Development of 27 novel cross-species microsatellite markers for the endangered Hucho bleekeri using next-generation seq
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Development of 27 novel cross-species microsatellite markers for the endangered Hucho bleekeri using next-generation sequencing technology Ke Wang • Shuhuan Zhang • Dengqiang Wang • Miaomiao Xin • Jinming Wu Qingliang Sun • Hao Du • Chengyou Wang • Jun Huang • Qiwei Wei
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Received: 11 September 2014 / Accepted: 27 September 2014 / Published online: 7 October 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract In this study, we developed 27 polymorphic microsatellite loci in Hucho bleekeri, a glacial relict and freshwater resident salmonid fish in China. The number of alleles varied from 2 to 8 for each primer set. The observed and expected heterozygosity ranged from 0.250 to 0.906 and 0.508 to 0.845, respectively. The polymorphic information content ranged from 0.371 to 0.808. These microsatellite loci should be useful to study population genetics, paternity identification, speciation and adaptive evolution of this lineage. Keywords species
Hucho bleekeri Microsatellite Endangered
Hucho bleekeri, a glacial relict and freshwater resident salmonid fish, is endemic to the Yangtze River drainage in China. It became a critically endangered species mainly due to over-fishing and habitat destruction since 1960s (Hu et al. 2008). A series of measures have been taken to protect this species such as listing it into China Red Data Book to ban on fishing and establishing a national nature reserve (Wang 1998). And recently it has been performed successfully controlled breeding, which has great significance to the species preservation. To better address the
Ke Wang and Shuhuan Zhang have contributed equally to this work. K. Wang Q. Wei (&) Wuxi Fisheries College, Nanjing Agricultural University, Wuxi 214081, China e-mail: [email protected]; [email protected] S. Zhang D. Wang M. Xin J. Wu Q. Sun H. Du C. Wang J. Huang Q. Wei Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Wuhan 430223, China
conservation status of the endangered species, a management programme based on genetics is being devised to define the efficiency of these conservation measures (Zhu et al. 2005). Despite the increasing demand for preservation and management plans, no suitable high resolution molecular markers have been available for this species. In current study, we report a set of polymorphic microsatellite loci for H. bleekeri that can serve as effective genetic markers for conducting further preservation and management in this endangered fish. We used the microsatellite makers isolated from closely related species Brachymystax lenok, that had been constructed DNA Library by Ion PGMTM sequencer with Ion 318 chip kit v2 (Life tech LTD) in previous study (unpublished). 72 primer pairs were designed using Primer Premier 5.0 based on the flanking sequence of the repeat motifs. The polymorphisms of the loci were analyzed using a population of 24 wild individuals collected from Taibai River in Baoji City, Shaanxi Province, China. Total genomic DNA of H. bleekeri was e
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