猕猴桃果实采后应答灰霉病发生的关键基因筛选和功能解析

kiwifruit is called as the “king of fruit”, due to its high content of vitamin C. Botrytis cinerea is one of the most serious postharvest fungal pathogens on fruits, including kiwifruit. Gray mold caused by B. cinerea can result in significant economic loss. Based on the establishment of the interaction system of kiwifruit and B. cinerea, we plan to employ kiwifruit cv. ‘Hongyang’ and B. cinerea (both genomes have been released) as research materials to invesitige the molecular response of kiwifruit to B. cinerea. This study includes disease symptom observation, cell structure and component analysis of kiwifruit infected with B. cinerea; utilization of transcriptomic analysis to investigate the expression profile of kiwifruit in response to B. cinerea and application of cluster analysis to discover genes involved in the pathogenesis; function characterization of the defense-related genes via transient expression and RNAi technique, and understanding the transcriptional regulatory network of crucial genes involved in the response of kiwifruit to Botrytis cinerea during postharvest storage by yeast two hybrid and genetic transformation. The information will provide basis for management of gray mold and molecular breeding for disease-resistance cultivars of kiwifruits.

猕猴桃因富含维生素C而被誉为“水果之王”。灰霉菌（Botrytis cinerea）是引起猕猴桃果实病变腐烂的重要致病菌，给采后储运带来巨大经济损失。明确猕猴桃果实采后应答灰霉菌的分子机理对灰霉病的预防以及猕猴桃的抗性育种至关重要。在建立了猕猴桃和灰霉菌互作研究体系的基础上，以遗传背景清晰的红阳猕猴桃和灰霉菌B. cinerea为材料，探索猕猴桃果实对灰霉菌应答的分子机理。本项目拟观测分析猕猴桃果实受到灰霉菌侵染后发病表形症状、寄主细胞结构与成分变化，明确发病规律；从转录调控网络分析灰霉菌侵染猕猴桃果实的差异基因表达，聚类关联分析并挖掘病程应答相关基因，通过瞬时表达和RNAi解析其功能；利用酵母双杂及遗传转化实验认知关键抗病基因的转录调控网络，为猕猴桃采后病害的防治提供理论依据，同时为猕猴桃品质改良分子育种提供基因储备.

猕猴桃因富含维生素C以及各种矿质元素深受消费者喜爱。然而，猕猴桃采后极易受到灰霉菌（Botrytis cinerea）侵染，进而引起果实病变腐烂，造成巨大的经济损失。项目组首先通过检测灰霉菌侵染对猕猴桃果实采后表型及其生理生化指标的影响，明确了寄主与灰霉菌的互作规律。其次，通过iTRAQ-LC-ESI-MS/MS蛋白组学技术，挖掘了响应灰霉菌侵染的猕猴桃蛋白，明确了猕猴桃应答灰霉菌侵染的信号通路；最后，通过VIGS沉默技术，对显著响应灰霉菌侵染的猕猴桃关键蛋白基因进行了生物学功能鉴定。.通过该项目的实施，明确了猕猴桃采后与灰霉菌的互作规律；阐明了猕猴桃果实采后应答灰霉侵染的信号转导通路；挖掘到了292个显著响应灰霉菌侵染的猕猴桃关键蛋白；证明了Myosin 10与TPR2基因在猕猴桃应答灰霉菌侵染中的生物学功能。项目成果共发表了SCI论文4篇，中文核心论文2篇；培养研究生6名；获批国家专利4项，其中国家发明专利1项；项目主持人刘奕清教授2017年入选国家“万人计划”领军人才。