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

长孢黄萎病（Verticillium longisporum）是一种土壤传播的维管病原体，主要侵染油菜（B. napus）作物的根系，导致发育迟缓和早期衰老。研究小组546的目标是鉴定真菌蛋白，这是长孢弧菌和甘蓝型酵母相互作用的潜在生物标志物。我们正在寻找寄主植物木质部汁液和根分泌物中存在的差异表达蛋白和相应的基因。在甘蓝型油菜木质部汁液诱导下，在胞内蛋白质组中鉴定出11种差异表达蛋白。目前，对过氧化氢酶过氧化物酶上调的同工基因进行了测序，并将通过RNA沉默来敲低。我们将研究长孢弧菌单倍体化的可能性，以便通过同源重组构建转基因菌株。我们将通过三种方法研究长孢弧菌的分泌组：我们将继续通过传统的2d凝胶电泳和2D-DIGE分析分泌组中的差异表达蛋白。est文库将被测序并分析分泌蛋白。在第三种方法中，酿酒酵母对长孢弧菌粘附蛋白的互补试验已经成功启动，并将继续进行。差异表达蛋白作为潜在的生物标志物，将被沉默，并作为报告融合物整合到长孢弧菌的基因组中。该系统将使我们能够研究这些潜在生物标志物在体内和植物中的表达和功能。

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.