title: Highly polymorphic microsatellite markers in Pulsatilla vulgaris (Ranunculaceae) using next-generation sequencing.
abstract: OBJECTIVE
  We developed novel microsatellite markers for the perennial plant Pulsatilla vulgaris (Ranunculaceae) to investigate the effects of fragmentation on gene flow in this imperiled species.
  
  
  RESULTS
  We identified microsatellites and developed primers based on 454 shotgun sequences. We identified 14 markers that were polymorphic and produced clean bands. Of these, eight could be analyzed as diploids. Genotyping of 97 individuals across two populations revealed these markers to be highly polymorphic with seven to 17 alleles per locus and observed heterozygosity from 0.41 to 0.83.
  
  
  CONCLUSIONS
  The markers are highly informative and will be used to test if the reintroduction of shepherding in southern Germany improves genetic connectivity among fragmented populations of P. vulgaris. The combination of diploid and tetraploid markers presented here will be useful in resolving the polyploidization history of this and related species.
authors.affiliation: Department of Biology, University of Toronto Mississauga, 3359 Mississauga Road, Mississauga, Ontario L5L 1C6, Canada.
authors.forename: Michelle F
authors.initials: MF
authors.lastname: DiLeo
authors.affiliation: WSL Swiss Federal Research Institute, Zürcherstrasse 111, CH-8903 Birmensdorf, Switzerland.
authors.forename: René
authors.initials: R
authors.lastname: Graf
authors.affiliation: WSL Swiss Federal Research Institute, Zürcherstrasse 111, CH-8903 Birmensdorf, Switzerland.
authors.forename: Rolf
authors.initials: R
authors.lastname: Holderegger
authors.affiliation: Department of Natural History, Royal Ontario Museum, 100 Queens Park, Toronto, Ontario M5S 2C6, Canada.
authors.forename: Yessica
authors.initials: Y
authors.lastname: Rico
authors.affiliation: Department of Biology, University of Toronto Mississauga, 3359 Mississauga Road, Mississauga, Ontario L5L 1C6, Canada.
authors.forename: Helene H
authors.initials: HH
authors.lastname: Wagner
keywords: 454 sequencing; Pulsatilla vulgaris; Ranunculaceae; microsatellites; simple sequence repeat (SSR) markers