AChE参与褐飞虱若虫中肠细胞凋亡进程的研究

项目名称： AChE参与褐飞虱若虫中肠细胞凋亡进程的研究

项目编号： No.31272050

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： 杨之帆

作者单位： 湖北大学

项目金额： 80万元

中文摘要： 褐飞虱是水稻的重要害虫之一。褐飞虱若虫中肠组织是其与水稻互作的重要界面，中肠组织存在发育过程的细胞凋亡现象，且抗性水稻能诱导凋亡加剧。乙酰胆碱酯酶(AChE)的非经典功能之一是参与细胞凋亡进程。本课题首先通过原位催化研究AChE在I型褐飞虱4龄若虫中肠凋亡细胞中的催化活性；并通过免疫组织化学法在中肠细胞中定位AChE；其次研究取食抗性水稻前后，饲喂amiRNA-AChE和amiRNA-IAP前后，中肠AChE表达变化，并检测凋亡细胞数量变化；进一步研究参与凋亡进程的AChE分子类型及其亚细胞分布规律；最后利用酵母双杂交寻找与AChE互作的蛋白，通过RNAi的方法验证该蛋白与中肠细胞凋亡的关联性，尝试构建AChE参与的凋亡信号通路。研究结果将拓展人们对AChE参与褐飞虱中肠细胞凋亡进程，及其参与褐飞虱与水稻互作的新功能的认识，为培育抗性水稻新品种提供参考，为寻找新的治理方案提供线索。

中文关键词： 褐飞虱；乙酰胆碱酯酶；中肠；细胞凋亡；水稻品种抗性

英文摘要： The brown planthopper (BPH) (Nilaparvata lugens Hemiptera: Delphacidae) is one of the most destructive pests of rice (Oryza sativa L.). The BPH midgut is the most important interface during the interaction of BPH with rice. Midgut apoptosis occurs in BPH nymphs during normal development. Host rice resistance can induce midgut apoptosis. One of the non-canonical functions of acetylcholinesterase (AChE) is involved in apoptosis. In this study, in situ analysis of acetylcholinesterase catalytic activity will be carried out firstly by biochemical staining. AChE will be located in apoptotic cells by immunohistochemistry. Secondly, AChE expression level will be investigated and the number of the apoptotic cells will be determined in the midgut of BPH nymphs ingested resistant rice or not, or fed with amiRNA-AChE, amiRNA-IAP, respectively, or not. Thirdly, molecular types of AChE involved in apoptotic progress and the distribution characteristics of AChE in subcellular level will be studied. Finally, yeast two-hybrid assay will be employed to screen putative proteins interacted with AChE during apoptosis. Relevance of the proteins to apoptosis will be studied, and then construction of the signal pathway will be attempted. This study will help in intensifying the knowledge of novel functions of AChE involved in apoptoti

英文关键词： Nilaparvata lugens；acetylcholinesterase (AChE)；midgut；apoptosis；host rice resistance