Development of microsatellite loci in the common reef starfish Linckia laevigata and Linckia multifora
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TECHNICAL REPORT
Nina Yasuda • Coralie Taquet • Satoshi Nagai Gert Wo¨rheide • Kazuo Nadaoka
Development of microsatellite loci in the common reef starfish Linckia laevigata and Linckia multifora
Received: 25 June 2012 / Accepted: 9 September 2012 / Published online: 25 September 2012 The Ecological Society of Japan 2012
Abstract We isolated seven polymorphic microsatellite loci from the coral reef starfish Linckia laevigata—a common coral reef sea star distributed widely in the Indo-Pacific. The number of alleles ranged from 4 to 13, with an average of 7.4. The ranges of observed and expected heterozygosities were 0.367/0.933 and 0.332/ 0.840 for L. laevigata and 0.182/0.818 and 0.329/0.697 for L. multifora, respectively. All of these markers can also be used in the sister species Linckia multifora. These microsatellite loci will be useful for population and conservation genetic studies in coral reef ecosystems. Keywords Coral reef Æ Genetic structure Æ Linckia Æ SSR Æ Mutiplex PCR N. Yasuda (&) Faculty of Agriculture, University of Miyazaki, Gakuenkibana-dai Nishi 1-1, Miyazaki 889-2192, Japan E-mail: [email protected] Tel.: +81-985-587233 Fax: +81-985-587233 C. Taquet Æ K. Nadaoka Graduate School of Information Science and Engineering, Tokyo Institute of Technology, O-okayama 2-12-1, Meguro-ku, Tokyo 152-8552, Japan C. Taquet USR 3278 CNRS-EPHE, Centre de Biologie et Ecologie Tropicale et Me´diterrane´enne, Universite´ de Perpignan, 52 avenue Paul Alduy, 66860 Perpignan Cedex, France S. Nagai National Research Institute of Fisheries and Environment of Inland Sea, National Fishery Research Institute, Fisheries Research Agency, 2-17-5 Maruishi, Hatsukaichi, Hiroshima 739-0452, Japan G. Wo¨rheide Department of Earth and Environmental Sciences, Palaeontology and Geobiology, and GeoBio-Center, Ludwig-Maximilians-Universita¨t (LMU), Munich, Germany G. Wo¨rheide Bayerische Staatssammlung fu¨r Pala¨ontologie und Geologie, Munich, Germany
Introduction Coral reef ecosystems are currently under heavy pressure from climate change and anthropogenic stresses across their distributions in the tropics (Hughes et al. 2003). Designing marine reserves is beneficial for their conservation; reserves are useful not only for protecting particular areas but also for replenishing neighboring populations via larval dispersal (Halpern and Warner 2002; Salm et al. 2006). Therefore, knowledge of larval dispersal patterns for a wide variety of coral reef organisms that disperse via a pelagic larval stage is important to design effective marine protected areas (Salm et al. 2006). To examine larval dispersal, we developed microsatellite markers for one of the most common coral reef organisms: Linckia laevigata.
Materials and methods Compound microsatellite markers were developed using the method described in Lian et al. (2006). Extracted genomic DNA was digested separately with 6 blunt-end restriction enzymes (AccII, DraI, HincII, SspI, NruI and BalI). The DNA fragments were then ligated with a blunt adaptor (Lian et al. 2006) using a DNA
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