Department of Agricultural Sciences, University of Helsinki, Helsinki, Finland.
J Bacteriol. 2012 Sep;194(18):4810-22. doi: 10.1128/JB.00611-12. Epub 2012 Jun 29.
When analyzing the secretome of the plant pathogen Pseudomonas syringae pv. tomato DC3000, we identified hemolysin-coregulated protein (Hcp) as one of the secreted proteins. Hcp is assumed to be an extracellular component of the type VI secretion system (T6SS). Two copies of hcp genes are present in the P. syringae pv. tomato DC3000 genome, hcp1 (PSPTO_2539) and hcp2 (PSPTO_5435). We studied the expression patterns of the hcp genes and tested the fitness of hcp knockout mutants in host plant colonization and in intermicrobial competition. We found that the hcp2 gene is expressed most actively at the stationary growth phase and that the Hcp2 protein is secreted via the T6SS and appears in the culture medium as covalently linked dimers. Expression of hcp2 is not induced in planta and does not contribute to virulence in or colonization of tomato or Arabidopsis plants. Instead, hcp2 is required for survival in competition with enterobacteria and yeasts, and its function is associated with the suppression of the growth of these competitors. This is the first report on bacterial T6SS-associated genes functioning in competition with yeast. Our results suggest that the T6SS of P. syringae may play an important role in bacterial fitness, allowing this plant pathogen to survive under conditions where it has to compete with other microorganisms for resources.
在分析植物病原体丁香假单胞菌 pv.番茄 DC3000 的分泌组时,我们鉴定出了溶血素调节蛋白(Hcp)作为分泌蛋白之一。Hcp 被认为是 VI 型分泌系统(T6SS)的细胞外成分。在丁香假单胞菌 pv.番茄 DC3000 基因组中存在两个 hcp 基因副本,hcp1(PSPTO_2539)和 hcp2(PSPTO_5435)。我们研究了 hcp 基因的表达模式,并测试了 hcp 敲除突变体在宿主植物定植和微生物间竞争中的适应性。我们发现 hcp2 基因在静止生长阶段表达最活跃,Hcp2 蛋白通过 T6SS 分泌,并以共价连接的二聚体形式出现在培养基中。hcp2 的表达在植物体内不会被诱导,也不会对番茄或拟南芥植物的毒力或定植产生贡献。相反,hcp2 对于与肠杆菌和酵母的生存竞争是必需的,其功能与抑制这些竞争者的生长有关。这是关于细菌 T6SS 相关基因在与酵母竞争中起作用的第一个报道。我们的结果表明,丁香假单胞菌的 T6SS 可能在细菌适应性中发挥重要作用,使这种植物病原体能够在与其他微生物竞争资源的条件下生存。