Development of microsatellite markers for the spiny-bellied frog Quasipaa boulengeri (Anura: Dicroglossidae) through tra
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MICROSATELLITE LETTERS
Development of microsatellite markers for the spiny-bellied frog Quasipaa boulengeri (Anura: Dicroglossidae) through transcriptome sequencing Siqi Yuan • Yun Xia • Yuchi Zheng Xiaomao Zeng
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Received: 26 June 2014 / Accepted: 27 September 2014 / Published online: 10 October 2014 Ó Springer Science+Business Media Dordrecht 2014
Abstract We detected and isolated thirty-two microsatellite loci from transcriptome-derived microsatellite DNA for Quasipaa boulengeri. The isolated loci were all polymorphic and the number of alleles per locus ranged from 2 to 10. The observed and expected heterozygosity per locus varied from 0.1493 to 0.9375 and from 0.1563 to 0.9688, respectively. By multiple comparisons with Bonferroni correction, seven loci were deviated from Hardy–Weinberg equilibrium and significant linkage disequilibrium was detected among twenty-five pairs of loci. Those variable microsatellite sites will be applied to assess gene flow between populations and investigate the evolutionary process of chromosomal polymorphism. Moreover, microsatellite loci were served as a valuable resource for drawing up criterion for the conservation management of this threatened species. Keywords Transcriptome Microsatellite Quasipaa boulengeri Quasipaa boulengeri, the spiny-bellied frog, is widely distributed in low mountain ranges of southern China (Fei et al. 2009). It was recently recorded as a de novo case of Electronic supplementary material The online version of this article (doi:10.1007/s12686-014-0344-z) contains supplementary material, which is available to authorized users. S. Yuan Y. Xia Y. Zheng X. Zeng (&) Department of Herpetology, Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, People’s Republic of China e-mail: [email protected] S. Yuan University of Chinese Academy of Sciences, Beijing 100049, People’s Republic of China
chromosomal rearrangements (Qing et al. 2012). The unexpected chromosomal polymorphism that caused by translocation was found within and between natural populations, which has never been found in amphibians before. This species have declined dramatically as a consequence of over-harvesting, water pollution and human consumption, especially estimated decline over 50 % over last three generations and ongoing habitat degradation and loss (Xie et al. 2007). This frog was listed as an ‘‘endangered’’ species in China Species Red List (Wang and Xie 2009), and in IUCN Red List of Threatened Species. Microsatellites usually provide adequate genetic information in the study of gene flow, genetic structure and diversity. Eleven polymorphic microsatellite loci have been developed for the species using AFLP of sequences containing repeats (FIASCO) protocol (Xia et al. 2013). However, the method is restricted in time-consuming. Furthermore, such a few microsatellite loci cannot offer enough information to understand the population structuring processes, especially involving in the chromosomal rearrangements. In order to prompt the rapid microsatellite i
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