Knock-out And Functional Characterization Of Several Genes Regulated By MoSom1in Magnaporthe Oryzae

In our previous research, a novel transcriptional regulator, MoSom1, which was proved to act on the downstream of cAMP-PKA signaling pathway and impact the morphodifferentiation of M. oryzae and plant infection. To study the expression of genes regulated by MoSoml, we performed global gene expression profile sequencing to Δmosoml and discovered that many genes expression was affected remarkably due to inactivation of MoSom1. In this study, we chose4significant differentially expressed genes to perform gene deletion and functional characterization, so as to find new MoSom1-related and pathogenicity-related genes. Meanwhile, according to bioinformatics analysis, we selected two transcription factors containing zinc finger structure and one harbouring ARID DNA-domain from M. oryzae genome to further perform targeted gene replacement, in the hope of discovering new transcription factors regulating important functional biology of M. oryzae. However, all the above genes deletion didn’t result in obvious biological phenotype variation in morphogenesis and virulence.In addition, another previous study identified two genes MoMET13and MoMET12encoding methylenetetrahydrofolate reductase （MTHFR） and particpating in the folate cycle of methionine syntheses. Deletion of MoMET13led to morphogenesis defect and no pathogenicity to barely and rice inoculated with mutant Δmome13cultured on methionine-free medium. To investigate the impact of MTHFR disruption to other genes involved in methionine pathway, we carried out qRT-PCR to analyse the expression difference of relative genes. The result indicated all the targeting genes are differentially expressed in mutant strains. Additionally, to examine the functional conservation of MTHFR in fungi, we introduced a homologous gene FGSG₀9572from F, graminearum to M. oryzae mutant of Δmomet13, but there is no complementary mutant founded until...