基于线粒体COⅠ基因鉴别云斑尖塘鳢和线纹尖塘鳢的分子标记

A Molecular Mark Based on CO Ⅰ Gene for Species Identification of Marbled Goby(Oxyeleotris marmoratus) and Sleepy Cod(Oxyeleotris lineolatus)

云斑尖塘鱼鳢(Qxyleotris marmoratus)原产于泰国、越南等东南亚国家线纹尖塘鳢(Oxyeletris lineolatus)原产于澳大利亚,目前两者已成为中国的养殖品种。由于其同属塘鳢属(Oxyeleotris)形态相似,特别是在幼苗阶段难以识别,在苗种市场上常出现混淆,因此本研究研发了一种分子鉴定技术。通过对mtDNA中COⅠ(cytochrome C oxidase subunitⅠ)基因特异性位点比对和分析,设计并筛选出一对特异性引物(XW1)。特异性引物XW1可在线纹尖塘鳢中扩增获得170 by的单一产物,而在云斑尖塘鳢中无产物。通过PCR扩增产物的有无,可以区分两个物种。并在随机选择的30个样本中得到验证。本研究提供一种可以快速且不需要测序的方法来鉴别云斑尖塘鳢和线纹尖塘鳢。

Marbled goby（Qxyeleotris marmoratus） is native to Southeast Asia,such as Thailand and Vietnam.The sleepy cod Oxyeleotris lineolatus is native to Queensland in northeastern of Australia.Currently,both of them are the species of aquaculture breeding in China.We found the morphology of their seedling stage is difficult to identify in seeding market.Via the alignment and analysis of CO I sequences of the Oxyeleotris marmoratus and Oxyeletris lineolatus,many primers were designed basing on the difference sites and one specific primers（XW1） were selected.Amplified with XW1,there was the 170 bp band in the Oxyeletris lineolatus,while nothing in the Oxyeleotris marmoratus.Then 30 individuals per fish were randomized to verify the primers,the results showed that the specific primers have specificity for species.The specific primers can distinguish two species simply and rapidly.This method can provide a molecular marker identification method for two species.