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Development and characterization of microsatellite loci in the ...

Development and characterization of microsatellite loci in the ...

Development and characterization of microsatellite loci in the

Molecular Ecology Notes (2004) 4, 649–652 doi: 10.1111/j.1471-8286.2004.00762.x Blackwell Publishing, Ltd. PRIMER NOTE Development and characterization of microsatellite loci in the endangered oyster mussel Epioblasma capsaeformis (Bivalvia: Unionidae) JESS W. JONES,* MELANIE CULVER,† VICTOR DAVID,‡ JENNIFER STRUTHERS,* NATHAN A. JOHNSON* RICHARD J. NEVES,§ STEPHEN J. O’BRIEN‡ and ERIC M. HALLERMAN* *College of Natural Resources, Department of Fisheries and Wildlife Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24061, USA, †School of Renewable Natural Resources, University of Arizona, Tucson, AR 85721, USA, ‡Laboratory of Genomic Diversity and National Cancer Institute–FCRDC, Frederick, MD 21702–1201, USA, §Virginia Cooperative Fish and Wildlife Research Unit, U.S. Geological Survey, Department of Fisheries and Wildlife Sciences, Virginia Polytechnic Institute and State, University, Blacksburg, VA 24061, USA Abstract Primers for 10 polymorphic microsatellite loci were developed and characterized for the endangered oyster mussel Epioblasma capsaeformis from the Clinch River, Tennessee. Microsatellite loci also were tested in four other populations or species. Amplification was successful for most loci in these closely related endangered species or populations; therefore, a high level of flanking sequence similarity was inferred for this group of species and populations. Allelic diversity ranged from nine to 20 alleles/locus, and averaged 13.6/locus. This study demonstrated the feasibility of using polymerase chain reaction (PCR) primers to amplify microsatellite loci across freshwater mussel species to conduct population genetics studies. Keywords: DNA, Epioblasma, freshwater mussel, microsatellites Received 7 May 2004; revision received 9 July 2004; accepted 9 July 2004 North America contains the greatest diversity of freshwater mussels in the world, including nearly 300 species. However, the mollusk superfamily Unionoida is the most imperiled group of animals in the United States, with 213 species (72%) listed as endangered, threatened, or of special concern (Williams et al. 1993). Most of the endangerment is caused by habitat loss or destruction affecting the natural structure and function of free-flowing rivers. Without immediate efforts to recover imperiled species in U.S. watersheds, the extinction of additional species is likely. To address the threat of species losses, biologists have Correspondence: Jess Jones. Fax: 1-540-231-7580; E-mail: vtaquaculture@hotmail.com The Unit is supported jointly by the U.S. Geological Survey, Virginia Department of Game and Inland Fisheries, Wildlife Management Institute, and Virginia Polytechnic Institute and State University. The content of this publication does not necessarily reflect the views or policies of the Department of Health and Human Services, nor does mention of trade names, commercial products, or organizations imply endorsement by the U.S. Government. developed techniques to propagate and culture endangered freshwater mussels for release of juveniles into rivers to augment or restore populations. However, recovery activities of many species will require genetic analysis of source and recipient populations to help manage species recovery. Samples of mantle tissue were collected from the following species and locations: (i) Epioblasma capsaeformis in the Clinch River, Hancock Co., TN, and Duck River, Maury, Co., TN; (ii) Epioblasma florentina walkeri in the upper Clinch River, Tazewell Co., VA, and Big South Fork Cumberland River, Scott County, TN; and (iii) Epioblasma torulosa rangiana from the Allegheny River, Venango County, PA. A small piece of mantle tissue (20–30 mg) was collected nonlethally from six to 20 live mussels from each population. Tissues were preserved in 95% ethanol and stored at −20 °C prior to DNA extraction. Total genomic DNA was isolated from ∼20 mg of fresh mantle tissue using the Purgene DNA extraction kit (Gentra Systems, Inc., Minneapolis, MN, USA). DNA concentration was determined by flourescence assay © 2004 Blackwell Publishing Ltd

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