何首乌及其常见混淆品的matK基因序列分析及鉴别

Analysis and Authentication of Fallopia multiflora and Its Adulterants by matK Sequence

目的:比较何首乌与其常见混淆品的matK基因序列,从分子水平对中药材何首乌进行鉴定。方法:改良的CTAB法提取基因组DNA,用一对matK通用引物进行PCR扩增,TA克隆后进行测序和序列分析。结果:除毛脉蓼之外,其他混淆品与正品何首乌间的matK基因序列差异较大,不论是核苷酸替代数还是遗传距离,都远远大于不同居群的何首乌之间;进一步比对分析,发现了可以快速鉴别何首乌及其同属混淆品的鉴别位点;运用matK序列分析成功地对3种市售何首乌药材进行了真伪鉴定。结论:根据matK序列特征,能有效区分何首乌及其常见混淆品,matK序列可以作为何首乌药材鉴定的分子标记。

Objective：To identify Fallopia multiflora from its adulterants by comparing their matK sequences.Methods： Genomic DNA of different materials was extracted using modified cetyltrimethyl ammonium bromide（CTAB） method.The double-strand matK genes were amplified using PCR method and then sequenced.The data were analyzed in Clustral W and MEGA 4.0 software package.Results： Besides F.multiflora var.ciliinerve,the matK sequences of other adulterants show distinct differences with F.multiflora,whether for nucleotides substitutions or genetic distances;and the specific identifying sites for distinguishing F.multiflora and other Fallopia adulterants were found through further comparative analysis.Moreover,the 3 inspected materials were successfully authenticated by comparing the matK sequences.Conclusion：matK sequences can be used for the molecular identification between F.multiflora and its adulterant species.