Q型烟粉虱RNAi过程中dsRNA吸收途径研究

项目名称： Q型烟粉虱RNAi过程中dsRNA吸收途径研究

项目编号： No.31272103

项目类型： 面上项目

立项/批准年度： 2013

项目学科： 农业科学

项目作者： 余昊

作者单位： 河南科技学院

项目金额： 70万元

中文摘要： 最新研究发现昆虫细胞对dsRNA的吸收主要有两种不同的路径：跨膜通道吸收和内吞作用。本研究基于前期烟粉虱RNAi的实验结果，以及对烟粉虱RNAi核心机制和传递路径的关键基因比对结果，首先采用烟粉虱羽化后成虫饲喂dsRNA，透过中肠等组织去沉默目标基因- - 雌虫卵巢特异表达的卵黄原蛋白受体基因的方法，来验证烟粉虱的RNAi系统性现象。然后采用抑制系统RNAi途径中关键跨膜通道基因sid1或者抑制环境RNAi内吞作用关键蛋白Vacuolar H+ ATPase的表达，检测路径关键基因沉默条件下烟粉虱RNAi效果变化的方法，验证烟粉虱的dsRNA吸收路径问题。烟粉虱是否具有系统性RNAi及其吸收路径，将决定通过植物表达dsRNA来控制烟粉虱的效果，也决定烟粉虱RNAi基因功能研究应采取的技术手段。本研究将丰富RNAi机制理论，并指导烟粉虱RNAi基因功能研究和控制研究。

中文关键词： 烟粉虱；双链RNA；RNAi；吸收途径；害虫防治

英文摘要： At least two pathways for dsRNA uptake in insects are described by latest research: the transmembrane channel-mediated uptake mechanism based on sid-1 protein and an 'alternative' endocytosis-mediated uptake mechanism. Based on pre-experimental results of Bemisia tabaci RNAi and genome blast research, we have identified the core RNAi genes, as well as genes potentially involved in transmembrane channel-mediated uptake mechanism and the endocytosis-mediated uptake mechanism. First, feed gene-specific dsRNA on emergence of B. tabaci female adults to post-transcriptional silencing other organizations of the female ovary expression vitellogenin receptor gene, this phenomenon would promote identification of systemic RNA interference of B. tabaci. Second, sid1 gene associated with the transmembrane channel-mediated uptake pathway would be identified which involved in dsRNA uptake using an 'RNAi of RNAi' approach in B. tabaci adults. The role of endocytosis in environmental RNAi was checked by blocking the endocytosis with pharmaceuticals against endosome maturation (bafilomycin A1 against vacuolar H+ ATPase) in B. tabaci cell line. Understanding different uptake pathways and systemic RNAi would highlighting the achievement of implementing RNAi in B. tabaci control with transgenic plants, and would assist with selectio

英文关键词： Bemisia tabaci；dsRNA；RNA interference；Uptake Pathway；Pest Control