福建农业学报 ›› 2017, Vol. 32 ›› Issue (7): 730-733.doi: 10.19303/j.issn.1008-0384.2017.07.008

• 园艺科学 • 上一篇    下一篇

基于rbcL序列的半边莲与通泉草的分子鉴别方法

陈建雄, 魏艺聪, 黄泽豪, 徐惠龙, 卢伟, 梁一池   

  1. 福建中医药大学药学院, 福建 福州 350122
  • 收稿日期:2017-04-07 修回日期:2017-06-01 出版日期:2017-07-28 发布日期:2017-07-28
  • 通讯作者: 魏艺聪(1981-),男,博士,讲师,研究方向:中药生物技术,中药药理(E-mail:yicongwei@126.com) E-mail:yicongwei@126.com
  • 作者简介:陈建雄(1991-),男,硕士研究生,研究方向:生药学,中药药理(E-mail:1258747617@qq.com)
  • 基金资助:

    中医药公益性行业专项(201407002)

Rapid PCR Differentiation Between Lobelia chinensis and Mazus pumilus

CHEN Jian-xiong, WEI Yi-cong, HUANFG Ze-hao, XU Hui-long, LU Wei, LIANG Yi-chi   

  1. College of Pharmacy, Fujian University of Traditional Chinese Medicine, Fuzhou, Fujian 350122, China
  • Received:2017-04-07 Revised:2017-06-01 Online:2017-07-28 Published:2017-07-28

摘要: 市场流通中半边莲容易与通泉草相互混淆,拟基于rbcL序列建立半边莲与通泉草的分子鉴别方法。采集不同产地的半边莲样品9份,通泉草样品7份,所有受试样品提取总DNA。对其叶绿体DNA中rbcL片段进行扩增、测序,用ClustalX 2.1软件进行多重序列比对后,应用Meg 5.0软件构建NJ系统聚类树进行聚类分析,根据二组样品间的SNP位点设计相互鉴别的特异性引物,建立特异PCR鉴别方法,并应用SYBR Green I染料法建立对2种中草药进行快速检测的方法。本研究建立了对半边莲与通泉草2种中草药进行分子鉴别的方法,并建立了荧光快速检测方法。

关键词: 半边莲, 通泉草, 特异性PCR, 荧光检测

Abstract: Molecular identifications of Lobelia chinensis and Mazus pumilus, as well as a specific PCR rapid method based on the SNP in rbcL sequence to differentiate between the two rather similar Chinese medicinal material were establish in this study. Commercial samples of 9 L. chinensis and 7 M. pumilus from different origins were collected to extract total DNA for rbcL gene sequencing. SNPs in the rbcL sequences of the samples were isolated using the ClustulX 2.1 program. The primers for identifying the two material were designed based on the SNP site. A specific PCR method was subsequently developed to rapidly and effectively detect the markers by the addition of SYBR Green I dye.

Key words: Lobelia chinensis, Mazus pumilus, SNP, molecular identification

中图分类号: 

  • R284