摘要
效应蛋白是病原菌操纵植物免疫过程和建立定殖的重要因素,研究山田胶锈菌Gymnosporangium yamadae分泌用于调控宿主防御反应的关键效应蛋白,对解析其致病机制具有重要意义。本研究基于山田胶锈菌性、锈孢子阶段的转录组数据,通过效应蛋白的结构特征和表达量等条件共预测得到100个候选效应蛋白,并克隆出43条序列正确的片段。本氏烟瞬时表达结果显示GyS-2、GyS-3、GyS-16、GyS-20、GyA-4共5个候选效应蛋白能够诱导本氏烟叶片细胞坏死,含致病结构域的GyS-44不引起烟草坏死。亚细胞定位观察到GyS-2和GyS-44定位在细胞质,GyA-4聚集在叶绿体周围,GyS-3、GyS-16、GyS-20定位在细胞核及其周围。酵母信号肽分泌功能验证证明了这6个效应蛋白的信号肽均具有分泌功能。研究结果对进一步深入开展山田胶锈菌效应蛋白分子机制研究奠定基础。
Effectors are essential for pathogens to manipulate plant immunity and colonize host.Elucidating the key effectors secreted by Gymnosporangium yamadae that regulate host defense responses is pivotal for understanding its pathogenic mechanism.In this study,100 candidate effectors were predicted based on the comparative transcriptome analysis of spermogonial and aecial stages of G.yamadae,combined with structural characteristics,and 43 fragments with accurate sequences were cloned.Transient expression in Nicotiana benthamiana showed that five candidate effectors(GyS-2,GyS-3,GyS-16,GyS-20,and GyA-4)triggerred cell death,while GyS-44 with a pathogenic domain had no impact on N.benthamiana’s immune defense.Subcellular localization assays revealed that GyS-2 and GyS-44 were localized in the cytoplasm,GyA-4 aggregated around the chloroplasts,and GyS-3,GyS-16,and GyS-20 were localized in and around the nucleus.Yeast signal peptide secretion assays verified that the signal peptides of these six key effectors possessed secretory capabilities.The results provide a basis for further in-depth exploration of the molecular interactions between G.yamadae effectors and the host.
作者
金之鑫
张晓漫
梁英梅
JIN Zhixin;ZHANG Xiaoman;LIANG Yingmei(The Key Laboratory for Silviculture and Conservation of Ministry of Education,Beijing Forestry University,Beijing 100083,China;Yanching Institute of Technology,Langfang 065201,Hebei,China;Museum of Beijing Forestry University,Beijing 100083,China)
出处
《菌物学报》
北大核心
2025年第6期73-84,共12页
Mycosystema
基金
国家自然科学基金(32371885,31870628)。
关键词
效应子
异源表达
亚细胞定位
植物免疫
effectors
heterologous expression
subcellular localization
plant immunity
