Isolation and characterization of fourteen microsatellite loci from the endangered octocoral Eunicella verrucosa (Pallas
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TECHNICAL NOTE
Isolation and characterization of fourteen microsatellite loci from the endangered octocoral Eunicella verrucosa (Pallas 1766) Lyndsey P. Holland • Deborah A. Dawson Gavin J. Horsburgh • Andrew P. Krupa • Jamie R. Stevens
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Received: 22 March 2013 / Accepted: 26 March 2013 / Published online: 9 April 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract We isolated 165 unique microsatellite sequences from the threatened and IUCN red-listed octocoral Eunicella verrucosa (common name: pink sea fan). Two enriched genomic libraries were created from a single individual collected in Lyme Bay, Dorset, England. Following testing of 76 markers, 14 loci were then selected, multiplexed and characterised in 44 individuals sampled at the Mewstone Ledges, Plymouth Sound, Devon, England. The number of alleles per locus ranged from 2 to 10 and expected heterozygosities from 0.05 to 0.82. The presence of null alleles was suggested for four loci, with three showing significant deviations from Hardy–Weinberg equilibrium. No linkage disequilibrium was detected. Cross-species utility was assessed in Eunicella singularis (n = 20), E. cavolinii (n = 12) and Leptogorgia sarmentosa (n = 3) collected from the Mediterranean and southern Portugal. At least ten loci were polymorphic in each Eunicella species and five in L. sarmentosa. These markers should prove useful in exploring population structure in these as yet poorly studied animals. Keywords Gorgoniidae IUCN red list Microsatellite Octocoral Pink sea fan Simple tandem repeat (STR)
L. P. Holland J. R. Stevens (&) Department of Biosciences, College of Life and Environmental Sciences, University of Exeter, Exeter, Devon EX4 4QD, UK e-mail: [email protected] L. P. Holland D. A. Dawson G. J. Horsburgh A. P. Krupa NERC Biomolecular Analysis Facility, Department of Animal and Plant Sciences, University of Sheffield, Sheffield S10 2TN, UK
The octocoral Eunicella verrucosa Pallas 1766 (common name: pink sea fan) is a focal taxon of several marine conservation strategies currently being implemented in the UK, and is one of seven cnidarians targeted for protection by the forthcoming UK Marine and Coastal Access Act. In the eastern Atlantic its range is vast, stretching from Angola to western Ireland, where it is typically found at depths between 10 and 200 m. In Britain its distribution is limited to southwest England and south Wales (Grasshof 1992), where it has been detrimentally affected by fishing activity—as such it is classed as a ‘vulnerable’ species on the IUCN red list. All microsatellite markers were developed from the same individual sea fan (EverETR1) collected in September 2008 from East Tennents Reef, Lyme Bay, Dorset, England (N50°390 0900 , W02°520 4400 ), in accordance with collection licence conditions. Genomic DNA was extracted from a 3 cm long clipping using a QIAGEN Plant Mini Kit following the manufacturer’s protocols. With the exception of loci Ever001 and Ever009, microsatellites were isolated at the Evolutionary Geneti
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