药学学报, 2014, 49(2): 260-266
引用本文:
冯杉杉, 郑司浩, 李亚康, 黄林芳. 中药材威灵仙及其伪品DNA条形码鉴别研究[J]. 药学学报, 2014, 49(2): 260-266.
FENG Shan-shan, ZHENG Si-hao, LI Ya-kang, HUANG Lin-fang. Identification of Radix et Rhizoma Clematidis and its adulterants using DNA barcoding[J]. Acta Pharmaceutica Sinica, 2014, 49(2): 260-266.

中药材威灵仙及其伪品DNA条形码鉴别研究
冯杉杉, 郑司浩, 李亚康, 黄林芳
中国医学科学院、北京协和医学院药用植物研究所, 北京 100193
摘要:
利用DNA条形码技术对中药威灵仙及其伪品进行鉴别,为威灵仙分子鉴定提供更多依据与参考。本实验利用psbA-trnH序列对来自3个属10个物种的72份威灵仙及其伪品样本进行扩增测序,结合GenBank数据库中7个属21个物种的284条序列,分析ITS、ITS2、psbA-trnH、rbcL、matK各序列种内与种间变异、遗传距离、barcoding gap,并采用最近距离法(nearest distance)和相似性搜索算法(BLAST1)评价5条序列的鉴定成功率。结果显示psbA-trnH序列在两种DNA序列鉴别评价方法中的效率均较高,且该序列种间变异较大,在barcoding gap检验中,其序列重叠区较少,种间与种内变异分布呈两边分开的趋势,NJ树分析可将各物种分开,差异显著。综合鉴别效率和扩增成功率,推荐使用psbA-trnH序列作为威灵仙药材及其伪品的鉴别序列。
关键词:    威灵仙      伪品      DNA条形码      分子鉴别      psbA-trnH     
Identification of Radix et Rhizoma Clematidis and its adulterants using DNA barcoding
FENG Shan-shan, ZHENG Si-hao, LI Ya-kang, HUANG Lin-fang
Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100193, China
Abstract:
This study provides the candidate sequences in the identification of Radix et Rhizoma Clematidis and its adulterants using DNA barcoding. We amplified and sequenced the region psbA-trnH, with the data of 284 sequences from GenBank, the differential intra-and inter-specific divergences, genetic distance, barcoding gap were used to evaluate five barcodes, and the identification efficiency was assessed using BLAST1 and Nearest Distance methods. The results showed that psbA-trnH barcodes performed high identification efficiency and inter-specific divergences among the five different DNA barcodes. Analysis of the barcoding gap and NJ tree showed psbA-trnH was superior to other barcodes. Based on the identification and PCR amplification efficiency, psbA-trnH can be the ideal barcode to identify Radix et Rhizoma Clematidis and its adulterants accurately.
Key words:    Radix et Rhizoma Clematidis    adulterants    DNA barcoding    molecular identification    psbA-trnH   
收稿日期: 2013-10-14
基金项目: 国家自然科学基金资助项目(81274013);国家自然科学基金重点项目(81130069);人事部留学人员择优资助项目(2009-2012).
通讯作者: 黄林芳
Email: lfhuang@implad.ac.cn
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