Abstract
Generation times, acid production, carbon utilization, immunological properties, plasmid content, protein profile and symbiotic properties of 15 isolates of rhizobia nodulating Lotus subbiflorus were studied. Based on specific growth rates, carbon source utilization and acid production, 13 out of the 15 isolates could be assigned to the slow-growing group of rhizobia (bradyrhizobia). Using antisera against whole cells of three isolates, we separated the 15 isolates into three serogroups. Only the slow-growing isolate Ls4 and the fast-growers Ls5 and Ls552 lacked cross-reactivity with any of the sera tested. Electrophoretic mobilities of whole cell protein from seven out of the eight isolates included in the serogroup represented by strain Ls31 were identical. Similarly, isolates Ls1B3 and Ls1B4, both in serogroup Ls1B3, had the same pattern of cell proteins. In contrast, isolates Ls3 and Ls7, belonging to serogroup Ls7, differed in protein profile. Plant growth experiments carried out under bacteriologically controlled conditions revealed that all of the isolates effectively nodulated L. subbiflorus and L. pedunculatus, but were unable to form effective nodules on L. tenuis and L. corniculatus. All isolates showed similar effectiveness in symbiosis with L. subbiflorus, except isolate Ls7, which gave significantly higher plant dry weight.
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Abbreviations
- ELISA:
-
enzyme linked immuno-sorbent assay
- kDa:
-
kiloDalton
- MM:
-
mineral medium
- PBS:
-
phosphate-buffered saline
- RE:
-
relative efficiency
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
- YEM:
-
deyeast extract mannitol
References
Arias A, Cerveñansky C, Gardiol A and Martínez-Drets G 1979 Phosphoglucose isomerase mutant of Rhizobium meliloti. J. Bacteriol. 137, 409–414.
Asuaga A 1994 Lotus subbiflorus cv. El Rincón, a new alternative for extensive improvements of natural pastures. In Proceedings of the First International Lotus Symposium. Eds. P R Beuselinck and A R Craig. pp 147–150. St. Louis, Missouri, USA.
Baraibar A, Frioni L, Guedes M E, Pagliano D, Viera S and Casartelli R 1988 Relevamiento y caracterización de la población de Rhizobium loti en suelos del Uruguay. In Proceedings of the XIV Reunión Latinoamericana de Rhizobiología. Ed. H Urzúa. pp 46–51. Santiago, Chile.
Brito B, Palacios J M, Hidalgo E, Imperial J and Ruiz-Argüeso T 1994 Nickel availability to pea (Pisum sativum) plants limits hydrogenase activity of Rhizobium leguminosarum bv. viciae bacteroids by affecting the processing of the hydrogenase structural subunits. J. Bacteriol. 176, 5297–5303.
Broughton W J, Samrey U and Stanley J 1987 Ecological genetics of Rhizobium meliloti: Symbiotic plasmid transfer in the Medicago sativa rhizosphere. FEMS Microbiol. Lett. 40, 251–255.
Corbin D, Ditta G and Helinski D R 1982 Clustering of nitrogen fixation nif genes in Rhizobium meliloti. J. Bacteriol. 149, 221–228.
Eckhardt T 1978 A rapid method for the identification of plasmid deoxyribonucleic acid in bacteria. Plasmid 1, 584–588.
Evans H J, Harker A R, Papen H, Russell A, Hanus F J and Zuber M 1987 Physiology, biochemistry and genetics of the uptake hydrogenase in rhizobia. Annu. Rev. Microbiol. 41, 335–361.
Evans H J, Russell S A, Hanus F J and Ruiz-Argüeso T 1988 The importance of hydrogen recycling in nitrogen fixation by legumes. In World Crops: Cool Season Food Legumes. Ed. R JSummerfield. pp 777–791. Kluwer Academic Publications, Boston, USA.
Fabiano E and Arias A 1990 Identification of inoculants strains and naturalized population of Rhizobium leguminosarum bv. trifolii using complementary methodologies. World J. Microbiol. Biotechnol. 6, 127–133.
Hartmann A and Amarger N 1991 Genotypic diversity of an indigenous Rhizobium meliloti field population assessed by plasmid profiles, DNA fingerprinting and insertion sequence typing. Can. J. Microbiol. 37, 600–608.
Jarvis B D, Pankhurst C E and Patel J J 1982 Rhizobium loti, a new species of legume root nodule bacteria. Int. J. Sys. Bacteriol. 32, 378–380.
Jordan D C 1982 Transfer of Rhizobium japonicum Buchanam 1980 to Bradyrhizobium japonicum gen. nov., a genus of slow-growing root nodules bacteria. Int. J. Sys. Bacteriol. 32, 136–139.
Jordan D C 1984 Gram-negative aerobic rods and cocci, Family III. Rhizobiaceae. In Bergey's Manual of Systematic Bacteriology. Eds. NKrieg and J GHolt. pp 234–244. Williams and Wilkins, Baltimore, USA.
Laemmli U K 1970 Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.
Leyva A, Palacios J M, Mozo T and Ruiz-Argüeso T 1987 Cloning and characterization of hydrogen uptake genes from Rhizobium leguminosarum. J. Bacteriol. 169, 4929–4934.
Maier R J 1986 Biochemistry, regulation and genetics of hydrogen oxidation in Rhizobium. Crit. Rev. Biotechnol. 3, 17–38.
Martinez de Drets G, Arias A and Rovira de Cutinella M 1974 Fast- and slow-growing rhizobia: differences in sucrose utilization and invertase activity. Can. J. Microbiol. 20, 605–609.
Masterson R, Russi P R and Atherly A G 1982 Nitrogen fixation (nif) genes and large plasmids of Rhizobium japonicum. J. Bacteriol. 152, 938–931.
Michin F R, Witty J F and Mytton L 1994 Reply to “Measurement of nitrogenase activity in legume root nodules: In defence of the acetylene education assay” by J K Vessey. Plant and Soil 158, 163–167.
Monza J, DeFelipe R and Bedmar E J 1992a Nódulos Fix-formados por rhizobios de crecimiento rápido en Lotus subbiflorus. In Resúmenes de la XVI Reunión Latinoamericana de Rhizobiología: p. 42. Sta. Rosa, La Pampa, Argentina.
Monza J, Fabiano E and Arias A 1992b Characterization of an indigenous population of rhizobia nodulating Lotus corniculatus. Soil Biol. Biochem. 24, 241–247.
Murillo J, Villa A, Chamber M and Ruiz-Argüeso T 1989 Occurrence of H2-uptake hydrogenases in Bradyrhizobium sp. (Lupinus) and their expression in nodules of Lupinus spp. and Ornithopus compressus. Plant Physiol. 89, 78–85.
Pankhurst C E 1977 Symbiotic effectiveness of antibiotic-resistant mutants of fast- and slow-growing strains of Rhizobium nodulating Lotus species. Can. J. Microbiol. 23, 1026–1033.
Pankhurst C E, Craig A S and Jones W T 1979 Effectiveness of Lotus root nodules: I. Morphology and flavolan content of nodules formed on Lotus pedunculatus by fast-growing Lotus rhizobia. J. Exp. Bot. 30, 1085–1093.
Pankhurst C E, Broughton W J and Wieneke U 1983 Transfer of an indigenous plasmid of Rhizobium loti to other rhizobia and Agrobacterium tumefaciens. J. Gen. Microbiol. 129, 2535–2543.
Rigaud J and Puppo A 1975 Indole-3-acetic catabolism by soybean bacteroids. J. Gen. Microbiol. 88, 223–228.
Roberst G P, Leps W T, Silver L E and Brill W J 1980 Use of two-dimensional polyacrilamide gel electrophoresis to identify and classify Rhizobium strains. Appl. Environ. Microbiol. 39, 114–422.
Rosemberg C, Boistard P, Denarié J and Casse-Delbart F 1981 Genes controlling early and late functions in symbiosis are located on a megaplasmid in Rhizobium meliloti. Mol. Gen. Genet. 184, 326–333.
Russel H, Pastorini D, Labandera C and Curbelo S 1990 Comportamiento simbiótico de Lotus subbiflorus spp. subbiflorus. In Seminario de Campo Natural. Ed. Hemisferio Sur. pp 255–262. Montevideo, Uruguay.
Sambrook J, Fritsch E F and Maniatis T 1989 Molecular cloning: a laboratory manual, 2nd. ed. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA.
Schubert K R and Evans H J 1976 Hydrogen evolution — a major factor affecting the efficiency of nitrogen fixation in nodulated symbionts. Proc. Natl. Acad. Sci. USA 73, 1207–1211.
Stower M D 1985 Carbon metabolism in Rhizobium species. Annu. Rev. Microbiol. 39, 89–108.
Toro N and Olivares J 1986 Characterization of a large plasmid of Rhizobium meliloti involved in enhancing nodulation. Mol. Gen. Genet. 202, 331–335.
Vessey J K 1994 Measurement of nitrogenase activity in legume root nodules: In defense of the acetylene reduction assay. Plant and Soil 158, 151–162.
Vignais P M and Toussaint B 1994 Molecular biology of membranebound H2 uptake hydrogenases. Arch. Microbiol. 161, 1–10.
Vincent J M 1970 A manual for the Practical Study of the Root-Nodule Bacteria. I.B.P. Handbook No. 15. Blackwell, Oxford, UK.
Wright S F, Foster G J and Bennett O L 1986 Production and use of monoclonal antibodies for identification of strains of Rhizobium trifolii. Appl. Environ. Microbiol. 52, 119–123.
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Irisarri, P., Milnitsky, F., Monza, J. et al. Characterization of rhizobia nodulating Lotus subbiflorus from Uruguayan soils. Plant Soil 180, 39–47 (1996). https://doi.org/10.1007/BF00015409
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DOI: https://doi.org/10.1007/BF00015409