Deconstruction of the phytotoxic secretome to decipher the necrotrophic pathogenesis of Botrytis cinerea

解构植物毒性分泌组以破译灰葡萄孢的坏死营养发病机制

• 批准号: 497740939
• 负责人: Professor Dr. Matthias Hahn
• 金额: --
• 依托单位: Abteilung für Pflanzenbiochemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/497740939?language=en
• 关键词: Deconstruction; phytotoxic; secretome; decipher; necrotrophic

Botrytis cinerea is one of the economically most important plant pathogens worldwide, causing pre- and postharvest grey rot on numerous crops. During infection, the fungus secretes a complex mixture of lytic enzymes and phytotoxic proteins and metabolites, thereby inducing rapid necrosis of host plant cells. To better understand the key infection mechanism of this broad host range necrotroph, we are using a subtractive approach to serially eliminate all plant cell death-inducing proteins (CDIPs) and metabolites, until complete loss of necrosis induction of the resulting mutants. To this end, we have established a highly efficient CRISPR/Cas editing method to generate multiple mutants within short time. Our preliminary work has shown that each of the known CDIPs secreted by B. cinerea contributes only to a minor extent to infection. To fully address the high redundancy of phytotoxic compounds, we want to identify the as yet unknown number of remaining CDIPs and analyse their roles in different stages of necrotrophic infection, by investigating the phenotypes of single and multiple mutants. We are further studying the interaction of CDIPs with their plant receptors or targets. Several fungal CDIPs have been shown to activate pattern recognitions receptors (PRR) at the cell surface, which upon activation induce a basic plant defence called PTI (pattern triggered immunity). Notably, PTI-activation by Botrytis often results eventually in plant cell death, a rather unusual feature of this basic plant defence. Therefore, we want to investigate how Botrytis is using CDIPs to exploit PTI for its own benefit without being stopped by plant defence mechanisms. To identify plant CDIP receptors, screenings will be performed with Arabidopsis PRR mutants and with Arabidopsis and tomato ecotypes that vary in their genetic PRR equipment. Identification of receptor-ligand pairs will allow functional insight into plant cell death induction and might lead to a better understanding of the wide host range of Botrytis. We believe that our proposed project will shed light on a central aspect of microbial pathogenesis, and will pave the way towards a comprehensive understanding of the necrotrophic lifestyle of B. cinerea.

灰葡萄孢（Botrytis cinerea）是世界上最重要的经济植物病原体之一，在许多作物上引起采前和采后灰腐病。在感染期间，真菌分泌溶解酶和植物毒性蛋白质和代谢物的复杂混合物，从而诱导宿主植物细胞的快速坏死。为了更好地理解这种广泛宿主范围的坏死营养型的关键感染机制，我们正在使用消减方法来连续消除所有植物细胞死亡诱导蛋白（CDIP）和代谢物，直到所得突变体的坏死诱导完全丧失。为此，我们建立了一种高效的CRISPR/Cas编辑方法，可以在短时间内产生多个突变体。我们的初步工作表明，由B分泌的每一种已知的CDIP。灰霉病对感染的影响很小。为了充分解决植物毒性化合物的高冗余性，我们希望通过研究单个和多个突变体的表型来确定剩余CDIP的未知数量，并分析它们在坏死营养型感染的不同阶段中的作用。我们正在进一步研究CDIP与其植物受体或靶标的相互作用。几种真菌CDIP已显示激活细胞表面的模式免疫受体（PRR），其在激活后诱导称为PTI（模式触发免疫）的基本植物防御。值得注意的是，葡萄孢菌的PTI激活通常最终导致植物细胞死亡，这是这种基本植物防御的一个相当不寻常的特征。因此，我们想研究葡萄孢菌是如何利用CDIP来利用PTI为自己的利益而不被植物防御机制所阻止的。为了鉴定植物CDIP受体，将用拟南芥PRR突变体以及在其遗传PRR设备上不同的拟南芥和番茄生态型进行筛选。受体-配体对的鉴定将允许对植物细胞死亡诱导的功能性洞察，并可能导致更好地理解葡萄孢属的广泛宿主范围。我们相信，我们提出的项目将阐明微生物发病机制的一个核心方面，并将铺平道路，全面了解的坏死营养型生活方式的B。灰叶