Isolation and characterization of eleven microsatellite loci in the marbled rockfish, Sebastiscus marmoratus (Scorpaenid
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MICROSATELLITE LETTERS
Isolation and characterization of eleven microsatellite loci in the marbled rockfish, Sebastiscus marmoratus (Scorpaenidae) Qinghui Li • Zhongbao Li • Gang Dai • Yuanyu Cao • Xiaojiao Chen • Lina Chen Jingbo Shangguan • Yefeng Ning
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Received: 22 June 2013 / Accepted: 19 July 2013 Ó Springer Science+Business Media Dordrecht 2013
Abstract The marbled rockfish, Sebastiscus marmoratus, is an important commercially near-shore fish inhabiting littoral rocky bottoms from Japan to the East China Sea. Eleven polymorphic microsatellite loci were isolated from S. marmoratus and the polymorphism of these loci was observed in 32 samples from one wild population. The number of alleles ranged from 2 to 8. The polymorphism information content ranged from 0.155 to 0.752. The levels of the observed and expected heterozygosity were 0.186–0.969 and 0.170–0.782, respectively. Only one locus (Sm3-63) deviated from Hardy–Weinberg equilibrium. These microsatellite markers can provide a useful tool for further study on genetic diversity and conservation of S. marmoratus. Keywords Sebastiscus marmoratu Microsatellite Genetic markers FIASCO Conservation
The rockfish, S. marmoratus is an important commercially near-shore fish inhabiting littoral rocky bottoms, distributed from southern Japan to eastern Korea, and in the East China Sea (Kita et al. 1996). In recent decades, its natural resource has decreased by over-exploitation of marine resources (Wu 1999). Therefore, S. marmoratus was classified as a national protected animal in Marine Special Q. Li Z. Li (&) G. Dai Y. Cao X. Chen L. Chen J. Shangguan Y. Ning Fisheries College, Jimei University, Xiamen 361021, China e-mail: [email protected] Q. Li Z. Li G. Dai Y. Cao X. Chen L. Chen J. Shangguan Y. Ning Fujian Provincial Key Laboratory of Marine Fishery Resources and Eco-environment, Xiamen 361021, China
Reserve of Zhejiang province by Chinese Government in 2006. For the purpose of protecting resource and recovery strategy, studies on the germplasm resources of S. marmoratus are urgently required. Microsatellite loci are considered as a powerful genetic marker due to its highly polymorphic and co-dominant, and has been widely used in population genetics, population differentiation, linkage analysis and evolutionary studies (Li 2006). In this study, we developed 11 microsatellite loci and established the foundation for the population genetic analysis of S. marmoratus. The microsatellite loci were developed according to the amplified fragment length polymorphism of sequences containing repeats (FIASCO) protocol (Zane et al. 2002). Genomic DNA was extracted from musculature of a single wild S. marmoratus individual using genomic DNA extraction kit DP304-03(TIANGEN). The Genomic DNA was digested with restriction enzyme MseI (Fermentas) and the digested fragments were then ligated to MseI adapter A (50 -ACGATGAGTCCTGAG-30 )/MseI adapter B (50 -TAC TCAGGACTCAT-30 ) by T4 DNA ligase. Afterwards, two biotinylated probes [(GT)15,
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