Molecular and Cellular Biology of Biotrophic Interactions in Rice Blast Disease

稻瘟病生物营养相互作用的分子和细胞生物学

• 批准号: 0446315
• 负责人: Barbara Valent
• 金额: $ 48.59万
• 依托单位: Kansas State University
• 依托单位国家: 美国
• 项目类别: Continuing Grant
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-15 至 2007-12-31
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=0446315&HistoricalAwards=false
• 关键词: Molecular; Cellular; Biology; Biotrophic; Interactions

Title: Molecular and Cellular Biology of Biotrophic Interactions in Rice Blast Disease Successful pathogenesis by plant-infecting fungi often requires an initial period of plant-fungal coexistence, and yet little is known about how a fungus suppresses plant defense mechanisms and establishes itself inside the plant. Rice blast disease occurs when the fungus, Magnaporthe grisea, punctures through the outer plant surface and produces specialized bulbous invasive hyphae inside living rice cells. This project will focus on microscopy coupled with fungal and plant gene expression analyses of the first and second invaded rice cells. Live-cell fluorescence confocal microscopy will be used to define the nature of the plant cytoplasm-fungus interface as invasive hyphae fill the first epidermal cell, approximately 24 to 35 hours after inoculation. Laser Microdissection (LM) and whole genome microarrays, of both fungus and rice, will provide a deep analysis of biotrophic fungal gene expression and of rice cell responses. Similar coupled microscopy and LM-microarray analysis will be performed after 35 hrs when the fungus has completely filled the first epidermal cell and is sending invasive hyphae into all neighboring cells. Cellular and molecular interactions in a successful biotrophic infection will be compared to the near-isogenic interaction in which fungal growth is blocked by Pi-ta resistance gene-mediated hypersensitive resistance. This research will identify genes showing differential expression in biotrophic hyphae or in plant cells responding to these hyphae. Expression patterns will be confirmed for differentially-expressed fungal genes. Functional analysis of biotroph-specific fungal genes will include live-cell microscopy of infection by gene-disruption mutants to link molecular and cellular phenotypes.Broader Impacts: Understanding the cellular and molecular mechanisms that determine if crop plants succumb to or resist disease will lead the way to developing durable disease resistance. Food security in the coming century depends on decreasing the potential for rice blast disease to cause catastrophic losses to rice production. In addition, studying biotrophic blast infection will aid research on other hemibiotrophic and biotrophic fungi that cause many plant diseases threatening agriculture. Success in using LM to define temporal and spatial details early in development of rice blast disease has the potential to revolutionize studies of fungus-plant interactions. This technology has exciting potential to enable a deep molecular analysis of plant infection structures formed by obligate pathogens such as the rust fungi, which fail to grow outside of plant tissue. This project will provide a stimulating learning and research experience for graduate and undergraduate students at K-State. The strikingly visual picture of an important fungus-plant struggle that emerges from this research will be presented in lectures at high schools and community colleges. A special challenge lies in attracting kids from an increasingly urbanized population to careers in agricultural research on organisms that are critical for food security in the US and worldwide. This research will be published and made available on the Valent Lab Web Site (http://www.oznet.ksu.edu/plantpath/valentlab/) or on M. grisea community web sites such as the NSF-funded "MGOS" database (http://www.mgosdb.org/

职务名称：稻瘟病生物营养相互作用的分子和细胞生物学感染植物的真菌的成功致病通常需要植物-真菌共存的初始阶段，但对真菌如何抑制植物防御机制并在植物体内建立自己知之甚少。 稻瘟病菌是一种真菌，稻瘟病菌，穿透植物的外表面，在水稻细胞内产生特殊的球茎状侵入菌丝。 该项目将侧重于显微镜结合真菌和植物基因表达分析的第一和第二入侵水稻细胞。 活细胞荧光共聚焦显微镜将用于定义植物细胞质-真菌界面的性质，因为在接种后约24至35小时，侵入性菌丝填充第一个表皮细胞。 真菌和水稻的激光显微切割（LM）和全基因组微阵列将为活体营养真菌基因表达和水稻细胞反应提供深入分析。 类似的显微镜和LM-微阵列分析将在35小时后进行，此时真菌已完全填充第一个表皮细胞并将侵入性菌丝送入所有相邻细胞。 将成功的活体营养型感染中的细胞和分子相互作用与近等基因相互作用进行比较，其中真菌生长被Pi-ta抗性基因介导的过敏性抗性阻断。 这项研究将确定在活体营养菌丝或植物细胞对这些菌丝的反应中显示差异表达的基因。 将确认差异表达的真菌基因的表达模式。 生物营养型真菌基因的功能分析将包括基因破坏突变体感染的活细胞显微镜，以连接分子和细胞表型。更广泛的影响：了解决定作物植物是否屈服于或抵抗疾病的细胞和分子机制将导致发展持久的抗病性。 未来世纪的粮食安全取决于减少稻瘟病对水稻生产造成灾难性损失的可能性。 此外，研究生体营养型菌群感染将有助于对其他半生体营养型和生体营养型真菌的研究，这些真菌导致许多威胁农业的植物疾病。 成功地使用LM来定义时间和空间的细节在水稻稻瘟病的发展早期有可能彻底改变真菌-植物相互作用的研究。 这项技术具有令人兴奋的潜力，可以对由专性病原体（如锈菌）形成的植物感染结构进行深入的分子分析，这些病原体无法在植物组织外生长。 该项目将为K州的研究生和本科生提供刺激的学习和研究经验。 这项研究中出现的一个重要的真菌-植物斗争的惊人的视觉画面将在高中和社区大学的讲座中呈现。 一个特殊的挑战在于吸引来自日益城市化人口的孩子从事对美国和世界粮食安全至关重要的生物农业研究。 本研究将在Valent Lab网站（http：//www.oznet.ksu.edu/plantpath/valentlab/）或M.稻瘟病菌社区网站，如国家科学基金会资助的“MGOS”数据库（http：www.mgosdb.org/