Project description:RNAi is a conserved mechanism in eukaryotic organisms and has been the subject of recent attention pertaining to many animal and plant models. But research related to fungal systems is limited, especially in plant pathogenic models. The present study was undertaken to examine the involvement of RNAi components on the biogenesis of small RNAs and their impact on growth and pathogenicity of the plant pathogenic model organism Magnaporthe oryzae. Through genetic and genomic analyses, we demonstrate that an individual Dicer, an RdRP and an Argonaute are essential for the generation of sRNAs. Collectively, they are involved in biogenesis of sRNAs matching to coding regions, repeats and intergenic regions. The loss of a particular Argonaute made the fungus less virulent on barley leaves and was also essential for sRNA production. Furthermore, one specific RdRP that was not essential to sRNA generation nonetheless reduced fungal virulence. In total, ten deletion mutants were constructed in each of the putative sRNA-generating genes, as well as a double deletion mutant in the two Dicers, and comparison of sRNA and RNA-seq profiles of each mutant yielded new insights into the involvement of sRNAs in transcriptional regulation and intergenic regions in M. oryzae. Together, these data support that RNAi machinery in M. oryzae is not only involved in generation of sRNAs, but is involved in regulating various developmental processes including fungal growth and virulence.
Project description:This SuperSeries is composed of the following subset Series: GSE8517: Magnaporthe oryzae gene expression during biotrophic invasion of rice using version 2 of the Agilent Magnaporthe grisea Array (G4137B). GSE8518: Rice gene expression during biotrophic invasion by the rice blast fungus Magnaporthe oryzae using the Agilent Rice Array (G4138A). Keywords: SuperSeries Refer to individual Series
Project description:The rice blast fungus, Magnaporthe oryzae is a destructive pathogen of rice and other related crops, causing significant yield losses worldwide. Endogenous small RNAs (sRNAs), including small interfering RNAs (siRNAs) and microRNAs (miRNAs) are critical components of gene regulation in many eukaryotic organisms. Recently several new species of sRNAs have been identified in fungi. This fact along with the availability of genome sequence makes M. oryzae a compelling target for sRNA profiling. We have examined sRNA species and their biosynthetic genes in M. oryzae, and the degree to which these elements regulate fungal stress responses. To this end, we have characterized sRNAs under different physiological stress conditions, which had not yet been examined in this fungus. The resulting libraries are composed of more than 37 million total genome matched reads mapping to intergenic regions, coding sequences, retrotransposons, inverted, tandem, and other repeated regions of the genome with more than half of the small RNAs arising from intergenic regions. The 24 nucleotide (nt) size class of sRNAs was predominant. A comparison to transcriptional data of M. oryzae undergoing the same physiological stresses indicates that sRNAs play a role in transcriptional regulation for a small subset of genes. Support for this idea comes from generation and characterization of mutants putatively involved in sRNAs biogenesis; our results indicate that the deletion of Dicer-like genes and an RNA-Dependent RNA Polymerase gene increases the transcriptional regulation of this subset of genes, including one involved in virulence. Various physiological stressors and in planta conditions alter the small RNA profile of the rice blast fungus. Characterization of sRNA biosynthetic mutants helps to clarify the role of sRNAs in transcriptional control.
Project description:The rice blast fungus, Magnaporthe oryzae is a destructive pathogen of rice and other related crops, causing significant yield losses worldwide. Endogenous small RNAs (sRNAs), including small interfering RNAs (siRNAs) and microRNAs (miRNAs) are critical components of gene regulation in many eukaryotic organisms. Recently several new species of sRNAs have been identified in fungi. This fact along with the availability of genome sequence makes M. oryzae a compelling target for sRNA profiling. We have examined sRNA species and their biosynthetic genes in M. oryzae, and the degree to which these elements regulate fungal stress responses. To this end, we have characterized sRNAs under different physiological stress conditions, which had not yet been examined in this fungus. The resulting libraries are composed of more than 37 million total genome matched reads mapping to intergenic regions, coding sequences, retrotransposons, inverted, tandem, and other repeated regions of the genome with more than half of the small RNAs arising from intergenic regions. The 24 nucleotide (nt) size class of sRNAs was predominant. A comparison to transcriptional data of M. oryzae undergoing the same physiological stresses indicates that sRNAs play a role in transcriptional regulation for a small subset of genes. Support for this idea comes from generation and characterization of mutants putatively involved in sRNAs biogenesis; our results indicate that the deletion of Dicer-like genes and an RNA-Dependent RNA Polymerase gene increases the transcriptional regulation of this subset of genes, including one involved in virulence. Various physiological stressors and in planta conditions alter the small RNA profile of the rice blast fungus. Characterization of sRNA biosynthetic mutants helps to clarify the role of sRNAs in transcriptional control. Examination of small RNA populations of eight mycelial as well as In planta tissues of M. oryzae strain 70-15. Mycelia grown in complete medium (CM), minimal medium (MM), carbon starved medium (CS: MM lacking carbon source), nitrogen starved medium (NS: MM lacking nitrogen source) and a reactive oxygen species-rich environment generated by amendment of Paraquat (PQ) to the CM. In planta tissues include mock inoculated rice leaves (LMg0), 72 hours post-inoculation in rice leaves (LMg72) and 96 hours post-inoculation in rice leaves (LMg96).
Project description:Mycelium from the rice blast fungus Magnaporthe oryzae was grown in both rich medium and under nutrient limiting conditions. Genes were identified that were more highly expressed in one condition as compared to the other.
Project description:Mycelium from the rice blast fungus Magnaporthe oryzae was grown in both rich medium and under nutrient limiting conditions. Genes were identified that were more highly expressed in one condition as compared to the other.
Project description:We report that the homeobox trasncription factor called HOX7 controls hyphae-associated genes, autophagy and cell cycle related genes necessary for appressorium development in the rice blast fungus Magnaporthe oryzae. We also report that the ste transcription factor MST12 regulates gene functions involved in septin reorientation.