褐飞虱唾液miRNA调控取食行为和水稻免疫反应的分子机制

褐飞虱在刺吸水稻获取营养的过程中，唾液蛋白和其他物质也随着取食进入水稻中。申请人前期研究表明，miRNA也随着取食进入水稻(简称为B2R-miRNA)，发挥效应因子的作用，但此领域的研究目前仍为空白。本项目拟通过生物信息学预测和分子功能验证等手段，研究:1)通过对唾液小RNA文库和褐飞虱取食后的水稻进行全RNA测序，全面鉴定B2R-miRNA;2)预测B2R-miRNA在褐飞虱和水稻中的靶标，利用双荧光素酶实验等验证靶标关系;3)过表达或抑制B2R-miRNA，通过EPG试验观察取食行为的变化，检测水杨酸通路等水稻免疫反应相关基因的变化;4)通过转基因水稻过表达B2R-miRNA，监测褐飞虱取食行为及水稻免疫反应的改变。本项目研究将揭示miRNA作为褐飞虱唾液效应因子的功能，阐明昆虫miRNA跨物种调控寄主的分子机制，为研究水稻抗虫性和利用miRNA进行害虫控制提供理论基础。

While piercing and sucking rice phloem to obtain nutrients, Nilaparvata lugens also transfers saliva proteins and other substance into rice. Our previous studies have shown that miRNAs would enter into rice, acting as effectors (abbreviated as B2R-miRNA). However, the research in this field is still absent. Our project intends to use bioinformatics prediction and experimental verification to study: 1) Identifying B2R-miRNAs by small RNA sequencing of Nilaparvata lugens’ saliva and whole transcriptome sequencing of rice fed by Nilaparvata lugens; 2) predicting the targets of B2R-miRNAs in Nilaparvata lugens and rice, and verify the miRNA-target interactions by using dual luciferase experiments; 3) Over-expressing or inhibiting B2R-miRNAs in Nilaparvata lugens to observe changes of feeding behavior by EPG, and detect the expression of the immune response-related genes such as salicylic acid pathway in rice; 4) Overexpressing B2R-miRNAs in transgenic rice, and monitoring changes of feeding behavior of Nilaparvata lugens and the immune response of the transgenic rice. Our project will reveal the function of miRNAs as a salivary effector of Nilaparvata lugens and elucidate the molecular mechanism of miRNA cross-species regulation between insects and host plants, to provide a theoretical basis for studying insect resistance and using miRNA for pest control.