中药材薄荷的DNA条形码鉴定研究.docVIP

  中药材薄荷的 DNA 条形码鉴定研究# 庞晓慧，徐海滨，韩建萍，宋经元** （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：利用 DNA 条形码鉴定技术快速、准确地鉴别中药材薄荷及其密切相关种。方 法：提取薄荷药材及其易混品的 DNA、对 ITS2 序列进行 PCR 扩增和测序，应用 CodonCode Aligner V3.0 对测序峰图进行校对拼接，去除低质量序列及引物区, 获得 ITS2 序列。利用 MEGA5.0 软件计算物种种内种间 Kimura 2-parameter (K2P)遗传距离。采用相似性搜索法、 最近距离法、构建 NJ 系统聚类树等方法进行鉴定分析。结果：薄荷药材与其他密切相关种 之间的 K2P 遗传距离分布于 0.071～0.231 之间，大于薄荷种内 K2P 遗传距离（0～0.006）； 三种鉴定方法也表明 ITS2 序列适合鉴别薄荷药材与其密切相关种。结论：ITS2 条形码可有 效鉴别中药材薄荷及其易混品，为快速、准确地鉴定薄荷提供了科学依据。 关键词：DNA 条形码；ITS2；薄荷 中图分类号：R282.5 Applying DNA barcoding technique to identify Menthae Haplocalycis Herba PANG Xiaohui, XU Haibin, HAN Jianping, SONG Jingyuan (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 Menthae Haplocalycis Herba and its closely related species using DNA barcoding technique. Method: Total genomic DNA was isolated from Mentha Canadensis 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 V3.0. 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 Mentha Canadensis were ranged from 0 to 0.006, which were lower than inter-specific genetic distances between Mentha Canadensis and its closely related species (0.071-0.231). All the three methods showed that ITS2 could discriminate Mentha Canadensis from its closely related species correctly. Conclusion: The ITS2 region is an efficient barcode for identification of Menthae Haplocalycis Herba, which provides a scientific basis for fast and a