Development and characterization of eight polymorphic tetra-nucleotide microsatellite markers for the threatened leopard
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TECHNICAL NOTE
Development and characterization of eight polymorphic tetranucleotide microsatellite markers for the threatened leopard darter (Percina pantherina) Michael R. Schwemm • Anthony A. Echelle
Received: 25 July 2012 / Accepted: 6 August 2012 / Published online: 12 August 2012 Ó Springer Science+Business Media B.V. 2012
Abstract We describe eight tetra-nucleotide microsatellite markers for the leopard darter (Percina pantherina), a federally threatened percid fish endemic to Oklahoma and Arkansas. We tested these markers on 42 individuals from two localities and provide summary statistics on population variability. Eight loci yielded 2–12 alleles per locus. These markers contribute to the availability of markers for programs aimed at monitoring and managing the genetic resources of P. pantherina and related taxa. Keywords
Percina Microsatellite markers Darter
Introduction In this paper we describe primers for eight microsatellite DNA loci and provide summary statistics of variability for each locus in two populations of a federally threatened percid fish, the leopard darter, Percina pantherina. The species is endemic to southeastern Oklahoma and southwestern Arkansas where it is restricted to five tributaries of the Little River system (Jones et al. 1984; James and Maughan 1989; Zale et al. 1994). Its restricted geographic range, together with habitat modification by reservoir impoundment and population loss below reservoirs, led to federal listing as threatened in 1978 (USFWS 43 FR 3715, 1978). Subsequent monitoring by the United States Fish and Wildlife Service demonstrates the need for continued conservation concern.
M. R. Schwemm (&) A. A. Echelle Zoology Department, Oklahoma State University, 501 Life Science West, Stillwater, OK 74078, USA e-mail: [email protected]
A previous description of genetic diversity based on allozymes indicated low levels of genetic diversity within and among tributary populations of P. pantherina (Echelle et al. 1999). The loci reported here, together with advances in analytical approaches (Beaumont 1999, 2003; Waples 2008; Waples and Do 2010), should allow improved insight into population structure and demographic history of P. pantherina, including estimates of the timing of population bottlenecks and the potential effect of reservoir construction on effective population size. An initial screening indicated polymorphism at eight of 10 loci from libraries generated by Genetic Identification Services (GIS; www.genetic-id-services.com). We assayed the polymorphic loci for two populations of P. pantherina in isolated tributaries of the Little River, one from the geographic center (site 1: Mountain Fork River, N = 24) and one from the eastern limit of the range (site 2: Cossatot River, N = 18). The following PCR amplification parameters were used for all loci: 95 °C for 12 min, 35 cycles of 94 °C for 40 s, 57 °C for 40 s, 72 °C for 30 s, and 72 °C for 4 min. The reaction mix (15 lL total volume) contained 1–3 ng of template DNA in 1 lL ddH20, 0.5 lL of eac
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