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基于ITS2 序列鉴定中药材金沸草、旋覆花及其近缘混伪品
引用本文:郭力城,胡志刚,凃媛,张卫东,金慧子,孙伟,贺海波,马孝熙,黄必胜.基于ITS2 序列鉴定中药材金沸草、旋覆花及其近缘混伪品[J].世界科学技术-中医药现代化,2014,16(2):307-312.
作者姓名:郭力城  胡志刚  凃媛  张卫东  金慧子  孙伟  贺海波  马孝熙  黄必胜
作者单位:湖北中医药大学药学院 武汉 430065;广西民族医药研究院 南宁 530001;湖北中医药大学药学院 武汉 430065;中国中医科学院中药研究所 北京 100700;湖北中医药大学药学院 武汉 430065;中国中医科学院中药研究所 北京 100700;上海交通大学药学院 上海 200240;上海交通大学药学院 上海 200240;中国中医科学院中药研究所 北京 100700;中国中医科学院中药研究所 北京 100700;中国中医科学院中药研究所 北京 100700;湖北中医药大学药学院 武汉 430065
基金项目:国家自然科学基金委青年科学基金项目(81102778):新型抗自身免疫性心肌炎倍半萜类成分的发现及机制研究,负责人:金慧子。
摘    要:目的:应用ITS2 序列鉴定中药材金沸草、旋覆花及其近缘种混伪品,以确保药材质量,保障临床安全用药。方法:通过对金沸草、旋覆花等药材的3 个基原物种条叶旋覆花、旋覆花和欧亚旋覆花及其同属近缘物种进行DNA 提取、PCR 扩增ITS2 序列并双向测序,用CodonCode Aligner 对测序峰图进行拼接后,用MEGA5.0 软件进行相关数据分析,并构建NJ 系统聚类树以直观反映鉴定结果。结果:各基原物种ITS2 序列变异位点稳定,其种内平均K2P 距离均远小于各自药材以及同属近缘混伪品的种间平均K2P 距离;基于ITS2 序列的NJ 树可以将金沸草和旋覆花药材与同属其它近缘混伪品明显区分开。结论:ITS2 序列可准确鉴别中药材金沸草和旋覆花及其同属近缘混伪品。

关 键 词:金沸草  旋覆花  ITS2  序列  鉴定  近缘物种
收稿时间:2/7/2014 12:00:00 AM
修稿时间:2014/2/19 0:00:00

Identification of Inulae Herba, Inulae Flos and Their Closely Related Species Based on ITS2 Sequence
Guo Licheng,Hu Zhigang,Tu Yuan,Zhang Weidong,Jin Huizi,Sun Wei,He Haibo,Ma Xiaoxi and Huang Bisheng.Identification of Inulae Herba, Inulae Flos and Their Closely Related Species Based on ITS2 Sequence[J].World Science and Technology-Modernization of Traditional Chinese Medicine,2014,16(2):307-312.
Authors:Guo Licheng  Hu Zhigang  Tu Yuan  Zhang Weidong  Jin Huizi  Sun Wei  He Haibo  Ma Xiaoxi and Huang Bisheng
Institution:College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China;Guangxi Institute of Minority Medicine, Nanning 530001, China;;College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China;Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China;Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China;School of Pharmacy, Shanghai Jiao Tong University, Shanghai 200240, China;Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;Institute of Chinese Materia Medica, China Academy of Chinese Medical Sciences, Beijing 100700, China;College of Pharmacy, Hubei University of Chinese Medicine, Wuhan 430065, China
Abstract:This study aimed to identify Inulae Herba, Inulae Flos and their closely related species using the ITS2 barcode, and secure the quality and clinical curative effect of these medicinal materials. DNA was extracted from Inula linariifolia, I. japonica, I. britanica, which are the original species of Inulae Herba and Inulae Flos, together with their closely related species. The ITS2 sequence was amplified by PCR and sequenced bidirectionally. Sequence assembly and generation of consensus sequence were conducted by the CodonCode Aligner. The genetic distances were computed using MEGA 5.0 in accordance with the Kimura-2-parameter (K2P) model, and the phylogenetic tree constructed by the neighbor-joining (NJ) method. The results showed that the ITS2 sequences of the various species have stable variable sites. The intraspecific genetic distances among Inulae Herba and Inulae Flos were obviously lower than the interspecific genetic distance among the above two medicinal materials and its adulterants. The NJ tree based on ITS2 sequences can clearly differ from Inulae Herba, Inulae Flos and their closely related species. It is concluded that ITS2 sequence can be used as DNA barcode to identify Inulae Herba, Inulae Flos and their closely related species.
Keywords:Inulae Herba  Inulae Flos  ITS2 sequence  identification  closely related species
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