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

甘蓝型油菜-长孢轮枝菌系统宿主-寄生虫相互作用中差异表达生物标志物的鉴定

• 批准号: 13922738
• 负责人: Professor Dr. Gerhard H. Braus
• 金额: --
• 依托单位: Abteilung für Molekulare Mikrobiologie und Genetik
• 依托单位国家: 德国
• 项目类别: Research Units
• 财政年份: 2005
• 资助国家: 德国
• 起止时间: 2004-12-31 至 2011-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/13922738?language=en
• 关键词: Identification; differentially; expressed; biomarkers; host

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.

黄萎病是一种土传维管束病原菌，主要侵染油菜(B.napus)作物的根部，导致矮化和早衰。546研究组的目标是鉴定真菌蛋白，这些蛋白是长孢霉和甘蓝型油菜相互作用的潜在生物标记。我们正在寻找在寄主植物木质部汁液和根分泌物存在的情况下差异表达的蛋白质和相应的基因。经甘蓝型油菜木质部汁液诱导后，在胞内蛋白质组中鉴定出11个差异表达蛋白。目前，上调的过氧化氢酶过氧化物酶的同源基因已经被测序，并将被RNA沉默击倒。我们将探索单倍化长孢霉的可能性，以便于通过同源重组构建转基因菌株。长孢弧菌的分泌组将通过三种方法进行研究：我们将继续用常规的2D-Gel电泳法和2D-DGE法分析分泌组中差异表达的蛋白质。将对EST文库进行测序并分析分泌蛋白。在第三种方法中，在酿酒酵母中成功地启动了长孢弧菌黏附蛋白的互补试验，并将继续进行。差异表达的蛋白质作为潜在的生物标志物，将被沉默，并作为报告融合整合到长孢弧菌的基因组中。这一系统将使我们能够研究这些潜在的生物标志物在体内和植物中的表达和功能。

项目成果

The plant host Brassica napus induces in the pathogen Verticillium longisporum the expression of functional catalase peroxidase which is required for the late phase of disease.

• DOI: 10.1094/mpmi-08-11-0217
• 发表时间: 2012-03
• 期刊: Molecular plant-microbe interactions : MPMI
• 作者: Seema Singh;S. Braus-Stromeyer;Christian Timpner;O. Valerius;A. von Tiedemann;P. Karlovsky;Christine Druebert;A. Polle;G. Braus