水稻类病斑基因OsSPL35的生物学功能及抗病性调控机制研究

项目名称： 水稻类病斑基因OsSPL35的生物学功能及抗病性调控机制研究

项目编号： No.31471758

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 农业科学

项目作者： 雷财林

作者单位： 中国农业科学院作物科学研究所

项目金额： 80万元

中文摘要： 植物类病斑突变体因其表型与抗病过程中的过敏反应类似而被广泛应用于植物抗病机理研究。我们从粳稻品种Kinmaze转空载体后代中发现一个类病斑突变体，经深入研究确认其类病斑表型是由于T-DNA插入干扰了一个编码包含CUE结构域蛋白的基因OsSPL35 (暂命名）的正常功能而造成的。进一步的研究证实，OsSPL35是一个新型的水稻类病斑基因，负向调控水稻PCD和类病斑的形成、水稻防卫反应及对稻瘟病/白叶枯病的抗性。本项目拟在前期研究工作基础上，通过分子生物学（基因过表达、基因表达谱及转录谱分析等）、生理生化（类病斑发育、酵母双杂、免疫共沉淀、蛋白质质谱分析等）、分子育种（差异表达基因等的遗传转化等）及植物病理（病原菌接种、抗性机制等）的多种研究手段全面、深入解析OsSPL35基因的生物学功能，以期明确该基因在水稻PCD、蛋白泛素化和抗病信号途径中的地位及作用机制，并发掘广谱抗病的水稻基因资源。

中文关键词： 水稻类病斑；突变基因；基因功能；抗病机制

英文摘要： Lesion mimic mutants with the phentypes resembling hypersensitive response are widely used to reveal the mechanisms of plant disease resistance. A lesion mimic mutant was once selected from transgenic progenies of cv. Kinmaze by transferring a empty vector. The previous studies confirmed that the lesion mimic phenotype of the mutant was caused by a T-DNA insertion which disturbed the normal founction of OsSPL35 gene. This gene encodes an unknown protein with one CUE (Couupling of Ubiquitin conjugation to Endoplasmic reticulum degradation) domain.Further studies verified that the OsSPL35 gene is a novel lesion-mimic gene, which could negatively regulate both programmed cell death (PCD)and resistance to rice blast and bacterial blight in rice. In this project, we will perform over-expression and induced expression of the OsSPL35 gene under different biotic or abiotic treatments, determine differential expression genes in the spl35 muant and wild type by using the RNA-seq technique, select interacting proteins with OsSPL35 protein by using H2Y and Co-IP techniques, investigate the functions of differentail expression geness by using transgenic techniques, and evaluate the spl35 mutant and transgenic progenies for their resistance to rice blast and bacterial blight, and so on. The objectives are to comprehensively dissect bilogical functions of the OsSPL35 gene, determine the role and position of the OsSPL35 gene in PCD, protein ubiquitination and plant defense pathways, and explore some braod-spectrum resistance genes effecive to rice diseases.

英文关键词： Rice lesion mimic;mutant gene;gene function;disease resistance