棉铃虫CYP6B6 2-十三烷酮响应转录因子的筛选及其调控机制研究

项目名称： 棉铃虫CYP6B6 2-十三烷酮响应转录因子的筛选及其调控机制研究

项目编号： No.31471781

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 刘小宁

作者单位： 新疆大学

项目金额： 80万元

中文摘要： 棉铃虫是重要的世界性农业害虫。细胞色素P450是棉铃虫代谢植物次生物质及杀虫药剂等外源化合物的重要酶系，对P450 CYP6B6基因的转录调控机制进行深入研究具有重要的理论和实践意义。近年来，国内外对农业昆虫解毒酶基因功能进行了大量研究，但涉及解毒酶调控机制的研究很少。本项目拟在前期研究的基础上，以CYP6B6 基因启动子中的2-十三烷酮应答核心区域中顺式作用元件为诱饵，利用酵母单杂交系统克隆与之结合的候选转录因子，然后用凝胶阻滞及转录激活等实验筛选转录因子；最后通过RNAi及qRT-PCR技术对得到的转录因子进行功能验证，研究结果可望阐明在2-十三烷酮胁迫下转录因子调控棉铃虫CYP6B6表达的调控机制。以期通过挖掘调控2-十三烷酮胁迫反应的开关，最终找到引领全局的调控总开关，为揭示棉铃虫响应2-十三烷酮胁迫信号转导途径及其应答机制，探索棉铃虫等重要害虫的基因调控抗虫技术手段奠定理论基础。

中文关键词： 棉铃虫CYP6B6；2-十三烷酮；转录因子；筛选；调控机制

英文摘要： Helicoverpa armigera is one of the impeotant agricutural pest in the world. The cytochrome P450 monooxygenases have important roles in the metabolism of endogenous and exogenous compounds. CYP6B6 is a cytochrome P450 isolated from cotton bollworm, which not only participates in endogenous metabolism of some substances which have important physiological functions, such as hormones, fatty acid, but also metabolizes some xenobiotics such as allelochemicals and insecticides. Transcription of CYP6B6 can be enhanced by toxic compounds, leading to increased expression of the protein and thus increased detoxi?cation, to make the cotton bollworm possess high tolerance to insecticides.In recent year, the fouctional genes of the detoxic enzyems have been widely investigated, but its regulatory mechanism has not been. On the basis of the previous work we intend to use yeast one-hybrid system to clone the transcription factor that can interacting with the bait sequence, the cis-acting element, which is in 2-tridecanone-responsive region in the promoter of cytochrome P450 CYP6B6 gene from H. armigera. And then confirm the interaction between bait sequence and the transcription factor by the electropjoretic mobility shift，and also validate the transcription activation of the transcription factor gene by the yeast one-hybrid system. To evaluate the function of the transcription factor, we will investigate the CYP6B6 expression profile, tolerance to insecticides and disturbance to the growing development of the cotton bollworm treated with the dsRNA, from the corresponding transcription factor. These results will elucidate the expression mechanism of CYP6B6, from the cotton bollworm treated with 2-tridecanone, regulated by the transcription factor, then dredge the on/off of 2-tridecanone-stress response and finally find main switch of regulation in the whole stress, which will helped us not only to draw out 2-tridecanone-stress signal transduction pathway and its responsive mechanism, but also provide a new technical method for control of Helicoverpa armigera and other pests.

英文关键词： Helicoverpa armigera CYP6B6;2-tridecanone;transcription factor;Screening;regulatory mechanism