基于ITS2序列的维吾尔药材蜀葵及其近缘种鉴定研究

Identification of Uyghur Medicine Althaea rosea and Its Closely Related Species Based on ITS2 Sequence

目的：研究蜀葵与药蜀葵特征DNA序列的差异，为鉴定维吾尔药材蜀葵及药蜀葵提供依据。方法：采用试剂盒法提取蜀葵及药蜀葵基因组DNA，PCR扩增ITS2片段，双向测序，应用Mega6．0软件分析序列，计算种内及种间遗传距离，构建NJ鉴别树。结果：蜀葵ITS2序列长度为233bp，与药蜀葵231—232bp存在差异；蜀葵种内最大K2P遗传距离为0．004，药蜀葵种内K2P遗传距离为0，蜀葵与药蜀葵种间平均K2P遗传距离为0．0732；NJ树结果表明蜀葵与药蜀葵均单独聚为一支。结论：ITS2序列可用于维吾尔药材蜀葵真伪鉴定的DNA条形码，为保障临床正确用药提供了新的方法。

Objective： To explore a new method to identify Uygur Medicine Althaea rosea and its closely related species A. officinalis by the difference of ITS2 regions. Methods ： Genomic DNA of A. rosea and A. offwinalis were extracted by Reagent kit, ITS2 sequences were obtained by PCR amplification and sequenced bidirectionally. Mega 6. 0 software was used to analyze sequence, calculate intra- and inter-specific genetic distance and construct neighbor joining （NJ） tree. Results ： The ITS2 sequence length of A. rosea was 233 bp and A. officinalis was 231 to 232 bp differences. The maximum K2P genetic distance of A. rosea was 0. 004, when A. officinalis was zero. The average interspecific K2P genetic distance between A. rosea and Althaea officinalis was 0. 0732. NJ tree results showed that A. rosea and A. officinalis were clustered, respectively. Conclusion： The ITS2 sequences can be used to identify Uygur A. rosea with A. officinalis as an effective DNA barcoding, which provides a new methods to ensure its clinical safety.