基于基因组水平的细胞色素P450基因农药响应元件的预测和验证

项目名称： 基于基因组水平的细胞色素P450基因农药响应元件的预测和验证

项目编号： No.31301691

项目类型： 青年科学基金项目

立项/批准年度： 2014

项目学科： 农业科学

项目作者： 刘金定

作者单位： 南京农业大学

项目金额： 23万元

中文摘要： 细胞色素P450基因增强表达是导致农业害虫对农药产生代谢抗性的重要机制，但P450基因响应农药诱导的机制尚不清楚。研究表明，果蝇、蝴蝶、蚊子等昆虫P450基因的启动子区存在植物次生代谢物质的响应调控元件。本项目拟对农业害虫二化螟基因组序列进行挖掘，获得完整P450基因序列，用NNPP等软件获得P450基因启动子序列；综合生测数据、P450基因表达谱芯片数据和P450基因酶活力数据，确定抗药性相关的P450基因；用MDScan等软件和关联分析方法，筛选出P450基因启动子区与农药诱导相关的顺式作用元件（简称农药响应元件）；采用定点突变技术和表达载体技术对这些农药响应元件进行验证。在此基础上，开发基因组层面的P450基因农药响应元件预测算法，并提供在线预测服务平台。本项目的完成不仅在害虫抗药性机理深入研究方面有重要的科学意义，而且在害虫化学防治方面具有重要的应用价值。

中文关键词： 细胞色素P450；基因组注释；二化螟；调控元件；抗性

英文摘要： The over-expression of cytochrome P450 gene is one of the important reasons conferring insecticide resistance. However, the mechanism of insecticide induced over-expression of P450 genes remains unclear. In fruit fly, butterflies and mosquitoes, it has been shown that plant secondary metabolites induce the over-expression of P450 genes by the Cis-elements in the promoter regions. This proposal intends to find insecticide response element (IRE) in the resistance associated P450 genes in rice stem borer. First, P450 gene sequences will be obtained from the genome sequences. Then, we will use promoter prediction software, such as NNPP, to predict the promoter regions. Second, we will collect pests in the fields from different place. The bioassay analysis will be carried out to determine the resistance ratio of the pests to conventional used insecticides. Third, the expression levels of P450 genes will be detected by using microarray. The enzyme activities will be measured by using kinetic method. By these methods, we can determine the resistance associated P450 genes at large scale level. Fourth, we will use MDScan to find the motifs in the promoters of resistance-associated or non-resistance associated P450 genes. The insecticide response elements will be found by further bioinformatics analysis. Fifth, we will u

英文关键词： cytochrome P450；genome annotation；chilo suppressalis；regulation element；resistance