中国中药杂志

2014, v.39(12) 2180-2183

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基于ITS2序列鉴别牡丹皮药材及其混伪品
Identification of Moutan Cortex and its adulterants by ITS2 sequence

魏蒙;邬兰;凃媛;任伟超;向丽;孙伟;张林碧;胡志刚;
WEI Meng;WU Lan;TU Yuan;REN Wei-chao;XIANG Li;SUN Wei;ZHANG Lin-bi;HU Zhi-gang;College of Pharmacy,Hubei University of Chinese Medicine;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences;

摘要(Abstract):

该研究应用ITS2序列鉴别牡丹皮药材及其混伪品,以期探索牡丹皮药材准确鉴别的新方法。提取牡丹皮及其混伪品的DNA,目标片段经PCR扩增后测序,通过CodonCode Aligner V3.7.1对测序序列进行质量分析和拼接,基于MEGA5.0中的K2P模型计算牡丹皮与其混伪品的种内、种间遗传距离及构建系统聚类树。牡丹皮ITS2序列长度为227 bp,种内最大K2P距离为0,它与各混伪品的种间最小K2P距离为0.041,种间平均K2P距离为0.222。由NJ树可知:不同产地来源的牡丹皮药材个体聚为一支,自展支持率为99%,呈现出明显的单系性,能很好地与其混伪品芍药、川赤芍、白鲜皮、朱砂根区分开来。应用ITS2序列可以准确鉴别牡丹皮药材及其混伪品,该方法可以作为传统鉴别方法的有效补充。
To explore a new method to identify Moutan Cortex to guarantee its safe use,internal transcribed spacer 2(ITS2)sequence was used to identify Moutan Cortex and its adulterants. DNA was extracted and target fragments were amplified. Sequences were analyzed and assembled by CodonCode Aligner V3. 7. 1. Genetic distances were computed and phylogenetic tree was constructed based on kimura 2-parameter(K2P) model by MEGA 5. 0. The length of the 20 ITS2 sequences of Moutan Cortex from nine different places is 227 bp,and no variation site was detected. The maximum inter-specificK2P distance of Moutan Cortex is 0,the minimum intra-specific K2P distance is 0. 041,the average intra-specific K2P distance is 0. 222. According to NJ analysis,Moutan Cortex from different places can get together as one branch with bootstrap support values 99%,which indicates Moutan Cortex can be easily distinguished from its adulterants. Using ITS2 sequence can accurately identify Moutan Cortex and its adulterants,it is an effective supplementary to traditional identification methods.

关键词(KeyWords): 牡丹皮;混伪品;ITS2;鉴别
Moutan Cortex;adulterants;ITS2;identification

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作者(Author): 魏蒙;邬兰;凃媛;任伟超;向丽;孙伟;张林碧;胡志刚;
WEI Meng;WU Lan;TU Yuan;REN Wei-chao;XIANG Li;SUN Wei;ZHANG Lin-bi;HU Zhi-gang;College of Pharmacy,Hubei University of Chinese Medicine;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences;

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