基于12S rRNA序列的梵净山地区泽陆蛙群体遗传多样性分析  

Genetic Diversity Analysis of Fejervarya multistriata Population in Fanjingshan Region Based on 12S rRNA Sequence

作　　者：吴震洋[1] 李丽[1] 张雷[1] 邱发根[1] 杨石龙 俄广鑫[2] WU Zhen-Yang;LI Li;ZHANG Lei;QIU Fa-Gen;YANG Shi-Long;E Guang-Xin(Guizhou Provincial Key Laboratory for Biodiversity Conservation and Utilization in the Fanjing Mountain Region,Tongren University,Tongren 554300,China;College of Animal Science and Technology,Southwest University,Chongqin 400715,China)

机构地区：[1]铜仁学院贵州省梵净山地区生物多样性保护与利用重点实验室,铜仁554300 [2]西南大学动物科学技术学院,重庆400715

出　　处：《农业生物技术学报》2020年第12期2209-2220,共12页Journal of Agricultural Biotechnology

基　　金：国家自然科学基金(31702097);贵州省重点实验室项目(黔科合平台人才[2020]2003);贵州省科技厅联合基金(黔科合LH字[2016]7311号);铜仁学院高端人才创新团队项目(CXTD[2020-19])。

摘　　要：梵净山是武陵山脉的主峰,因其多样的气候类型和优越的水热条件而拥有丰富的两栖动物多样性。但是,相关研究主要集中在1980年代,且多基于野外调查。因此,利用线粒体基因作为分子标记对梵净山地区两栖动物的遗传多样性进行评估十分迫切。为评估梵净山周边地区泽陆蛙(Fejervarya multistriata)遗传多样性及梵净山特殊的地理环境对其生存现状的影响,为进一步保护和利用泽陆蛙遗传资源提供科学依据,本研究以线粒体DNA(mitochondrial DNA,mtDNA)的12S rRNA部分序列作为分子标记,对梵净山周边地区10个区县共185只泽陆蛙样本进行遗传多样性分析。结果表明,梵净山地区泽陆蛙核苷酸多样性指数Pi为0.02942±0.00034,单倍型多样性(haplotype diversity,Hd)为0.563±0.017;共发现10个单倍型,优势单倍型为单倍型1、2和8,分别占样本总数的36.76%、44.32%和12.43%。单倍型1和8主要由梵净山以东地区的泽陆蛙种群构成,而单倍型2则主要存在于梵净山以西的种群中。单倍型网络图显示单倍型1和8聚为一类,单倍型2聚为另一类。以GenBank中虎纹蛙(Hoplobatrachus chinensis)和腹斑倭蛙(Nanorana ventripunctata)为外群构建单倍型系统发生树,单倍型聚类分析表明,腹斑倭蛙构成单倍型聚类的外群,而虎纹蛙和泽陆蛙单倍型聚在一起,其中,泽陆蛙又分为2个亚支。这与单倍型网络图及种群系统发生树的结果相符,即泽陆蛙10个种群分为梵净山东部和西部地区2个支系,可能为梵净山及其所属武陵山脉的地理隔离所致。固定系数Fst表明泽陆蛙种群分化程度明显。分子变异分析(analysis of molecular variance,AMOVA)显示,泽陆蛙的遗传变异主要来自种群之间(86.14%),种群内部的遗传变异较低(13.86%)。综合分析认为,泽陆蛙不同群体间遗传多样性较低,遗传分化明显,主要原因可能是复杂的地形地貌和山脉的地理隔离作用。本研究结果�Fanjing mountain is the main peak of Wuling mountain range.It has rich amphibian diversity due to its diverse climate types and superior hydrothermal conditions.However,the related research mainly concentrated in the 1980’s,and mostly based on field investigation.Therefore,it is urgent to evaluate the genetic diversity of amphibians in Fanjingshan region by using mitochondrial genes as analytical markers.To evaluate the genetic diversity of Fejervarya multistriata in the surrounding area of Fanjing mountain,the influence of the specific geographical environment of Fanjing mountain on its survival status,and to provide scientific basis for further protection and utilization of genetic resources of F.multistriata,the present study used partial sequences of mitochondrial DNA(mtDNA)12 S rRNA as molecular markers,185 samples from 10 districts and counties in Fanjingshan area were sequenced and the genetic diversity was analyzed.The results showed that nucleotide diversity index Pi and haplotype diversity(Hd)were 0.02942±0.00034 and 0.563±0.017,respectively.Ten haplotypes were found,and the dominant haplotypes were haplotype 1 and 2,followed by haplotype 8,accounting for 36.76%,44.32%and 12.43%of the total sample,respectively.Haplotype 1 and haplotype 8 were mainly composed of the populations east of Fanjing mountain,while haplotype 2 mainly existed in the population west of Fanjing mountain.Haplotype network mediation diagram showed that dominant haplotype 1 and haplotype 8 gathered into one class,and haplotype 2 gathered into another class.Hoplobatrachus chinensis and Nanorana ventripunctata were used as extranet to construct a haplotype systemic-occurrence tree,and the haplotype clustering analysis indicated that the haplotype cluster of Hoplobatrachus chinensis and F.multistriata were together,and the haplotype cluster of F.multistriata was divided into 2 subbranches,which was consistent with the results of haplotype network map and population phylogenetic tree,that was,the 10 populations of the F.multistriata

关 键 词：泽陆蛙 遗传多样性 线粒体12S rRNA 遗传结构 地理隔离 

分 类 号：S813.9[农业科学—畜牧学]