Summary
Mutagenesis provoked by exposure at elevated temperature of the cold-adapted, arctic Rhizobium strain N31 resulted in the generation of five deletion mutants, which exhibited loss of their smaller plasmid (200 kb), whereas the larger plasmid (> 500 kb) was still present in all mutants. Deletion mutants did not show differences from the wild type in the antibiotic resistance pattern, the carbohydrates and organic acids utilization, and the growth rate at low temperature. However, deletion mutants differed from the wild type and among themselves in the ex planta nitrogenase activity, the nodulation index, and the symbiotic effectiveness. The deletion mutant N31.6rif r showed higher nodulation index and exhibited higher nitrogenase activity and symbiotic efficiency than the other deletion mutants and the wild type. The process of deletion mutation resulted in the improvement of an arctic Rhizobium strain having an earlier and higher symbiotic nitrogen fixation efficiency than the wild type.
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References
Antoun H, Bordeleau LM, Sauvageau R (1984) Utilization of the tricarboxylic acid cycle intermediates and symbiotic effectiveness in Rhizobium meliloti. Plant Soil 77:29–38
Banfalvi Z, Sakanyan V, Konez C, Kiss A, Dusha I, Kondorosi A (1981) Location of nodulation and nitrogen fixation genes on a high molecular weight plasmid of R. meliloti. Mol Gen Genet 184:318–325
Beringer JE (1974) R-factor transfer in Rhizobium leguminosarum. J Gen Microbiol 84:188–198
Bordeleau LM, Antoun H, Lachance RA (1977) Effets des souches de Rhizobium meliloti et des coupes successives de la luzerne (Medicago sativa) sur la fixation symbiotique d'azote. Can J Plant Sci 57:433–439
Cannon FC, Beynon J, Hankinson T, Kwiatkowski R, Legocki RP, Ratcliffe H, Ronson C, Szeto W, Williams M (1988) Increased biological nitrogen fixation by genetic manipulation. In: Bothe H, Bruijn FJ de, Newton WE (eds) Nitrogen fixation: hundred years after. Gustav Fisher, Stuttgart New York, pp 735–740
Caudry-Reznick S, Prévost D, Schulman HM (1986) Some properties of arctic rhizobia. Arch Microbiol 146:12–18
Cho MJ, Yang MS, Yun HD, Choe ZR, Kang KY (1985) Genetic engineering of biological nitrogen fixation and its application to agronomy: selection of Rhizobium japonicum mutants having greater symbiotic nitrogen fixation activity with soybean. Korean J Appl Microbiol Bioeng 13:79–86
Gueye M, Bordeleau LM (1988) Nitrogen fixation in bambara groundnut, Voandzeia subterranea (L.) Thouars. MIRCEN J 4:365–375
Hanson RS, Phillips JA (1981) Chemical composition. In: Gerhardt P, Murray RGE, Costilow RN, Nester EW, Wood WA, Krieg NR, Phillips GB (ed) Manual of methods for general bacteriology. American Society for Microbiology, Washington, D.C., pp 328–364
Jain DK, Prévost D, Bordeleau LM (1990) Role of bacterial polysaccharides in the derepression of ex-planta nitrogenase activity with rhizobia. FEMS Microbiol Ecol 73:167–174
Maier RJ, Brill WJ (1978) Mutant strains of Rhizobium Japonicum with increased ability to fix nitrogen for soybean. Science 201:448–450
May JW, Houghton RH, Perret CJ (1964) The effect of growth at elevated temperatures on some heritable properties of Staphylococcus aureus. J Gen Microbiol 37:157–169
Morrison NA, Hau CY, Trinick MJ, Shine J, Rolfe BG (1983) Heat curing of a sym plasmid in a fast-growing Rhizobium sp. that is able to nodulate legumes and the nonlegume Parasponia sp. J Bacteriol 153:527–531
Paau AS (1989) Improvement of Rhizobium inoculants. Appl Environ Microbiol 55:862–865
Prévost D, Bordeleau LM, Caudry-Reznick S, Schulman HM, Antoun H (1987a) Characteristics of rhizobia isolated from three legumes indigenous to the Canadian high arctic: Astragalus alpinus, Oxytropis maydelliana, and Oxytropis arctobia. Plant Soil 98:313–324
Prevost D, Antoun H, Bordeleau LM (1987b) Effects of low temperatures on nitrogenase activity in sainfoin (Onobrychis viciifolia) nodulated by arctic rhizobia. FEMS Microbiol Ecol 45:205–210
Prévost D, Bordeleau LM, Antoun H (1987c) Symbiotic effectiveness of indigenous arctic rhizobia on a temperate forage legume: Sainfoin (Onobrychis viciifolia). Plant Soil 104:63–69
Rosenberg C, Casse-Delbart F, Dusha I, David M, Boucher C (1982) Megaplasmids in the plant-associated bacteria Rhizobium meliloti and Pseudomonas solanaceaurum. J Bacteriol 150:402–406
Shukla RS, Singh CB, Dubey JN (1989) Induced genetic variability in Rhizobium leguminoserum for nitrogen fixation parameters in Vicia faba L. Theor Appl Genet 78:433–435
Simon R, Priefer V, Pühler A (1983) A broad host range mobilization system for in vivo genetic engineering: transposon mutagenesis is gram negative bacteria. Biotechnology 1:784–791
Terawaki Y, Takayasu H, Akiba T (1967) Thermosensitive replication of a kanamycin resistance factor. J Bacteriol 94:687–690
Vincent JM (1970) A manual for the practical study of root nodule bacteria. IBP Handbook No. 15. Blackwell Scientific Publications. Oxford, England
Ward CM, Johnston FB (1962) Chemical methods of plant analysis. Res Branch Agric Canada Publ No. 1064
Zurkowski W, Lorkiewicz Z (1978) Effective method for the isolation of nonnodulating mutants of Rhizobium trifolii. Genet Res 32:311–314
Zurkowski W, Lorkiewicz Z (1979) Plasmid-mediated control of nodulation in Rhizobium trifolii. Arch Microbiol 123:195–201
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Jain, D.K., Bordeleau, L.M. Enhanced N2-fixing ability of a deletion mutant of arctic rhizobia with sainfoin (Onobrychis viciifolia). Theoret. Appl. Genetics 80, 795–800 (1990). https://doi.org/10.1007/BF00224194
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DOI: https://doi.org/10.1007/BF00224194