DNA条形码在西藏水系裂腹鱼亚科鱼类鉴定中的研究  

Studies on identification of Schizothoracinae population in Tibet water system by DNA barcode

作　　者：周建设 张驰[1] 刘海平 马波 王万良[1] 曾本和 牟振波 ZHOU Jian-she;ZHANG Chi;LIU Hai-ping;MA Bo;WANG Wan-liang;ZENG Ben-he;MOU Zhen-bo(Institute of Fisheries Science,Tibet Academy of Agricultural and Animal Husbandry Sciences,Lhasa 850032,China;Heilongjiang Fisheries Research Institute,Chinese Academy of Fishery Sciences,Harbin 150000,China;College of Animal Science and Veterinary Medicine,Henan Agricultural University,Zhengzhou 450002,China)

机构地区：[1]西藏自治区农牧科学院水产科学研究所,拉萨850032 [2]河南农业大学牧医工程学院,郑州450002 [3]中国水产科学研究院黑龙江水产研究所,哈尔滨150000

出　　处：《淡水渔业》2019年第1期33-41,共9页Freshwater Fisheries

基　　金：西藏自治区自然科学基金项目(2016ZR-NK-14);农业部公益性行业专项(201403012)

摘　　要：为确定DNA条形码技术在西藏水系裂腹鱼亚科(Schizothoracinae)物种鉴定中的可行性,利用西藏水系所采集260尾裂腹鱼亚科样本,测定其线粒体细胞色素C氧化酶亚基1(COI)基因片段序列,计算遗传距离,构建系统发育树,并与GenBank中相关序列进行比对。260个样本经检测均获得有效COI基因扩增片段,COI基因碱基组成偏倚明显,A+T含量为54.74%,显著高于G+C含量(45.26%)。基于Kimura双参数模型计算遗传距离,遗传距离阈值设置为0.02时,260个样本可被鉴定到种的为249尾样本,11尾由于样本量少,数据库中存在多个命名,只鉴定到属,其中与形态学鉴定一致的179尾样本,一致率为68.8%,裂腹鱼亚科裸鲤属(Gymnocypris)不能通过COI基因鉴定到种,拉萨裂腹鱼(Schizothoracinae)、异齿裂腹鱼(S.schizothorax oʼconnori)、巨须裂腹鱼(S.macropogon)3个种之间的遗传距离阈值以0.02不能有效鉴别,而遗传距离阈值以0.01作为这3个种的鉴定标准,可达到有效鉴别的目的。基于邻接法构建系统发育树,裂腹鱼亚科鱼类形成一个支持率较高的单系群,Bootstrap检验支持率为99%,系统进化树聚类方式与以遗传距离值为标准的鉴定结果一致,很好地反映了水系物种间的地理和历史联系。There are more than 40 species of Schizothoracinae in Tibet water system.For the similitude of morphologically between individual within the genus,it is restricted to identified them by traditional morphology.The COI gene fragments from260 individuals of Schizothoracinae were amplified and sequenced to test the applicability of DNA barcode technology in the species identification of schizothoracinae in Tibet water system.Meanwhile,the genetic distance was calculated,the phylogenetic tree was constructed,and the sequences alignment was carried out through the databases of GenBank.This research obtained available gene fragments of the COI from 260 individuals,the base compositions of COI genes were significantly biased.The contents of A+T were 54.74%,which were significantly higher than that of G+C ( 45.26%).The species identification standard of Schizothroacinae were calculated based on Kimura two parameter model as the genetic distance was set in 0.02.The results showed that 249 individuals among 260 could be identified to the species,however,11 individuals of them were identified as genus because of small sample sizes and multiple naming.The identification results of 179 individuals accorded with morphology,the accordant rate was 68.8%.The individuals in the genus of Gymnocypris of Schizothroacinae could not be identified through DNA barcode technology based on COI gene.Among S . racoma waltoni , S . schizothorax o connori and S . macropogon could not be identified based on the genetic distance value of 0.02,but the value of 0.01 could effective identified.The phylogenetic tree based on Neighbor-Joining method indicated that schizothroachinae was a simple group with 99% approval rating of Bootstrap test.The cluster pattern of phylogenetic tree had the consistent results of identification by standard of genetic distance value.The relationship of geography and history between water system were primely responded by the phylogenetic tree.

关 键 词：西藏 裂腹鱼 DNA条形码 COI 物种鉴定 

分 类 号：S917.4[农业科学—水产科学]