Characterization of Blumeria graminis peptides as potential effectors targeting barley RAC/ROP proteins

小麦白粉菌肽作为针对大麦 RAC/ROP 蛋白的潜在效应物的表征

• 批准号: 178859977
• 负责人: Professor Dr. Ralph Hückelhoven
• 金额: --
• 依托单位: Lehrstuhl für Phytopathologie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2010
• 资助国家: 德国
• 起止时间: 2009-12-31 至 2013-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/178859977?language=en
• 关键词: Characterization; Blumeria; graminis; peptides; potential

Small monomeric RAC/ROP GTPases are molecular switches in cell polarity and plant defence responses. In barley RACB and related RAC/ROPs function in susceptibility to the biotrophic ascomycete pathogen Blumeria graminis f.sp. hordei (Bgh). To identify RACBinteracting proteins, we have carried out yeast two hybrid screenings of a cDNA library from infected barley. This identified six fungal clones coding for to the same class of small peptides. Protein-protein interaction was confirmed in independent assays, and a GFP-fusion protein of a fungal RACB-interacting peptide labelled small structures in the cytoplasm and cell cortex and accumulated at the site of pathogen entry. Over-expression or host-induced gene silencing of two Bgh-peptides, enhanced or reduced susceptibility to fungal entry, respectively. By the use of cell-biological and genetic tools, we would like now to provide evidence that the peptides represent fungal virulence effectors that target barley RAC/ROPs for effector-triggered susceptibility.

小单体 RAC/ROP GTP 酶是细胞极性和植物防御反应中的分子开关。在大麦中，RACB 和相关的 RAC/ROP 在对生物营养子囊菌病原体 Blumeria graminis f.sp 的易感性中起作用。大麦 (Bgh)。为了鉴定 RACB 相互作用蛋白，我们对受感染大麦的 cDNA 文库进行了酵母两种杂交筛选。这鉴定出六个编码同一类小肽的真菌克隆。蛋白质-蛋白质相互作用在独立测定中得到证实，真菌 RACB 相互作用肽的 GFP 融合蛋白标记了细胞质和细胞皮层中的小结构，并在病原体进入位点积累。两种 Bgh 肽的过度表达或宿主诱导的基因沉默，分别增强或降低对真菌侵入的敏感性。通过使用细胞生物学和遗传工具，我们现在希望提供证据，证明这些肽代表真菌毒力效应子，以大麦 RAC/ROP 为目标，实现效应子触发的易感性。