当归药材及其混伪品ITS2条形码鉴定  

Authentication of Angelicae Sinensis Radix and Its Adulterants by Analysis of ITS2 Barcode

作　　者：辛天怡[1] 姚辉[1] 韩建萍[1] 宋经元[1] 

机构地区：[1]中国医学科学院北京协和医学院药用植物研究所濒危药材繁育国家工程实验室,北京100193

出　　处：《科研信息化技术与应用》2013年第3期76-82,共7页E-science Technology & Application

基　　金：国家自然科学基金重点项目(81130069);国家高技术研究发展计划(863计划)(2012AA021602);长江学者和创新团队发展计划(IRT1150)

摘　　要：为了确保该药材的质量及临床安全用药,需要对常用中药当归及其混伪品进行分子鉴定。本文介绍了利用ITS2条形码进行鉴定的方法:对ITS2序列进行PCR扩增,并将纯化后的PCR产物进行双向测序,运用CodonCode Aligner拼接后,基于HMMer注释方法去掉两端的5.8S区和28S区获得ITS2条形码,用MEGA5.0软件进行相关数据分析,构建NJ树。结果表明:当归与其混伪品ITS2条形码间存在明显差异,基于ITS2条形码构建的NJ树能够将当归及其混伪品区分开。因此,ITS2条形码可有效地鉴别中药当归及其混伪品,为保证其临床用药安全提供了新的技术手段。In this paper, a method using ITS2 barcode to identify Angelicae sinensis Radix and its adulterants is introduced. Firstly, the PCR technique was performed to get the ITS2 barcodes and purified PCR products were sequenced bi-directionally. Then sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. To remove the 5.8S and 28S region, the hidden Markov model （HMM）-based annotation methods were used. Finally, the ITS2 barcodes were aligned with Clustal-W and the genetic distances were computed by MEGA 5.0 according to the Kimura-2-Parameter （K2P） model. The Neighbor-joining （NJ） phylogenetic tree was constructed. The results showed that there is obvious genetic divergence in the ITS2 sequences between Angelicae sinensis Radix and its adulterants. NJ tree based on ITS2 barcodes can distinguish Angelicae sinensis Radix and its adulterants. This concludes that ITS2 region as a DNA barcode can effectively provided a new technique for identification ofAngelicae sinensis Radix.

关 键 词：当归 ITS2条形码 鉴定 

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