Development of 12 microsatellite markers for Bombina orientails based on RNA-Seq and their usefulness in population genetic diversity

Abstract

Bombina orientalis is widely used due to bombesin which isolated from its skin. But in recent years, the population of B. orientalis has become declining distinctly because of human activities, environmental pollution, drought climatic conditions and other factors. In order to provide the molecular basis for the proposal of biodiversity conservation, we report the development of 12 microsatellite markers for B. orientalis based on RNA-Seq. We test polymorphism against in 48 B. orientalis individuals which randomly selected from 182 individuals take advantage of polyacrylamide gel electrophoresis (PAGE). These markers will be useful in the research on the genetic diversity, population genetic structure and other studies. For B. orientalis, all of these loci showed polymorphism, and in line with the H-W equilibrium law. The number of alleles per locus ranged from 3 to 21. The observed and expected heterozygosities ranged from 0.0118 to 0.7795 and from 0.1612 to 0.8703, respectively. The polymorphism information content ranged from 0.153 to 0.857. And the genetic diversity of B. orientalis in Lushui Rivers is significantly higher than that in the Maoer Mountains.

Evolutionary history of the oriental fire‐bellied toad (Bombina orientalis) in Northeast China

“…The protocol of Shi et al (2018) was followed for microsatellite polymorphism primer information and PCR amplification process to obtain relevant data of 12 microsatellite loci in all individuals. The microsatellite loci were verified by three methods: (a) using Genepop v.4.0.10 (Raymond & Rousset, 1995) software to estimate null allele frequencies, (b) p-Value of Hardy-Weinberg equilibrium (H we ), and…”

Dispersal and mating patterns determine the fate of naturally dispersed populations: evidence from Bombina orientalis

“…All 258 sequences were aligned using BioEdit 7.0.9 [ 47 ]. We also genotyped 515 individuals at 12 polymorphic loci following the methods described in Shi et al [ 13 ].…”

“…However, as a naturally dispersed population, it is unclear what determined the fate of the northern B. orientalis population. Shi et al (2018) did basic analysis on genetic diversity of the northern population while developing microsatellite markers and found high level genetic diversity [13]. In this study, population genetics methods were used to explore whether the dispersal and mating patterns of B. orientalis population are beneficial to maintaining population stability.…”