DNA条形码序列对9种常见蒿属药用植物的鉴定.docVIP

  DNA 条形码序列对 9 种常见蒿属药用植物 的鉴定# 刘美子1,2，宋经元1，罗焜1，林余霖1，刘萍2，姚辉1** 5 10 15 20 25 30 35 （1. 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 部重点实验室，北京 100193； 2. 天津中医药大学，天津 300193） 摘要：目的：检验 DNA 条形码序列对 9 种常见的蒿属药用植物的鉴定效果，为常见蒿属药用 植物的鉴定提供分子依据。方法：对 9 种常见蒿属药用植物的 4 条候选 DNA 条形码序列（ITS2、 rbcL、matK、psbA-trnH）进行 PCR 扩增和测序，比较各序列的扩增和测序效率，应用 BLAST、 Distance 方法来评估各序列的鉴定效率。此外，基于 MEGA5 分析 9 种常见蒿属药用植物 ITS2 序列种间 K2P 距离并构建 NJ 树。结果：除 matK 外，其余三条片段的 PCR 扩增后测序效率均 为 100%，ITS2 序列对 9 种蒿属药用植物的物种水平鉴定成功率最高，为 100%，而 psbA-trnH、 rbcL、matK、rbcL+matK 的鉴定成功率分别为 75%、66.7%、45.5%和 75%。通过 ITS2 序列的 种间 K2P 遗传距离及 NJ 树均能将不同物种全部区分。结论：ITS2 序列可以作为鉴定蒿属药 用植物的潜在条形码。 关键词：蒿属；DNA 条形码；ITS2；鉴定；药用植物 中图分类号：R Testing DNA barcoding regions in identification of the nine common medicinal plants of the genus Artemisia LIU Meizi1,2, SONG Jingyuan1, LUO Kun1, LIN Yulin1, LIU Ping2, YAO Hui1 (1. 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; 2. Tianjin University of Traditional Chinese Medicine, TianJin 300193) Abstract: Objective: To test DNA barcoding regions in identification of nine common medicinal plants of Artemisia and to provide molecular basis for the identification of Artemisia. Methods: In this study, four candidate sequences (ITS2, rbcL, matK, psbA-trnH) of nine species of Artemisia were amplified and sequenced. The PCR amplification and sequencing efficiency were used to evaluate different loci. The identification efficiency was assessed using BLAST and Nearest Distance methods. The genetic distances of ITS2 region were analyzed and NJ tree was constructed by MEGA5. Results: Except matK, the amplification and sequencing efficiency of the other three sequences were 100%. The Identification success rate of ITS2 was 100%, while the regions of psbA-trnH, rbcL, matK and rbcL+matK were 75%, 67%, 46% and 75%, respectively. The nine Artemisi