摘要:
为满足鳗鱼养殖、加工、贸易企业及执法部门的需求,建立我国主要养殖鳗鱼美洲鳗(Anguilla rostrata)、欧洲鳗(Anguilla anguilla)、日本鳗(Anguilla japonica)的物种DNA条形码鉴定方法.以扩增16SrRNA基因的通用引物扩增美洲鳗、欧洲鳗、日本鳗的16S rRNA基因片段,各获得1条16S rDNA片段,测序结果表明,前二者的长度均为638 bp、日本鳗的长度为640 bp,为了使物种鉴定方法更简便,结果更准确,从各自片段中截取具有物种特异序列的片段(243 bp),作为3种鳗鱼的标准DNA条形码,设计两端碱基数分别为22 bp与23 bp的正向与反向引物,建立聚合酶链式反应-DNA条形码检测方法.3年来的应用结果表明:建立的美洲鳗、欧洲鳗、日本鳗3种鳗鱼的DNA条形码鉴定方法,操作简便、准确、稳定,可应用于3种鳗鱼的物种鉴定.%To meet the demands of enterprises engaged in eel farming,processing and trading and law enforcements,a DNA barcoding method for the identification of three main farmed eel species,Anguilla rostrata,A.Anguilla and A.japonica,was developed.Universal 16S rRNA primers were designed and used to amplify partial 16S rRNA gene fragments ofA.rostrata,A4.Anguilla and A.japonica and their PCR products were sequenced to be 638,638 and 640 bp in length,respectively.The specific DNA fragment (243 bp) was selected as standard DNA barcode with eel species specificity to identify eel species conveniently,and a forward primer (22 bp) and a reversed primer (23 bp) were designed and used to establish the PCR-DNA barcoding method.The results of the application of the proposed method over the past three years showed that it is convenient,accurate,stable,and useful to identify the three eel species.