title: Development of microsatellite markers for the clonal shrubOrixa japonica(Rutaceae) using 454 sequencing.
abstract: OBJECTIVE
  Microsatellite markers were developed for a dioecious shrub,Orixa japonica(Rutaceae). BecauseO. japonicavigorously propagates by vegetative growth, microsatellite markers can be used to identify clonal relationships among its ramets.
  
  
  RESULTS
  Sixteen polymorphic microsatellite markers were identified by 454 next-generation sequencing. The number of alleles and expected heterozygosity for each locus among four populations ranged from two to 10 and from 0.140 to 0.875, respectively. Five of the 16 loci showed a low null allele frequency. BecauseOrixais a monotypic genus, cross-amplification in a consubfamilial species,Skimmia japonica, was tested, and only one locus showed polymorphism.
  
  
  CONCLUSIONS
  These microsatellite markers developed forO. japonicacontribute to clone identification for studies examining the clonal structure and true sex ratio in the wild. Moreover, five markers that have a low null allele frequency can also be used for estimating mating systems or performing parentage analysis.
authors: Gifu Academy of Forest Science and Culture, 88 Sodai, Mino, Gifu 501-3714, Japan.
authors: Ichiro
authors: I
authors: Tamaki
authors: Department of Tree Molecular Genetics, Forestry and Forest Products Research Institute, 1 Matsunosato, Tsukuba, Ibaraki 305-8687, Japan.
authors: Suzuki
authors: S
authors: Setsuko
authors: Department of Wildlife Biology, Forestry and Forest Products Research Institute, 1 Matsunosato, Tsukuba, Ibaraki 305-8687, Japan.
authors: Kyoko
authors: K
authors: Sugai
authors: Gifu Academy of Forest Science and Culture, 88 Sodai, Mino, Gifu 501-3714, Japan.
authors: Nao
authors: N
authors: Yanagisawa
keywords: Orixa; Rutaceae; clonal structure; dioecious plant; next-generation sequencing; sex ratio