牛蒡子及其伪品的ITS序列分子鉴定研究  

Identification of traditional medicine "Fructus Arctii" by nuclear ribosomal DNA ITS sequences

作　　者：许亮[1] 窦德强[1] 王冰[1] 杨燕云[1] 康廷国[1] 刘阳楠[1] 

机构地区：[1]辽宁中医药大学药学院,辽宁大连116600

出　　处：《中国中药杂志》2011年第3期338-341,共4页China Journal of Chinese Materia Medica

基　　金：国家"十一五"科技支撑计划项目(2006BAIO09B02-6);科技部基础条件平台项目(2005DKA21004);辽宁省教育厅项目(2009A496);辽宁中医药大学青年药学人才基金项目(YXRC0920)

摘　　要：目的:研究牛蒡子与伪品之间的DNA分子鉴别方法。方法:采集全国26个不同产地的牛蒡子药材26份和4种伪品毛头牛蒡、大翅蓟、紫穗槐、水飞蓟10份样品进行总DNA的提取,PCR扩增,对扩增产物克隆并测序,计算机软件统计分析。结果:测得了36份样品的ITS序列全长,分别为牛蒡子641 bp,毛头牛蒡为641 bp;大翅蓟为639 bp;水飞蓟为630～631bp;紫穗槐为625～626 bp,在GenBank中注册,获得登记号。牛蒡子样品间的相似度大于99%,牛蒡子与大翅蓟、水飞蓟、紫穗槐间的相似度低于95%,毛头牛蒡与牛蒡子的相似度为98.29%～99.22%。通过以ITS序列重建系统树进行的聚类分析可以将牛蒡子与伪品有效的区分开。结论:根据ITS序列,能够有效的区分牛蒡子与伪品。Objective： To identify the traditional medicine Fructus Arctii from its adulterants by ITS. Method： Twenty-six samples of the different Fructus Arctii materials and 10 samples of the adulterants of the fruits of A. tomentosum, Onopordum acantlum, Silybum marianum, and Amorpha fruticosa were collected. The total DNA of the samples were extracted, amplified cloned and se- quenced. Result： ITS sequences were obtained from 26 samples respectively, there were Fructus Arctii 641 bp, A. tomentosum 641 bp, Onopordum acantium 639 bp, Silybum marianum 630-631 bp, Amorphafruticosa 625-626 bp, which were registered in the Gen- Bank. The similarity in ITS sequences between Fructus Arctii and the adulterants was less than 95%. In contrast, the similarity be- tween any pair of Fructus Arctii was greater than 99%. The similarity was 98.29% -99.22% between Fructus Arctii and A. tomento- sum. Phylogeny tree reconstruction using UPGMA analysis based on ITS nucleotide sequences can effectively distinguish Fructus Arctii from adulterants. Conclusion： ITS sequences can be used as a reliable molecular marker for the identification of Fructus Arctii.

关 键 词：牛蒡子 ITS序列 分子鉴定 

分 类 号：S567.239[农业科学—中草药栽培]