腐烂病（Valsa canker）是我国乃至亚洲苹果产区的重大真菌性病害。类受体激酶（Receptor Like Kinase, RLK）在植物识别病原菌信号和激发寄主免疫反应中起重要的调控作用。其中，CrRLK1L亚家族的发现较晚，但部分成员对植物抗病反应的调控作用已得到证实。在苹果上，我们发现4个CrRLK1L基因的表达在腐烂病发生前后存在极为显著的差异。本项目拟以此4个基因为研究对象，在‘皇家嘎啦’中过表达和RNA干扰（RNAi），并比较野生型和转化株系的抗病性，筛选1个抗腐烂病CrRLK1L（R-MdCrRLK1L）基因。之后，基于定量蛋白组学分析，比较R-MdCrRLK1L野生型和转化株系抗病反应的差异，探讨其调控的分子网络。结合酵母双杂交筛选和双分子荧光互补验证，鉴定与其互作的蛋白。通过预测互作蛋白的功能，初步明确其调控的分子路径。

Valsa canker is a fungal disease of apple growing area in China even Asia. Receptor like kinase (RLK) takes crucial roles in plant recognition of the pathogen-drived signals and subsequent activation of host proper immune responses. Even though the subfamily CrRLK1L was found recently, several members have been confirmed to regulate plant immune responses against pathogens. In Apple, we demonstrated that 4 CrRLK1L genes showed significantly differential expression during the onset and progression of Valsa canker. In this project, we will focus on the functional analysis of the above 4 genes in ‘Royal Gala’. Taking advantage of over-expression analysis and RNA interference (RNAi) study to compare the resistance of wild types and transgenic lines, we expect to identify disease-resistant CrRLK1L (R-MdCrRLK1L). Furthermore, through comparing the resistant responses of wild types and transgenic lines by using Quantitative Proteomic Analysis, we will investigate the molecular networks regulated by R-MdCrRLK1L. Next, the potential interaction proteins of R-MdCrRLK1L will be screened by using a Yeast Two-Hybrid system and further confirmed by using Bimolecular Fluorescence Complementation (BiFC). Finally, through functional prediction of the interaction potential proteins, we looking forward to clarify the molecular pathway regulated by R-MdCrRLK1L.