CAREER: Dissecting the Molecular Regulation of Septin-Mediated Plant Invasion by the Blast Fungus Magnaporthe Oryzae

该奖项全部或部分由2021年美国救援计划法案（公法117- 2）资助。真菌Magnaporthe pasta导致种植水稻和小麦的高度破坏性疾病，称为稻瘟病，对世界各地的粮食安全构成持续威胁。在田间，使用有限范围的杀真菌剂来对抗稻瘟病，对这些化学物质的抗性的出现是一个迫在眉睫的威胁。因此，迫切需要开发新的控制爆炸的策略，但这需要对M。传染病生物学。感染植物，M.它产生一种专门的压力产生感染细胞，称为附着胞，它用来打破叶子内部。这项研究的重点是更好地了解这些感染细胞是如何构建的以及它们是如何工作的，因此其结果可能有助于保障全球粮食安全和提高农业生产力。拟议的活动将为植物、微生物及其相互作用的研究提供更好的生物成像培训和教育，并将为低收入和第一代高中生提供更多的机会，以获得尖端真菌细胞生物学的实践研究经验，从而促进他们参与高等教育和科学。在当地活动中传播这些研究成果将为公众参与提供更多的机会，从而提高科学素养和意识，并增强对科学和科学家的信任。它需要一个环形结构的调节装配，该环形结构由一类称为septins的细胞骨架毒性形成GTP结合蛋白组成。这种感染特异性的隔蛋白环如何在正确的地方，在正确的时间，在发育中的附着胞内形成，使植物能够入侵，在很大程度上仍然是个谜。事实上，了解septins如何在不同的细胞类型中组装和动态重塑为不同的功能架构是该领域的当前前沿。重要的是，提高了对隔膜环组装和M中功能的机械理解。真菌毒素可能会为开发新的杀真菌剂以控制稻瘟病和其他真菌作物疾病提供信息，特别是考虑到植物不含septins。本项目的目标是获得新的洞察力的分子机制，其中初期隔蛋白盘状结构形成在附着胞的基础上，并重塑成环，以及这些皮层结构如何组织渗透界面植物入侵。该项目结合使用时间分辨邻近蛋白质组学，活细胞成像，和反向遗传学，以功能性地映射隔蛋白组织的附着胞基底皮层和穿透界面，并采用新的基于荧光显微镜的基因组，广泛的化学诱变筛选，以确定Septin环组装的新型调节剂。该奖项反映了NSF的法定使命，并被认为值得通过使用基金会的知识价值和更广泛的影响审查标准。

This award is funded in whole or in part under the American Rescue Plan Act of 2021 (Public Law 117- 2) The fungus Magnaporthe oryzae causes a highly destructive disease of cultivated rice and wheat, called blast, which poses an ongoing threat to food security around the world. In the field, blast is combatted using a limited range of fungicides, and the emergence of resistance to these chemistries is a looming threat. Thus, there is an urgent need to develop new strategies to control blast, but this demands a more complete mechanistic understanding of M. oryzae infection biology. To infect plants, M. oryzae produces a specialized pressure-generating infection cell, called an appressorium, which it uses to break inside of leaves. This research focuses on gaining a better understanding of how these infection cells are built and how they work, and therefore the outcomes may help to safeguard global food security and enhance agricultural productivity. The proposed activities will provide improved training and education in bioimaging for the study of plants, microbes and their interactions, and will provide enhanced opportunities for low-income, and first-generation high school students to gain hands-on research experience in cutting-edge fungal cell biology, therein promoting their participation in higher-education and science. Communication of these research outcomes at local events will provide enhanced opportunities for public engagement, leading to increased scientific literacy and awareness and greater trust in science and scientists.Appressorium-mediated plant invasion by M. oryzae requires the regulated assembly of a toroidal ring structure, composed of a class of cytoskeletal filament-forming GTP-binding proteins called septins, within its base. How this infection-specific septin ring forms in the right place, and at the right time, within the developing appressorium to enable plant invasion, remains largely mysterious. Indeed, understanding how septins are assembled and dynamically remodeled into different functional architectures in diverse cell types is a current frontier in the field. Critically, improved mechanistic understanding of septin ring assembly and function in M. oryzae may inform the development of new fungicides to control blast, and other fungal crop diseases, especially given that plants do not contain septins. The goals of this project are to gain new insight into the molecular mechanisms by which incipient septin disc-like structures form in the base of appressoria and are remodeled into rings, and how these cortical structures organize the penetration interface for plant invasion. This project combines the use of time-resolved proximity proteomics, live cell imaging, and reverse genetics to functionally map the septin-organized appressorium basal cortex and penetration interface and employs a novel fluorescence microscopy-based genome-wide chemical mutagenesis screen to identify novel regulators of septin ring assembly.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.