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为应用ITS2序列对前胡、紫花前胡药材及其混伪品进行DNA条形码鉴定研究,该文依据国家药典委员会公布的《中药材DNA条形码分子鉴定指导原则》,PCR扩增前胡、紫花前胡及其混伪品的ITS2序列并进行双向测序。经CodonCode Aligner V 3.7.1对测序峰图进行校对拼接,采用MEGA5.0软件分析相关数据。结果表明:前胡药材的ITS2序列长度为229~230 bp,种内变异较小,平均K2P遗传距离为0.005。紫花前胡药材的ITS2序列长度为227 bp,种内无变异。前胡、紫花前胡药材与其混伪品的种间平均K2P遗传距离分别为0.044,0.065。紫花前胡的种间最小K2P遗传距离明显大于其种内的最大K2P遗传距离。最小距离法和NJ树鉴定结果表明前胡、紫花前胡均能与其混伪品明显区分。因此,ITS2序列可有效鉴别前胡、紫花前胡药材及其混伪品。
In order to study the DNA barcoding of Peucedanum petal and Peucedanum mongolicus var. Mariqueter and its adulterants using ITS2 sequence, according to the “Guiding principles of DNA barcode molecular identification of Chinese herbal medicines” promulgated by the State Pharmacopoeia Commission, ITS ITS2 Sequences of Hu and Its Adulterant and Bidirectional Sequencing. The CodonCode Aligner V 3.7.1 on the sequencing peak map proofing stitching, using MEGA5.0 software analysis of the relevant data. The results showed that the length of ITS2 sequence from R. solanacearum was 229-230 bp, and the intraspecific variation was small with an average K2P genetic distance of 0.005. The length of ITS2 sequence of Z. przewalskii was 227 bp with no variation in the species. The average K2P genetic distances between Qianhu and Zaofanqinhu and their adulterants were 0.044 and 0.065, respectively. The minimum K2P genetic distance between species of Viola purpurea was significantly greater than the maximum K2P genetic distance within its species. The minimum distance method and the NJ tree identification results show that the former Hu, Zizania caduceus can be clearly distinguished from its counterfeit products. Therefore, ITS2 sequence can effectively identify the former Hu, Peucedanum and its adulterants.