title: Isolation and characterization of novel microsatellite loci for the endangered orchid Cypripedium japonicum (Orchidaceae).
abstract: OBJECTIVE
  Twenty-six microsatellite markers were developed for the endangered orchid Cypripedium japonicum (Orchidaceae) to estimate the clonal diversity and genetic structure of the remaining populations in Japan.
  
  
  RESULTS
  Microsatellite loci of C. japonicum were isolated using Ion Personal Genome Machine (PGM) sequencing. The primer sets were tested on 55 ramets sampled from two populations in Japan. Sixteen loci showed polymorphism in at least one population, with two to five alleles per locus. Observed and expected heterozygosities for the two populations ranged from 0.00 to 0.92 and 0.00 to 0.71, respectively.
  
  
  CONCLUSIONS
  The microsatellite markers developed here provide a useful tool to analyze clonal structure and sexual regeneration status and will help to manage the remaining genetic variation within C. japonicum.
authors.affiliation: Graduate School of Symbiotic Systems Science and Technology, Fukushima University, Kanayagawa 1, Fukushima, Fukushima 960-1296, Japan.
authors.forename: Yumi
authors.initials: Y
authors.lastname: Yamashita
authors.affiliation: Laboratory of Forest Biology, Division of Forest and Biomaterials Science, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Ayako
authors.initials: A
authors.lastname: Izuno
authors.affiliation: Laboratory of Forest Biology, Division of Forest and Biomaterials Science, Graduate School of Agriculture, Kyoto University, Kitashirakawa, Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan.
authors.forename: Yuji
authors.initials: Y
authors.lastname: Isagi
authors.affiliation: Graduate School of Symbiotic Systems Science and Technology, Fukushima University, Kanayagawa 1, Fukushima, Fukushima 960-1296, Japan.
authors.forename: Takahide
authors.initials: T
authors.lastname: Kurosawa
authors.affiliation: Graduate School of Symbiotic Systems Science and Technology, Fukushima University, Kanayagawa 1, Fukushima, Fukushima 960-1296, Japan.
authors.forename: Shingo
authors.initials: S
authors.lastname: Kaneko
keywords: Cypripedium japonicum; Ion PGM sequencing; Orchidaceae; clonal analysis; conservation genetics; microsatellites