川木通与其混伪品和近缘种的ITS2条形码分子鉴定

摘要

Objective To identify Clematidis Armandii Caidis and its adulterants and closely related species using ITS2 barcode, and study the feasibility of this new method. Methods ITS2 regions were amplified by PCR technology and sequenced bidirectionally. Sequence assembly and consensus sequence generation were performed using the CodonCode Aligner. The Interspecific and intraspecific genentic distances of Clematidis Armandii Akebiae and its adulterants and closely related species were computed by MEGA5. Predict the ITS2 secondary structure was predicted using the ITS2 website founded by Koetschan et al. BLAST method was used for identification of Clematidis Armandii Caidis and its closely related species. Results The interspecific genetic distance of Clematidis Armandii Caidis and its adulterants was higher. ITS2 secondary structure can distinguish Clematidis Armandii Caulis from its adulterants. The success identification rate was 100% between Clematidis Armandii Caidis and its closely related species by BLAST. Conclusion ITS2 barcode could be used to identify Clematidis Armandii Caulis and its adulterants and closely related species, and provide a basis for molecular identification of traditional Chinese medicine.%目的 分析川木通及其混伪品和近缘种的ITS2条形码序列,探讨川木通及其混伪品和近缘种鉴定的新方法.方法 对样品进行DNA提取,PCR扩增ITS2片段并进行双向测序,CodonCode Aligner拼接序列,MEGA5分析川木通的种内遗传距离及与混伪品和近缘物种的种间遗传距离,预测ITS2二级结构,应用BLAST方法进行鉴定分析.结果 川木通与混伪品的种间变异较大,ITS2二级结构的差异分析能够区分川木通与其主要混伪品开,用BLAST方法可将川木通与同属混伪品和近缘物种区分开.结论 ITS2条形码序列可用于区分中药川木通及其混伪品和近缘种,为中药材及其混伪品和近缘物种的分子鉴定提供依据.