Isolation and characterization of fourteen polymorphic microsatellite loci for Odontobutis sinensis
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Isolation and characterization of fourteen polymorphic microsatellite loci for Odontobutis sinensis Ping Qiu • Xuefen Yang • Kai-Jian Wei Zhihong Ma • Ruibin Yang
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Received: 24 March 2014 / Accepted: 3 April 2014 Springer Science+Business Media Dordrecht 2014
Abstract Odontobutis sinensis is an indigenous freshwater fish in China. The wild population size of this species has declined sharply in inland waters in recent years. For the purpose of the conservation of natural resources of O. sinensis, 14 polymorphic microsatellite loci were isolated and characterized in 30 individuals collected from Liangzi Lake in Hubei Province. The number of alleles per locus ranged from 2 to 14 with an average of 5.07. The expected and observed heterozygosities ranged from 0.291 to 0.887 and from 0.067 to 0.700, respectively. These newly isolated loci are currently being used for population genetic diversity analysis and will be valuable for the conservation of O. sinensis. Keywords Odontobutis sinensis Microsatellite loci Polymorphism Odontobutis sinensis, an indigenous freshwater fish species in China, is mainly distributed in shallow lakes along the middle reaches of the Yangtze River (Wu et al. 2002). Among the community of indigenous freshwater fishes of China, it is peculiar that the embryonic development of O. sinensis is very long and most of the organ developments is basically completed before its larva get out of the egg membranes. In recent years, the wild resources of this species have declined sharply in inland waters due to P. Qiu X. Yang K.-J. Wei Z. Ma R. Yang (&) Key Laboratory of Freshwater Animal Breeding, Ministry of Agriculture, College of Fisheries, Huazhong Agricultural University, Wuhan 430070, People’s Republic of China e-mail: [email protected] X. Yang K.-J. Wei R. Yang Freshwater Aquaculture Collaborative Innovation Center of Hubei Province, Wuhan 430070, People’s Republic of China
overfishing and water environment deterioration. In order to conserve natural resources of O. sinensis, it is essential to understand its genetic diversity and population genetic structure. However, there is limited genetic information about this species except for little studies on its karyotype analysis, phylogeny and morphological characteristics. In this study, fourteen polymorphic microsatellite loci were isolated in O. sinensis in order to further estimate its genetic diversity and population genetic structure. The genomic DNA was extracted from fin tissue digested by a traditional proteinase-K. Microsatellite loci were isolated using the fast isolation by AFLP of sequences containing repeats (FIASCO) protocol (Zane et al. 2002) with magnetic beads enrichment. The methods of constructing microsatellite enriched libraries and screening positive clones were referred to Li et al. (2009). In total, 145 clones possibly with microsatellite fragments were sent for sequencing. Eighty-seven of the sequenced clones were detected to contain simple sequence repeats. Seventyeight pri
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