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Transcriptional Analysis of the Global Regulatory Networks Active in Pseudomonas syringae during Leaf Colonization

机译：丁香假单胞菌在叶片定殖过程中活跃的全球调控网络的转录分析。

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The plant pathogen Pseudomonas syringae pv. syringae B728a grows and survives on leaf surfaces and in the leaf apoplast of its host, bean (Phaseolus vulgaris). To understand the contribution of distinct regulators to B728a fitness and pathogenicity, we performed a transcriptome analysis of strain B728a and nine regulatory mutants recovered from the surfaces and interior of leaves and exposed to environmental stresses in culture. The quorum-sensing regulators AhlR and AefR influenced few genes in planta or in vitro. In contrast, GacS and a downstream regulator, SalA, formed a large regulatory network that included a branch that regulated diverse traits and was independent of plant-specific environmental signals and a plant signal-dependent branch that positively regulated secondary metabolite genes and negatively regulated the type III secretion system. SalA functioned as a central regulator of iron status based on its reciprocal regulation of pyoverdine and achromobactin genes and also sulfur uptake, suggesting a role in the iron-sulfur balance. RetS functioned almost exclusively to repress secondary metabolite genes when the cells were not on leaves. Among the sigma factors examined, AlgU influenced many more genes than RpoS, and most AlgU-regulated genes depended on RpoN. RpoN differentially impacted many AlgU- and GacS-activated genes in cells recovered from apoplastic versus epiphytic sites, suggesting differences in environmental signals or bacterial stress status in these two habitats. Collectively, our findings illustrate a central role for GacS, SalA, RpoN, and AlgU in global regulation in B728a in planta and a high level of plasticity in these regulators’ responses to distinct environmental signals. >IMPORTANCE Leaves harbor abundant microorganisms, all of which must withstand challenges such as active plant defenses and a highly dynamic environment. Some of these microbes can influence plant health. Despite knowledge of individual regulators that affect the fitness or pathogenicity of foliar pathogens, our understanding of the relative importance of various global regulators to leaf colonization is limited. Pseudomonas syringae strain B728a is a plant pathogen and a good colonist of both the surfaces and interior of leaves. This study used global transcript profiles of strain B728a to investigate the complex regulatory network of putative quorum-sensing regulators, two-component regulators, and sigma factors in cells colonizing the leaf surface and leaf interior under stressful in vitro conditions. The results highlighted the value of evaluating these networks in planta due to the impact of leaf-specific environmental signals and suggested signal differences that may enable cells to differentiate surface versus interior leaf habitats.
机译：植物病原菌 Pseudomonas syringae pv。丁香香蒲B728a在其寄主豆（ Phaseolus vulgaris ）的叶片表面和叶片质外体中生长并存活。为了了解不同调节剂对B728a适应性和致病性的贡献，我们对B728a菌株和从叶片表面和内部回收并暴露于培养环境压力的9个调节突变体进行了转录组分析。群体感应调节剂AhlR和AefR影响植物 或体外的少数基因。相比之下，GacS和下游调节剂SalA形成了一个大型的调节网络，该网络包括一个调节多种性状且独立于植物特异性环境信号的分支，以及一个植物信号依赖性分支，该分支正向调节次生代谢产物基因而对负向调节。 III型分泌系统。 SalA基于其对嘧啶和无色菌素基因的相互调节以及对硫的吸收，从而起着铁状态中央调节器的作用，表明其在铁硫平衡中的作用。当细胞不在叶子上时，RetS几乎只具有抑制次级代谢产物基因的功能。在所检查的sigma因子中，AlgU影响的基因比RpoS多得多，并且大多数受AlgU调节的基因都依赖RpoN。 RpoN差异影响从质生性和附生性部位恢复的细胞中许多AlgU和GacS激活的基因，表明这两个栖息地的环境信号或细菌应激状态存在差异。总的来说，我们的发现说明了GacS，SalA，RpoN和AlgU在植物B728a中的全球调节中的核心作用，并且在这些调节剂对独特环境信号的响应中具有高度可塑性。 >重要意义：叶子含有丰富的微生物，所有这些微生物都必须经受住有效的植物防御和高度动态的环境等挑战。这些微生物中的一些会影响植物健康。尽管了解影响叶病原体适应性或致病性的单个调节剂的知识，但我们对各种全球调节剂对叶片定殖的相对重要性的了解仍然有限。丁香假单胞菌B728a株是一种植物病原体，是叶片表面和内部的良好定植者。本研究使用菌株B728a的整体转录本概况研究了在体外压力下定殖在叶片表面和叶片内部的细胞中推定的群体感应调节剂，两组分调节剂和sigma因子的复杂调节网络。条件。结果表明，由于叶片特有的环境信号的影响，在植物中评估这些网络的价值，并提出了可能使细胞区分表面和内部叶片生境的信号差异。

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