1887

Abstract

Pseudomonas fluorescens strains are important candidates for use as biological control agents to reduce fungal diseases on crop plants. To understand the ecological success of these bacteria and for successful and stable biological control, determination of how these bacteria colonize and persist in soil environments is critical. Here we show that P. fluorescens Pf0-1 is negatively impacted by reduced water availability in soil, but adapts and persists. A pilot transcriptomic study of Pf0-1 colonizing moist and dehydrated soil was used to identify candidate genetic loci, which could play a role in the adaptation to dehydration. Genes predicted to specify alginate production were identified and chosen for functional evaluation. Using deletion mutants, predicted alginate biosynthesis genes were shown to be important for optimal colonization of moist soil, and necessary for adaptation to reduced water availability in dried soil. Our findings extend in vitro studies of water stress into a more natural system and suggest alginate may be an essential extracellular product for the lifestyle of P. fluorescens when growing in soil.

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2019-05-03
2019-10-19
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References

  1. Weller DM. Pseudomonas biocontrol agents of soilborne pathogens: looking back over 30 years. Phytopathology 2007;97:250–256 [CrossRef]
    [Google Scholar]
  2. Fravel DR. Commercialization and implementation of biocontrol. Annu Rev Phytopathol 2005;43:337–359 [CrossRef]
    [Google Scholar]
  3. Chet I, Inbar J. Biological control of fungal pathogens. Appl Biochem Biotechnol 1994;48:37–43 [CrossRef]
    [Google Scholar]
  4. Rodriguez F, Pfender WF. Antibiosis and antagonism of Sclerotinia homoeocarpa and Drechslera poae by Pseudomonas fluorescens Pf-5 in vitro and in planta. Phytopathology 1997;87:614–621 [CrossRef]
    [Google Scholar]
  5. Loper JE, Gross H. Genomic analysis of antifungal metabolite production by Pseudomonas fluorescens Pf-5. Eur J Plant Pathol 2007;119:265–278 [CrossRef]
    [Google Scholar]
  6. Kraus J, Loper JE. Lack of evidence for a role of antifungal metabolite production by Pseudomonas fluorescens Pf-5 in biological control of Pythium damping-off of cucumber. Phytopathology 1992;82:264–271 [CrossRef]
    [Google Scholar]
  7. Hamdan H, Weller DM, Thomashow LS. Relative importance of fluorescent siderophores and other factors in biological control of Gaeumannomyces graminis var. tritici by Pseudomonas fluorescens 2-79 and M4-80R. Appl Environ Microbiol 1991;57:3270–3277
    [Google Scholar]
  8. Thomashow LS, Weller DM. Role of a phenazine antibiotic from Pseudomonas fluorescens in biological control of Gaeumannomyces graminis var. tritici. J Bacteriol 1988;170:3499–3508 [CrossRef]
    [Google Scholar]
  9. Gross H, Loper JE. Genomics of secondary metabolite production by Pseudomonas spp. Nat Prod Rep 2009;26:1408–1446 [CrossRef]
    [Google Scholar]
  10. Paterson J, Jahanshah G, Li Y, Wang Q, Mehnaz S et al. The contribution of genome mining strategies to the understanding of active principles of PGPR strains. FEMS Microbiol Ecol 2017;93:fiw249 [CrossRef]
    [Google Scholar]
  11. Rainey PB. Adaptation of Pseudomonas fluorescens to the plant rhizosphere. Environ Microbiol 1999;1:243–257 [CrossRef]
    [Google Scholar]
  12. Gal M, Preston GM, Massey RC, Spiers AJ, Rainey PB. Genes encoding a cellulosic polymer contribute toward the ecological success of Pseudomonas fluorescens SBW25 on plant surfaces. Mol Ecol 2003;12:3109–3121 [CrossRef]
    [Google Scholar]
  13. Silby MW, Cerdeño-Tárraga AM, Vernikos GS, Giddens SR, Jackson RW et al. Genomic and genetic analyses of diversity and plant interactions of Pseudomonas fluorescens. Genome Biol 2009;10:R51 [CrossRef]
    [Google Scholar]
  14. Silby MW, Nicoll JS, Levy SB. Requirement of polyphosphate by Pseudomonas fluorescens Pf0-1 for competitive fitness and heat tolerance in laboratory media and sterile soil. Appl Environ Microbiol 2009;75:3872–3881 [CrossRef]
    [Google Scholar]
  15. Varivarn K, Champa LA, Silby MW, Robleto EA. Colonization strategies of Pseudomonas fluorescens Pf0-1: activation of soil-specific genes important for diverse and specific environments. BMC Microbiol 2013;13:92 [CrossRef]
    [Google Scholar]
  16. Ghiglione JF, Richaume A, Philippot L, Lensi R. Relative involvement of nitrate and nitrite reduction in the competitiveness of Pseudomonas fluorescens in the rhizosphere of maize under non-limiting nitrate conditions. FEMS Microbiol Ecol 2002;39:121–127 [CrossRef]
    [Google Scholar]
  17. Rediers H, Vanderleyden J, De Mot R. Nitrate respiration in Pseudomonas stutzeri A15 and its involvement in rice and wheat root colonization. Microbiol Res 2009;164:461–468 [CrossRef]
    [Google Scholar]
  18. Philippot L, Clays-Josserand A, Lensi R. Use of Tn5 mutants to assess the role of the dissimilatory nitrite reductase in the competitive abilities of two Pseudomonas strains in soil. Appl Environ Microbiol 1995;61:1426–1430
    [Google Scholar]
  19. Bojanovič K, D'Arrigo I, Long KS. Global transcriptional responses to osmotic, oxidative, and imipenem stress conditions in Pseudomonas putida. Appl Environ Microbiol 2017;83:e03236–16 [CrossRef]
    [Google Scholar]
  20. Freeman BC, Chen C, Yu X, Nielsen L, Peterson K et al. Physiological and transcriptional responses to osmotic stress of two Pseudomonas syringae strains that differ in epiphytic fitness and osmotolerance. J Bacteriol 2013;195:4742–4752 [CrossRef]
    [Google Scholar]
  21. Sledjeski DD, Gottesman S. Osmotic shock induction of capsule synthesis in Escherichia coli K-12. J Bacteriol 1996;178:1204–1206 [CrossRef]
    [Google Scholar]
  22. Roberson EB, Firestone MK. Relationship between desiccation and exopolysaccharide production in a soil Pseudomonas sp. Appl Environ Microbiol 1992;58:1284–1291
    [Google Scholar]
  23. Miller KJ, Wood JM. Osmoadaptation by rhizosphere bacteria. Annu Rev Microbiol 1996;50:101–136 [CrossRef]
    [Google Scholar]
  24. Niu X, Song L, Xiao Y, Ge W. Drought-tolerant plant growth-promoting rhizobacteria associated with foxtail millet in a semi-arid Agroecosystem and their potential in alleviating drought stress. Front Microbiol 2018;8: [CrossRef]
    [Google Scholar]
  25. Gülez G, Dechesne A, Workman CT, Smets BF. Transcriptome dynamics of Pseudomonas putida KT2440 under water stress. Appl Environ Microbiol 2012;78:676–683 [CrossRef]
    [Google Scholar]
  26. Compeau G, Al-Achi BJ, Platsouka E, Levy SB. Survival of rifampin-resistant mutants of Pseudomonas fluorescens and Pseudomonas putida in soil systems. Appl Environ Microbiol 1988;54:2432–2438
    [Google Scholar]
  27. Marshall B, Robleto EA, Wetzler R, Kulle P, Casaz P et al. The adnA transcriptional factor affects persistence and spread of Pseudomonas fluorescens under natural field conditions. Appl Environ Microbiol 2001;67:852–857 [CrossRef]
    [Google Scholar]
  28. Silby MW, Levy SB. Use of in vivo expression technology to identify genes important in growth and survival of Pseudomonas fluorescens Pf0-1 in soil: discovery of expressed sequences with novel genetic organization. J Bacteriol 2004;186:7411–7419 [CrossRef]
    [Google Scholar]
  29. Bertani G. Lysogeny at mid-twentieth century: P1, P2, and other experimental systems. J Bacteriol 2004;186:595–600 [CrossRef]
    [Google Scholar]
  30. Kirner S, Krauss S, Sury G, Lam ST, Ligon JM et al. The non-haem chloroperoxidase from Pseudomonas fluorescens and its relationship to pyrrolnitrin biosynthesis. Microbiology 1996;142:2129–2135 [CrossRef]
    [Google Scholar]
  31. Kolter R, Inuzuka M, Helinski DR. Trans-complementation-dependent replication of a low molecular weight origin fragment from plasmid R6K. Cell 1978;15:1199–1208 [CrossRef]
    [Google Scholar]
  32. Simon R, Priefer U, Puhler A. A broad host range mobilisation system for in vivo engineering: Transposon mutagenesis in gram-negative bacteria. Biotechnology 1983;1:748–751
    [Google Scholar]
  33. Matthews M, Roy CR. Identification and subcellular localization of the Legionella pneumophila IcmX protein: a factor essential for establishment of a replicative organelle in eukaryotic host cells. Infect Immun 2000;68:3971–3982 [CrossRef]
    [Google Scholar]
  34. Mastropaolo MD, Silby MW, Nicoll JS, Levy SB. Novel genes involved in motility and biofilm formation in Pseudomonas fluorescens Pf0-1. Appl Environ Microbiol 2012;78:4318–4329
    [Google Scholar]
  35. Green MR, Sambrook J. Molecular Cloning: a Laboratory Manual, 4th ed. Cold Spring Harbor, New York: Cold Spring Harbor Laboratory Press; 2012
    [Google Scholar]
  36. Horton RM, Hunt HD, Ho SN, Pullen JK, Pease LR. Engineering hybrid genes without the use of restriction enzymes: gene splicing by overlap extension. Gene 1989;77:61–68 [CrossRef]
    [Google Scholar]
  37. Benjamini Y, Hochberg Y. Controlling the false discovery rate: a practical and powerful approach to multiple testing. J R Stat Soc Series B Stat Methodol 1995;57:289–300 [CrossRef]
    [Google Scholar]
  38. Seaton SC, Silby MW, Levy SB. Pleiotropic effects of GacA on Pseudomonas fluorescens Pf0-1 in vitro and in soil. Appl Environ Microbiol 2013;79:5405–5410 [CrossRef]
    [Google Scholar]
  39. Silby MW, Rainey PB, Levy SB. IVET experiments in Pseudomonas fluorescens reveal cryptic promoters at loci associated with recognizable overlapping genes. Microbiology 2004;150:518–520 [CrossRef]
    [Google Scholar]
  40. Chang WS, van de Mortel M, Nielsen L, Nino de Guzman G, Li X et al. Alginate production by Pseudomonas putida creates a hydrated microenvironment and contributes to biofilm architecture and stress tolerance under water-limiting conditions. J Bacteriol 2007;189:8290–8299 [CrossRef]
    [Google Scholar]
  41. Li X, Nielsen L, Nolan C, Halverson LJ. Transient alginate gene expression by Pseudomonas putida biofilm residents under water-limiting conditions reflects adaptation to the local environment. Environ Microbiol 2010;12:1578–1590 [CrossRef]
    [Google Scholar]
  42. Remminghorst U, Rehm BHA. Alg44, a unique protein required for alginate biosynthesis in Pseudomonas aeruginosa. FEBS Lett 2006;580:3883–3888 [CrossRef]
    [Google Scholar]
  43. Moradali MF, Donati I, Sims IM, Ghods S, Rehm BHA. Alginate polymerization and modification are linked in Pseudomonas aeruginosa. MBio 2015;6:e00453–00415 [CrossRef]
    [Google Scholar]
  44. Merighi M, Lee VT, Hyodo M, Hayakawa Y, Lory S. The second messenger bis-(3′-5′)-cyclic-GMP and its PilZ domain-containing receptor Alg44 are required for alginate biosynthesis in Pseudomonas aeruginosa. Mol Microbiol 2007;65:876–895 [CrossRef]
    [Google Scholar]
  45. Deretic V, Gill JF, Chakrabarty AM. Gene algD coding for GDPmannose dehydrogenase is transcriptionally activated in mucoid Pseudomonas aeruginosa. J Bacteriol 1987;169:351–358 [CrossRef]
    [Google Scholar]
  46. Wozniak DJ, Ohman DE. Pseudomonas aeruginosa AlgB, a two-component response regulator of the NtrC family, is required for algD transcription. J Bacteriol 1991;173:1406–1413 [CrossRef]
    [Google Scholar]
  47. Goldberg JB, Dahnke T. Pseudomonas aeruginosa AlgB, which modulates the expression of alginate, is a member of the NtrC subclass of prokaryotic regulators. Mol Microbiol 1992;6:59–66 [CrossRef]
    [Google Scholar]
  48. Chew SC, Kundukad B, Seviour T, van der Maarel JRC, Yang L et al. Dynamic remodeling of microbial biofilms by functionally distinct exopolysaccharides. MBio 2014;5:e01536-14 [CrossRef]
    [Google Scholar]
  49. Colvin KM, Irie Y, Tart CS, Urbano R, Whitney JC et al. The Pel and Psl polysaccharides provide Pseudomonas aeruginosa structural redundancy within the biofilm matrix. Environ Microbiol 2012;14:1913–1928 [CrossRef]
    [Google Scholar]
  50. Stapper AP, Narasimhan G, Ohman DE, Barakat J, Hentzer M et al. Alginate production affects Pseudomonas aeruginosa biofilm development and architecture, but is not essential for biofilm formation. J Med Microbiol 2004;53:679–690 [CrossRef]
    [Google Scholar]
  51. Ghafoor A, Hay ID, Rehm BHA. Role of exopolysaccharides in Pseudomonas aeruginosa biofilm formation and architecture. Appl Environ Microbiol 2011;77:5238–5246 [CrossRef]
    [Google Scholar]
  52. Periasamy S, Nair HAS, Lee KWK, Ong J, Goh JQJ et al. Pseudomonas aeruginosa PAO1 exopolysaccharides are important for mixed species biofilm community development and stress tolerance. Front Microbiol 2015;6:851 [CrossRef]
    [Google Scholar]
  53. Slauch JM, Mahan MJ, Mekalanos JJ. In vivo expression technology for selection of bacterial genes specifically induced in host tissues. Methods Enzymol 1994;235:481–492
    [Google Scholar]
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