Identification of differentially expressed biomarkers in the host-parasite interaction in the Brassica napus - Verticillium longisporum system

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Verticillium longisporum is a soil-borne vascular pathogen that infects roots of predominantly rapeseed (B. napus) crops causing stunting and early senescence. The goal within the Research Group 546 is to identify fungal proteins, which are potential biomarkers for the interaction between V. longisporum and B. napus. We are looking for differentially expressed proteins and the corresponding genes in the presence of xylem sap and root exudates of host-plants. 11 differentially expressed proteins were identified in the intracellular proteome upon induction by xylem sap of B. napus. Currently the isogenes for an up-regulated catalase peroxidase are sequenced and will be knocked-down by RNA silencing. We will investigate in the possibility to haplodise V. longisporum in order to facilitate the construction of transgenic strains by homologous recombination. The secretome of V. longisporum will be investigated by three approaches: We will continue analyzing differentially expressed proteins in the secretome by conventional 2D-gel electrophoresis and by 2D-DIGE. The EST-library will be sequenced and analyzed for secreted proteins. In the third approach a complementation assay in S. cerevisiae for adhesive proteins of V. longisporum was successfully initiated and will be continued. Differentially expressed proteins, being potential biomarkers, will be silenced and in addition integrated into the genome of V. longisporum as reporter fusions. This system will enable us to study the expression and function of such potential biomarkers in vivo and in planta.