的等位基因数从2到13不等, 所有位点平均的期望杂合度和PIC值分别为0.5780和0.53。中国和泰国红原鸡29个微卫星位点平均有效等位基因数分别为3.79和4.79, 平均基因杂合度为0.5379和0.6385, 两个红原鸡亚种均表现出较高的群体杂合度和丰富的遗传多样性。群体分化系数为19.4%(P<0.01), 两个红原鸡亚种间的Reynolds’遗传距离和Nm值分别为0.157和1.040。由此可见, Gallus gallus spadiceus亚种和Gallus gallus gallus亚种群体具有不同的群体遗传结构, 群体之间存在明显的遗传分化, 并不能将其认定为是同一亚种, 这也为中国家鸡具有独立的起源提供了一定的佐证。     

Multilocus approach reveals an incipient differentiation process in the Stone-curlew, <Emphasis Type="Italic">Burhinus oedicnemus</Emphasis> around the Mediterranean basin

The Stone-curlew Burhinus oedicnemus, a steppe bird species, is mainly distributed in the Mediterranean and Macaronesian regions, which are considered hotspots of biodiversity with priority for animal and plant species richness conservation. In this study, we investigated the genetic diversity of the Stone-curlew in the Mediterranean basin and in the Canary Islands by applying a multilocus approach. We analysed mitochondrial and nuclear markers in order to evaluate the genetic structure and the congruence between morphological subspecies and geographic samples. We found a significant level of genetic differentiation between Mediterranean and Canary Island populations with all markers. Both in the Mediterranean basin and in the Canary Islands, we found a significant level of genetic diversity with nuclear markers only. We identified seven population groups, including insular populations. The four subspecies described for the Western Palaearctic were confirmed with some changes in distribution range. In spite of habitat fragmentation and negative population trend, the Stone-curlew showed a significant level of genetic diversity and gene flow among continental populations. However, islands constitute important reservoirs of genetic diversity and a potential for the evolution of the species.     

Population genetic diversity and structure within and among disjunct populations of <Emphasis Type="Italic">Alnus maritima</Emphasis> (seaside alder) using microsatellites

Small, isolated populations are prone to genetic drift and high levels of inbreeding that can threaten their long-term survival. Alnus maritima persists exclusively in three groups of small, highly disjunct, regional populations in the Delmarva Peninsula, Georgia, and Oklahoma. Trees in the three regions are recognized as separate subspecies. Microsatellite markers were used to measure fine-scale population genetic diversity and structure (1) within and among regions and (2) within and among populations in each region. Compared to a previous study utilizing allozymes, microsatellite data show higher levels of variation, lower levels of inbreeding, but similar levels of genetic differentiation among regions. Significant genetic differentiation was detected among regions and among distinct populations within regions. Genetic differentiation was significantly correlated with geographic distance among regional populations, but not among populations within regions. Populations, therefore, likely represent fragments of formerly extensive networks of populations that have decayed and retracted due to competition with other species better adapted to the shadier habitats of late-succession environments. The unique genetic features of populations within different regions should be considered as part of future conservation efforts.