Nine original microsatellite loci in prickly sculpin ( Cottus asper ) and their applicability to other closely related C
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TECHNICAL NOTE
Nine original microsatellite loci in prickly sculpin (Cottus asper) and their applicability to other closely related Cottus species Jason Baumsteiger • Andres Aguilar
Received: 12 September 2012 / Accepted: 28 September 2012 / Published online: 9 October 2012 Ó Springer Science+Business Media Dordrecht 2012
Abstract Prickly sculpin (Cottus asper) are a widespread but largely understudied native freshwater fish in coastal and inland rivers of Western North America. Given the extreme anthropogenic changes in this region, prickly sculpin represent a model organism to study historical and contemporary changes. We present nine novel microsatellites and four additional loci developed on a distantly related Cottus species. Loci range in allelic size from one to eleven and expected heterozygosity from 0.08 to 0.65 within a single inland population. Most loci were genotyped on three different prickly sculpin populations and three closely related sympatric Cottus species allowing for future comparative studies between and within species. Keywords Conservation Population genetics Pyrosequencing
Contemporary rivers along the westernmost edge of North America are under intensive anthropogenic influence, needing improved information to conserve native freshwater species (Frissell 1993). Thanks to a wide distribution, prickly sculpin (Cottus asper) may shed light on past and present connectivity amongst watersheds (Page and Burr 1991). Early studies suggest the species is divided into coastal and inland forms (Krejsa 1965), with a suspected intergrade form (Moyle 2002). We present data from nine novel microsatellite loci and four previously developed loci on another species to sufficiently discern population level
J. Baumsteiger (&) A. Aguilar School of Natural Sciences and Sierra Nevada Research Institute, University of California Merced, 5200 N. Lake Rd., Merced, CA 95344, USA e-mail: [email protected]
differences for this species. Loci are also relatively effective in amplifying three closely related species of Cottus, an important factor given the sympatric distribution and potential hybridization of these species with C. asper (Moyle 2002). Three populations of 25–32 individuals of C. asper were collected from ‘‘inland’’ Kings River (Kings County, CA), ‘‘coastal’’ Redwood Creek (Humboldt Co., CA), and ‘‘intergrade’’ Suisun Bay (Contra Costa Co., CA) populations. Three additional species were cross-amplified: 25 riffle sculpin (C. gulosus) from the Kings River (Kings Co., CA), 30 Pit sculpin (C. pitensis) from the Pit River (Modoc Co., CA), and six coastrange sculpin (C. aleuticus) from the Smith River (Del Norte Co., CA). Tissue samples were taken from lower caudal fin clips and stored in 100 % ethanol. All genomic DNA was extracted using the DNeasy Tissue Kit (Qiagen). Microsatellite markers were developed via two methods: cloning and pyrosequencing. Cloning followed the enrichment protocol of Glenn and Schable (2005). Nuclear DNA from eight prickly sculpin was digested with restriction enzy
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