JNK信号传递途径在褐飞虱翅型发育中的作用机理研究

The rice brown planthopper(BPH) is an important migratory pests in China, and is a typical polyphenism insect. Long- and short-winged brown planthopper facilitates migration and reproduction switch, so that the population can adapt to the environment and be continuous and prosperous. Wing polyphenism in brown planthopper is affected by population density, host plant nutrients, and other factors. Therefore, the research on the mechanism of the development of brown planthopper wing form, is important for accurate prediction of the brown planthopper outbreaks and analysis of the insect source, as well as for timely and effective prevention and treatment. JNK signaling pathway is a highly conserved pathway of organisms in response to UV radiation, high temperature and osmotic stress and other factors. Our recent studies have found that brown planthopper JNK play a role in wing polyphenism. This is the first discovery that JNK signaling pathway plays a role in wing polyphenism development. However, to date, the molecular mechanism of JNK signaling in wing polyphenism remains unclear. Thus, the present study is to screen candidate genes that interact with JNK through yeast two-hybrid analysis, RNAi and phosporylation assay, further study by various means at the genetic, molecular and biochemical level, for resolving the mechanism of JNK signaling pathway in brown planthopper wing polyphenism development, will also provides an important theoretical basis for understanding the JNK signaling pathway and control the pest in the future.

褐飞虱是我国重要的迁飞性水稻害虫，其长、短翅型是一种典型的翅多型性现象。褐飞虱的长、短翅型有利于迁飞和生殖的切换, 使种群适应环境得以延续与繁荣。褐飞虱的翅型发育受到虫口密度，寄主植物营养等因素的影响。因此，研究褐飞虱翅型发育的机理，对于准确预报褐飞虱暴发和分析虫源，及时有效地进行防治具有重要意义。JNK信号传递途径是生物体对紫外线照射，高温和渗透胁迫等进行响应的一个高度保守途径。我们最近发现，JNK调控褐飞虱的翅型发育。这是首次发现JNK信号传递途径在翅型发育中起作用。但是，目前尚不明确JNK信号传递途径在翅型发育中的作用机理。因此，本研究拟通过RNA干扰,酵母双杂交及JNK对互作蛋白的磷酸化分析等多种手段在遗传、分子与生化水平上研究JNK和互作基因的相互作用，解析JNK信号传递途径在褐飞虱翅型发育中的作用机理，为我们理解JNK信号传递途径和今后的害虫防治提供重要的理论依据。

通过转录组测序数据、初步的RNA干扰实验和酵母双杂交实验筛选获得一批JNK互作基因。对互作基因进行RNA干扰实验，观察记录并分析RNA干扰后翅型等表型，研究JNK及互作基因在分子水平上的相互作用,通过定量PCR等技术检测细胞分裂及DNA复制等相关基因的表达，同时通过酵母双杂交、免疫共沉淀等等手段进行蛋白-蛋白相互作用研究。根据翅型等表型和蛋白-蛋白互作等结果，选择互作较强的JNK互作基因，着重进行其在分子水平、生化水平、遗传水平及细胞水平上与JNK及胰岛素信号途径的相互作用研究。综合以上研究结果，JNK互作基因与JNK及胰岛素信号途径之间存在着分子、生化、遗传和细胞水平上的相互作用。