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基于16S rRNA基因DNA条形码的美洲鳗、欧洲鳗、日本鳗的鉴定

陈文炳1,缪婷玉彭娟3,邵碧英3,陈彬3,江树勋3,张志灯1   

  • 收稿日期:2016-06-27 修回日期:2016-12-05 出版日期:2017-02-25 发布日期:2017-02-28
  • 通讯作者: 陈文炳 E-mail:621213wbc@163.com
  • 基金资助:
    福建省科技计划农业引导性项目

Identification of Anguilla rostrata, A. anguilla and A. japonica based on DNA-Barcode of 16S rRNA Gene

  • Received:2016-06-27 Revised:2016-12-05 Online:2017-02-25 Published:2017-02-28
  • Contact: CHEN Wen-Bing E-mail:621213wbc@163.com

摘要: 摘 要:由于鳗鱼加工如烧烤,或因鳗苗体积太小,无法从形态上辨别其所属种类,为满足企业与执法部门需求,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16S rRNA基因的通用引物扩增美洲鳗(A. rostrata)、欧洲鳗(A. anguilla) 、日本鳗(A. japonica)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,为了便于物种鉴别,再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PCR-DNA条形码检测方法。3年来的应用结果表明:建立美洲鳗(A. rostrata)、欧洲鳗(A. anguilla) 、日本鳗(A. japonica) 3种鳗鱼的DNA条形码鉴定方法,操作便利、准确、稳定,可应用于检验检疫实践。

关键词: 美洲鳗, 欧洲鳗, 日本鳗, 16S rRNA基因, DNA条形码, 鉴定

Abstract: Abstract: After processing, such as roasted eel, or eel seedling size to be too small, eel samples could not be identified its species from their morphological characters. To meet the demand of enterprises and law enforcement, the DNA barcode identification method for the main breeding eel species, Anguilla rostrata, Anguilla Anguilla, Anguilla japonica were developed. General primers amplifying the 16S rRNA were used to amplify 16S rDNA fragments of partial 16S rRNA Gene of A. rostrata, A. Anguilla and A. japonica and their PCR products were sequenced, 2 fragments with the length of 638 bp and one with 640 bp of 3 pieces was obtained, respectively. The specific DNA fragment (243 bp) was selected as standard DNA barcode with eel Species specificity to identify eel species conveniently, forward primer(22 bp) and reversed primer (23 bp) were designed to use for establishing PCR-DNA Barcode method. More than three years since, the application results show that DNA barcode identification methods for A. rostrata, A. Anguilla and A. japonica is convenient, accurate and stable, it can be applied to the practice of inspection and quarantine.

Key words: A. rostrata, A. Anguilla, A. japonica, 16S rRNA gene, DNA-Barcode, Identification

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