类caspase调控铝诱导花生根尖PCD作用机理的研究

项目名称： 类caspase调控铝诱导花生根尖PCD作用机理的研究

项目编号： No.31301249

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： 詹洁

作者单位： 广西大学

项目金额： 23万元

中文摘要： 以花生品种中花2号为材料，以类caspase在细胞死亡中的作用为核心，通过生理生化、分子生物学和形态结构三个方面研究类caspase在铝诱导花生根尖产生PCD中的作用及其机制。通过荧光底物及人工底物活性抑制剂测定获得与细胞程序性死亡（PCD）相关性最大的类casapse-N;构建Ahc-N、RNAi载体，转入烟草或花生，获取Ahmc-N表达上调、下调或沉默的转基因材料，利用转基因材料进一步分析类caspase、PCD和衰老基因（SAG）之间的关系；通过免疫电镜和荧光显微镜技术检测Ahmc-N在铝诱导PCD发生进程中的亚细胞定位。阐明铝处理导致活性氧迸发，激活类caspase，进而活化SAG等蛋白酶，导致液泡膜破裂，细胞器解体，引发PCD的可能机制。研究结果对明确类caspase在铝诱导PCD中的调控机制，阐明铝诱导PCD机制具有重要科学意义。

中文关键词： 铝胁迫；花生根尖；细胞程序性死亡；类caspase-3；调控机制

英文摘要： This project is to uncover functional mechansim of caspse-like in programme cell death (PCD) induced by aluminum (Al) in the roots of peanut cultivar Zhonghua 2, based on the description of caspase-like in cell death on physiology and biochemistry, molecular biology and morphostructure. The designed strategy mainly includes the obtainment of caspase-like-N that is the most relevant with PCD by characterizing using inhibitors associated with activity of fluorescent substrates and artificate substrates, the construction of vector Ahc-N and RNAi which are introduced into tabacco or peanut, in order to obtain transgenetic plants in which Ahc-N expression is up- or down-regulated, or silenced. Furthermore, the relationship between caspase-like,PCD and senescence-associated genes (SAGs) is determined and analysized using above obtained plants. Then, the location of caspase-like in cell by analysizing the results from the localization of Ahc-N under immnuo-electron microscope and fluorescence microscope during the process of Al-induced PCD. A possible mechanism of caspase-like controlling Al-induced PCD will be certified as follows. Al stress produces ROS burst, ROS activates caspase-like,activates other protease including SAG in turn, then vacuole breaks,cell organelle disintegrates, and produce PCD. The conclusion w

英文关键词： Al stress；root tips of peanut；programmed cell death；caspase-3-like；controlling mechanism