CYP6CM1基因RNA干扰载体的构建及转基因烟草对烟粉虱抗药性的影响

摘要

Bemisia tabaci is a worldwide pest and presents high level insecticide resistance. The pesticide resistance has brought great challenge on controlling. The mechanism of Bemisia tabaci resistance to insecticide is relative to the enhanced detoxification by cytochrome P450 monooxygenases. A RNAi cassette taxgetting a P450 gene (CYP6CM1) of Bemisia tabaci biotype Q was integrated into tobacco genome through Agrobacterium transformation. Six independent T0 lines were regenerated and confirmed by PCR, and individuals of T1 lines were subjected to Bemisia tabaci. After 5 d feeding, qRT-PCR (quantitative real-time polymerase chain reaction) result showed that the expression level of CYP6CM1 gene in Bemisia tabaci was only 13% -27% of the control (fed with non-transgenic tobacco) , which suggested that the CYP6CM1 -RNAi cassette expressed in transgenic tobacco was effective to decrease the expression of insecticide resistant P450 gene in Bemisia tabaci.%烟粉虱的抗药性与体内细胞色素P450单加氧酶基因的过量表达有关.根据Q型烟粉虱一个P450基因(CYP6CM1基因)的特异区域构建RNAi载体,通过农杆菌介导法转化烟草,PCR检测获得6个独立转基因株系；以非转基因烟草和T1代转基因烟草饲喂烟粉虱5d后,荧光定量PCR结果显示取食转基因烟草的烟粉虱体内CYP6CM1基因相对表达量仅为取食非转基因烟草烟粉虱的13％ ～27％,表明CYP6CM1基因RNAi表达框架可以干扰烟粉虱CYP6CM1基因的表达.