应用分子生物学和细胞生物学的研究手段和技术，在茶树自交不亲性机理前期研究的基础上，深入研究茶树自交不亲和过程中基于S-RNase的信号识别及转导机制。本项目拟开展工作：通过SAS-PCR鉴定试验品种茶树S基因型，分离相关雌蕊和花粉S基因并获得基因表达产物；利用酵母双杂交和荧光互补等分析S-RNase的识别机制；应用药理学试剂、荧光探针，共聚焦显微镜和电子显微镜技术，分析S-RNase对自交花粉管中细胞核DNA降解、线粒体结构变化和Cyt c释放的影响，以及S-RNase对自交花粉管中Ca2+浓度及梯度、细胞骨架聚合状态和顶端ROS浓度及分布的影响及其间的信号交叉，进而明确茶树自交不亲和过程中基于S-RNase-Ca2+-ROS的信号转导机制和程序性死亡（PCD）响应途径。该研究对揭示茶树自交不亲和的作用机理，获取纯系茶树资源，促进茶树遗传改良都具有重要科学意义。

According to the basis of previous study on the mechanism on Camellia sinensis self-incompatibility, signal recognition and transduction mechanism based on S-RNase will be deeply research using molecular biology and cell biology techniques。wherefore，this new project is designed to research the following works: Appraising the type of S-Gene of the test varieties C. sinensis, and cloning cDNA full length of S-Gene in the pistil and pollen using RACE; analyzing the recognition mechanism of S-RNase using yeast two hybrid and bimolecular fluorescence complementation technology；By using the related pharmacological reagents，fluorescent probe and microscope technology，analyzing the effects of S-RNase on degradation of nuclear DNA， structural changes of mitochondria， release of Cyt c，concentration and gradient of Ca2+，polymerization status of cytoskeleton and concentration and distribution of ROS，so as to clear the signal transduction mechanism and PCD response pathway based on S-RNase-Ca2+-ROS in the process of C. sinensis self-incompatibility，and then to determine the mechanism of C. sinensis self-incompatibility. The results from the research work mentioned above can also provide the scientific theory basis for the study on the genetic breeding of tea plant and the obtaining for pure line tea cultivars.