基于ITS2序列鉴别维吾尔药材薰衣草及其混伪品

Identification of Uighur Medicine Lavandula Angustifolia and Its Adulterants Based on Internal Transcribed Spacer 2

建立了基于ITS2序列鉴别维吾尔药材薰衣草及其混伪品(全叶青兰和夏枯草)的方法。对维吾尔药材薰衣草及其混伪品的ITS2序列进行PCR扩增和双向测序,使用CodonCode Aligner软件对测序峰图进行序列拼接,用MEGA 6.0软件对拼接后的序列进行多重比对。并计算种内、种间遗传距离,构建NJ系统聚类树,预测其ITS2二级结构。结果表明,经PCR扩增测序后,18份维吾尔药材薰衣草药材ITS2序列比对无差异,序列长度均为235 bp,GC含量为67.23%~69.36%;9份全叶青兰药材序列长度均为218 bp,GC含量为66.21%~69.51%;9份夏枯草药材序列长度均为234 bp,GC含量为66.24%~67.09%。并且维吾尔药材薰衣草的种内遗传距离明显小于种间遗传距离。ITS2序列可作为鉴定维吾尔药材薰衣草及其混伪品的DNA条形码,为维吾尔药材薰衣草的用药安全提供有效的科学技术手段。

A method was established to identify the Uighur medicine Lavandula Angustifolia and its adulterants(Dracocephalumn integrifolium and Prunella vulgaris)based on the internal transcribed spacer(ITS2)sequence.The ITS2 sequences of the Uighur Medicine Lavandula Angustifolia and its adulterants were subjected to PCR amplification and Bi-directional sequencing.CodonCode Aligner software was used to splice sequence of the sequence peak maps,and the MEGA 6.0 software was used to perform multiple alignment of the assembled sequences.In addition,the genetic distances of interspecies and intraspecies were calculated,the NJ system clustering tree was constructed,and ITS2 secondary structure of Uighur medicine Lavandula Angustifolia and its adulterants was predicted.The results showed that the ITS2 sequence comparison of eighteen Uighur Medicine Lavandula Angustifolia showed no difference after PCR amplification and sequencing,with the sequence lengths were all 235 bp,and the GC content was 67.23%-69.36%.The sequence lengths of nine Dracocephalumn integrifolium were all 218 bp,and the GC content was 66.21%-69.51%.And the sequence lengths of nine Prunella vulgaris were all 234 bp,and the GC content was 66.24%-67.09%.In addition,the intraspecific distance of Uighur Medicine Lavandula Angustifolia was more less than the interspecific distance.The ITS2 sequence could be used as a DNA barcode to identify the Uighur Medicine Lavandula Angustifolia and its adulterants,which provided an effective scientific and technological reference for the safety of the Uighur Medicine Lavandula Angustifolia.