Abstract
Plasmid pBS501, responsible for the resistance of the wild-type Pseudomonas sp. BS501(pBS501) to cobalt and nickel ions, was conjugatively transferred to the rhizosphere Pseudomonas aureofaciens strain BS1393, which is able to synthesize phenazine antibiotics and to suppress a wide range of phytopathogenic microorganisms. The transconjugant P. aureofaciens BS1393(pBS501) turned out to be resistant to cobalt and nickel with an MIC of 8 mM. When grown in a synthetic medium with 0.25 mM cobalt, the transconjugant accumulated 6 times more cobalt than the wild-type strain BS501(pBS501) (1.2 versus 0.2 μg Co/mg protein). Electron microscopic studies showed that cobalt accumulates on the surface of transconjugant cells in the form of electron-opaque granules. In a culture medium with 2 mM cobalt or nickel, strain BS1393 produced phenazine-1-carboxylic acid in trace amounts. The transconjugant P. aureofaciens BS1393(pBS501) produced this antibiotic in still smaller amounts. Unlike the parent strain BS1393, the transconjugant P. aureofaciens BS1393(pBS501) was able to suppress in vitro the growth of the phytopathogenic fungus Gaeumannomyces graminis var. tritici1818 in a medium containing 0.5 mM cobalt or nickel.
Similar content being viewed by others
REFERENCES
Chin-A-Woeng, T.F.C., Bloemberg, G.V., van der Bij, A.J., van der Drift, K.M.G.M., Scheffer, J., Keel, Ch., Baker, P.A.H.M., Tichy, H.-V., de Bruijn, F.J., Thomas-Oates, J.E., and Lugtenberg, J.J., Biocontrol by Phenazine-1-Carboxamide-producing Pseudomonas chlororaphis PCL 1391 of Tomato Root Rot Caused by Fusarium oxysporum f. sp. radicislycopersici, Mol. Plant-Microbe Interact., 1998, vol. 11, no. 11, pp. 1069–1077.
Slininger, P.J. and Jackson, M.A., Nutritional Factors Regulating Growth and Accumulation of Phenazine-1-Carboxylic Acid by Pseudomonas fluorescens 2-79, Appl. Microbiol. Biotechnol., 1992, vol. 37, pp. 388–392.
Duffy, B.K. and Defago, G., Environmental Factors Modulating Antibiotic and Siderophore Biosynthesis by Pseudomonas fluorescens Biocontrol Strains, Appl. Environ. Microbiol., 1999, vol. 65, no. 6, pp. 2429–2438.
Ivanov, A.Yu., Gavryushkin, A.V., Siunova, T.V., Khasanova, L.A., and Khasanova, Z.M., Investigation of Heavy Metal Resistance of Some Pseudomonas Strains, Mikrobiologiya, 1999, vol. 68, no. 3, pp. 366–374.
Boronin, A.M. and Kochetkov, V.V., Biopreparations Based on Pseudomonads, AGRO XXI, 2000, no. 3, pp. 3–5.
Maniatis, T., Fritsch, E.F., and Sambrook, J., Molecular Cloning: A Laboratory Manual, Cold Spring Harbor: Cold Spring Harbor Lab., 1982. Translated under the title Molekulyarnoe klonirovanie, Moscow: Mir, 1984.
King, E.O., Ward, M.K., and Raney, D.E., Two Simple Media for the Demonstration of Pyocyanin and Fluorescein, J. Lab. Clin. Med., 1954, vol. 44, pp. 301–307.
Mergeay, M., Nies, D., Schlegel, H.G., Gerits, J., Charles, P., and Van Gijsegem, F., Alcaligenes eutrophus CH34 Is Facultative Chemolithotroph with Plasmidbound Resistance to Heavy Metals, J. Bacteriol., 1985, vol. 162, pp. 328–334.
Hamdan, H., Weller, D.M., and Thomashow, L.S., 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, vol. 57, pp. 3270–3277.
Fernandez, R.O. and Pizarro, R.A., High Performance Liquid Chromatographic Analysis of Pseudomonas aeruginosa Phenazines, J. Chromatogr., 1977, vol. 771, pp. 99–104.
Lowry, O.H., Rosebrough, N.J., Farr, A.L., and Randall, R.J., Protein Measurement with the Folin Phenol Reagent, J. Biol. Chem., 1957, vol. 193, pp. 265–275.
Reynolds, E.S., The Use of Lead Citrate at High pH as an Electron-Opaque Stain in Electron Microscopy, J. Cell Biol., 1963, vol. 17, pp. 208–212.
Smirnov, V.V. and Kiprianova, E.A., Bakterii roda Pseudomonas (Bacteria of the Genus Pseudomonas), Kiev: Nauk. Dumka, 1990.
Gelmi, M., Apostoli, P., Porru, S., Alessio, L., and Turano, A., Resistance to Cadmium Salts and Metal Absorption by Different Microbial Species, Curr. Microbiol., 1994, vol. 29, no. 6, pp. 335–341.
Beveridg, T.J. and Fyfe, W.S., Metal Fixation by Bacterial Cell Walls, Can. J. Earth Sci., 1985, vol. 22, pp. 1893–1898.
Holmes, J.D., Smith, P.R., Evansgowing, R., Richardson, D.J., Russel, D.A., and Sodeau, J.R., Energy-Dispersive X-Ray Analysis of the Extracellular Cadmium Sulfide Crystallites of Klebsiella aerogenes, Arch. Microbiol., 1995, vol. 163, no. 2, pp. 143–147.
Lee, Y. and Tebo, B.M., Cobalt(II) Oxidation by Marine Manganese(II)-oxidizing Bacillus sp. Strain SG-1, Appl. Environ. Microbiol., 1994, vol. 60, no. 8, pp. 2949–2957.
Nies, D.H., Resistance to Cadmium, Cobalt, Zinc, and Nickel in Microbes, Plasmid, 1992, vol. 27, pp. 17–28.
Silver, S., Bacterial Heavy Metal Resistance: New Surprises, Annu. Rev. Microbiol., 1996, vol. 50, pp. 753–789.
Mordukhova, E.A., Sokolov, S.L., Kochetkov, V.V., Kosheleva, I.A., Zelenkova, N.F., and Boronin, A.M., Involvement of Naphthalene Dioxygenase in Indole-3-Acetic Biosynthesis by Pseudomonas putida, FEMS Microbiol. Lett., 2000, vol. 190, pp. 279–285.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Siunova, T.V., Kochetkov, V.V., Validov, S.Z. et al. The Production of Phenazine Antibiotics by the Pseudomonas aureofaciens Strain with Plasmid-Controlled Resistance to Cobalt and Nickel. Microbiology 71, 670–676 (2002). https://doi.org/10.1023/A:1021427806161
Issue Date:
DOI: https://doi.org/10.1023/A:1021427806161