基于DNA条形码技术的重楼药材疑似伪品基原鉴定  

Original identification of doubtful fake Paridis Rhizoma based on DNA barcoding technology

作　　者：刘杰[1,2] 过立农[2] 马双成[2] 高妍[2] 郑健 王俊丽[1] LIU Jie;GUO Li-nong;MA Shuang-cheng;GAO Yan;ZHENG Jian;WANG Jun-li(Minzu University of China,Beijing 100081,China;National Institutes for Food and Drug Control,Beijing 102629,China)

机构地区：[1]中央民族大学,北京100081 [2]中国食品药品检定研究院,北京102629

出　　处：《药物分析杂志》2020年第8期1437-1442,共6页Chinese Journal of Pharmaceutical Analysis

摘　　要：目的:基于DNA条形码技术鉴定重楼药材疑似伪品的基原。方法:通过聚合酶链式反应(PCR)对重楼的内部转录间隔区2(ITS2)序列进行扩增;通过与GenBank数据库进行Blast比对,以及利用建树法推断重楼药材疑似伪品的基原。结果:重楼药材疑似伪品经Blast比对,结果显示为狭叶重楼(变种)和黑籽重楼(原变种),一致率均达到99%;通过与前期研究的重楼ITS2序列进行NJ聚类分析,可以判定待鉴定样品的基原与西藏延龄草较为接近;通过将重楼药材疑似伪品与收集的吉林延龄草标本、西藏延龄草标本、宽瓣重楼标本、华重楼标本的ITS2序列进行NJ聚类分析,推断重楼药材疑似伪品的基原为吉林延龄草。结论:确定重楼药材疑似伪品为非2015年版《中华人民共和国药典》收载物种,并推断其基原为吉林延龄草,通过DNA条形码技术为鉴别药材正伪品开辟了一条新途径。Objective:To identify the origin of doubtful fake Paridis Rhizoma samples based on DNA barcoding technology. Methods:Internal transcribed spacer of nuclear ribosomal DNA2(ITS2) sequences of doubtful fake Paridis Rhizoma samples were amplified with polymerase chain reaction(PCR)and blasted with GenBank database to determine species. The origin of doubtful fake Paridis Rhizoma was evaluated by NJ phylogenetic analysis. Results:The Blast result of doubtful fake Paridis Rhizoma were Paris polyphylla var. stenophylla and Paris thibetica var. thibetica wtih consistencies of 99%. By performing NJ phylogenetic analysis with the ITS2 sequences of Paridis Rhizoma in the previous study,the doubtful fake samples were identified to be closer to Trillium Kamtschaticum. The NJ phylogenetic analysis result on ITS2 sequences of Trillium govanianum specimen,Trillium kamtschaticum specimen,Paris polyphylla var Yunnanensis specimen,Paris polyphylla var Chinensis specimen and the doubtful fake Paridis Rhizoma showed that the original species of doubtful fake Paridis Rhizoma was Trillium govanianum. Conclusion:The original species of doubtful fake Paridis Rhizoma are not according to the 2015 Version Pharmacopoeia of The People’s Republic of China,and inferred that the original species of doubtful fake Paridis Rhizoma were T. govanianum. DNA barcoding technology is a new method to identify the falsify.

关 键 词：重楼 聚合酶链式反应(PCR) 内部转录间隔区2(ITS2) 基原鉴定 植物鉴别 DNA条形码技术 

分 类 号：R917[医药卫生—药物分析学]