Development and characterization of twelve microsatellite loci for Amolops ricketti using next-generation sequencing
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Development and characterization of twelve microsatellite loci for Amolops ricketti using next‑generation sequencing Ning Wei1 · Xia Kan1 · Chen Shao1 · Yu Wang1,2 Received: 4 April 2020 / Accepted: 8 July 2020 © Springer Nature B.V. 2020
Abstract Amphibian populations have decreased dramatically because of human disturbance. To better conserve and manage native or wild populations of Amolops ricketti, 12 novel microsatellite loci were developed using high-throughput sequencing. The number of alleles ranged from 2 to 19, with an average of 7.67 alleles per locus. The observed and expected heterozygosity ranged from 0.42 to 1.00 (average 0.81) and from 0.51 to 0.95 (average 0.76), respectively. The polymorphism information content per locus ranged from 0.38 to 0.92 (average 0.69). These polymorphic microsatellite markers will contribute to the assessment of population structure, gene flow and genetic differentiation in A. ricketti. Keywords Microsatellites · Amolops ricketti · Next-generation sequencing · Population genetics
Introduction Amolops ricketti is a common species that inhabits rapidly flowing mountain streams in southern China and northern Vietnam [1]. As an excellent focal species, it attracts considerable attention from scientists. Its skin can secrete antimicrobial peptides that exhibit significant antimicrobial activities against Gram-positive bacteria [2] and secrete Bradykinin-related peptides that have a contractive effect on the smooth muscle of the ileum [3]. Additionally, the broad distribution of this species provides an opportunity to investigate the mechanisms causing speciation and the development of cryptic species [4] and to test specific biogeographic hypotheses [5]. In addition, genetic information for A. ricketti is helpful for clarifying the delimitation, taxonomy and phylogenetic relationships within the genus Amolops [5, 6]. Finally, similar to other frogs with relatively low vagility, this species can be used to understand the effects of ecological processes on genetic variables since it is easily influenced by geographical barriers and tends to * Yu Wang [email protected] 1
College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua, China
Key Laboratory of Wildlife Biotechnology, Conservation and Utilization of Zhejiang Province, Zhejiang Normal University, Jinhua, China
2
be highly genetically structured [5, 7]. However, its populations have declined recently because of threats from human disturbance, such as urbanization, ecological tourism, dam construction and potential overexploitation [8]. To conserve the species and utilize it responsibly, twelve novel polymorphic microsatellite loci are described here, which could be used to assess genetic diversity, genetic structure and population connectivity.
Materials and methods Genomic DNA (gDNA) was extracted using a proteinase K, phenol–chloroform approach [9]. The sequencing library was prepared and constructed following the manufacturer’s protocol and using gDNA from a single in
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