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Differential Expression Analysis of a Histidine phosphotransferase RNAi mutant in Magnaporthe oryzae

ABSTRACT: Differential Expression Analysis of a Histidine phosphotransferase RNAi mutant in Magnaporthe oryzae

PROVIDER: PRJNA793841 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR17410087_1.fastq.gz	Fastqsanger.gz
	SRR17410087_2.fastq.gz	Fastqsanger.gz
	SRR17410088_1.fastq.gz	Fastqsanger.gz
	SRR17410088_2.fastq.gz	Fastqsanger.gz

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Differential Expression Analysis of a Histidine phosphotransferase RNAi mutant in Magnaporthe oryzae

Project description:Magnaporthe oryzae is notorious for its efficient evolution as a plant pathogen. Analysis of well organised fungal pathogenic mechanisms would enable the scientific understanding to fight against pathogens. Histidine phosphotransferase in fungal systems are good targets for antifungals since they are component of SAPK pathway, which is essential for survival of the fungus. The histidine Phosphotransferase YPD1 in M. oryzae is known to be light responsive and important for both asexual reproduction as well as pathogenicity. In this study, the knock-down (KD) mutant of YPD1 was highly sensitive to cell wall perturbing agents and were highly deficient in laccase expression and activity. A RNA-seq transcriptome screening of KD mutant compared to wild type revealed altered expression of about 146 genes important in biotrophic and necrotrophic phases of the pathogen. About 96 genes were found to be down regulated, of which several plant cell wall degrading genes vital for pathogenicity and fungal cell wall maintenance genes important for growth and fungal development were identified. Most of the genes identified are involved in biotrophic invasion and metabolism. Our results explain the reason for the deficiency of the KD mutant in asexual reproduction as well as pathogenicity. Analysis of differentially expressed genes in the KD YPD1 mutant suggests that YPD1 can be a good target for development of novel antifungal compounds, as YPD1 expression affects genes required for pathogenicity.

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Characterization of RNAi genes and their impact on sRNA biogenesis, growth and development of the rice blast fungus, Magnaporthe oryzae

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.

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