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目的:利用ITS2条形码鉴定区分中药板蓝根与南板蓝根及其混淆品蓼蓝和大青。方法:提取4种中药基原植物基因组DNA扩增ITS2序列,隐马尔科夫模型HMMer进行序列注释,Clustal W法进行序列比对,K2P模型计算种内和种间遗传距离,邻接法构建进化树。结果:遗传距离分析显示,4个物种的最大种内遗传距离为0.034,最小种间遗传距离为0.347,最大种内遗传距离大于最小种间遗传距离。同时4个物种可以在进化树上明显区分开来。结论:ITS2序列可以用于板蓝根与南板蓝根及其混淆品蓼蓝和大青的分子鉴定,相较于传统鉴定方法,该方法更加高效与准确。
OBJECTIVE: To distinguish the Chinese medicinal plants Radix Isatidis and Southern Banlangen and their confused products Polygonatum odoratum and Daqing using ITS2 barcode identification. Methods: Genomic DNA of four kinds of Chinese medicinal plants were extracted to amplify ITS2 sequence, Hidden Markov Model HMMer sequence annotation, Clustal W method for sequence alignment, K2P model to calculate intraspecific and interspecific genetic distance, and adjacent method to construct phylogenetic tree . Results: The genetic distance analysis showed that the maximum genetic distance of the four species was 0.034, the minimum genetic distance was 0.347, the maximum genetic distance was greater than the minimum genetic distance. At the same time four species can be clearly distinguished in the evolutionary tree. CONCLUSION: The ITS2 sequence can be used for the molecular identification of Indigowoad and Daqing, which are the major constituents of Radix Isatidis and Southern Bananas and their adulterants. Compared with traditional identification methods, this method is more efficient and accurate.