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Development and Characterization of Microsatellite Markers for the Ornamented Pygmy Frog Microhyla fissipes and Transferability in Microhyla

Development and Characterization of Microsatellite Markers for the Ornamented Pygmy Frog Microhyla fissipes and Transferability in Microhyla

Lusha Liu1,*, Lei Cheng2, Xingqi Zhang3, Yulong Li1 and Jianping Jiang1,*

1Chengdu Institute of Biology, Chinese Academy of Sciences, Chengdu 610041, China
2Heilongjiang River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Haerbin, 150076, China
3Sichuan Normal University, Chengdu 610101, China

*      Corresponding authors: liuls@cib.ac.cn; jiangjp@cib.ac.cn

 

ABSTRACT

Microhyla fissipes, from family Microhylidae suborder Neobatrachia, widely distributes from western Myanmar eastward through Indochina and northward into southern China including Hainan and Taiwan. But the availability of microsatellites especially from expressed sequences is currently very limited in this species and the genus Microhyla. A total of 17,339 potential microsatellites were identified in 15,385 unigenes which were generated by Illumina paired-end sequencing in M. fissipes. Within all microsatellites, AG/CT, AAG/CTT, and AAAG/CTTT are most prevalent motif in each repeat class. We randomly selected 61 unigenes with the microsatellite to design primers and do genetic analysis in the Sichuan basin population of M. fissipes. Of all, 35 primer pairs (57.38%) successfully amplificated in M. fissipes, of which 14 (40.00%) were polymorphism. The observed and expected heterozygosity in the test population ranged from 0.02 to 0.92 and from 0.02 to 0.62, respectively. High transferability rates were detected in M. butleri (37.14%), M. heymonsi (45.71%), M. pulchra (57.14%)and M. mixture (71.43%).These results indicate that the Illumina paired-end sequencing system is of great value for identifying massive numbers of genic microsatellites in M. fissipes with high-efficiency. Furthermore, the described polymorphic loci in this study should be useful for population genetic and conservation genetic studies in M. fissipes and other closely related species from this important genus Microhyla.
 

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Pakistan Journal of Zoology

October

Vol. 51, Iss. 5, Pages 1599-1997

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