A2M: Exploring in-silico predicted arms-races at the plant-pathogen interface

A2M：探索植物-病原体界面的计算机预测军备竞赛

• 批准号: BB/Y000560/1
• 负责人: Renier Van Der Hoorn
• 金额: $ 75.27万
• 依托单位: University of Oxford
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2024
• 资助国家: 英国
• 起止时间: 2024 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FY000560%2F1
• 关键词: A2M; Exploring; silico; predicted; arms

The arms race at the plant-pathogen interface is a fascinating field of biology that can deliver important new, extracellular strategies for crop protection. We have used cutting edge artificial intelligence to predict novel protein-protein interactions at the plant-pathogen interface using Alphafold2 Multimer (A2M). Using an A2M screen for novel, pathogen-derived inhibitors of the secreted P69B immune protease of tomato, we discovered that P69B is targeted by (at least) five unrelated inhibitors produced by four different tomato pathogens: Epi1 from the oomycete late blight pathogen Phytophthora infestans; P3 from the bacterial spot pathogen Xanthomonas perforans; Ecp36 from the fungal leaf mould pathogen Cladosporium fulvum and TIL and Six15 from the fungal Fusarium wilt pathogen Fusarium oxysporum. That P69B is a major target for tomato pathogens is consistent with the facts that: i) P69B is induced and highly abundant in the apoplast of infected plants; ii) P69B is under positive selection in wild tomato at residues that probably interact with inhibitors; iii) P69B has nine paralogs encoded from a fast evolving gene cluster, that differ mostly in residues surrounding the substrate binding groove, where inhibitors interact. In this proposal, we aim to elucidate this arms-race and use this knowledge to engineer extracellular immunity. We will first resolve the inhibition mechanisms and determine the specificities of interactions with P69B paralogs and homologs, also from non-host plants. Second, we will determine the role of P69s in immunity and the role of inhibitors in pathogen virulence using reverse genetics on tomato and the pathogens. Third, we will elucidate the evolution of the P69 gene family in solanaceous plants and engineer inhibitor-insensitive P69s to build a strategy for durable extracellular resistance to apoplastic pathogens.

植物病原界面的军备竞赛是一个引人入胜的生物学领域，可以提供重要的新的细胞外作物保护策略。我们使用尖端人工智能使用Alphafold2多聚体（A2M）来预测植物病原体界面处的新型蛋白质 - 蛋白质相互作用。使用A2M筛选用于番茄的分泌P69B免疫蛋白酶的新颖，病原体衍生的抑制剂，我们发现P69B是（至少）五种由四种不同的番茄病原体产生的（至少）五个无关的抑制剂：EPI1从Oomycete the Oomycete the Oomycete the Oomycete the Oomycete the Oomycete晚期病原体病原体phytophthora phytophthora inferstansans产生。来自细菌斑点病原体Xanthomonas Perforans的P3； ECP36来自真菌叶片病原体cladosporium fulvum和TIL，以及来自真菌镰刀菌病原体oxysarium oxysporum的六15。 p69b是番茄病原体的主要靶标与以下事实一致：i）p69b在感染植物的凋亡中被诱导并且高度丰富； ii）p69b在可能与抑制剂相互作用的残基中呈阳性选择； iii）p69b具有从快速发展的基因簇编码的九个旁系同源物，这些基因簇主要在底物结合凹槽周围的残基上有所不同，抑制剂相互作用。在此提案中，我们旨在阐明这种武器竞赛，并利用这些知识来设计细胞外免疫。我们将首先解决抑制机制，并确定与p69b旁系同源物以及非主持人植物相互作用的特异性。其次，我们将确定p69在免疫中的作用以及使用反向遗传学在番茄和病原体上使用反向遗传学在病原体毒力中的作用。第三，我们将阐明在静电植物和工程师抑制剂不敏感的p69中p69基因家族的演变，以建立一种策略，以持续耐受性塑性病原体的细胞外耐药性。