High genetic diversity and differentiation of an extremely narrowly distributed and critically endangered decaploid rose (<Emphasis Type="Italic">Rosa praelucens</Emphasis>): implications for its conservation

Rosa praelucens is a critically endangered decaploid alpine rose with an extremely narrow geographic distribution in Northwestern Yunnan, China. We sampled almost all the extant individuals (527 individuals in 31 natural locations and 56 individuals preserved in three local living collections) to assess the genetic variation and to probe the genetic connectivity among the individuals and populations based on three cpDNA intergenic spacers and six fluorescent amplified fragment length polymorphism (AFLP) markers. The morphological traits from seven populations were also measured. R. praelucens exhibited high levels of morphological variation, genetic diversity, and differentiation. The extant individuals were clustered into eight groups in neighbor-net networks, and subsequent Bayesian analysis assigned them into three larger gene pools, both in accordance with their morphological traits, especially flower color. The living collections embraced two private cpDNA haplotypes and included three out of the species’ total eight AFLP genotypes. Rhizome clonal growth, decaploid, and mixed breeding system may largely contribute to high genetic diversity and differentiation in R. praelucens. We concluded that the endangered status of R. praelucens may mainly be due to habitat fragmentation and loss and inherent reproductive difficulties, rather than low genetic diversity. The populations contributing higher cpDNA genetic diversity, representing more AFLP genotypes, and encompassing private cpDNA haplotypes should be given conservation priority by creating plant-micro reserves. The living collections should also be targeted for further ex situ conservation, population recovery, and reintroduction of R. praelucens plants.     

Admixture of two phylogeographic lineages of the Eurasian beaver in Poland

The Eurasian beaver (Castor fiber) represents an uncommon example of an endangered species in which the restoration programs proved a spectacular success and led to enormous spatial and demographic expansion. Documented reintroduction of beavers in Poland has been conducted using animals of the eastern European origin, most likely derived from the eastern mtDNA lineage. However demographic and spatial expansion of beavers from Germany, which represent the western lineage, may have led to admixture of these two genetically distinct entities in Poland. We detected significant genetic differentiation between the populations from W and NE Poland both in mitochondrial DNA control region and microsatellites, but also substantial admixture including apparent first-generation migrants between regions. Our results indicate that beavers from the western mtDNA lineage have contributed considerably to the Polish population, particularly in W Poland. As there have been no adequately documented translocations of beavers from the western European populations to Poland, the observed situation appears to result from natural migration or range expansion from the west. In contrast to previous findings we detected a substantial diversity in mtDNA control region, which indicates that either the variation in relict populations has been underestimated, or that additional relict beaver populations survived at the end of the 19th century in Poland and Germany as indicated by considerable similarity of ancient and extant mtDNA haplotypes. The implications of our findings for beaver conservation and management are discussed.     

Genetic diversity and biogeography of the traditional Chinese medicine, Gardenia jasminoides, based on AFLP markers

Gardenia jasminoides Ellis is used in traditional Chinese medicine (TCM) in China. Levels of genetic variation and patterns of population structure within and among eight wild or cultivated populations of G. jasminoides Ellis in China were investigated using amplified fragment length polymorphism (AFLP) markers. Of the 11 primers screened, four produced highly reproducible AFLP bands. Using these primers, 244 discernible DNA fragments were generated with 165 bands (67.6%), were polymorphic, indicating considerable genetic variation at the species level. In contrast, there were relatively low levels of polymorphism at the population level with the percentage of polymorphic bands (PPB) ranging from 36.89% to 59.43%. Genetic diversity within populations ranged from 0.2086 to 0.3108, averaging 0.2392 at the species level. A high level of genetic differentiation among populations was detected based on Nei's genetic diversity analysis (76.59%), Shannon's index analysis (64.8%) and AMOVA analysis (72.75%). No significant statistical differences (analysis of molecular variance [AMOVA], p = 0.0639) in AFLP variation were found between regions. However, the variance among populations and within populations differed significantly (p < 0.001). An indirect estimate of historical levels of gene flow (N_m = 1.7448) was consistent with the high mean genetic identity (mean I = 0.9263) found among populations. There is an association between geographic and genetic distances between populations. Presently gene change exists between populations.     

Genetic variation in zinc-tolerant populations of Glyceria fluitans

Amplified fragment length polymorphism (AFLP) was used to conduct a study of the genetic diversity of zinc-tolerant populations of Glyceria fluitans from 10 sites from across Europe. Six different primer combinations were used on five to nine plants from each of the 10 sites to generate a total of 796 bands, of which 670 were polymorphic. These data were then used to calculate a dendrogram by agglomerative clustering using the unweighed pair group method with average linkage (UPGMA). The dendrogram contained two distinct clusters, with little overlap between populations. Genetic diversity between populations of G. fluitans did not always correlate with geographical distances, for example, plants from the Navan population from Ireland were more genetically similar to populations from Poland than other populations from Ireland. In other instances, geographical origin was significant, for example, all Polish populations were genetically similar to each other. Populations from two English sites only 160 km apart, showed such a high degree of genetic diversity that they were placed in different clusters in the dendrogram. They were more closely related to Irish and Polish populations than to each other. Plants from different zinc-contaminated sites were found not to cluster together. The conclusion was that the cluster groupings were not related to exposure to zinc at the sites of origin, and that the drive to generate distinct metal-tolerant populations may not occur in this species due to the existence of a constitutive tolerance to metals.     

Genetic assessment of population restorations of the critically endangered Silene hifacensis in the Iberian Peninsula

In order to preserve endangered plant populations and recover their evolutionary potential and ecological behavior, some restoration measures generally involve the reinforcement of the population size in existing natural populations or the reintroduction of new populations. 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. The highly threatened Spanish species Silene hifacensis (Caryophyllaceae) has only three natural reduced mainland populations in the Iberian Peninsula, following decline and extinction that occurred during the late 20th century. Preterit restoration strategies were essentially based on the implantation of new populations and reinforcement of certain existing populations using transplants mostly cultivated in greenhouses. In the present contribution, levels and patterns of genetic variability within natural and restored populations of Silene hifacensis were assessed using the molecular technique AFLP. Our results pointed out significant genetic diversity differences across the three existing natural populations though their population fragmentation and progressive loss of individuals have not had an impact on the global genetic diversity of this species. For restored populations, their levels of genetic diversity were similar and even higher than in natural populations. As a result, the past restoration protocols were successful in capturing similar and even higher levels of genetic diversity than those observed within natural pools. However, inbreeding processes have been detected for two restored populations. Finally, the main source of plant material for the long-time restored transplants appears to be the natural population of Cova de les Cendres. This study demonstrates, once again, how genetic markers are useful tools to be taken in consideration for endangered plant species conservation plans.     

Increased inbreeding and strong kinship structure in Taxus baccata estimated from both AFLP and SSR data

Habitat fragmentation can have severe genetic consequences for trees, such as increased inbreeding and decreased effective population size. In effect, local populations suffer from reduction of genetic variation, and thus loss of adaptive capacity, which consequently increases their risk of extinction. In Europe, Taxus baccata is among a number of tree species experiencing strong habitat fragmentation. However, there is little empirical data on the population genetic consequences of fragmentation for this species. This study aimed to characterize local genetic structure in two natural remnants of English yew in Poland based on both amplified fragment length polymorphism (AFLP) and microsatellite (SSR) markers. We introduced a Bayesian approach that estimates the average inbreeding coefficient using AFLP (dominant) markers. Results showed that, in spite of high dispersal potential (bird-mediated seed dispersal and wind-mediated pollen dispersal), English yew populations show strong kinship structure, with a spatial extent of 50–100 m, depending on the population. The estimated inbreeding levels ranged from 0.016 to 0.063, depending on the population and marker used. Several patterns were evident: (1) AFLP markers showed stronger kinship structure than SSRs; (2) AFLP markers provided higher inbreeding estimates than SSRs; and (3) kinship structure and inbreeding were more pronounced in denser populations regardless of the marker used. Our results suggest that, because both kinship structure and (bi-parental) inbreeding exist in populations of English yew, gene dispersal can be fairly limited in this species. Furthermore, at a local scale, gene dispersal intensity can be more limited in a dense population.     

Conservation genetics of endangered leaf-beetle Cheilotoma musciformis populations in Poland

Steppe-like habitats in Europe are seriously threatened as a result of fragmentation and anthropogenic degradation, at least in western and central parts. Considering the dramatic loss of steppe-like habitats, the evaluation of genetic variation in populations of steppe species is of immediate importance if appropriate conservation measures are to be undertaken. In this paper, we examine the genetic diversity of the highly endangered populations of the leaf-beetle Cheilotoma musciformis, which inhabits only a limited area in south-central Poland, which is geographically isolated from the continuous range of this species. Both mitochondrial and nuclear markers show that the Polish populations are distinct from Slovakian and Ukrainian ones. These regional populations should be considered independent conservation units. On the other hand, very little (mtDNA) or no (nuclear DNA) diversity has been found among the Polish subpopulations. This leads to the conclusion that this species has gone through a strong bottleneck leading to a drastic reduction in its genetic diversity prior to the establishment of present-day populations. Host plants have been identified for this species using barcodes, and the only hosts for the Polish and Ukrainian samples are sainfoins Onobrychis spp. while for the Slovakian sample it is either Dorycnium pentaphyllum or Lotus spp. (all Fabaceae). All of these data can be very valuable for the conservation of C. musciformis populations (e.g. for reintroductions).     

Epigenetic variation associated with responses to different habitats in the context of genetic divergence in Phragmites australis

The mechanisms underlying heritable phenotypic divergence associated with adaptation in response to environmental stresses may involve both genetic and epigenetic variations. Several prior studies have revealed even higher levels of epigenetic variation than genetic variation. However, few population‐level studies have explored the effects of epigenetic variation on species with high levels of genetic diversity distributed across different habitats. Using AFLP and methylation‐sensitive AFLP markers, we tested the hypothesis that epigenetic variation may contribute to differences in plants occupying different habitats when genetic variation alone cannot fully explain adaptation. As a cosmopolitan invasive species, Phragmites australis (common reed) together with high genetic diversity and remarkable adaptability has been suggested as a model for responses to global change and indicators of environmental fluctuations. We found high levels of genetic and epigenetic diversity and significant genetic/epigenetic structure within each of 12 studied populations sampled from four natural habitats of P. australis. Possible adaptive epigenetic variation was suggested by significant correlations between DNA methylation‐based epigenetic differentiation and adaptive genetic divergence in populations across the habitats. Meanwhile, various AMOVAs indicated that some epigenetic differences may respond to various local habitats. A partial Mantel test was used to tease out the correlations between genetic/epigenetic variation and habitat after controlling for the correlation between genetic and epigenetic variations. We found that epigenetic diversity was affected mostly by soil nutrient availability, suggesting that at least some epigenetic differentiation occurred independently of genetic variation. We also found stronger correlations between epigenetic variation and phenotypic traits than between genetic variation and such traits. Overall, our findings indicate that genetically based differentiation correlates with heterogeneous habitats, while epigenetic variation plays an important role in ecological differentiation in natural populations of P. australis. In addition, our results suggest that when assessing global change responses of plant species, intraspecific variation needs to be considered.     

Assessment of the domestication state of ackee (Blighia sapida K.D. Koenig) in Benin based on AFLP and microsatellite markers

Ackee (Blighia sapida) is a native multipurpose species important for the livelihoods of the rural populations in Benin. Trees are found in natural forests or are managed by farmers in different traditional agroforestry systems. Genetic variation at amplified fragment length polymorphism (AFLP) markers, four nuclear microsatellites (nSSRs) and one chloroplast microsatellite (cpSSR) were investigated in 279 individuals from six wild and eight cultivated populations from Benin. The AFLP data revealed moderate levels of diversity of ackee in Benin (mean diversity values are proportion of polymorphic loci = 52.8% and Nei??s gene diversity = 0.157, for 375 AFLP fragments). The mean diversity values based on nSSR-markers are expected heterozygosity = 0.286, allelic richness = 2.77. Genetic variation of wild and cultivated populations did not differ markedly. AMOVA revealed that only 7.3 and 5.2% of the variation was partitioned among populations for nSSR- and AFLP-markers, respectively. A Mantel test based on these both marker-types revealed significant correlations between population pairwise geographic distance and genetic differentiation. Differentiation among cultivated populations was higher than among wild populations. The only polymorphic chloroplast microsatellite marker (ccmp7) showed three haplotypes. Cultivated populations from northeastern Benin were fixed on one haplotype which was not observed elsewhere indicating a different origin of these populations possibly from neighboring Nigeria. Farmer-led domestication had an impact on the spatial distribution of genetic variation but did not result in significant losses of diversity within populations. Measures to conserve genetic resources of ackee in each of the three main bioclimatic zones in Benin are proposed.     

Temporal changes in population genetic diversity and structure in red and white clover grown in three contrasting environments in northern Europe

Backgound and Aims

Extending the cultivation of forage legume species into regions where they are close to the margin of their natural distribution requires knowledge of population responses to environmental stresses. This study was conducted at three north European sites (Iceland, Sweden and the UK) using AFLP markers to analyse changes in genetic structure over time in two population types of red and white clover (Trifolium pratense and T. repens, respectively): (1) standard commercial varieties; (2) wide genetic base (WGB) composite populations constructed from many commercial varieties plus unselected material obtained from germplasm collections.

Methods

At each site populations were grown in field plots, then randomly sampled after 3–5 years to obtain survivor populations. AFLP markers were used to calculate genetic differentiation within and between original and survivor populations.

Key Results

No consistent changes in average genetic diversity were observed between original and survivor populations. In both species the original varieties were always genetically distinct from each other. Significant genetic shift was observed in the white clover ‘Ramona’ grown in Sweden. The WGB original populations were more genetically similar. However, genetic differentiation occurred between original and survivor WGB germplasm in both species, particularly in Sweden. Regression of climatic data with genetic differentiation showed that low autumn temperature was the best predictor. Within the set of cold sites the highest level of genetic shift in populations was observed in Sweden.

Conclusions

The results suggest that changes in population structure can occur within a short time span in forage legumes, resulting in the rapid formation of distinct survivor populations in environmentally challenging sites.     

Population structure and genetic diversity of the threatened quillwort Isoëtes malinverniana and implication for conservation

The goal of this research was to investigate genetic variation in Isoëtes malinverniana (Isoëtaceae) to select candidate populations for future conservation efforts. To this aim, ISSR and AFLP analyses, carried out using six and four primer combinations, respectively, produced a total of 425 bands, 97.18% of which were polymorphic.Our results suggest that I. malinverniana shows medium to high genetic diversity (mean Nei's genetic diversity: H = 0.1491 for ISSR data; H = 0.2289 for AFLP data) and a substantial amount of gene flow between the analysed populations (N_m = 1.768, with combined ISSR and AFLP data). The moderate levels of population differentiation support the hypothesis that the fragmentation and isolation of I. malinverniana occurred only recently, probably due to the intensive agriculture practice and water pollution.These results will be used to focus further studies aimed at supporting reintroduction programs within suitable sites of the distribution area.     

Conservation genetics of Heritiera littoralis (Sterculiaceae), a threatened mangrove in China,based on AFLP and ISSR markers

AFLP and ISSR markers were used to determine the genetic variations in eight mangrove and non-mangrove populations of Heritiera littoralis (Sterculiaceae), a threatened species in China. Our results showed a moderate to high level of genetic variation in this species (P = 63.69%, H_T = 0.20 for AFLP; P = 76.07%, H_T = 0.22 for ISSR), and a relatively high level of genetic differentiation among populations (G_ST = 0.24 for AFLP and 0.27 for ISSR). Life history traits, long-distance dispersal of floating seeds, and local environments may play important roles in shaping the genetic diversity and genetic structure of this species. Investigation of the plant’s reproductive capacity showed that the natural seed production of H. littoralis was low, as was the germination rate and the transformation rate from juvenile to adult. H. littoralis is seriously threatened and is in urgent need of conservation in China.     

AFLP and RAPD based genetic diversity assessment of industrially important reed bamboo (Ochlandra travancorica Benth)

A high level of genetic diversity was recorded in molecular analysis of random collections of industrially important reed bamboos (O. travancorica) from Kerala region of India. Fifty primers (8 AFLP & 42 RAPD) detected 914 polymorphic loci. Cluster and Principal Coordinate Analysis (PCA) based on combined AFLP & RAPD data grouped all the random accessions into three different populations. Further, AMOVA revealed a moderate to high level of genetic variation. Most of the variation occurred within population (54 %) than among population (46 %). Highly significant and high PhiPT estimate (Fst value; 0.456) indicated that these populations are not panmictic and are significantly isolated. Relatively large number of polymorphism obtained could be useful in finding the elite germplasm resources for industrial use.     

Population genetics and conservation of critically small cycad populations: a case study of the Albany Cycad,Encephalartos latifrons (Lehmann)

Declining populations of less than 250 mature individuals are symptomatic of many Critically Endangered cycads, which, globally, comprise the most threatened group of organisms as a result of collecting and habitat loss. Survival plans focus on law enforcement, reintroduction, and augmentation programmes using plants from the wild and botanical gardens. Augmentation is one of the few remaining options for cycad populations, although the assumed benefits remain untested and there is a possibility that augmentation from different sources could compromise the genetic integrity of existing populations, especially when garden plants have no provenance data. We studied Encephalartos latifrons, a South African endemic, which is a typical Critically Endangered cycad. We studied the extent and structure of genetic diversity in wild and ex situ populations to assess the potential benefits and risks associated with augmentation programmes. We examined 86 plants using amplified fragment length polymorphisms (AFLPs). The 417 AFLP markers thus generated yielded a unique DNA ‘fingerprint’ for each plant. Wild populations retain high levels of genetic diversity and this is reflected among the ex situ holdings at the Kirstenbosch Botanical Garden. No population differentiation is evident, indicating a single panmictic population, consistent with moderately high levels of gene flow between subpopulations and a sexual mode of reproduction. Bayesian clustering identified four genotype groups in the wild, as well as a genotype group only found in ex situ collections. Our results indicate that E. latifrons would benefit from augmentation programmes, including the use of undocumented collections, and careful management of breeding plants would increase the heterogeneity of propagules. © 2011 The Linnean Society of London, Biological Journal of the Linnean Society, 2012, 105 , 293–308.     

Comparative molecular studies on the genetic diversity of an ex situ garden collection and its source population of the critically endangered polish endemic plant <Emphasis Type="Italic">Cochlearia polonica</Emphasis> E. Fröhlich

Cochlearia polonica (Brassicaceae) is a narrow endemic plant extinct in the wild, known only from one transplanted population in southern Poland. The suitable habitats for this species are confined to shallow water courses and springs on sandy ground. During the second half of twentieth century the natural populations of C. polonica declined due to anthropogenic effects (primarily due to a major change of ground water levels). Consequently, 14 individuals were transplanted in the 1970 s into a secondary locality of similar habitat conditions of Centuria river. In the late 1980s five individuals from the transplanted population were used to establish an artificial ex situ site in the Botanical Garden of the Polish Academy of Sciences in Warsaw. In order to investigate the effect of 18 years of ex situ conservation, the level of genetic diversity and genetic structure of the ex situ conserved botanical garden population and its source population were sampled and analyzed using inter simple sequence repeat markers. The percentage values of polymorphic bands and Nei’s gene diversity indicated average genetic variation at species level, whereas at the population level it was relatively low, especially for the garden population. The results suggested differences in genetic composition of the analyzed populations and implied homogenization of the genetic structure of the ex situ conserved population. Ex situ conservation resulted in a decrease of the species’ genetic diversity, implying that the artificial population only partly represents the primary genetic variability found in the source population.     

Genetic diversity from pre-bottleneck to recovery in two sympatric pinniped species in the Northwest Atlantic

Conservation successes of the past several decades provide natural settings to study post-bottleneck evolutionary processes in species undergoing recovery. Here, we study the impact of demographic change on genetic diversity in parallel natural experiments of historical decline and subsequent recovery in two sympatric pinniped species in the Northwest Atlantic, the gray seal (Halichoerus grypus atlantica) and harbor seal (Phoca vitulina concolor). We compare genetic diversity at the mitochondrial control region today to diversity in archaeological specimens, which represent the populations prior to the regional bounties of the late 1800s to mid-1900s that drastically reduced population sizes and led to local extirpations. We further assess genetic diversity throughout recovery, using biological collections from ongoing long-term studies of both species. Overall, the genetic data are consistent with the historical presence of large, genetically diverse populations of pinnipeds prior to human exploitation, and suggest that gray seals were more dramatically impacted by historical bottlenecks than harbor seals in the Northwest Atlantic. Current mitochondrial diversity in both species is relatively high, and we observe little change over the past several decades during a period of roughly parallel rapid population increases. However, there remain large differences in haplotype composition between pinniped populations of pre-exploitation and today, a lasting genetic signature of historical exploitation that is likely to persist into the future.     

Microsatellite diversity and mating system of Sinojackia xylocarpa (Styracaceae), a species extinct in the wild

Sinojackia xylocarpa is a Chinese endemic species that is extinct in the wild but extant in botanical gardens. Microsatellites were used to investigate the genetic diversity and mating system of this species for future use in a reintroduction program. Ex situ conserved populations of S. xylocarpa maintain intermediate levels of genetic diversity (H_E = 0.570–0.640). However, a general and significant heterozygote excess was found, with a mean F_IS of −0.103. S. xylocarpa was determined to be predominantly outcrossing (t_m = 0.992; t_s = 1.092). Population size and genetic diversity were found to be positively correlated (r = 0.991; P = 0.084). Principal coordinate analysis (PCA) suggests that all extant individuals are derived from two source populations. Reintroduction strategies of S. xylocarpa were proposed on the basis of these results.     

DNA fingerprinting of Leonurus cardiaca L. germplasm in Iran using amplified fragment length polymorphism and inter-retrotransposon amplified polymorphism

In the present study, the extent of inter and intra-population genetic variation was evaluated in Leonurus cardiaca accessions naturally growing in Iran by AFLP and IRAP markers. The fingerprints corresponding to AFLP and IRAP markers revealed high levels of heterozygosity, indicating that L. cardiaca is predominantly an out-crossing species. The average percentage polymorphism was detected as 58% and 90.8% on utilizing AFLP and IRAP data, respectively. Gene diversity values within populations varied 0.14 to 0.20 for AFLP and 0.12 to 0.21 for IRAP. The overall levels of genetic variation present in the L. cardiaca germplasm in Iran were finally determined by combining the AFLP and IRAP datasets to ensure wide genome coverage. The phenogram depicted that the accessions of Dargaz population were genetically distinct from other populations. Based on AFLP and IRAP analysis, it is concluded that L. cardiaca maintains high levels of genetic variation at inter and intra-population level.     

Conservation genetics of highly isolated populations of the xerothermic beetle Crioceris quatuordecimpunctata (Chrysomelidae)

Xerothermic species are rare and threatened in central and eastern Europe. In light of the continuing loss of steppe‐like habitats due to anthropogenic fragmentation and degradation, the evaluation of genetic variation in populations inhabiting them is of immediate importance if appropriate conservation measures are to be undertaken. Here we report on the genetic diversity of the rare leaf beetle Crioceris quatuordecimpunctata, whose populations in central and eastern Europe inhabit highly geographically isolated areas. All of the studied populations (in Poland, Ukraine, and Slovakia) were differentiated at the mitochondrial marker COI. However, with respect to the nuclear marker ITS1, Polish populations were monomorphic, but distinct from all other populations. The distinctiveness of the studied populations was confirmed by Wolbachia screening, which showed that all populations carried different strains (one or two), which were probably transferred independently from other insects. On the other hand, no diversity was found in any marker within particular populations, which could be caused (at least for mtDNA) by a Wolbachia selective sweep. Crioceris quatuordecimpunctata probably consists of isolated populations, which went through narrow bottlenecks leading to a drastic reduction in their genetic diversity. As these populations are reciprocally monophyletic for mtDNA haplotypes and show a significant divergence of allele frequencies at nuclear loci, they could be classified as evolutionarily significant units (ESUs). In addition, DNA barcodes were used to identify Asparagus officinalis as the host plant for members of all studied populations. These data should be valuable in efforts to conserve populations of C. quatuordecimpunctata (e.g., for guiding reintroductions).     

Homogeneity and distinctiveness of Polish paternal lineages revealed by Y chromosome microsatellite haplotype analysis

Different regional populations from Poland were studied in order to assess the genetic heterogeneity within Poland, investigate the genetic relationships with other European populations and provide a population-specific reference database for anthropological and forensic studies. Nine Y-chromosomal microsatellites were analysed in a total of 919 unrelated males from six regions of Poland and in 1,273 male individuals from nine other European populations. AMOVA revealed that all of the molecular variation in the Polish dataset is due to variation within populations, and no variation was detected among populations of different regions of Poland. However, in the non-Polish European dataset 9.3% ( P<0.0001) of the total variation was due to differences among populations. Consequently, differences in R(ST)-values between all possible pairs of Polish populations were not statistically significant, whereas significant differences were observed in nearly all comparisons of Polish and non-Polish European populations. Phylogenetic analyses demonstrated tight clustering of Polish populations separated from non-Polish groups. Population clustering based on Y-STR haplotypes generally correlates well with the geography and history of the region. Thus, our data are consistent with the assumption of homogeneity of present-day paternal lineages within Poland and their distinctiveness from other parts of Europe, at least in respect to their Y-STR haplotypes. Electronic supplementary material to this paper can be obtained by using the Springer LINK server located at http://dx.doi.org/10.1007/s00439-002-0728-0.