The role of fungal Argonautes and other types of RNA-binding proteins in Botrytis-plant cross-kingdom RNAi

真菌 Argonautes 和其他类型的 RNA 结合蛋白在 Botrytis-植物跨界 RNAi 中的作用

• 批准号: 447515952
• 负责人: Dr. Arne Weiberg
• 金额: --
• 依托单位: Institut für Genetik
• 依托单位国家: 德国
• 项目类别: Research Units
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/447515952?language=en
• 关键词: role; fungal; Argonautes; other; types

Botrytis cinerea is an aggressive fungal plant pathogen. For successful infection, B. cinerea secretes cross-kingdom small RNAs (ck-sRNAs) into plant cells that hijack the plant Argonaute/RNA-induced silencing complex (AGO/RISC) to suppress important host defence genes. This fascinating virulence strategy is called cross-kingdom RNA interference (ck-RNAi). Arabidopsis thaliana, host plant of Botrytis, transfers its ck-sRNAs into the fungus to suppress pathogen genes, too. Although the underlying mechanisms of host-induced gene silencing and ck-sRNA transport bear great potential for plant protection purposes, they are unfortunately, largely unknown. With this subproject, we propose to shed light on the roles of the Botrytis BcAGOs and other types of RNA-binding proteins (RBPs) in bidirectional plant-fungal ck-RNAi. i) We aim to identify tomato ck-sRNAs that bind to BcAGOs and direct host-induced gene silencing. We will characterize main pathogen target genes of tomato ck-sRNAs, which will inform on the host-induced gene silencing mechanism and on promising fungal target genes to improve plant protection. ii) We propose to investigate the role BcAGOs in sorting ck-sRNAs for extracellular RNA transport. In plants and animals, AGO proteins associate with endogenous membrane structures of the Endoplasmic Reticulum and multivesicular bodies. We aim to elucidate the inter-connection of BcAGOs with endogenous secretory pathways. This will broaden our understanding of the sorting and secretion mechanism of ck-sRNAs. Moreover, extracellular vesicles (EVs) are suggested to mediate RNA transport in plant-fungal interaction. Our laboratory recently discovered that Botrytis EVs contain ck-sRNAs and putative RBPs. iii) We propose to investigate the role and function of EV-RBPs in the ck-sRNA transport process. This information is highly valuable for implementation into existing RNA-mediated plant protection strategies to improve efficiency, a major goal of the RU5116. Knowledge gained by this subproject is highly complementary to the other projects of the RU5116, not only by providing new information on the ckRNAi mechanism, but also by delivering valuable information that can be translated into innovative RNA-based plant protection strategies.

灰霉病菌是一种侵袭性较强的植物真菌病原菌。为了成功感染，灰霉菌将跨王国的小RNA(ck-sRNAs)分泌到植物细胞中，劫持植物ArgAerte/RNA诱导的沉默复合体(AGO/RISC)，以抑制重要的宿主防御基因。这种令人着迷的毒力策略被称为跨王国RNA干扰(ck-RNAi)。拟南芥是番茄灰霉病菌的寄主植物，它将Ck-sRNAs转移到真菌体内以抑制病原菌基因。尽管寄主诱导的基因沉默和Ck-SRNA转运的潜在机制在植物保护方面具有巨大的潜力，但不幸的是，它们在很大程度上是未知的。通过这个子项目，我们打算阐明Botrytis BcAGos和其他类型的RNA结合蛋白(RBPs)在双向植物-真菌Ck-RNAi中的作用。I)我们的目标是识别与BcAGO结合的番茄Ck-sRNAs，并直接诱导宿主诱导的基因沉默。我们将对番茄Ck-sRNAs的主要致病靶基因进行特征分析，这将有助于了解寄主诱导的基因沉默机制和有望提高植物保护的真菌靶标基因。Ii)我们建议研究BcAGO在分离Ck-sRNA进行细胞外RNA运输中的作用。在植物和动物中，AGO蛋白与内质网和多囊泡体的内源性膜结构有关。我们的目标是阐明BcAGO与内源性分泌途径之间的相互联系。这将拓宽我们对ck-sRNA分选和分泌机制的理解。此外，细胞外小泡(EVS)被认为在植物-真菌相互作用中介导了RNA的运输。我们实验室最近发现Botrytis EV含有Ck-sRNA和推测的限制性商业惯例。Iii)我们建议研究EV-RBPs在Ck-sRNA转运过程中的作用和功能。这些信息对于实施现有的RNA介导的植物保护策略以提高效率非常有价值，这是RU5116的主要目标。这一分项目获得的知识与RU5116的其他项目具有很强的互补性，不仅提供了有关ck RNAi机制的新信息，而且提供了可转化为创新的基于RNA的植物保护战略的宝贵信息。