Development of an eDNA metabarcoding tool for surveying the world’s largest amphibian  

作　　者：Jie Wang Ping Liu Jiang Chang Cheng Li Feng Xie Jianping Jiang 

机构地区：[1]CAS Key Laboratory of Mountain Ecological Restoration and Bioresource Utilization&Ecological Restoration Biodiversity Conservation Key Laboratory of Sichuan Province,Chengdu Institute of Biology,Chinese Academy of Sciences,Chengdu 610041,China,and State Key Laboratory of Environmental Criteria and Risk Assessment,Chinese Research Academy of Environmental Sciences,Beijing 100012,China

出　　处：《Current Zoology》2022年第5期608-614,共7页动物学报（英文版）

基　　金：the National Key Program of Research and Development(2016YFC0503200);the National Natural Science Foundation of China(31570391,31200411);the Biodiversity Survey and Assessment Project of the Ministry of Ecology and Environment(2019HJ2096001006);the Construction of Basic Conditions Platform of Sichuan Science and Technology Department(2019JDPT0020);the China Biodiversity Observation Networks(Sino BON-Amphibian&Reptile).

摘　　要：Due to the overexploitation of farming,as well as habitat destruction,the wild population of Chinese giant salamander(CGS)Andrias davidianus,a species with seven genetically distinct lineages,has decreased by over 80%in the past 70 years.Traditional survey methods have proven to be unsuitable for finding this rare and elusive species.We evaluated the efficacy of environmental DNA(eDNA)sampling to detect CGS indirectly from its aquatic environment.We developed several species-specific primer sets;validated their specificity and sensitivity;and assessed their utility in silico,in the laboratory,and at two field sites harboring released farm-bred CGS.We detected the presence of CGS DNA by using polymerase chain reaction and Sanger sequencing.We also sequenced an amplicon mixture of seven haplotype-represented samples using high-throughput sequencing.Our eDNA methods could detect the presence of CGS at moderate densities reported across its range,proving them as a cost-effective way to establish broad-scale patterns of occupancy for CGS.In addition,our primers enabled the detection of mitochondrial lineage mixture or introduced individuals from geographically isolated populations of CGS.

关 键 词：Andrias davidianus Chinese giant salamander EDNA metabarcoding population survey 

分 类 号：Q95[生物学—动物学]