Integrated Analyses of the Sites- and Mode-of-Action of the ToxA Virulence Determinant from the Fungal Wheat Pathogen, Pyrenophora Tritici-Repentis

小麦黑斑病是由小麦黑粉病病原真菌Prenophora tritici-prepentis引起的一种具有破坏性和重大经济意义的病害，其特点使其非常适合于研究病原菌的致病力和植物对病害的敏感性。例如，真菌产生多种蛋白质毒素，其中每一种都是其致病能力所必需的，具体取决于宿主的基因构成。了解这些毒素是如何发挥作用的，应该能为病原体如何导致疾病以及植物如何变得容易受到病原体的攻击提供实质性的见解。该项目将评估其中一种名为PTR Toxa的毒素，并调查这种蛋白质在细胞水平上如何以及在哪里相互作用。实验方案是高度集成的，并结合了多种方法，包括生化、细胞学、分子和遗传学分析，旨在识别相互作用的植物伙伴。这些伙伴包括宿主蛋白，这些蛋白介导其摄取，促进细胞内转运，并构成毒素的作用部位。这些目标的成功完成不仅有助于理解病原菌毒力的基本生物学和控制寄主疾病易感性的机制，而且还有助于理解对植物生物学至关重要的过程，包括受体生物学和功能、细胞内蛋白质运输和叶绿体结构和功能。该项目将通过多种方式支持培训和外联活动，包括：在传统大学举办一个关于植物-微生物分子相互作用的研讨会；传统大学是一所位于华盛顿州顶端的Yakama印第安人保留地的非宗派私立附设高等教育机构；在俄勒冈州立大学开展为期11周的实习计划，招待选定的传统大学学生；以及在对植物科学具有广泛影响的领域为博士后助理或研究助理、研究生和本科生提供直接培训经验。

The plant disease, tanspot, caused by the pathogenic fungus, Pyrenophora tritici-repentis, is a destructive and economically significant disease of wheat, which exhibits characteristics that make it ideally suited for investigating properties of pathogen virulence and plant disease susceptibility. For example, the fungus produces multiple protein toxins each of which is required for its ability to cause disease depending on the genetic makeup of the host. Understanding how these toxins function should provide substantial insight into how pathogens cause disease and how plants become vulnerable to pathogens. This project will evaluate one of these toxins called Ptr ToxA and investigates how and where this protein interacts at the cellular level. The experimental protocol is highly integrative and incorporates a diversity of approaches including biochemical, cytological, molecular, and genetical analyses designed to identify interacting plant partners. Such partners include host proteins that mediate its uptake, facilitate intracellular transport, and comprise the toxin's site-of-action. Successful completion of the objectives promises to be transformative not only for its contribution to understanding the basic biology of pathogen virulence and mechanisms that govern host disease susceptibility, but also for understanding processes fundamentally important to plant biology including, receptor biology and function, intracellular protein transport and chloroplast structure and function. The project will support training and outreach activities in several ways including: the implementation of a workshop in molecular plant-microbe interactions at Heritage University, a nondenominational private, accreted institution of higher education sited on the Yakama Indian Nation Reservation in Toppenish Washington; conducting an eleven-week internship program at Oregon State University hosting select Heritage University students; and direct training experiences in an area with broad implications to the plant sciences for a postdoctoral associate or a research assistant and graduate and undergraduate students.