应用ITS2条形码鉴定中药材麻黄.docVIP

  应用 ITS2 条形码鉴定中药材麻黄# 庞晓慧，宋经元，徐海滨，姚辉** （中国医学科学院 北京协和医学院 药用植物研究所，中草药物质基础与资源利用教育部重 5 10 15 20 25 30 35 40 点实验室，北京 100193） 摘要：目的：应用 ITS2 条形码快速、准确地鉴定中药材麻黄，为其用药安全、有效提供保 障。方法：提取麻黄药材及其同属密切相关种的 DNA、扩增 ITS2 序列并测序，采用软件 CodonCode Aligner V3.0 对测序峰图进行校对拼接，并进行质量控制。应用 MEGA5.0 软件 计算种内种间 Kimura 2-parameter (K2P)遗传距离。应用相似性搜索法、最近距离法进行鉴定 分析，并构建 NJ 系统聚类树直观反映鉴定结果。结果：麻黄药材基原物种种内 K2P 遗传距 离分布于 0～0.002 之间，基原物种与其他密切相关种之间的 K2P 遗传距离分布于 0.004～ 0.034 之间；几种鉴定方法分析结果表明 ITS2 序列能将麻黄药材与其他密切相关种鉴别开。 结论： ITS2 条形码适用于鉴别中药材麻黄，进一步验证了 DNA 条形码技术鉴定中药材的 有效性。 关键词：DNA 条形码；ITS2；麻黄 中图分类号：R282.5 Using the ITS2 barcode to identify Ephedrae Herba PANG Xiaohui, SONG Jingyuan, XU Haibin, YAO Hui (The Key Laboratory of Bioactive Substances and Resources Utilization of Chinese Herbal Medicine, Ministry of Education, Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences Peking Union Medical College, Beijing 100193) Abstract: Objective: To identify Ephedrae Herba using the ITS2 barcode and to secure its quality and safety in medication. Method: Total genomic DNA was isolated from Ephedrae Herba and its closely related species. Nuclear DNA ITS2 sequences were amplified, and purified PCR products were sequenced. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. The Kimura 2-Parameter (K2P) distances were calculated using software MEGA 5.0. Identification analyses were performed using BLAST1, Nearest Distance and neighbor-joining (NJ) methods. Result: The intra-specific genetic distances of Ephedrae Herba were ranged from 0 to 0.002. The inter-specific genetic distances between Ephedrae Herba and its closely related species were ranged from 0.004 to 0.034. All the three methods showed that ITS2 could discriminate Ephedrae Herba from its closely related species correctly. Conclusion: The ITS2 region is suitable to be used for authentication of Ephedrae Herba, and our study further confirmed the effectiveness of ITS2 to identify traditional Chinese medicina