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.
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. Comparison of altered gene expression in Arabidopsis WT and wrky33 mutant plants 14 hours post inoculation with Botrytis cinerea 2100.
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.