Dataset Information

Proteomic analysis of several non pathogenic mutants from Botrytis cinerea.

ABSTRACT: 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.

INSTRUMENT(S): Orbitrap Fusion Lumos

ORGANISM(S): Botryotinia Fuckeliana (noble Rot Fungus) (botrytis Cinerea)

SUBMITTER: Dupuy Jean-William

LAB HEAD: Nathalie Poussereau

PROVIDER: PXD013359 | Pride | 2021-11-03

REPOSITORIES: Pride

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Publications

A Similar Secretome Disturbance as a Hallmark of Non-pathogenic Botrytis cinerea ATMT-Mutants?

de Vallée Amélie A Bally Pascal P Bruel Christophe C Chandat Lucie L Choquer Mathias M Dieryckx Cindy C Dupuy Jean William JW Kaiser Sophie S Latorse Marie-Pascale MP Loisel Elise E Mey Géraldine G Morgant Guillaume G Rascle Christine C Schumacher Julia J Simon Adeline A Souibgui Eytham E Viaud Muriel M Villalba François F Poussereau Nathalie N

Frontiers in microbiology 20191206

The gray mold fungus Botrytis cinerea is a necrotrophic pathogen able to infect hundreds of host plants, including high-value crops such as grapevine, strawberry and tomato. In order to decipher its infectious strategy, a library of 2,144 mutants was generated by random insertional mutagenesis using Agrobacterium tumefaciens-mediated transformation (ATMT). Twelve mutants exhibiting total loss of virulence toward different host plants were chosen for detailed analyses. Their molecul ...[more]

PMID: 31866989

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Project description:'Mutagenomics' is a combination approach of random mutagenesis, phenotypic screening, and whole-genome re-sequencing to uncover all tagged and untagged mutations linked with phenotypic changes in an organism. In this study, we performed a mutagenomics screen on the wheat pathogenic fungus Zymoseptoria tritici for altered morphogenetic switching and stress sensitivity phenotypes using Agrobacterium-mediated “random” T-DNA mutagenesis (ATMT). Biological screening identified four mutants which were strongly reduced in virulence on wheat. Whole genome re-sequencing defined the positions of the T-DNA insertion events and revealed several unlinked mutations potentially also affecting gene functions. Remarkably, two independent reduced virulence mutant strains, with similarly altered stress sensitivities and aberrant hyphal growth phenotypes, were found to have distinct loss of function mutations in the ZtSSK2 MAPKKK gene. One mutant strain had a direct T-DNA insertion affecting the N-terminus of the predicted protein, whilst the other possessed an unlinked frameshift mutation towards the C-terminus. We used genetic complementation to restore both strains' wild-type (WT) function (virulence, morphogenesis, and stress response). We demonstrated that ZtSSK2 has a non-redundant function with ZtSTE11 in virulence through the biochemical activation of the stress-activated HOG1 MAPK pathway. Moreover, we present data suggesting that SSK2 has a unique role in activating this pathway in response to specific stresses. Finally, dual RNAseq-based transcriptome profiling of WT and SSK2 mutant strains revealed many HOG1-dependent transcriptional changes in the fungus during early infection and suggested that the host response does not discriminate between WT and mutant strains during this early phase. Together these data define new genes implicated in the virulence of the pathogen and emphasise the importance of a whole genome sequencing step in mutagenomic discovery pipelines.

Dataset Information

Proteomic analysis of several non pathogenic mutants from Botrytis cinerea.

Publications

A Similar Secretome Disturbance as a Hallmark of Non-pathogenic <i>Botrytis cinerea</i> ATMT-Mutants?

Similar Datasets

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