Skip to main content
NCBI home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1997 Dec;179(24):7663–7670. doi: 10.1128/jb.179.24.7663-7670.1997

N-acyl-homoserine lactone-mediated regulation of phenazine gene expression by Pseudomonas aureofaciens 30-84 in the wheat rhizosphere.

D W Wood 1, F Gong 1, M M Daykin 1, P Williams 1, L S Pierson 3rd 1
PMCID: PMC179727  PMID: 9401023

Abstract

Pseudomonas aureofaciens 30-84 is a soilborne bacterium that colonizes the wheat rhizosphere. This strain produces three phenazine antibiotics which suppress take-all disease of wheat by inhibition of the causative agent Gaeumannomyces graminis var. tritici. Phenazines also enhance survival of 30-84 within the wheat rhizosphere in competition with other organisms. Expression of the phenazine biosynthetic operon is controlled by the phzR/phzI N-acyl-homoserine lactone (AHL) response system (L. S. Pierson III et al., J. Bacterial 176:3966-3974, 1994; D. W. Wood and L. S. Pierson III, Gene 168:49-53, 1996). By using high-pressure liquid chromatography coupled with high-resolution mass spectrometry, the AHL produced by PhzI has now been identified as N-hexanoyl-homoserine lactone (HHL). In addition, the ability of HHL to serve as an interpopulation signal molecule in the wheat rhizosphere has been examined by using isogenic reporter strains. Disruption of phzI reduced expression of the phenazine biosynthetic operon 1,000-fold in the wheat rhizosphere. Coinoculation of an isogenic strain which produced the endogenous HHL signal restored phenazine gene expression in the phzI mutant to wild-type levels in situ. These results demonstrate that HHL is required for phenazine expression in situ and is an effective interpopulation signal molecule in the wheat rhizosphere.

Full Text

The Full Text of this article is available as a PDF (675.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bassler B. L., Greenberg E. P., Stevens A. M. Cross-species induction of luminescence in the quorum-sensing bacterium Vibrio harveyi. J Bacteriol. 1997 Jun;179(12):4043–4045. doi: 10.1128/jb.179.12.4043-4045.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Boettcher K. J., Ruby E. G. Detection and quantification of Vibrio fischeri autoinducer from symbiotic squid light organs. J Bacteriol. 1995 Feb;177(4):1053–1058. doi: 10.1128/jb.177.4.1053-1058.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Chhabra S. R., Stead P., Bainton N. J., Salmond G. P., Stewart G. S., Williams P., Bycroft B. W. Autoregulation of carbapenem biosynthesis in Erwinia carotovora by analogues of N-(3-oxohexanoyl)-L-homoserine lactone. J Antibiot (Tokyo) 1993 Mar;46(3):441–454. doi: 10.7164/antibiotics.46.441. [DOI] [PubMed] [Google Scholar]
  4. Eberhard A. Inhibition and activation of bacterial luciferase synthesis. J Bacteriol. 1972 Mar;109(3):1101–1105. doi: 10.1128/jb.109.3.1101-1105.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Eberhard A., Widrig C. A., McBath P., Schineller J. B. Analogs of the autoinducer of bioluminescence in Vibrio fischeri. Arch Microbiol. 1986 Oct;146(1):35–40. doi: 10.1007/BF00690155. [DOI] [PubMed] [Google Scholar]
  6. Eberl L., Winson M. K., Sternberg C., Stewart G. S., Christiansen G., Chhabra S. R., Bycroft B., Williams P., Molin S., Givskov M. Involvement of N-acyl-L-hormoserine lactone autoinducers in controlling the multicellular behaviour of Serratia liquefaciens. Mol Microbiol. 1996 Apr;20(1):127–136. doi: 10.1111/j.1365-2958.1996.tb02495.x. [DOI] [PubMed] [Google Scholar]
  7. Engebrecht J., Nealson K., Silverman M. Bacterial bioluminescence: isolation and genetic analysis of functions from Vibrio fischeri. Cell. 1983 Mar;32(3):773–781. doi: 10.1016/0092-8674(83)90063-6. [DOI] [PubMed] [Google Scholar]
  8. Fuqua C., Winans S. C., Greenberg E. P. Census and consensus in bacterial ecosystems: the LuxR-LuxI family of quorum-sensing transcriptional regulators. Annu Rev Microbiol. 1996;50:727–751. doi: 10.1146/annurev.micro.50.1.727. [DOI] [PubMed] [Google Scholar]
  9. Hanzelka B. L., Greenberg E. P. Quorum sensing in Vibrio fischeri: evidence that S-adenosylmethionine is the amino acid substrate for autoinducer synthesis. J Bacteriol. 1996 Sep;178(17):5291–5294. doi: 10.1128/jb.178.17.5291-5294.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Kaplan H. B., Greenberg E. P. Diffusion of autoinducer is involved in regulation of the Vibrio fischeri luminescence system. J Bacteriol. 1985 Sep;163(3):1210–1214. doi: 10.1128/jb.163.3.1210-1214.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kuo A., Blough N. V., Dunlap P. V. Multiple N-acyl-L-homoserine lactone autoinducers of luminescence in the marine symbiotic bacterium Vibrio fischeri. J Bacteriol. 1994 Dec;176(24):7558–7565. doi: 10.1128/jb.176.24.7558-7565.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. LEVITCH M. E., STADTMAN E. R. STUDY OF THE BIOSYNTHESIS OF PHENAZINE-1-CARBOXYLIC ACID. Arch Biochem Biophys. 1964 Jul 20;106:194–199. doi: 10.1016/0003-9861(64)90175-4. [DOI] [PubMed] [Google Scholar]
  13. Latifi A., Winson M. K., Foglino M., Bycroft B. W., Stewart G. S., Lazdunski A., Williams P. Multiple homologues of LuxR and LuxI control expression of virulence determinants and secondary metabolites through quorum sensing in Pseudomonas aeruginosa PAO1. Mol Microbiol. 1995 Jul;17(2):333–343. doi: 10.1111/j.1365-2958.1995.mmi_17020333.x. [DOI] [PubMed] [Google Scholar]
  14. Lindgren P. B., Frederick R., Govindarajan A. G., Panopoulos N. J., Staskawicz B. J., Lindow S. E. An ice nucleation reporter gene system: identification of inducible pathogenicity genes in Pseudomonas syringae pv. phaseolicola. EMBO J. 1989 May;8(5):1291–1301. doi: 10.1002/j.1460-2075.1989.tb03508.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Marsh J. L., Erfle M., Wykes E. J. The pIC plasmid and phage vectors with versatile cloning sites for recombinant selection by insertional inactivation. Gene. 1984 Dec;32(3):481–485. doi: 10.1016/0378-1119(84)90022-2. [DOI] [PubMed] [Google Scholar]
  16. Mazzola M., Cook R. J., Thomashow L. S., Weller D. M., Pierson L. S., 3rd Contribution of phenazine antibiotic biosynthesis to the ecological competence of fluorescent pseudomonads in soil habitats. Appl Environ Microbiol. 1992 Aug;58(8):2616–2624. doi: 10.1128/aem.58.8.2616-2624.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Milton D. L., Hardman A., Camara M., Chhabra S. R., Bycroft B. W., Stewart G. S., Williams P. Quorum sensing in Vibrio anguillarum: characterization of the vanI/vanR locus and identification of the autoinducer N-(3-oxodecanoyl)-L-homoserine lactone. J Bacteriol. 1997 May;179(9):3004–3012. doi: 10.1128/jb.179.9.3004-3012.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Moré M. I., Finger L. D., Stryker J. L., Fuqua C., Eberhard A., Winans S. C. Enzymatic synthesis of a quorum-sensing autoinducer through use of defined substrates. Science. 1996 Jun 14;272(5268):1655–1658. doi: 10.1126/science.272.5268.1655. [DOI] [PubMed] [Google Scholar]
  19. Nealson K. H., Platt T., Hastings J. W. Cellular control of the synthesis and activity of the bacterial luminescent system. J Bacteriol. 1970 Oct;104(1):313–322. doi: 10.1128/jb.104.1.313-322.1970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Peet R. C., Lindgren P. B., Willis D. K., Panopoulos N. J. Identification and cloning of genes involved in phaseolotoxin production by Pseudomonas syringae pv. "phaseolicola". J Bacteriol. 1986 Jun;166(3):1096–1105. doi: 10.1128/jb.166.3.1096-1105.1986. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Pierson L. S., 3rd, Gaffney T., Lam S., Gong F. Molecular analysis of genes encoding phenazine biosynthesis in the biological control bacterium. Pseudomonas aureofaciens 30-84. FEMS Microbiol Lett. 1995 Dec 15;134(2-3):299–307. doi: 10.1111/j.1574-6968.1995.tb07954.x. [DOI] [PubMed] [Google Scholar]
  22. Pierson L. S., 3rd, Keppenne V. D., Wood D. W. Phenazine antibiotic biosynthesis in Pseudomonas aureofaciens 30-84 is regulated by PhzR in response to cell density. J Bacteriol. 1994 Jul;176(13):3966–3974. doi: 10.1128/jb.176.13.3966-3974.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pierson L. S., 3rd, Thomashow L. S. Cloning and heterologous expression of the phenazine biosynthetic locus from Pseudomonas aureofaciens 30-84. Mol Plant Microbe Interact. 1992 Jul-Aug;5(4):330–339. doi: 10.1094/mpmi-5-330. [DOI] [PubMed] [Google Scholar]
  24. Schaefer A. L., Val D. L., Hanzelka B. L., Cronan J. E., Jr, Greenberg E. P. Generation of cell-to-cell signals in quorum sensing: acyl homoserine lactone synthase activity of a purified Vibrio fischeri LuxI protein. Proc Natl Acad Sci U S A. 1996 Sep 3;93(18):9505–9509. doi: 10.1073/pnas.93.18.9505. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Shaw P. D., Ping G., Daly S. L., Cha C., Cronan J. E., Jr, Rinehart K. L., Farrand S. K. Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography. Proc Natl Acad Sci U S A. 1997 Jun 10;94(12):6036–6041. doi: 10.1073/pnas.94.12.6036. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Swift S., Karlyshev A. V., Fish L., Durant E. L., Winson M. K., Chhabra S. R., Williams P., Macintyre S., Stewart G. S. Quorum sensing in Aeromonas hydrophila and Aeromonas salmonicida: identification of the LuxRI homologs AhyRI and AsaRI and their cognate N-acylhomoserine lactone signal molecules. J Bacteriol. 1997 Sep;179(17):5271–5281. doi: 10.1128/jb.179.17.5271-5281.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Swift S., Stewart G. S., Williams P. The inner workings of a quorum sensing signal generator. Trends Microbiol. 1996 Dec;4(12):463–466. doi: 10.1016/s0966-842x(97)82904-6. [DOI] [PubMed] [Google Scholar]
  28. Swift S., Throup J. P., Williams P., Salmond G. P., Stewart G. S. Quorum sensing: a population-density component in the determination of bacterial phenotype. Trends Biochem Sci. 1996 Jun;21(6):214–219. [PubMed] [Google Scholar]
  29. Winson M. K., Camara M., Latifi A., Foglino M., Chhabra S. R., Daykin M., Bally M., Chapon V., Salmond G. P., Bycroft B. W. Multiple N-acyl-L-homoserine lactone signal molecules regulate production of virulence determinants and secondary metabolites in Pseudomonas aeruginosa. Proc Natl Acad Sci U S A. 1995 Sep 26;92(20):9427–9431. doi: 10.1073/pnas.92.20.9427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Wirth R., Muscholl A., Wanner G. The role of pheromones in bacterial interactions. Trends Microbiol. 1996 Mar;4(3):96–103. doi: 10.1016/0966-842X(96)81525-3. [DOI] [PubMed] [Google Scholar]
  31. Wood D. W., Pierson L. S., 3rd The phzI gene of Pseudomonas aureofaciens 30-84 is responsible for the production of a diffusible signal required for phenazine antibiotic production. Gene. 1996 Feb 2;168(1):49–53. doi: 10.1016/0378-1119(95)00754-7. [DOI] [PubMed] [Google Scholar]
  32. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  33. Zhang L., Murphy P. J., Kerr A., Tate M. E. Agrobacterium conjugation and gene regulation by N-acyl-L-homoserine lactones. Nature. 1993 Apr 1;362(6419):446–448. doi: 10.1038/362446a0. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES