title: Transcriptional analysis of the global regulatory networks active in Pseudomonas syringae during leaf colonization.
abstract: 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.
  
  
  OBJECTIVE
  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.
authors.affiliation: Department of Plant Pathology and Microbiology, Iowa State University, Ames, Iowa, USA.
authors.forename: Xilan
authors.initials: X
authors.lastname: Yu
authors.forename: Steven P
authors.initials: SP
authors.lastname: Lund
authors.affiliation: Department of Plant Pathology and Microbiology, Texas A&M University, College Station, Texas, USA.
authors.forename: Jessica W
authors.initials: JW
authors.lastname: Greenwald
authors.affiliation: Department of Plant Pathology and Microbiology, Texas A&M University, College Station, Texas, USA.
authors.forename: Angela H
authors.initials: AH
authors.lastname: Records
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Russell A
authors.initials: RA
authors.lastname: Scott
authors.affiliation: Department of Statistics, Iowa State University, Ames, Iowa, USA.
authors.forename: Dan
authors.initials: D
authors.lastname: Nettleton
authors.affiliation: Department of Plant and Microbial Biology, University of California, Berkeley, Berkeley, California, USA.
authors.forename: Steven E
authors.initials: SE
authors.lastname: Lindow
authors.affiliation: Department of Plant Pathology and Microbiology, Texas A&M University, College Station, Texas, USA.
authors.forename: Dennis C
authors.initials: DC
authors.lastname: Gross
authors.affiliation: Department of Plant Pathology and Microbiology, Iowa State University, Ames, Iowa, USA gbeattie@iastate.edu.
authors.forename: Gwyn A
authors.initials: GA
authors.lastname: Beattie
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism; D017353:Gene Deletion; D020869:Gene Expression Profiling; D015964:Gene Expression Regulation, Bacterial; D053263:Gene Regulatory Networks; D005809:Genes, Regulator; D010935:Plant Diseases / Q000382:microbiology; D018515:Plant Leaves / Q000382:microbiology*; D044224:Pseudomonas syringae / Q000235:genetics* / Q000254:growth & development; D053038:Quorum Sensing / Q000235:genetics*; D012329:RNA, Bacterial / Q000235:genetics / Q000302:isolation & purification; D018085:Regulon / Q000235:genetics*; D012808:Sigma Factor / Q000235:genetics / Q000378:metabolism; D013312:Stress, Physiological
chemical_list: D001426:Bacterial Proteins; D004268:DNA-Binding Proteins; C085855:HrpL protein, Pseudomonas syringae; D012329:RNA, Bacterial; D012808:Sigma Factor