Development of eighteen microsatellite loci in walleye ( Sander vitreus )
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MICROSATELLITE LETTERS
Development of eighteen microsatellite loci in walleye (Sander vitreus) D. K. Coykendall • C. L. Morrison W. Stott • M. J. Springmann
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Received: 25 June 2014 / Accepted: 7 July 2014 / Published online: 20 July 2014 Ó Springer Science+Business Media Dordrecht (outside the USA) 2014
Abstract A suite of tri- and tetra-nucleotide microsatellite loci were developed for walleye (Sander vitreus) from 454 pyrosequencing data. Eighteen of the 50 primer sets tested amplified consistently in 35 walleye from two lakes on Isle Royale, Lake Superior: Chickenbone Lake and Whittlesey Lake. The loci displayed moderate levels of allelic diversity (average 5.5 alleles/locus) and heterozygosity (average 35.8 %). Levels of genetic diversity were sufficient to produce unique multi-locus genotypes and detect phylogeographic structuring as individuals assigned back to their population of origin. Cross-species amplification within S. canadensis (sauger) was successful for 15 loci, and 11 loci were diagnostic to species. The loci characterized here will be useful for detecting fine-scale spatial structuring, resolving the taxonomic status of Sander species and sub-species, and detecting walleye/sauger hybrids. Keywords Sander Walleye Sauger Blue pike Microsatellites Walleye (Sander vitreus) remains ecologically and economically important in Lake Erie despite population declines due to overfishing, habitat loss, and impacts of invasive species. The congenerics, sauger (S. canadensis) and blue pike (S. vitreus glaucus), have disappeared from the lake, although catches are reported. Microsatellite loci D. K. Coykendall C. L. Morrison (&) M. J. Springmann United States Geological Survey, Leetown Science Center, Kearneysville, WV, USA e-mail: [email protected] W. Stott United States Geological Survey, Great Lakes Science Center, Cleveland, OH, USA
for Sander species will provide efficient multi-species markers for: verifying taxonomic status; detecting genetic structure at fine spatial scales; clarifying the taxonomic status of blue pike using archived scale samples; determining the best source of sauger or walleye for broodstock development; and for tracking the impacts of sauger stocking in inland waters (e.g., hybridization with walleye). DNA was extracted from dried fin samples and frozen liver samples using DNeasy columns (Qiagen). Separate genomic libraries were constructed for two lakes (Erie and Superior) using DNA from four individuals per lake. The libraries were sequenced on a 454 GS Junior (Roche) following the manufacturer’s protocol. Raw sequencing reads were used as input for QDD (Megle´cz et al. 2010), permitting designs A and B. Primer pairs were selected based on the lowest penalty ranking for 50 putative microsatellite loci. Universal forward primers contained a 50 oligonucleotide that hybridized with one of four fluorescent probes: VIC, FAM, HEX, or NED (Blacket et al. 2012). A test panel of seven walleye individuals was screened at the 50 initial loci. Loci were amplified singly via PC
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