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Gene expression profiles of 4 non-pathogenic mutants in the phytopathogenic fungus Botrytis cinerea

ABSTRACT: Gene expression profiles of 4 non-pathogenic mutants in the phytopathogenic fungus Botrytis cinerea

PROVIDER: PRJNA553802 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR9668420.fastq.gz	Fastqsanger.gz
	SRR9668430.fastq.gz	Fastqsanger.gz
	SRR9668421.fastq.gz	Fastqsanger.gz
	SRR9668431.fastq.gz	Fastqsanger.gz
	SRR9668422.fastq.gz	Fastqsanger.gz

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Gene expression profiles of 4 non-pathogenic mutants in the phytopathogenic fungus Botrytis cinerea

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Unravelling the initial triggers of Botrytis cinerea infections. Botrytis cinerea surfactome

Project description:Botrytis cinerea, the causal agent of the gray mold, is one of the most important phytopathogenic fungi due to its ability to affect more than 1400 cultivable vegetal species. Signal transduction cascades mediate the dialogue between the environment, the plant and the fungus, which is essential in the infection process of B. cinerea. In this context, surface proteins (surfactome) have an important role as the first receptors in the signaling cascades, connecting the fungus response to the environment changes. Moreover, the relevant role of surfactome in infection process of pathogenic microorganism has been described, but nothing has been reported in B. cinerea. Therefore, in order to unravel new proteins and to complete the whole view of signal transduction cascades during infection of B. cinerea, surfactome of this fungus has been analyzed under two plant-based elicitors previously validated by the research group: glucose (GLU) as a constitutive stage and deproteinized tomato cell wall (TCW) as a virulence inductor. To identify the B. cinerea surfactome, the trypsin shaving of intact living cells approach has been optimized. Subsequently, all the peptide mixture obtained in surfactome isolation was analyzed using LC-MS/MS to identify their components. Our results shows that none of the used protocols disturbs the hyphae integrity, obtaining the best when PBS buffer plus 30% sucrose was used. In conclusion, in this work it has been carried out for the first time the optimization of a surface protein extraction protocol in Botrytis cinerea giving rise to a new subproteome; the surfactome. Integration of surfactome data inside the previous proteomics data collected by the research group has improved the complete view of signal transduction cascades during infection of B. cinerea, helping to unravel key points in the regulation of this process and therefore to further develop new and better discriminatory fungicides from a molecular approach.

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Proteomic analysis of several non pathogenic mutants from Botrytis cinerea.

Project description:The grey mould fungus Botrytis cinerea is a typical necrotrophic fungus that can infect hundreds of host plants, including high-value crops such as grapevine, strawberry and tomato. In order to find new important genes involved in the fungal virulence, a mutant library was generated by random insertional mutagenesis, using Agrobacterium tumefaciens mediated transformation. Ten mutants exhibiting total loss of virulence towards different host plants were considered. Their molecular characterization revealed a single T-DNA insertion in unique and different loci. Using a proteomics approach, the secretome of four of these strains was compared to that of the parental strain and a common profile of reduced lytic enzymes was recorded. Significant variations in this profile, notably proteases and hemicellulases deficiencies, were observed and validated by biochemical tests. They were also a hallmark of the remaining six non pathogenic strains, suggesting the importance of these secreted proteins in the infectious process.

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Project description:Light and BcLTF3-dependent gene expression profiles in the phytopathogenic fungus Botrytis cinerea

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Project description:Light and BcLTF1-dependent gene expression profiles in the phytopathogenic fungus Botrytis cinerea

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Project description:Full transcriptomes of the Botrytis cinerea wild-type strain B0510 and the null-mutant deltaBcLTF2, cultured in continuous darkness or submitted to a light stimulation, were compared to identify light- or/and BcLTF2-dependent genes. The Botrytis cinerea wild-type strain and the null-mutant deltaBcLTF2 were cultured for 54h in continuous darkness ; then, cultures were exposed for 60 min to light before harvest (L) or were harvested after additional 60 min darkness (D). 4 replicates were performed. The 12 total-RNA samples (4 conditions* 3 replicates) were used for hybridization on NimbleGen 4plex gene expression arrays (20,885 gene models from Botrytis cinerea with three 60-mer probes per gene).

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Project description:The Arabidopsis thaliana mutant wrky33 is highly susceptible to the necrotrophic fungus Botrytis cinerea. Comparing the expression profiles of B. cinerea-infected wrky33 and WT plants we identified 2765 differentially expressed genes dependent on WRKY33, of which 1675 were up-regulated in the mutant (termed WRKY33-repressed genes) and 1090 were down-regulated in the mutant. Combined with ChIP-seq data 318 genes were identified as direct functional targets of WRKY33 at 14 h post inoculation with spores of Botrytis cinerea 2100.

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