Isolation and characterization of microsatellite loci in Pulsatilla patens (L.) Mill. (Ranunculaceae) a rare and endangered plant species in Europe

Pulsatilla patens is a rare and endangered plant species in many areas of Europe and protected under the Bern Convention and it is listed in Annex II and Annex IV to the Habitats Directive. In this study we developed 12 novel microsatellite loci using via 454 sequencing. We determined 11,220 contigs with a length of 156–11,384 bp. Within this dataset, we identified 319 SSR motifs in 301 contigs. All markers were genotyped on 56 individuals from three populations located in Poland. The number of alleles and expected heterozygosity were 2–12 (mean 3.7) and 0.142–0.820 (0.541 on average) respectively. The markers described in this study will be useful for evaluating genetic diversity of P. patens populations, could be applied to investigate the biological aspects and to develop effective conservation programs for the European populations of this species.