冬虫夏草及虫草类产品DNA条形码鉴定研究

Rapid identification of Cordyceps and its products using DNA barcoding method

冬虫夏草为我国名贵中药材,替代、掺伪、造假等现象普遍,为实现其准确鉴别,该研究采用DNA barcoding方法对6种116份虫草菌和146份寄主分别进行鉴定,并收集了6种29份虫草类产品,基于ITS序列设计虫草菌特异引物,结合虫体COⅠ序列鉴定,对冬虫夏草及其类似产品进行更精确的鉴定。结果表明,基于ITS序列和COⅠ序列均可实现冬虫夏草与虫草类物种及伪品的鉴别;通过2对ITS特异引物ITSSF1/ITSSR1和ITSSF2/ITSSR2,可分别特异扩增到297 bp和136 bp的冬虫夏草菌短片段,实现了冬虫夏草菌与易混淆物种的快速鉴别;而结合COⅠ序列可应用于粉末状态下冬虫夏草全草与人工培育菌丝的准确区分,并实现虫草类产品的快速检测。因此,ITS特异引物结合COⅠ序列,冬虫夏草与发酵菌丝及虫草类产品可快速、准确鉴别。

Cordyceps is one of the most valuable traditional Chinese medicines. There are various counterfeits in markets because of high price and limited output. In this study,116 Cordyceps,146 hosts and 29 related products were collected and detected by using normal DNA barcoding technology and specific PCR method. The results indicated that Cordyceps and its adulterants could be distinguished from each other through DNA barcoding technology based on ITS and COⅠsequences. Two pairs specific primers ITSSF1/ITSSR1 and ITSSF2/ITSSR2 were developed to amplify 297 bp and 136 bp ITS regions of Cordyceps sinensis,respectively. It could be used to identify C. sinensis specifically and rapidly. Furthermore,specific primers ITSSF1/ITSSR1 and ITSSF2/ITSSR2 combined with ITS and COⅠsequences could differentiate powder Cordyceps from fermentation mycelia and could identify related products. Therefore,the method developed from this study could be applied to identify the powder of Cordyceps from fermentation mycelia and related products efficiently.