Analysis of Magnaporthe grisea pathogenicity by insertional mutagenesis and hierarchical metabolomics

该项目将利用高通量分析生物化学技术，提供有关谷物被一种具有重要经济意义的植物病原真菌感染的新信息。真菌导致世界上最严重和最具破坏性的作物疾病，该项目将提供关于这些微生物病原体感染植物过程的新信息。我们将调查一种名为稻瘟菌的真菌，它会引起稻瘟病，这种疾病每年摧毁足够养活6000万人的大米。我们已经产生了在植物感染中受损的真菌突变，这些突变将受到代谢组分析的影响。代谢组学技术将使我们能够分解真菌细胞中的许多低分子化合物(有机酸、氨基酸、糖和多元醇)，并为所研究的每个突变菌株产生代谢物指纹。我们将研究一组在糖原、脂肪和海藻糖代谢方面受到影响的突变株，这些突变株是在真菌产生的特殊感染结构中形成膨压所必需的。为了侵入稻叶，这种真菌会产生巨大的压力，使谷类作物叶片的角质层破裂。我们还将研究一组新的突变体，这些突变体在其定植活植物组织的能力方面受到影响，导致小的或不合时宜的疾病损伤。我们计划使用的技术允许同时对细胞内包含的所有小分子(数百种代谢物)进行全局分析。这一全面的分析将确保研究新陈代谢的所有主要领域，这将使我们能够对正常的野生型真菌和每个突变株之间存在的确切差异提出新的想法。这些复杂的数据需要专门的数据分析方法来确定菌株之间的代谢差异。在未来，我们预计可能会开发出新的杀菌剂和疾病控制策略，以扰乱本研究中确定的与毒力相关的代谢过程。

The project will utilize high throughput analytical biochemistry techniques to provide new information concerning the infection of cereals by an economically significant plant pathogenic fungus. Fungi cause the world's most serious and devastating crop diseases and this project will provide new information about the process of plant infection by these microbial agents of disease. We will investigate a fungus called Magnaporthe grisea which causes rice blast disease, a disease that destroys enough rice each year to feed 60 million people. We have produced mutants of the fungus that are impaired in plant infection, and these will be subject to metabolome analysis. Metabolomics techniques will allow us to resolve many of the low molecular weight compounds in the cells of the fungus (organic acids, amino acids, sugars, and polyols) and produce metabolite fingerprints for each mutant strain investigated. We will investigate a set of mutants affected in glycogen, lipid and trehalose metabolism that are required for development of turgor pressure in the specialised infection structures produced by the fungus. To break into a rice leaf, the fungus generates enormous pressure that allows the cuticle of the cereal leaf to be broken. We will also investigate a novel set of mutants that are affected in their ability to colonize living plant tissue, resulting in small, or mis-timed disease lesions. The techniques we propose to use allow the simultaneous global analysis of all the small molecules (several hundreds of metabolites) contained within a cell. This comprehensive analysis will ensure that all major areas of metabolism are studied which will allow us to develop new ideas of the precise differences exist between the normal, wild type fungus and each mutant. These complex data require specialised methods for data analysis to determine metabolic differences between strains. In the future, we expect that novel fungicides and disease-control strategies may be developed that disrupt the virulence-associated metabolic processes identified in this study.