9种检疫性实蝇PCR-RFLP快速鉴定研究

Rapid identification among 9 species of quarantine fruit flies(Diptera:Tephritidae)by PCR-RFLP.

应用PCR RFLP技术 ,对我国口岸截获频率较高的 9种检疫性实蝇开展了快速鉴定方法的研究 ,结果表明 ,设计的 2对引物对 9种供试实蝇线粒体DNA(mtDNA)的PCR扩增片断大小分别约为 350bp和 4 50bp。用限制性内切酶MSE1和DRAI对PCR扩增产物进行酶切 ,得到的酶切位点可将供试的 9种实蝇区分开来。该方法不受供试实蝇地理来源及食物源的影响 ,对各种虫态 (卵、幼虫、蛹 )和不同性别的成虫均适用 ,可用于口岸截获实蝇的快速检疫鉴定。

The method of rapid identification among 9 species o f quarantine fruit flies intercepted frequently was conducted based on PCR-RFLP of the mitochondrial DNA(mtDNA).The results showed that DNAs,about 350bp and 4 50bp respectively,were amplified by means of PCR with two pairs of primers desi gned from mtDNA of target pests,The PCR-products could be digested by restrict ion Enzymes(MSE I and DRA I).All 9 species can be discriminated according to t he relevant banding patterns.The method was not affected by individuals,which collected from different places and different host plants.The results indicated that the method is reliable for the discrimination of intercepted fruit fly at port of entry.