Morphology,molecules and taxonomy: extreme incongruence in pleurocerids (Gastropoda,Cerithioidea, Pleuroceridae)

Pronounced mitochondrial heterogeneity within putative species of Pleuroceridae has prevented meaningful systematic revisions of this critically imperilled freshwater family. Previous studies have demonstrated that this mitochondrial diversity often produces polyphyletic species on mitochondrial gene trees, but its significance is unclear. Hypotheses advanced to explain this pattern have included cryptic species, retained ancestral polymorphisms and introgression; other possible explanations such as doubly uniparental inheritance or the presence of pseudogenes have not been given due consideration. Previous analyses have not included adequate sampling, neither in terms of number of individuals nor in geographic coverage, to adequately test any of these hypotheses. To fully characterize mitochondrial heterogeneity in pleurocerids and robustly assess possible causal explanations, we collected 239 individuals representing four putative species from seven sites and sequenced the COI and 16S rRNA mitochondrial genes and the H3 nuclear gene for all individuals. We also used whole‐genome shotgun sequencing to construct and annotate a mitochondrial genome for one individual. Characters with demonstrated utility in morphospecies delineation of gastropods (head–foot coloration, shell and radular morphology, pallial oviduct anatomy) were examined for a subset of individuals to determine whether morphology co‐varied by haplotype clade. We found pronounced mitochondrial heterogeneity at both the population level and species level in three of the species examined, but our data reject paralogous nuclear copies of mitochondrial genes (NUMTs) and doubly uniparental inheritance as causal mechanisms, and there was no evidence of cryptic diversity. Mutation rates were found to differ significantly among mitochondrial lineages, and population genetic statistics revealed a signature of balancing selection that could be acting to maintain this diversity. The observed pattern is similar to that seen in lineages with inherited endosymbionts like Wolbachia infections, which merits further investigation. Although questions remain concerning the precise cause(s) of intraspecific mitochondrial diversity in pleurocerids, nuclear and/or genomic data, combined with anatomical and life history investigations in an integrative phylogenetic context, is the most promising avenue for resolving pleurocerid systematics.     

Contrasting genetic diversity patterns in two sister kelp species co‐distributed along the coast of Brittany,France

We investigated patterns of genetic structure in two sister kelp species to explore how distribution width along the shore, zonation, latitudinal distribution and historical factors contribute to contrasting patterns of genetic diversity. We implemented a hierarchical sampling scheme to compare patterns of genetic diversity and structure in these two kelp species co‐distributed along the coasts of Brittany (France) using a total of 12 microsatellites, nine for Laminaria hyperborea and 11 for Laminaria digitata, of which eight amplified in both species. The genetic diversity and connectivity of L. hyperborea populations were greater than those of L. digitata populations in accordance with the larger cross‐shore distribution width along the coast and the greater depth occupied by L. hyperborea populations in contrast to L. digitata populations. In addition, marginal populations showed reduced genetic diversity and connectivity, which erased isolation‐by‐distance patterns in both species. As L. digitata encounters its southern range limit in southern Brittany (SBr) while L. hyperborea extends down to mid‐Portugal, it was possible to distinguish the effect of habitat continuity from range edge effects. We found that L. digitata did not harbour high regional diversity at its southern edge, as expected in a typical rear edge, suggesting that refuges from the last glacial maximum for L. digitata were probably not located in SBr, but most likely further north. For both species, the highest levels of genetic diversity were found in the Iroise Sea and Morlaix Bay, the two regions in which they are being currently harvested. Preserving genetic diversity of these two foundation species in these areas should, thus, be a priority for the management of this resource in Brittany.     

Genetic variation and bidirectional gene flow in the riparian plant Miscanthus lutarioriparius,across its endemic range: implications for adaptive potential

Miscanthus lutarioriparius is an endemic species that grows along the middle and lower reaches of the Yangtze River and is a valuable source of germplasm for the development of second‐generation energy crops. The plant that propagates via seeds, stem nodes, and rhizomes shows high phenotypic variation and strong local adaptation. Here, we examined the magnitude and spatial distribution of genetic variation in M. lutarioriparius across its entire distributional range and tested underlying factors that shaped its genetic variation. Population genetic analyses were conducted on 644 individuals from 25 populations using 16 microsatellite markers. M. lutarioriparius exhibited a high level of genetic variation (H_E = 0.682–0.786; A_r = 4.74–8.06) and a low differentiation (F_ST = 0.063; D_est = 0.153). Of the total genetic variation, 10% was attributed to the differences among populations (df = 24, P < 0.0001), whereas 90% was attributed to the differences among individuals (df = 619, P ≤ 0.0001). Genetic diversity did not differ significantly across longitudes and did not increase in the populations growing downstream of the Yangtze River. However, significant associations were found between genetic differentiation and spatial distance. Six genetic discontinuities were identified, which mostly distributed among downstream populations. We conclude that anthropogenic factors and landscape features both contributed to shaping the pattern of gene flow in M. lutarioriparius, including long‐distance bidirectional dispersal. Our results explain the genetic basis of the high degree of adaptability in M. lutarioriparius and identify potential sources of new germplasm for the domestication of this potential second‐generation energy crop.     

Genetic variation and effective population size in isolated populations of coastal cutthroat trout

Following glacial recession in southeast Alaska, waterfalls created by isostatic rebound have isolated numerous replicate populations of coastal cutthroat trout (Oncorhynchus clarkii clarkii) in short coastal streams. These replicate isolated populations offer an unusual opportunity to examine factors associated with the maintenance of genetic diversity. We used eight microsatellites to examine genetic variation within and differentiation among 12 population pairs sampled from above and below these natural migration barriers. Geological evidence indicated that the above-barrier populations have been isolated for 8,000–12,500 years. Genetic differentiation among below-barrier populations (F _ST = 0.10, 95% C.I. 0.08–0.12) was similar to a previous study of more southern populations of this species. Above-barrier populations were highly differentiated from adjacent below-barrier populations (mean pairwise F _ST = 0.28; SD 0.18) and multiple lines of evidence were consistent with asymmetric downstream gene flow that varied among streams. Each above-barrier population had reduced within-population genetic variation when compared to the adjacent below-barrier population. Within-population genetic diversity was significantly correlated with the amount of available habitat in above-barrier sites. Increased genetic differentiation of above-barrier populations with lower genetic diversity suggests that genetic drift has been the primary cause of genetic divergence. Long-term estimates of N _e based on loss of heterozygosity over the time since isolation were large (3,170; range 1,077–7,606) and established an upper limit for N _e if drift were the only evolutionary process responsible for loss of genetic diversity. However, it is likely that a combination of mutation, selection, and gene flow have also contributed to the genetic diversity of above-barrier populations. Contemporary above-barrier N _e estimates were much smaller than long-term N _e estimates, not correlated with within-population genetic diversity, and not consistent with the amount of genetic variation retained, given the approximate 10,000-year period of isolation. The populations isolated by waterfalls in this study that occur in larger stream networks have retained substantial genetic variation, which suggests that the amount of habitat in headwater streams is an important consideration for maintaining the evolutionary potential of isolated populations.     

GENETIC DIVERSITY AND CLONAL STRUCTURE IN A COLUMNAR CACTUS,LOPHOCEREUS SCHOTTII

In southern Arizona the columnar cactus, Lophocereus schottii, inhabits desert riparian environments. Reproduction in this part of its range is predominantly asexual and occurs by either the dispersal of stems in the immediate vicinity of parents or the long-distance transport of detached stem pieces downstream by floodwaters. Genetic diversity and clonal structure of eight populations of L. schottii in Organ Pipe Cactus National Monument, Arizona, were examined. In all populations ramets were mapped and stem tissue from each ramet was examined electrophoretically. At the species level, 44.4% of the loci were polymorphic and the genetic diversity was 0.145. Within populations, the mean proportion of polymorphic loci and genetic diversity were 34.4% and 0.126%, respectively. Although most of the allozyme variation was within populations, appreciable heterogeneity was found among populations (G_ST = 0.130). Genetic and genotypic diversity was greatest in three of the four populations located in the principal area of L. schottii occurrence in the monument. Genotypic diversity was lowest in the smallest population and in the most isolated population. Ramets in all populations were spatially aggregated. Plant pairs with identical multilocus genotypes were usually ≤ 10 m apart, but some widely separated individuals had identical genotypes. Occasional long-distance dispersal of stems and the periodic recruitment of seedlings have caused genets to intermingle, promoting outcrossing and maintaining genetic diversity.     

Genetic variation within the endangered mangrove species Sonneratia paracaseolaris (Sonneratiaceae) in China detected by inter-simple sequence repeats analysis

Sonneratia paracaseolaris, is a critically endangered mangrove species in China. Using inter-simple sequence repeats (ISSR) markers, we compared the genetic variation of introduced populations with that of natural populations to check whether the genetic diversity has been conserved. At the species level, genetic diversity was relatively high (P = 81.37%, He = 0.2241, and SI = 0.3501). Genetic variation in introduced populations (P = 75.78%, He = 0.2291, and SI = 0.3500) was more than that in natural populations (P = 70.81%, He = 0.1903, and SI = 0.2980). Based on Nei's G_ST value, more genetic differentiation among natural populations was detected (G_ST = 0.3591). Our data show that the genetic diversity of S. paracaseolaris was conserved in introduced populations to some extent, however, owing to the small natural populations and the threats they encountered, more plants should be planted to enlarge and restore the populations.     

Inter-populational variation,but no-annual variation within populations,in terms of reproductive size and genetic structure in a monocarpic perennial herb,Cardiocrinum cordatum var. glehnii

Cardiocrinum cordatum var. glehnii (Liliaceae) is a monocarpic perennial herb living in temperate broad-leaved deciduous forests. In the present study we examined the sizes (basal diameter of the stem and flower numbers) of flowering individuals and genetic diversity using microsatellite loci of 23 populations of C. cordatum var. glehnii in Hokkaido, Japan, over 2 years (2009 and 2010). As a result, we found both the basal stem diameter and the number of flowers varied widely among the populations. However, although the sizes of flowering individuals differed among the populations, these were very stable in each population and in each year. In addition, for genetic diversity, the same trends (i.e. wide variation among the populations but non-annual variation) were detected.     

Disentangling the effects of geographic peripherality and habitat suitability on neutral and adaptive genetic variation in Swiss stone pine

It is generally accepted that the spatial distribution of neutral genetic diversity within a species’ native range mostly depends on effective population size, demographic history, and geographic position. However, it is unclear how genetic diversity at adaptive loci correlates with geographic peripherality or with habitat suitability within the ecological niche. Using exome‐wide genomic data and distribution maps of the Alpine range, we first tested whether geographic peripherality correlates with four measures of population genetic diversity at > 17,000 SNP loci in 24 Alpine populations (480 individuals) of Swiss stone pine (Pinus cembra) from Switzerland. To distinguish between neutral and adaptive SNP sets, we used four approaches (two gene diversity estimates, F_ST outlier test, and environmental association analysis) that search for signatures of selection. Second, we established ecological niche models for P. cembra in the study range and investigated how habitat suitability correlates with genetic diversity at neutral and adaptive loci. All estimates of neutral genetic diversity decreased with geographic peripherality, but were uncorrelated with habitat suitability. However, heterozygosity (H_e) at adaptive loci based on Tajima's D declined significantly with increasingly suitable conditions. No other diversity estimates at adaptive loci were correlated with habitat suitability. Our findings suggest that populations at the edge of a species' geographic distribution harbour limited neutral genetic diversity due to demographic properties. Moreover, we argue that populations from suitable habitats went through strong selection processes, are thus well adapted to local conditions, and therefore exhibit reduced genetic diversity at adaptive loci compared to populations at niche margins.     

Quantifying phenotypic gradients in freshwater snails: a case study in Lithasia (Gastropoda: Pleuroceridae)

Many authors have described a pattern of morphological variation in freshwater bivalves where shells taken from lentic and lotic environments, or headwaters and main stem reaches, appear to exhibit phenotypic gradients in size and shape. For example, mussels taken from headwater reaches tend to possess smooth, less inflated shells compared to the more obese, sculptured individuals downstream. Others observed similar relationships in certain freshwater gastropods, but this variation has not been quantified nor its existence explained in an ecological or evolutionary context. Geometric morphometrics indicated freshwater snails shells from the pleurocerid genus Lithasia from the Duck River, Tennessee, USA, show phenotypic gradients similar to those in freshwater mussels. Shells from upstream areas were narrow and less sculptured on the posterior portions of their body whorls, while downstream shells were more inflated and possessed significantly more sculpture. This phenotypic variation may reduce predation or damage due to dislodging. The nature of the observed plasticity suggests an unidirectional environment similar to that proposed by the river continuum concept. Handling editor: K. Martens     

Postglacial climate changes and rise of three ecotypes of harbour porpoises,Phocoena phocoena,in western Palearctic waters

Despite no obvious barriers to gene flow in the marine realm, environmental variation and ecological specializations can lead to genetic differentiation in highly mobile predators. Here, we investigated the genetic structure of the harbour porpoise over the entire species distribution range in western Palearctic waters. Combined analyses of 10 microsatellite loci and a 5085 base‐pair portion of the mitochondrial genome revealed the existence of three ecotypes, equally divergent at the mitochondrial genome, distributed in the Black Sea (BS), the European continental shelf waters, and a previously overlooked ecotype in the upwelling zones of Iberia and Mauritania. Historical demographic inferences using approximate Bayesian computation (ABC) suggest that these ecotypes diverged during the last glacial maximum (c. 23–19 kilo‐years ago, kyrbp ). ABC supports the hypothesis that the BS and upwelling ecotypes share a more recent common ancestor (c. 14 kyrbp ) than either does with the European continental shelf ecotype (c. 28 kyrbp ), suggesting they probably descended from the extinct populations that once inhabited the Mediterranean during the glacial and post‐glacial period. We showed that the two Atlantic ecotypes established a narrow admixture zone in the Bay of Biscay during the last millennium, with highly asymmetric gene flow. This study highlights the impacts that climate change may have on the distribution and speciation process in pelagic predators and shows that allopatric divergence can occur in these highly mobile species and be a source of genetic diversity.     

Crayfish populations genetically fragmented in streams impounded for 36–104 years

1. Dams and their associated impoundments may restrict dispersal and gene flow among populations of numerous freshwater species within stream networks, leading to genetic isolation. This can reduce effective population sizes and genetic diversity, increasing the risk of local extinction.
2. We studied crayfishes from multiple up- and downstream sites in three impounded and two unimpounded streams in the Bear Creek and Cahaba River drainages, Alabama, U.S.A. Using mitochondrial DNA (cytochrome oxidase subunit I gene) sequence data generated from population-level sampling of two abundant native crayfishes, Faxonius validus and Faxonius erichsonianus (Decapoda: Cambaridae), we assessed species’ spatial genetic structure and genetic diversity, estimated the magnitude and directionality of gene flow, and compared results between the species.
3. For both species, levels of genetic diversity (number of haplotypes, and haplotypic and nucleotide diversity) were the same or higher in impounded compared to unimpounded streams. Conversely, crayfish populations in up- and downstream sections of unimpounded streams displayed high genetic similarity and bidirectional gene flow, whereas in impounded streams, crayfish populations typically had greater up- and downstream genetic differentiation and predominantly unidirectional, downstream gene flow.
4. Although impoundments were associated with lower connectivity between up- and downstream sections for F. validus and F. erichsonianus, the magnitude of genetic effects was species-specific, with greater differentiation between F. validus populations up- and downstream of impoundments.
5. In an ecologically short timeframe, impoundments appear to have fragmented stream crayfish populations, and even species with relatively high abundances and large ranges had lower gene flow among populations in impounded streams compared to unimpounded streams. In addition, feedbacks between genetic and demographic effects on fragmented populations may decrease the probability of long-term persistence.

     

Genetic wealth,population health: Major histocompatibility complex variation in captive and wild ring‐tailed lemurs (Lemur catta)

Across species, diversity at the major histocompatibility complex (MHC) is critical to individual disease resistance and, hence, to population health; however, MHC diversity can be reduced in small, fragmented, or isolated populations. Given the need for comparative studies of functional genetic diversity, we investigated whether MHC diversity differs between populations which are open, that is experiencing gene flow, versus populations which are closed, that is isolated from other populations. Using the endangered ring‐tailed lemur (Lemur catta) as a model, we compared two populations under long‐term study: a relatively “open,” wild population (n = 180) derived from Bezà Mahafaly Special Reserve, Madagascar (2003–2013) and a “closed,” captive population (n = 121) derived from the Duke Lemur Center (DLC, 1980–2013) and from the Indianapolis and Cincinnati Zoos (2012). For all animals, we assessed MHC‐DRB diversity and, across populations, we compared the number of unique MHC‐DRB alleles and their distributions. Wild individuals possessed more MHC‐DRB alleles than did captive individuals, and overall, the wild population had more unique MHC‐DRB alleles that were more evenly distributed than did the captive population. Despite management efforts to maintain or increase genetic diversity in the DLC population, MHC diversity remained static from 1980 to 2010. Since 2010, however, captive‐breeding efforts resulted in the MHC diversity of offspring increasing to a level commensurate with that found in wild individuals. Therefore, loss of genetic diversity in lemurs, owing to small founder populations or reduced gene flow, can be mitigated by managed breeding efforts. Quantifying MHC diversity within individuals and between populations is the necessary first step to identifying potential improvements to captive management and conservation plans.     

Mitochondrial DNA variation in natural populations of endangered Indian Feather-Back Fish, Chitala chitala

Genetic variation at mitochondrial cytochrome b (cyt b) and D-loop region reveals the evidence of population sub-structuring in Indian populations of highly endangered primitive feather-back fish Chitala chitala. Samples collected through commercial catches from eight riverine populations from different geographical locations of India were analyzed for cyt b region (307 bp) and D-loop region (636–716 bp). The sequences of the both the mitochondrial regions revealed high haplotype diversity and low nucleotide diversity. The patterns of genetic diversity, haplotypes networks clearly indicated two distinct mitochondrial lineages and mismatch distribution strongly suggest a historical influence on the genetic structure of C. chitala populations. The baseline information on genetic variation and the evidence of population sub-structuring generated from this study would be useful for planning effective strategies for conservation and rehabilitation of this highly endangered species.     

Genetic variation of Carthamus tinctorius L. and related species revealed by SRAP analysis

Genetic diversity of 23 populations of Carthamus tinctorius L. and two populations of Carthamus lanatus L. in China was investigated using Sequence-related Amplified Polymorphism (SRAP). All populations could be uniquely distinguished by 30 primer combinations with 483 bands and 274 polymorphic bands which generated 57% of polymorphic ratio. Unweighed pair-group method of with arithmetical averages (UPGMA) cluster analysis enabled construction of a dendrogram for estimating genetic distances among different populations. The extreme variation was observed when No. 4 cultivated and No. 13 wild population of C. lanatus were grouped at GS = 0.58, and separated from 23 populations of C. tinctorius at GS = 0.10. The result suggested that the cultivated and wild populations of C. lanatus had close relationship with each other and far relationship with C. tinctorius. Dendrogram also revealed a large genetic variation in 23 C. tinctorius populations; different primer combinations allowed them distinctly distinguished one from others with relatively low genetic similarity. Furthermore, five typical representative fragments in C. lanatus were obtained by four most informative primer combinations, which provided a possibility to distinguish C. lanatus from the C. tinctorius evidently.     

Mechanisms that influence sex ratio variation in the invasive hymenopteran Sirex noctilio in South Africa

Sirex noctilio is an economically important invasive pest of commercial pine forestry in the Southern Hemisphere. Newly established invasive populations of this woodwasp are characterized by highly male‐biased sex ratios that subsequently revert to those seen in the native range. This trend was not observed in the population of S. noctilio from the summer rainfall regions in South Africa, which remained highly male‐biased for almost a decade. The aim of this study was to determine the cause of this persistent male bias. As an explanation for this pattern, we test hypotheses related to mating success, female investment in male versus female offspring, and genetic diversity affecting diploid male production due to complementary sex determination. We found that 61% of females in a newly established S. noctilio population were mated. Microsatellite data analysis showed that populations of S. noctilio from the summer rainfall regions in South Africa are far less genetically diverse than those from the winter rainfall region, with mean Nei's unbiased gene diversity indexes of 0.056 and 0.273, respectively. These data also identified diploid males at low frequencies in both the winter (5%) and summer (2%) rainfall regions. The results suggest the presence of a complementary sex determination mechanism in S. noctilio, but imply that reduced genetic diversity is not the main driver of the male bias observed in the summer rainfall region. Among all the factors considered, selective investment in sons appears to have the most significant influence on male bias in S. noctilio populations. Why this investment remains different in frontier or early invasive populations is not clear but could be influenced by females laying unfertilized eggs to avoid diploid male production in populations with a high genetic relatedness.     

Genetic diversity and population structure of a cyprinid fish (Ancherythroculter nigrocauda) in a highly fragmented river

In this study, samples of Ancherythroculter nigrocauda, an endemic fish in the upper Yangtze River, were collected above and below dams in the Longxi River, a tributary of the upper Yangtze River, China, to investigate the genetic impacts of dams. The mitochondrial cytochrome b gene (cyt b) and 13 microsatellite (SSR) loci were used to analyze whether dams have resulted in loss of genetic diversity of the two fragmented populations or caused genetic differentiation between them. The results showed that the haplotype diversity (0.488; 0.486), nucleotide diversity (0.084%; 0.082%) and average expected heterozygosity (0.652; 0.676) of the two populations were all at a low level, and recent bottlenecks were detected. However, there was no genetic differentiation detected by the low genetic differentiation index (F_st, cyt b: ?0.1677, p = 0.99707; SSR: 0.00259, p = 0.81427). Besides, 11 pairs of half‐sibling relationship were found between the two populations indicating that there were individual movements and gene flow between them. This could be the larvae moving from upstream to downstream when water spilled over dams in flooding season. Therefore, our analysis showed that the dams have caused a loss of genetic diversity of the populations of A. nigrocauda in the Longxi River, blocked the active upstream movement but allowing passive downstream drift of larvae.     

Genetic variation and plant performance in fragmented populations of globeflowers (Trolliuseuropaeus) within agricultural landscapes

The management of remnant populations in highly fragmented landscapes requires a thorough understanding of the processes shaping population persistence. We investigated relationships between population characteristics (i.e. size, density and pollinator abundance), offspring performance, genetic diversity and differentiation in Trollius europaeus, a plant with a nursery pollination system. In 19 populations of different sizes and located in north-east Switzerland, an area which has undergone widespread land use changes over the last decades, we assessed neutral genetic diversity (N _total = 383) using AFLPs and plant performance in a greenhouse experiment (N _total = 584) using competition and control treatments. Overall genetic differentiation was low (F _ST = 0.033) with a marginal significant isolation by distance effect (P = 0.06) indicating (historical) genetic connectivity among the populations. Mean expected heterozygosity was H _E of 0.309 (0.0257–0.393) while inbreeding coefficients (F _IS) were significant in only three populations. Genetic diversity was not related to population size, plant density or pollinator abundance. Plant performance was reduced under competition (P < 0.001) but the severity of competition was independent of genetic diversity and population size. In summary, remnant populations of T. europaeus retain genetic diversity and seem capable of persisting under the present conditions within an agricultural matrix. T. europaeus is a perennial herb, thus it may require several generations for the negative effects of fragmentation and isolation to manifest. Our findings indicate that small populations are as important as large populations for the conservation and management of genetic resources.     

Functional variation in a disease resistance gene in populations of Arabidopsis thaliana

Analyses of functional genetic diversity in natural populations may provide important new insights into gene function and are necessary to understand the evolutionary processes maintaining diversity itself. The importance of including diversity within and between local populations in such studies is often ignored although many of the processes affecting genetic diversity act on this scale. Here we examine the molecular diversity in RPW8 (Recognition of Powdery Mildew), a gene conferring broad‐spectrum resistance to powdery mildews in Arabidopsis thaliana stock‐center accessions. Our eight UK study populations of the weedy A. thaliana were from locations judged to be subject to a minimum of anthropogenic disturbance and potentially long established. The majority of populations comprised considerable variation both in disease phenotype and RPW8 genotype. Although resistant individuals shared a major RPW8 genotype, no single allele was uniquely associated with resistance. It is concluded that RPW8 is an essential component of resistance to powdery mildews in A. thaliana, but not the only genetic factor involved in this process. No signature of selection was detected at RPW8 with a microsatellite multilocus test using an empirical null model. Unlike many previous studies of this model plant species, we found high levels of genetic diversity and relatively low differentiation (F_ST = 0.31) between populations at 14 microsatellite markers. This is judged to be due to our sampling being aimed at potentially long established populations and highlights the importance of population choice for studies of genetic diversity within this species.     

Genetic variation in lowland and mountain populations of Tofieldia calyculata and their ability to survive within low levels of genetic diversity

Loss of genetic diversity is expected to be a reason behind the decline of populations of many rare species. To what extent this is true for populations at the range periphery remains to be explored. Alpine species with peripheral lowland populations are an ideal but little-known model system to address this issue. We used 17 microsatellite markers to investigate the genetic diversity and structure of populations of Tofieldia calyculata, a common species in central European mountains, but highly endangered in lowlands. We showed that lowland populations have lower genetic diversity than mountain populations and that the two groups of populations are not clearly differentiated genetically. The species probably survived the last glaciation in refugia in the margins of the Alps and the western Carpathians and some lowland populations likely originated by postglacial colonisation. Some lowland populations may be relictual, but our data did not unequivocally confirm this. Low genetic diversity of lowland populations is likely the result of the reduction of population sizes, limited gene flow, and selfing. Based on data from herbarium specimens from extinct lowland populations, within-population genetic diversity has not changed over the last century suggesting that, under suitable habitat conditions, these populations are able to survive with low levels of genetic diversity. This idea is also supported by the presence of large viable extant populations with very low genetic diversity. Comparisons between modern and historic collection also showed that a large proportion of genetic diversity was lost, due mainly to the extinction of whole populations. Our results provided detailed insight into the recent past of the populations of Tofieldia calyculata, but the genetic diversity of the populations before the twentieth century remains unknown due to the poor quality of old DNA from herbaria samples. Overall, the study indicates that despite reduced genetic diversity, the lowland populations harbour some unique alleles and, with the current levels of genetic diversity, have a chance to survive in the long-term, and thus deserve conservation.