Mean genetic diversity parameters for three clusters identified by the STRUCTURE analysis.

<p>Mean genetic diversity parameters for three clusters identified by the STRUCTURE analysis.</p

Population structure of 598 samples of <i>Pulsatilla patens</i> s.s. uncovered by STRUCTURE (K = 3); colors represent the groups identified; each individuals is represented by vertical line, each population is represented by one chart, showing the proportion of membership to the different clusters.

<p>a) admixture analysis; b) non admixture analysis.</p

Genetic Diversity and Population Structure of the Rare and Endangered Plant Species <i>Pulsatilla patens (L</i>.<i>) Mill</i> in East Central Europe - Fig 1

<p>a) General distribution of <i>P</i>. <i>patens</i> in Europe (redrawn and slightly modified after Meusel et. al 1965). b) The location of 29 populations analyzed in this study. Each locations are represented by circles, and colors represent the genetic groups to which individuals within each site were assigned based on STRUCTURE results using K = 3 and a prior that takes sampling location into account–admixture analysis c) non admixture analysis.</p

Geographical location of analyzed populations of <i>Pulsatilla patens</i> s.s.

<p>Geographical location of analyzed populations of <i>Pulsatilla patens</i> s.s.</p

The relationschip between genetic distances (pairwise <i>F</i>_st) and geographic distances for a 29 sampled populations of <i>Pulsatilla patens</i>.

<p>P. values were derived from Mantel tests.Note log scales for geographic distances.</p

Genetic Diversity and Population Structure of the Rare and Endangered Plant Species <i>Pulsatilla patens (L</i>.<i>) Mill</i> in East Central Europe

<div><p><i>Pulsatilla patens</i> s.s. is a one of the most endangered plant species in Europe. The present range of this species in Europe is highly fragmented and the size of the populations has been dramatically reduced in the past 50 years. The rapid disappearance of <i>P</i>. <i>patens</i> localities in Europe has prompted the European Commission to initiate active protection of this critically endangered species. The aim of this study was to estimate the degree and distribution of genetic diversity within European populations of this endangered species. We screened 29 populations of <i>P</i>. <i>patens</i> using a set of six microsatellite primers. The results of our study indicate that the analyzed populations are characterized by low levels of genetic diversity (<i>H</i>_<i>o</i> = 0.005) and very high levels of inbreeding (<i>F</i>_<i>IS</i> = 0.90). These results suggest that genetic erosion could be partially responsible for the lower fitness in smaller populations of this species. Private allelic richness was very low, being as low as 0.00 for most populations. Average genetic diversity over loci and mean number of alleles in <i>P</i>. <i>patens</i> populations were significantly correlated with population size, suggesting severe genetic drift. The results of AMOVA point to higher levels of variation within populations than between populations.The results of Structure and PCoA analyses suggest that the genetic structure of the studied <i>P</i>. <i>patens</i> populations fall into three clusters corresponding to geographical regions. The most isolated populations (mostly from Romania) formed a separate group with a homogeneous gene pool located at the southern, steppic part of the distribution range. Baltic, mostly Polish, populations fall into two genetic groups which were not fully compatible with their geographic distribution.Our results indicate the serious genetic depauperation of <i>P</i>. <i>patens</i> in the western part of its range, even hinting at an ongoing extinction vortex. Therefore, special conservation attention is required to maintain the populations of this highly endangered species of European Community interest.</p></div

DNA barcoding, ecology and geography of the cryptic species of <i>Aneura pinguis</i> and their relationships with <i>Aneura maxima</i> and <i>Aneura mirabilis</i> (Metzgeriales, Marchantiophyta)

<div><p><i>Aneura pinguis</i> is a thalloid liverwort species with broad geographical distribution. It is composed of cryptic species, however, the number of cryptic species within <i>A</i>. <i>pinguis</i> is not known. Five cpDNA regions (<i>matK</i>, <i>rbcL</i>, <i>rpoC1</i>, <i>trnH-psbA</i> and <i>trnL-trnF</i>) and the entire nuclear ITS region were studied in 130 samples of <i>A</i>. <i>pinguis</i> from different geographical regions. The relationships between the cryptic species of <i>A</i>. <i>pinguis</i>, <i>A</i>. <i>maxima</i> and <i>A</i>. <i>mirabilis</i> were analyzed. All of the examined samples were clustered into 10 clades corresponding to 10 cryptic species of <i>A</i>. <i>pinguis</i> (marked A to J). <i>Aneura mirabilis</i> and <i>A</i>. <i>maxima</i> were nested among different cryptic species of <i>A</i>. <i>pinguis</i>, which indicates that <i>A</i>. <i>pinguis</i> is a paraphyletic taxon. Subgroups were found in cryptic species A, B, C and E. As single barcodes, all tested DNA regions had 100% discriminant power and fulfilled DNA barcode criteria for species identification; however, the only combination detected in all subgroups was <i>trnL</i>-<i>trnF</i> with <i>trnH</i>-<i>psbA</i> or ITS2. The distances between cryptic species were 11- to 35-fold higher than intraspecific distances. In all analyzed DNA regions, the distances between most pairs of cryptic <i>A</i>. <i>pinguis</i> species were higher than between <i>A</i>. <i>maxima</i> and <i>A</i>. <i>mirabilis</i>. All cryptic species of <i>A</i>. <i>pinguis</i> clearly differed in their habitat preferences, which suggests that habitat adaptation could be the main driving force behind cryptic speciation within this taxon.</p></div

Average genetic divergences (%) for <i>Aneura pinguis</i> (A-J) cryptic species, <i>A</i>. <i>maxima</i> and <i>A</i>. <i>mirabilis</i>, based on the combined data set K2P (below diagonal) and uncorrected p-distance (above diagonal).

<p>Average genetic divergences (%) for <i>Aneura pinguis</i> (A-J) cryptic species, <i>A</i>. <i>maxima</i> and <i>A</i>. <i>mirabilis</i>, based on the combined data set K2P (below diagonal) and uncorrected p-distance (above diagonal).</p

Mean intraspecific and interspecific K2P distances of individual loci and their combinations in <i>Aneura pinguis</i>.

<p>Mean intraspecific and interspecific K2P distances of individual loci and their combinations in <i>Aneura pinguis</i>.</p

Ultrametric tree obtained by UPGMA analysis of the studied <i>Aneura</i> species generated from the combined dataset.

<p>The cryptic species of <i>A</i>. <i>pinguis</i> and the results of the ABGD analysis for the examined individual loci and their combinations were marked in different colors. The numbers below the diagram represent the number of groups detected as recursive partitions in ABGD.</p