Abstract
The kinetics of asymbiotic nitrogenase activity in three strains of the actinomycete Frankia were studied. Decay rates for enzyme activity were determined by adding chloramphenicol to active acetylene-reducing cells and measuring the time required for all activity to cease. Synthesis rates were measured by bubbling oxygen through actively-reducing cells (which totally destroyed all activity) and then measuring the time required for activity to return to normal. Decay rates (t 1/2) for these three strains were approximately 30 to 40 min. Synthesis rates were slower and initial nitrogenase activities were recorded about 110 min (DDB 011610) or 210 min (DDB 020210 and WgCc1.17) after return to air-equilibrated cultures. Frankia strain WgCc1.17 showed a greater sensitivity to oxygen and nitrogenase activity was totally lost when cells were bubbled only with atmospheric concentrations of oxygen. The results presented here indicate that nitrogenase activity turnover time is relatively rapid, on the order of minutes rather than hours or days. However, regulation of nitrogenase activity will differ from one strain to another and asmmbiotic characterization will be useful for understanding nitrogenase regulation in the bacterial-plant symbiosis.
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Baker D, O'Keefe D (1984) A modified sucrose fractionation procedure for the isolation of frankiae from actinorhizal root nodules and soil samples. Plant Soil 78:23–28
Bergersen FJ (1984) Oxygen and the physiology of diazotrophic microorganisms. In: Veeger C, Newton WE (eds) Advances in nitrogen fixation research. Nijhoff/Junk. The Hague, pp 171–180
Bisseling T, Staveren W van, Kammen A van (1980a) The effect of waterlogging on the synthesis of the nitrogenase components in bacteroids of Rhizobium leguminosarum in root nodules of Pisum sativum. Biochem Biophys Res Commun 93:687–693
Bisseling T, Straten J van, Houwaard F (1980b) Turnover of nitrogenase and leghemoglobin in root nodules of Pisum sativum. Biochim Biophys Acta 610:360–370
David KAV, Apte SK, Thomas J (1978) Stimulation of nitrogenase by acetylene: fresh synthesis or conformation change? Biochem Biophys Res Commun 82:39–45
Eady R, Isaac R, Kennedy C, Postgate JR, Ratcliffe HD (1978) Nitrogenase synthesis in Klebsiella pneumoniae: comparison of ammonium and oxygen regulation. J Gen Microbiol 104:277–285
Huss-Danell K, Sellstedt A (1985) Nitrogenase activity in response to darkening and defoliation in Alnus incana. J Exp Bot 36:1352–1358
Huss-Danell K, Sellstedt A, Flower-Ellis A, Sjöström M (1982) Ammonium effects on function and structure of nitrogen-fixing root nodules of Alnus incana (L.) Moench. Planta 156:332–340
Murry M, Hallenbeck PC, Esteva D (1983) Nitrogenase inactivation by oxygen and enzyme turnover in Anabaena cylindrica. Can J Microbiol 29:1286–1294
Murry M, Fontaine MS, Tjepkema JD (1984) Oxygen protection of nitrogenase in Frankia sp. HFP Ar13. Arch Microbiol 139:162–166
Scherer S, Kerfin W, Böger P (1980) Increase of nitrogenase activity in the blue-green alga Nostoc muscorum (Cyanobacterium). J Bacteriol 144:1017–1023
Skeffington RA, Stewart WDP (1976) Evidence from inhibitor studies that the endophyte synthesises nitrogenase in the root nodules of Alnus glutinosa L. Gaertn. Planta 129:1–6
Tjepkema JD, Ormerod W, Torrey JG (1980) Vesicle formation and acetylene reduction activity in Frankia sp. CpI1 cultured in defined media. Nature 287:633–635
Torrey JG, Tjepkema JD, Turner GL, Bergersen FJ, Gibson AH (1981) Dinitrogen fixation by cultures of Frankia sp. CpI1 demonstrated by 15N2 incorporation. Plant Physiol 68:983–984
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Contribution no. 879 from the Battelle-Kettering Laboratory
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Baker, D., Huss-Danell, K. Effects of oxygen and chloramphenicol on Frankia nitrogenase activity. Arch. Microbiol. 144, 233–236 (1986). https://doi.org/10.1007/BF00410953
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DOI: https://doi.org/10.1007/BF00410953