Analysis Of The Genetic Diversity Of Lesser Salamander In Zhoushan Area

Hynobius yiwuensis is one of the endemic species in China,which distributed in Yiwu,Zhenhai,Zhoushan,Xiaoshan of Zhejiang province.It has been classified as a vulnerable species by the IUCN(World Conservation Union).Zhoushan archipelago belongs to Zhejiang province,which is one of the main distributed lacation of Hynobius yiwuensis.In recent years,Great changes have taken place in the ecological environment with the rapid development of the Zhoushan regional economic.The transformation of human activities has caused many fragmentation and even fragmentation of many local continuous habitats.Amphibians have poor diffusion capacity and have high regional stability of habitats.Therefore,changes in habitat environment will result in a sharp decline in the number of Hynobius yiwuensis.In this study,microsatellite markers developed in recent years have been used to screen for polymorphic microsatellite loci in Hynobius yiwuensis by using interspecies amplification of near-source species.And then we also analyzed the genetic diversity and genetic structure of Hynobius yiwuensis in Zhoushan archipelago.It provides an important theoretical basis for the protection of Hynobius yiwuensis while we detected the the success rate and reliability of the method.The main results of this paper are listed as below:1?40.0%loci can be amplification(about 50%loci from Hynobius amjiensis and 15.4%loci from Hynobius chinensis)and 54.5%loci had no amplification products by used 113 pairs of microsatellite loci which belong to other affinitive species.The kinship between H.yiwuensis and H.amjiensis is closer than that of H.chinensis.And that cross-species amplification success rate is related to the genetic relationship of species.Therefore,the success rate of cross-species amplification is considered to be related to the kinship of species.In addition,42 pairs of primers were found to be homozygous or only 2 alleles in the polymorphism assay.2?10 pairs of polymorphic microsatellite loci of H.yiwuensis was isolated from Silver staining.In the subsequent PCR process,there were 3 sites,and the sample size successfully amplified was insufficient for follow-up genetic analysis.Therefore,these 3 pairs of primers were abandoned.For other 7 sites,the allelic number per locus varied from 3 to 4(average 3.43).The number of effective alleles is 3.01.The observed heterozygosity and expected heterozygosity varied from 2.250 to 0.537 and from 0.613 to 0.712.3?7 pairs of primers were used to analyze the genetic diversity and genetic structure of H.yiwuensis in Zhoushan area.The results showed that the expected heterozygosity at each site reached a high level of genetic diversity(p>0.5).And FIS indicated that there was a more serious inbred phenomenon within the population.The detection of the genetic bottleneck showed that there was a bottleneck effect in the whole Zhoushan population.Only the No.38 site under the TPM mode test,and the No.38,No.55,No.56,and No.97 sites under the SMM mode had no bottleneck effect.4?Cluster analysis was performed on the kinship among the 120 individuals of H.yiwuensis,and the average coupling between groups was used to measure the genetic distance between individuals using Euclidean distance.Cluster analysis was also performed to remove samples 38,47,91 and 78 individuals which can be clustered into 3 categories at the distance coefficient of 16.065.The possible causes may be the cause of the species itself or human factors.The problem can be explored through further research.