基于核糖体DNA ITS序列鉴定传统药材“牛蒡子”

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

作者信息

Xu Liang, Dou Deqiang, Wang Bing, Yang Yanyun, Kang Tingguo, Liu Yangnan

机构信息

Liaoning University of Traditional Chinese Medicine, Dalian 116600, China.

出版信息

Zhongguo Zhong Yao Za Zhi. 2011 Feb;36(3):338-41.

PMID:21585039

Abstract

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 acantium, Silybum marianum, and Amorpha fruticosa were collected. The total DNA of the samples were extracted, amplified cloned and sequenced.

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, Amorpha fruticosa 625-626 bp, which were registered in the GenBank. The similarity in ITS sequences between Fructus Arctii and the adulterants was less than 95%. In contrast, the similarity between any pair of Fructus Arctii was greater than 99%. The similarity was 98.29%-99.22% between Fructus Arctii and A. tomentosum. 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.

摘要

目的

通过ITS序列鉴别中药牛蒡子及其伪品。

方法

收集26份不同牛蒡子材料样品以及10份绒毛蓟、大翅蓟、水飞蓟和紫穗槐果实伪品样品。提取样品总DNA，进行扩增、克隆和测序。

结果

分别从26份样品中获得ITS序列，其中牛蒡子641 bp，绒毛蓟641 bp，大翅蓟639 bp，水飞蓟630 - 631 bp，紫穗槐625 - 626 bp，并已提交至GenBank。牛蒡子与其伪品的ITS序列相似度小于95%。相比之下，任意两份牛蒡子之间的相似度大于99%。牛蒡子与绒毛蓟之间的相似度为98.29% - 99.22%。基于ITS核苷酸序列采用UPGMA分析构建系统发育树，可有效区分牛蒡子与其伪品。