Genome-wide functional analysis of pathogenicity genes in the rice blast fungus

Abstract

Rapid translation of genome sequences into meaningful biological information hinges on the integration of multiple experimental and informatics methods into a cohesive platform. Despite the explosion in the number of genome sequences available, such a platform does not exist for filamentous fungi. Here we present the development and application of a functional genomics and informatics platform for a model plant pathogenic fungus, Magnaporthe oryzae. In total, we produced 21,070 mutants through large-scale insertional mutagenesis using Agrobacterium tumefaciens-mediated transformation. We used a high-throughput phenotype screening pipeline to detect disruption of seven phenotypes encompassing the fungal life cycle and identified the mutated gene and the nature of mutation for each mutant. Comparative analysis of phenotypes and genotypes of the mutants uncovered 202 new pathogenicity loci. Our findings demonstrate the effectiveness of our platform and provide new insights on the molecular basis of fungal pathogenesis. Our approach promises comprehensive functional genomics in filamentous fungi and beyond.

Inhibition of histone deacetylase causes reduction of appressorium formation in the rice blast fungus Magnaporthe oryzae

“…In recent years, numerous studies have focused on the molecular mechanisms of appressorium formation and many genes involved in this process have been reported (Wilson and Talbot, 2009), for example by SAGE analyses (Kronstad, 2006;Matsumura and Terauchi, 2007) and T-DNA insertional mutagenesis (Jeon et al, 2007). This study tested the effects of TSA and other HDAC inhibitors at the early infection stage in M. oryzae.…”

- Enhanced Disease Susceptibility 1 and Salicylic Acid Act Redundantly to Regulate Resistance Gene-Mediated Signaling

“…A T-DNA insertion mutant (ATMT0144A2) showing reduced virulence was identified from the M. oryzae ATMT mutant library [26]. This mutant developed restricted resistant-type lesions on a susceptible rice cultivar, Nakdongbyeo, and the number of lesions was much less than in the wild-type strain (Fig.…”

“…Complete genome sequence information is available for both the host and the pathogen, and various molecular functional genomics approaches have been initiated [25]. To investigate pathogenicity genes on a genome scale, our research group has generated .20,000 insertional mutants using Agrobacterium tumefaciens-mediated transformation (ATMT) and has evaluated the characteristics of each mutant in the essential steps for disease development [26]. The disease cycle of this pathogen consists of several steps that are essential for successful disease development.…”

Identification of new pathogenicity related to the gene encoding hypothetical protein in the gray mold fungus (Botrytis cinerea)

“…However, it is important to investigate each gene in a more descriptive way, because it cannot be considered to prove that the integration of T-DNA is almost associated with the real phenotypic expression, which is also previously manifested by scientist (Blaise 2007;Jeon 2007;Michielse et al, 2009). The alteration of gene function via T-DNA integration led to analysis of phenotypes than the targeted deletions of the relevant genes (Idnurm 2004.).…”