Isolation and characterization of fifteen microsatellite loci for the giant clam Hippopus hippopus (family Tridacnidae)
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MICROSATELLITE LETTERS
Isolation and characterization of fifteen microsatellite loci for the giant clam Hippopus hippopus (family Tridacnidae) Josina Tiavouane • The´a Jacob • Pascal Paul Dumas Ce´cile Fauvelot
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Received: 3 April 2014 / Accepted: 5 April 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract Fifteen polymorphic microsatellite markers were developed for Hippopus hippopus in order to assess the effectiveness of population replenishment within marine protected areas in New Caledonia. Number of alleles varied from 2 to 11 per locus, observed and expected heterozygosities ranged from 0.300 to 0.866 and 0.495 to 0.858 respectively. Significant deviations from HWE were detected in two loci. Cross-amplifications were tested in four other species of Tridacnidae. Keywords Hippopus hippopus Tridacnidae Microsatellite loci Marine protected area Paternity analyses New Caledonia Giant clams are an important though declining resource for the Indo-Pacific countries. Because of the overexploitation for its meat and shell, Hippopus hippopus (Linne´ 1758) is already extinct from several island countries and is listed on the IUCN Red List of Threatened Species since 2004. Alerted by recent stock declines in New Caledonia, in agreement and following local communities wish, World Wildlife Fund with the support of the Department of J. Tiavouane C. Fauvelot (&) LabEx CORAIL, UR 227 CoReUs 2, Institut de Recherche pour le De´veloppement (IRD), 101 Promenade Roger Laroque, BPA5, 98848 Noume´a Cedex, New Caledonia, France e-mail: [email protected] T. Jacob WWF France bureau Nouvelle-Cale´donie, World Wide Fund for Nature (WWF), Parc Forestier Michel Corbasson, BP692, 98845 Noume´a Cedex, New Caledonia, France P. P. Dumas UR 227 CoReUs 2, Fisheries Department of Vanuatu, Institut de Recherche pour le De´veloppement (IRD), Port-Vila, Vanuatu
Fisheries and Aquaculture of the Northern Province initiated and coordinated a restocking event of Hippopus hippopus in September 2009 within the co-managed Marine Protected Areas (MPAs) of the North-East Lagoon: Hyabe´/LeJao (Poue´bo) and Yeega (Hienghe`ne). Microsatellite loci were developed to provide genetic tools for assessing the effectiveness of this replenishment and estimating spatial scales of larvae dispersal of this species from these MPAs. Approximately 20 ng of genomic DNA was isolated from muscle tissue of one individual conserved in 80 % EtOH. Size-selected fragments from genomic DNA were enriched for SSR content by using magnetic streptavidin beads and biotin-labeled CT and GT repeat oligonucleotides. The SSR-enriched library was analyzed on a Roche 454 platform using the GS FLX Titanium reagents. From the 28’526 reads, 1’689 contained a microsatellite insert with a tetra- or a trinucleotide of at least 6 repeat units or a dinucleotide of at least 10 repeat units. Suitable primer design was possible in 4141 reads and 72 were tested for polymorphism on 8 individuals. Genomic DNA was isolated from mantel biopsies using DNeasy Blood and Tissue K
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