Abstract
Pseudomonas fluorescens 2P24 is a biocontrol agent isolated from a wheat take-all decline soil in China. This strain produces several antifungal compounds, such as 2,4-diacetylphloroglucinol (2,4-DAPG), hydrogen cyanide and siderophore(s). Our recent work revealed that strain 2P24 employs a quorum-sensing system to regulate its biocontrol activity. In this study, we identified a quorum-sensing system consisting of PcoR and PcoI of the LuxR–LuxI family from strain 2P24. Deletion of pcoI from 2P24 abolishes the production of the quorum-sensing signals, but does not detectably affect the production of antifungal metabolites. However, the mutant is significantly defective in biofilm formation, colonization on wheat rhizosphere and biocontrol ability against wheat take-all, whilst complementation of pcoI restores the biocontrol activity to the wild-type level. Our data indicate that quorum sensing is involved in regulation of biocontrol activity in P. fluorescens 2P24.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
References
Abbas A., Morrissey J.P., Marquez P.C., Sheehan M.M., Delany I.R. and O’Gara F. (2002). Characterization of interactions between the transcriptional repressor PhlF and its binding site at the phlA promoter in Pseudomonas fluorescens F113. J. Bacteriol. 184:3008–3016
Andrews H.L., Vogel J.P. and Isberg R.R. (1998). Identification of linked Legionella pneumophila genes essential for intracellular growth and evasion of the endocytic pathway. Infect. Immun. 66:950–958
von Bodman S.B., Bauer W.D. and Coplin D.L. (2003). Quorum sensing in plant-pathogenic bacteria. Annu. Rev. Phytopathol. 41:455–482
Brelles-Mariño G. and Bedmar E.J. (2001). Detection, purification and characterisation of quorum-sensing signal molecules in plant-associated bacteria. J. Biotechnol. 91:197–209
Brint J.M. and Ohman D.E. (1995). Synthesis of multiple exoproducts in Pseudomonas aeruginosa is under control of RhlR-RhlI, another set of regulators in strain PAO1 with homology to the autoinducer-responsive LuxR-LuxI family. J. Bacteriol. 177:7155–7163
Brodhagen M., Henkels M.D. and Loper J.E. (2004). Positive autoregulation and signaling properties of pyoluteorin, an antibiotic produced by the biological control organism Pseudomonas fluorescens Pf-5. Appl. Environ. Microbiol. 70:1758–1766
Cámara M., Daykin M. and Chhabra S.R. (1998). Detection, purification and synthesis of N-acyl homoserine lactone quorum sensing molecules. Methods Microb. Bacterial Pathogen. 27:319–330
Carl B., Arnokd A., Hauer B. and Fetzner S. (2004). Sequence and transcriptional analysis of a gene cluster of Pseudomonas putida 86 involved in quinoline degradation. Gene 331:177–188
Cha C., Gao P., Chen Y.C., Shaw P.D. and Farrand S.K. (1998). Production of acyl-homoserine lactone quorum-sensing signals by Gram-negative plant-associated bacteria. Mol. Plant Microbe Interact. 11:1119–1129
Chilton M.D., Currier T.C., Farrand S.K., Bendich A.J., Gordon M.P. and Nester E.W. (1974). Agrobacterium tumefaciens and PS8 bacteriophage DNA not found in crown gall tumors. Proc. NatI. Acad. Sci. USA 71:3672–3676
Collado-Vides J., Magasanik B. and Gralla J.D. (1991). Control site location and transcriptional regulation in Escherichia coli. Microbiol. Rev. 55:371–394
Cooper S.M., Laosripaiboon W., Rahman A.S., Hothersall J., El-Sayed A.K., Winfield C., Crosby J., Cox R.J., Simpson T.J. and Thomas C.M. (2005). Shift to Pseudomonic acid B production in P. fluorescens NCIMB10586 by mutation of mupirocin tailoring genesmupO, mupU, mupV, and macpE. Chem Biol. 12:825–833
Delany I., Sheehan M.M., Fenton A., Bardin S., Aarons S. and O’Gara F. (2000). Regulation of production of the antifungal metabolite 2,4-diacetylphloroglucinol in Pseudomonas fluorescens F113: genetic analysis of phlF as a transcriptional repressor. Microbiology 146:537–546
Del Sal G., Manfioletti G. and Schneider C. (1988). A one-tube plasmid DNA mini-preparation suitable for sequencing. Nucleic Acids Res. 16:9878
Duffy B.K. and Défago G. (1997). Zinc improves biocontrol of Fusarium crown and root rot of tomato by Pseudomonas fluorescens an represses the production of pathogen metabolites inhibitory to bacterial antibiotic biosynthesis. Phytopathology 87:1250–1257
Egland K.A. and Greenberg E.P. (2000). Conversion of the Vibrio fischeri transcriptional activator, LuxR, to a repressor. J. Bacteriol. 182:805–811
El-Sayed K.A., Hothersall J., Cooper S.M., Stephens E., Simpson T.J. and Thomas C.M. (2003). Characterization of the mupirocin biosynthesis gene cluster from Pseudomonas fluorescens NCIMB 10586. Chem. Biol. 10:419–430
El-Sayed K.A., Hothersall J. and Thomas C.M. (2001). Quorum-sensing-dependent regulation of biosynthesis of the polyketide antibiotic mupirocin in Pseudomonas fluorescens NCIMB 10586. Microbiology 147:2127–2139
Engebrecht J., Nealson K. and Silverman M. (1983). Bacterial bioluminescence: isolation and genetic analysis of functions from Vibrio fischeri. Cell 32:773–781
Finan T.M., Kunkel B., de Vos G.F. and Signer E.R. (1986). Second symbiotic megaplasmid in Rhizobium meliloti carrying exopolysaccharide and thiamine synthesis genes. J. Bacteriol. 167:66–72
Gambello M.J., and Iglewski B.H. (1991). Cloning and characterization of the Pseudomonas aeruginosa lasR gene, a transcriptional activator of elastase expression. J. Bacteriol. 173:3000–3009
Gray K.M., Garey J.R. (2001). The evolution of bacterial LuxI and LuxR quorum sensing regulators. Microbiology 147:2379–2387
Hanahan D. (1983) Studies on transformation of Escherichia coli with plasmids. J. Mol. Biol. 166:557–580
Hanzelka B.L. and Greenberg E.P. (1995). Evidence that the N-terminal region of the Vibrio fischeri LuxR protein constitutes an autoinducer-binding domain. J. Bacteriol. 177:815–817
Hayase N., Taira K. and Furukawa K. (1990). Pseudomonas putida KF715 bphABCD operon encoding biphenyl and polychlorinated biphenyl degradation: cloning, analysis and expression in soil bacteria. J. Bacteriol. 172:1160–1164
Hoben H.J. and Somasegaran P. (1982). Comparison of the pour, spread, and drop plate methods for enumeration of Rhizobium spp. in inoculants made from presterilized peat. Appl. Environ. Microbiol. 44:1246–1247
Hwang I., Li P.L., Zhang L., Piper K.R., Cook D.M., Tate M.E. and Farrand S.K. (1994). TraI, a LuxI homologue, is responsible for the production of conjugation factor, the Ti plasmid N-acylhomoserine lactone autoinducer. Proc. Natl. Acad. Sci. USA 91:4639–4643
Keen N.T., Tamaki S., Kobayashi D. and Trollinger D. (1988). Improved broad-host-range plasmids for DNA cloning in Gram-negative bacteria. Gene 70:191–197
King E.O., Ward M.K. and Raney D.E. (1954). Two simple media for the demonstration of pyocyanin and fluorescein. J. Lab. Clin. Med. 44:301–307
Laue R.E., Jiang Y., Chhabra S.R., Jacob S., Stewart G.S.A.B., Hardman A., Downie J.A., O’Gara F., Williams P. (2000). The biocontrol strain Pseudomonas fluorescens F113 produces the Rhizobium small bacteriocin, N-(3-hydroxy-7-cis-tetradecenoyl) homoserine lactone, via HdtS, a putative novel N-acylhomoserine lactone synthase. Microbiology – UK 146:2469–2480
Lewenza S., Conway B., Greenberg E.P., Sokol P.A. (1999). Quorum sensing in Burkholderia cepacia: identification of the LuxRI homologs CepRI. J. Bacteriol. 181:748–756
de Lorenzo V. and Timmis K.N. (1994). Analysis and construction of stable phenotype in gram-negative bacteria with Tn5- and Tn10-derived minitransposons. Methods Enzymol. 235:387–405
Luo Z.Q. and Farrand S.K. (1999). Signal-dependent DNA binding and functional domains of the quorum-sensing activator TraR as identified by repressor activity. Proc. Natl. Acad. Sci. USA 96:9009–9014
Maurhofer M., Baehler E., Notz R., Martinez V. and Keel C. (2004). Cross talk between 2,4-diacetylphloroglucinol-producing biocontrol Pseudomonads on wheat roots. Appl. Environ. Microbiol. 70:1990–1998
Miller M.B. and Bassler B.L. (2001). Quorum sensing in bacteria. Annu. Rev. Microbiol. 55:165–199
Nardini M. and Dijkstra B.M. (1999). α/β hydrolase fold enzymes: the family keeps growing. Curr. Opin. Struct. Biol. 9:732–737
Ownley B.H., Weller D.M., Thomashow L.S. (1992). Influence of in situ and in vitro pH on suppression of Gaeumannomyces graminis var. tritici by Pseudomonas fluorescens 2–79. Phytopathology 82:178–184
Parsek M.R., Schaefer A.L., Greenberg E.P. (1997). Analysis of random and site-directed mutations in rhlI, a Pseudomonas aeruginosa gene encoding an acylhomoserine lactone synthase. Mol. Microbiol. 26:301–310
Passador L., Cook J.M., Gambello M.J., Rust L. and Iglewski B.H. (1993). Expression of Pseudomonas aeruginosa virulence genes requires cell-to-cell communication. Science 260:1127–1130
Pearson J.P., Gray K.M., Passador L., Tucker K.D., Eberhard A., Iglewski B.H. and Greenberg E.P. (1994). Structure of the autoinducer required for expression of Pseudomonas aeruginosa virulence genes. Proc. Natl. Acad. Sci. USA 91:197–201
Pierson L.S., Gaffney T., Lam S., and Gong F. (1995). Molecular analysis of genes encoding phenazine biosynthesis in the biological control bacterium Pseudomonas aureofaciens 30–84. FEMS Microbiol. Lett. 134:299–307
Pierson III L.S. and Thomashow L.S. (1992). Cloning and heterologous expression of the phenazine biosynthetic locus from Pseudomonas aureofaciens 30–84. Mol. Plant Microbe Interact. 5:330–339
Pierson E.A., Wood D.W., Cannon J.A., Blachere F.M. and Pierson III L.S. (1998). Interpopulation signaling via N-acyl-homoserine lactones among bacteria in the wheat rhizosphere. Mol. Plant Microbe Interact. 11:1078–1084
Sambrook J., Fritsch E.F., Maniatis T. (1989). Molecular Cloning: A Laboratory Manual, 2nd edn. Cold Spring Harbor Laboratory, Cold Spring Harbor, NY
Schnider-Keel U., Seematter A., Maurhofer M., Blumer C., Duffy B., Gigot-Bonnefoy C., Reimmann C., Notz R., Défago G., Haas D., Keel C. (2000). Autoinduction of 2,4-diacetylphloroglucinol biosynthesis in the biocontrol agent Pseudomonas fluorescens CHA0 and repression by the bacterial metabolites salicylate and pyoluteorin. J. Bacteriol. 182:1215–1225
Schwyn B., and Neilands J.B. (1987). Universal chemical assay for the detection and determination of siderophores. Anal. Biochem. 160:47–56
Shaw P.D., Ping G., Daly S., Cha C., Cronan Jr J.E., Rinehart K.L. and Farrand S.K. (1997). Detecting and characterizing N-acyl-homoserine lactone signal molecules by thin-layer chromatography. Proc. Natl. Acad. Sci. USA 94:6036–6041
Steidle A., Allesen-Holm M., Riedel K., Berg G., Givskov M., Molin S. and Eberl L. (2002). Identification and characterization of an N-acylhomoserine lactone-dependent quorum-sensing system in Pseudomonas putida Strain IsoF. Appl. Environ. Microbiol. 68:6371–6382
Stepanovic S., Vukovic D., Dakic I., Savic B. and Svabic-Vlahovic M. (2000). A modified microtiter-plate test for quantification of staphylococcal biofilm formation. J. Microbiol. Methods 40:175–179
Wei H.L., Wang Y., Zhang L.Q. and Tang W. (2004a). Identification and characterization of biocontrol bacterial strain 2P24 and CPF-10. Acta Phytopathol. Sin. 34:80–85
Wei H.L. and Zhang L.Q. (2005). Cloning and functional characterization of the gacS gene of the biocontrol strain Pseudomonas fluorescens 2P24. Acta Microbiol. Sin. 45:368–372
Wei H.L., Zhou H.Y., Zhang L.Q., Wang Y. and Tang W.H. (2004b). Experimental evidence on the functional agent of 2,4-diacetylphloroglucinol in biocontrol activity of Pseudomonas fluorescens 2P24. Acta Microbiol. Sin. 44:663–666
Weller D.M. (1983). Colonization of wheat roots by a fluorescent pseudomonad suppressive to take-all. Phytopathology 73:1548–1553
Wood D.W., Gong F., Daykin M.M., Williams P. and Pierson L.S. (1997). N-acyl-homoserine lactone-mediated regulation of phenazine gene expression by Pseudomonas aereofaciens 30-84 in the wheat rhizosphere. J. Bacteriol. 179:7663–7670
Zhang Z. and Pierson III L.S. (2001). A second quorum-sensing system regulates cell surface properties but not phenazine antibiotic production in Pseudomonas aureofaciens. Appl. Environ. Microbiol. 67:4305–4315
Zhou H.Y., Wei H.L., Liu X.L., Wang Y., Zhang L.Q., Tang W.H. (2005). Improving biocontrol activity of Pseudomonas fluorescens through chromosomal integration of 2,4-diacetylphloroglucinol biosynthesis genes. Chin. Sci. Bull. 50:775–781
Zhu J., Beaber J.W., Moré M. I., Fuqua C., Eberhard A., and Winans S.C. (1998). Analogs of the autoinducer 3-oxooctanoyl-homoserine lactone strongly inhibit activity of the trar protein of Agrobacterium tumefaciens. J. Bacteriol. 180:5398-5405
Acknowledgements
The authors thank Dr Stephen Farrand for providing the A. tumefaciens autoinducer detection system. We also thank Dr Ralph Isberg for reagents and Dr Zhao-Qing Luo for reagents, helpful suggestions and for critical reading of the manuscript. This work was supported by Chinese National Natural Science Found (30100120, 30370952) and Chinese National Programs for High Technology Research and Development (2003AA241170).
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Wei, HL., Zhang, LQ. Quorum-sensing system influences root colonization and biological control ability in Pseudomonas fluorescens 2P24. Antonie Van Leeuwenhoek 89, 267–280 (2006). https://doi.org/10.1007/s10482-005-9028-8
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10482-005-9028-8