Development and characterization of polymorphic microsatellite marker for East Asian species of the genus Plestiodon
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TECHNICAL NOTE
Development and characterization of polymorphic microsatellite marker for East Asian species of the genus Plestiodon Kazuki Kurita • Tsutomu Hikida • Mamoru Toda
Received: 21 October 2012 / Accepted: 29 October 2012 / Published online: 10 November 2012 Ó Springer Science+Business Media Dordrecht 2012
Abstract We isolated and characterized nine microsatellite markers for Plestiodon kishinouyei and Plestiodon marginatus, which are of conservation concern, with sever population decline. The number of alleles and expected heterozygosity at each locus in P. kishinouyei were 1–15 and 0.000–0.910, respectively, and those in P. marginatus were 1–9 and 0.000–0.842, respectively. Cross-amplification in five other congeners in East Asia confirmed that these markers are applicable to almost all of those species and widley useful for genetic analyses of the East Asian Plestiodon. Keywords Plestiodon East Asia Microsatellite Multiplex PCR Fluorescently labelled universal primer
The scincid lizard genus Plestiodon comprises 45 species belonging to 11 species groups in East Asia, and North and Middle America (Brandley et al. 2012; Feria-Ortiz and Garcı´a-Va´zquez 2012; Okamoto and Hikida 2012). Of these, six species belonging to two species groups are distributed in the Ryukyu Archipelago, Japan and its surronding regions, and almost all of them are listed in Red List of Japan [P. elegans as endangered; P. barbouri as vulnerable; P. kishinouyei, P. marginatus, and P. stimpsonii as near threatened (Ota 2000; WDNCBJ 2010)]. P. kishinouyei is also on the IUCN Red List as lower risk/near K. Kurita (&) T. Hikida Department of Zoology, Graduate School of Science, Kyoto University, Sakyo, Kyoto 606-8502, Japan e-mail: [email protected] M. Toda Tropical Biosphere Research Center, University of the Ryukyus, Nishihara, Okinawa 903-0213, Japan
threatened (IUCN 2012). Sever population declines were reported or suggested for P. marginatus and P. kishinouyei most probably due to impacts by itroduced carnivorous mammals and/or human mediated habitat destruction (Ota 2000; Toyama 2005; Kurita and Kadota 2010). On the other hand, they are sometimes found in even very small islets where almost no other reptiles occur. Because a small population is prone to extinction due to loss of the genetic variation to adapt to environmental changes (Frankham et al. 2002), population genetic research toward understanding how these populations maintain themselves in the small islets would serve some insights into the conservation management of these species. Microsatellite loci in Plestiodon have ever been reported for a few species in Nearctic region (P. fasciatus: Howes et al. 2004, 2006; P. longirostris: Coughlan et al. 2004; P. reynoldsi: Reid et al. 2004). We tested several of these primers on Asian Plestiodon, but could not obtain good amplification in most of the loci (unpublished data). Therefore, we newly developed nine microsatellite markers for two East Asian species of Plestiodon belonging to different species
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