斑翅果蝇的PCR-RFLP快速鉴定方法  

Rapid identification of Drosophila suzukii by PCR-RFLP

作　　者：廖太林[1] 崔平福[1] 吴军[1] 袁克[1] 孙鹏[1] 师振华[1] 陈集翰[1] 

机构地区：[1]昆山出入境检验检疫局,江苏昆山215301

出　　处：《植物检疫》2013年第1期25-29,共5页Plant Quarantine

基　　金：江苏出入境检验检疫局科技资助项目(2011KJ62)

摘　　要：应用PCR-RFLP技术分析斑翅果蝇及其相关种间的差异。选用果蝇COI基因通用引物并通过PCR扩增斑翅果蝇、亚艳丽果蝇等11种15个样品COI基因序列,对PCR产物进行回收和测序,根据测序结果及NCBI上序列比对结果,选用限制性内切酶ApaⅠ和HinfⅠ分别对果蝇COI基因PCR产物进行酶切,酶切产物经琼脂糖电泳分析。结果表明:同时使用ApaⅠ和HinfⅠ酶,可以将斑翅果蝇与其他种相区分。根据果蝇COI基因建立的PCR-RFLP技术可对斑翅果蝇快速、准确进行鉴定。A polymerase chain reaction-restriction fragment length polymorphism（PCR-RFLP） analysis was used for distinguishing of Drosophila suzukii and the related species.Using a pair of conserved primers,the sequences of the mitochondrial cytochrome C oxidaseⅠ（COI） of D.suzukii,D.subpulchrella,etc.were amplified by PCR.The am-plicons were purified,reclaimed,cloned,and digested by ApaⅠ and HinfⅠ selected according to the analysis of COI sequences,followed by agarose gel electrophoresis analysis.It showed that D.suzukii and the related species could be distinguished by using ApaⅠand HinfⅠ.It had been proved that the PCR-RFLP technique based on the COI gene sequences was a simple,specific method for the identification of D.suzukii.

关 键 词：斑翅果蝇 COI PCR-RFLP 鉴定 

分 类 号：S41-30[农业科学—植物保护]