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Denitrifying ability of thirteen Rhizobium meliloti strains

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Abstract

The denitrifying ability of thirteen strains of Rhizobium meliloti was tested. Most of the strains were able to reduce nitrate to nitrous oxide or dinitrogen. However, they failed to use nitrate as electron acceptor for ATP generation or growth at low oxygen tensions. Under micro-aerobic conditions, free-living cells of R. meliloti 102-F-51 strain exhibited a constitutive nitrate reductase activity independent of the presence of nitrate. On the other hand, nitrite reductase activity was dependent not only on low levels of oxygen but also on the presence of a high nitrate concentration in the medium. Denitrification activity proceeded immediately once a threshold level of nitrite was accumulated in the medium or in cells incubated with 1mM nitrite. However, a lag period was required when cells were incubated with nitrate.

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References

  • Arrese-Igor C, Estavillo J M, Peña J I, Gonzalez-Murua C and Aparicio-Tejo P M 1989 Effect of low nitrate supply on nitrogen fixation in alfalfa root nodules induced by Rhizobium meliloti strains with varied nitrate reductase activity. J. Plant Physiol. 135, 207–211.

    CAS  Google Scholar 

  • Arrese-Igor C, Garcia-Plazaola J I, Diaz A and Aparicio-Tejo P M 1991 Distribution of nitrate reductase activity in nodulated lucerne plants. Plant and Soil 131, 107–113.

    CAS  Google Scholar 

  • Arrese-Igor C, Garcia-Plazaola J I, Hernandez A and Aparicio-Tejo P M 1990 Effect of low nitrate supply to nodulated lucerne on time course of activities of enzymes involved in inorganic nitrogen metabolism. Physiol. Plant. 80, 185–190.

    Article  CAS  Google Scholar 

  • Bazylinski D A and Blakemore R P 1983 Denitrification and assimilatory nitrate reduction in Aquaspirillum magnetotacticum. Appl. Environ. Microbiol. 46, 1118–1124.

    PubMed  CAS  Google Scholar 

  • Betlach M R and Tiedje J M 1981 Kinetic explanation for accumulation of nitrite, nitric oxide, and nitrous oxide during bacterial denitrification. Appl. Environ. Microbiol. 42, 1074–1084.

    PubMed  CAS  Google Scholar 

  • Bhandari B and Nicholas D J D 1984 Denitrification of nitrate to nitrogen gas by washed cells of Rhizobium japonicum and by bacteroids from Glycine max. Planta 161, 81–85.

    Article  CAS  Google Scholar 

  • Bourguignon C 1987 Activité dénitrifiante et temps de génération chez 4 espèces de Rhizobium. Ann. Inst. Pasteur/Microbiol. 138, 449–455.

    CAS  Google Scholar 

  • Bradford M M 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

    Article  PubMed  CAS  Google Scholar 

  • Casella S, Shapleigh J P, Lupi F and Payne W J 1988 Nitrite reduction in bacteroids of Rhizobium “hedysari” strain HCNT1. Arch. Microbiol. 149, 384–388.

    Article  CAS  Google Scholar 

  • Cawse P A 1969 The determination of nitrate in soil solutions by ultraviolet spectrophotometry. Analyst 92, 311–315.

    Article  Google Scholar 

  • Chan Y K, Barran L R and Bromfield S P 1989 Denitrification activity of phage types representative of two populations of indigenous Rhizobium meliloti. Can. J. Microbiol. 35, 737–740.

    Article  CAS  Google Scholar 

  • Coyne M S and Tiedje J M 1990 Induction of denitrifying enzymes in oxygen-limited Achromobacter cycloclastes continuous culture. FEMS Microbiol. Ecol. 73, 263–270.

    Article  CAS  Google Scholar 

  • Daniel R M and Appleby C A 1972 Anaerobic-nitrate symbiotic growth of Rhizobium japonicum: effects of cytochrome P-450, other haemoproteins, nitrate and nitrite reductases. Biochim. Biophys. Acta 275, 347–354.

    Article  PubMed  CAS  Google Scholar 

  • Daniel R M, Limmer A W, Steele K W and Smith I M 1982 Anaerobic growth, nitrate reduction and denitrification in 46 Rhizobium strains. J. Gen. Microbiol. 128, 1811–1815.

    CAS  Google Scholar 

  • Daniel R M, Smith I M, Phillip J A D, Ratcliffe H D, Drozd J W and Bull A T 1980 Anaerobic growth and denitrification by Rhizobium japonicum and other rhizobia. J. Gen. Microbiol. 120, 517–521.

    CAS  Google Scholar 

  • Evans H J 1981 Symbiotic nitrogen fixation in legume nodules. In Research Experiences in Plant Physiology. Ed. T CMoore. p 294. Springer-Verlag, New York.

    Google Scholar 

  • Jaworek D, Gruber W and Bergmeyer H U 1974 Methods of Enzymatic Analysis, Vol. 4. Academic Press, London. 2097 p.

    Google Scholar 

  • Kennedy I R, Rigaud J and Trinchant J C 1975 Nitrate reductase from bacteroids of Rhizobium japonicum: Enzyme characteristics and possible interaction with nitrogen fixation. Biochim. Biophys. Acta 397, 24–35.

    PubMed  CAS  Google Scholar 

  • Knowles R 1982 Denitrification. Microbiol. Rev. 46, 43–70.

    PubMed  CAS  Google Scholar 

  • Körner H and Zumft W G 1989 Expression of denitrification enzymes in response to the dissolved oxygen level and respiratory substrate in continuous culture of Pseudomonas stutzeri. Appl. Environ. Microbiol. 55, 1670–1676.

    PubMed  Google Scholar 

  • Manhart J R and Wong P P 1979 Nitrate reductase activities of rhizobia and the correlation between nitrate reduction and nitrogen fixation. Can. J. Microbiol. 25, 1169–1174.

    PubMed  CAS  Google Scholar 

  • Myshkina V L and Bonartseva G A 1990 Testing of a group of rapidly growing Rhizobium cultures for the ability to reduce nitrate and acetylene. Microbiol. 59, 26–28.

    Google Scholar 

  • Nicholas D J D and Nason A 1957 Determination of nitrate and nitrite. Methods Enzymol. 3, 981–984.

    Article  Google Scholar 

  • O'Hara G W and Daniel R M 1985 Rhizobial denitrification: A review. Soil Biol. Biochem. 17, 1–9.

    Article  Google Scholar 

  • O'Hara G W, Daniel R M and Steele K W 1983 Effect of oxygen on the synthesis, activity, and breakdown of Rhizobium denitrification system. J. Gen. Microbiol. 129, 2405–2412.

    Google Scholar 

  • Rigaud J and Puppo A 1977 Effect of nitrite upon leghemoglobin and interaction with nitrogen fixation. Biochim. Biophys. Acta 497, 702–706.

    PubMed  CAS  Google Scholar 

  • Stephens B D and Neyra C A 1983 Nitrate and nitrite reduction in relation to nitrogenase activity in soybean nodules and Rhizobium japonicum bacteroids. Plant Physiol. 71, 731–735.

    Article  PubMed  CAS  Google Scholar 

  • Trinchant J C and Rigaud J 1980 Nitrite inhibition of nitrogenase from soybean bacteroids. Arch. Microbiol. 124, 49–54.

    Article  CAS  Google Scholar 

  • Trinchant J C and Rigaud J 1982 Nitrite and nitric oxide as inhibitors of nitrogenase from soybean bacteroids. Appl. Environ. Microbiol. 44, 1385–1388.

    PubMed  CAS  Google Scholar 

  • Vairinhos F, Wallace W and Nicholas D J D 1989 Simultaneous assimilation and denitrification of nitrate by Bradyrhizobium japonicum. J. Gen. Microbiol. 135, 189–193.

    CAS  Google Scholar 

  • Vincent J M 1970 A Manual for the Practical Study of Root Nodule Bacteria. Blackwell Scientific Publications, Oxford, UK. 165 p.

    Google Scholar 

  • Yoshinari T and Knowles R 1976 Acetylene inhibition of nitrous oxide reduction by denitrifying bacteria. Biochim. Biophys. Res. Commun. 69, 705–710.

    Article  CAS  Google Scholar 

  • Zablotowicz R M, Eskew D L and Focht D D 1978 Denitrification in Rhizobium. Can J. Microbiol. 24, 757–760.

    Article  PubMed  CAS  Google Scholar 

  • Zablotowicz R M and Focht D D 1979 Denitrification and anaerobic, nitrate-dependent acetylene reduction in cowpea Rhizobium. J. Gen. Microbiol. 111, 445–448.

    CAS  Google Scholar 

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Garcia-Plazaola, J.I., Becerril, J.M., Arrese-Igor, C. et al. Denitrifying ability of thirteen Rhizobium meliloti strains. Plant Soil 149, 43–50 (1993). https://doi.org/10.1007/BF00010761

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  • DOI: https://doi.org/10.1007/BF00010761

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