基于DNA条形码的藏药洪连鉴定研究 |
投稿时间:2019-07-18 点此下载全文
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引用本文:尼玛潘多,李婷,胡小松,刘春生.基于DNA条形码的藏药洪连鉴定研究[J].中国现代中药,2020,22(4):542-545 |
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基金项目:国家药典委员会2009—2010年度“国家药品标准提高”项目(765) |
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中文摘要:目的:对藏区常使用的洪连药材进行DNA条形码鉴定,为准确鉴定藏区洪连药材提供方法。方法:从5个藏区收集12批洪连药材,提取其DNA,采用ITS以及psbA-trnH序列进行聚合酶链式反应(PCR)引物扩增并测序,所得序列从GenBank数据库进行相似性同源查询,采用DNAMAN、Editseq、Mega等软件,寻找其特异位点,计算样品Kimura2-parameter(K2-P)遗传距离,并构建N-J系统构建树。结果:12批样品来源于3个种,ITS序列长度为552 bp,变异位点为34个,在107 bp处能将3种兔耳草区分;psbA-trnH序列长度约为250 bp,存在12个变异位点,在136 bp处能将3种兔耳草区分。根据遗传距离构建N-J树,3种兔耳草明显分为3支。结论:ITS序列以及psbA-trnH序列可以作为兔耳草属鉴别的1种方法,为藏药的准确鉴别提供了方法。 |
中文关键词:洪连 兔耳草 DNA条形码 鉴定 |
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Identification of Tibetan Medicine Honglian Based on DNA Barcoding |
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Abstract:Objective:To identify the DNA barcodes of Honglian medicinal materials commonly used in Tibetan areas,and to provide a method for accurately identifying Honglian medicinal materials in Tibetan areas.Methods:12 batches of Honglian medicinal materials were collected from 5 Tibetan areas,and the DNA was extracted. PCR primers were amplified and sequenced using ITS and psbA-trnH sequences. The sequence was similarly homologously searched from GenBank database,using DNAMAN,Editseq,Software such as Mega searches for its specific site,calculates the genetic distance of the sample Kimura2-parameter (k2-P),and constructs the NJ system building tree.Results:12 batches of samples were derived from three species. The length of ITS sequence was 552 bp and the number of mutation sites was 34. 3 kinds of rabbit ear grasses could be distinguished at 107 bp. The length of psbA-trnH sequence was about 250 bp,and there were 12 mutation sites,3 kinds of Lagotis Gaertn can be distinguished at 136 bp. According to the genetic distance,N-J trees were constructed,and 3 kinds of Lagotis Gaertn were clearly divided into three branches.Conclusion:ITS sequence and psbA-trnH sequence can be used as a method to identify the ear genus of Aurora,providing a method for accurate identification of Tibetan medicine. |
keywords:Honglian Lagotis Gaertn DNA barcoding identification |
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