RESUMO
Transcription factors play a critical regulatory role in development by binding DNA and initiating alterations in gene transcription. The transcript of the putative Magnaporthe oryzae transcription factor-encoding gene TRA1 accumulates during germination and this accumulation was previously found to depend on the transcription factor Con7p. In the current work tra1â» mutants were generated and these strains were found to exhibit a reduced attachment, germination, appressorium formation and virulence. Adhesion to artificial and plant surfaces was affected, and FITC-labelled concanavalin A, a lectin which inhibits attachment of Magnaporthe spores, showed a reduced affinity for mutant spore tip where it normally preferentially binds. We used microarray analysis to identify Tra1p-dependent genes from two different sources: aerial structures and conidia. Mutation of 11 Tra1p-dependent genes showed that the predicted transcription factor encoding gene TDG2 is required for normal adhesion and virulence, that the genes TDG7 and TDG4 are required for normal sporulation and that TDG6 is required for wild-type levels of spore adhesion.
Assuntos
Proteínas Fúngicas/genética , Magnaporthe/genética , Esporos/genética , Fatores de Transcrição/genética , Regulação Fúngica da Expressão Gênica , Proteínas de Fluorescência Verde , Magnaporthe/crescimento & desenvolvimento , Magnaporthe/patogenicidade , Mutação , Oryza/genética , Oryza/crescimento & desenvolvimento , Oryza/microbiologia , Esporos/crescimento & desenvolvimento , Esporos/patogenicidade , Esporos Fúngicos/genética , Esporos Fúngicos/crescimento & desenvolvimento , VirulênciaRESUMO
A strain harbouring an insertion within the promoter of the CON7 gene of Magnaporthe grisea was isolated. This gene was previously shown to be essential for appressorium formation and growth in planta and is predicted to encode a transcription factor. Microarray-based gene expression analysis was used to identify several genes whose transcription during germination depends on Con7p. These include the pathogenicity factor-encoding gene PTH11 and several other genes which like PTH11 are predicted to encode G protein-coupled receptors. Microarray analysis also revealed several Con7p-dependent genes which may encode factors determining cell wall structure or function, either through the synthesis/degradation of cell wall components or by association with the cell exterior. One Con7p-dependent gene predicted to encode a class VII chitin synthase was deleted, leading to dramatic consequences on the pathogenic development of the resultant strain. Within the con7(-) mutant, a 29% reduction in chitin content of germinated spores was found and the mutant was hypersensitive to the chitin synthase inhibitor nikkomycin Z. A green fluorescent protein-tagged Con7p was found to have nuclear localization within spores. Taken together, these observations suggest that Con7p encodes a transcription factor required for the transcription of several genes which participate in disease-related morphogenesis in M. grisea.