药用昆虫金环胡蜂及其混伪品DNA条形码鉴别研究

Identification of Medicinal Insect Vespa mandarinia from Its Adulterants Using DNA Barcode

目的:以COI序列作为DNA条形码对药用昆虫金环胡蜂及其混伪品进行物种鉴别,探讨快速、准确鉴别金环胡蜂及其混伪品的方法。方法:以COI条形码序列为基础,对金环胡蜂及其近缘种混伪品黄纹大胡蜂进行总DNA提取、PCR扩增和双向测序,并比对Gen Bank中金环胡蜂及其混伪品的COI序列,继而用MEGA6.06软件对所有序列进行分析、计算种内及种间遗传距离,并用邻接(Neighbor-Joining,NJ)法构建出系统进化树。结果:金环胡蜂及黄纹大胡蜂COI序列扩增成功。金环胡蜂与其混伪品COI序列种间最小遗传距离为0.152±0.017,远大于金环胡蜂种内的最大遗传距离0.009±0.004。构建出的系统进化树图也明确显示,各物种都形成了独立的分支。结论:基于COI序列的DNA条形码技术可有效鉴别药用昆虫金环胡蜂及其混伪品,为其质量控制和市场监管提供了新的技术手段,保证金环胡蜂相关药材的安全性。

This study aimed at exploring a fast method to accurately identify the medicinal insect Vespa mandarinia Smith from its adulterants using DNA barcode and COI sequences.The extracted DNAs from V.mandarinia and its adulterants V.soror were amplified by polymerase chain reaction（PCR）and sequenced bilaterally based on COI barcode sequence investigation.The information of the COI sequences of V.mandarinia and V.soror were gathered from Gen Bank.All the sequences were compared and analyzed,and their intraspecific and interspecific genetic distances were calculated using MEGA 6.06.In addition,the phylogenetic tree was established with neighbor-joining（NJ）method.As a result,the COI sequences of V.mandarinia and V.soror were successfully amplified.The minimum interspecific distance between V.mandarinia and its adulterants was 0.152±0.017,being considerably larger than the maximal intraspecific distance between V.mandarinia,0.009±0.004.The constructed phylogenetic tree showed an independent branch for each species.It was concluded that the DNA barcode based on COI sequence can efficiently identify V.mandarinia and its adulterants.This study provided an innovative tool for the quality control and market regulation of Chinese materia medica,securing the safe medication of V.mandarinia.