放线菌新型激发子BAR11诱导苹果植株抗腐烂病的分子靶标及其功能研究

微生物产生的激发子（elicitor）可诱发植物防卫机制从而增强植物抗病害能力。新激发子的筛选及其作用机制的研究可为开发新型环保生物农药奠定基础。项目前期从内生放线菌Hhs.015的外泌蛋白中筛选到一新型蛋白激发子BAR11，可显著促进苹果植株抗腐烂病能力，这一发现为研究内生放线菌的诱抗机理提供了材料，有必要对BAR11诱导苹果抗病性的分子靶标及其功能展开研究。本项目拟通过酵母双杂交、双分子荧光互补和GST pull-down等技术鉴定苹果中BAR11的互作蛋白，通过CRISPR/Cas9基因编辑技术构建苹果中互作蛋白的突变体验证靶标功能。利用RNA-seq技术分析在苹果内过表达BAR11基因后苹果转录组的变化及相应的信号通路，从而揭示BAR11促进苹果抗腐烂病的机理，为研究生防菌诱导抗病的机制奠定基础。

Elicitors produced by microorganisms can enhance plant autoimmunity to prevent and control plant diseases through inducing defense mechanism. The study on the screening of new elicitors and action mechanism has laid a foundation for the development and utilization of this kind of new environmental biological pesticides. In the past, elicitors were mainly found in bacteria and fungi, none was found in actinomycetes. In the early stage of the project, a new type of protein elicitor BAR11which can significantly promote apple resistance to apple Valsa canker was screened from the secretory proteins of the endophytic Saccharothrix strain Hhs.015, which was obtained in our lab. The function of this class of proteins has not been reported, and the relevant mechanisms are still poorly understood. The discovery of BAR11 provides necessary materials for the study on the mechanism of inducing resistance of endophytic actinomycetes. In order to study the molecular target and its function of elicitor BAR11, GFP-Trap beads Immunoprecipitation, Yeast Double Hybridization and Immunoprecipitation Co-IP will be used to screen and identify target proteins in apple. CRISPR/Cas9 gene editing techniques are to be used build apple mutant to verify the target function. The RNA-seq technique was used to analyze the transcriptome changes of apples after the overexpression of BAR11 gene, so as to reveal the mechanism of BAR11 promoting plant immunity. Thus, the mechanism of BAR11 promoting apple immunity will be revealed, providing a theoretical basis for the production and application of elicitors.