Abstract
O2 protection of nitrogenase in a cultured Frankia isolate from Alnus rubra (HFPArI3) was studied in vivo. Evidence for a passive gas diffusion barrier in the vesicles was obtained by kinetic analysis of in vivo O2 uptake and acetylene reduction rates in response to substrate concentration. O2 of NH +4 -grown cells showed an apparent K m O2 of approximately 1μM O2. In N2-fixing cultures a second K m O2 of about 215 μM O2 was observed. Thus, respiration remained unsaturated by O2 at air-saturation levels. In vivo, the apparent K m for acetylene was more than 10-fold greater than reported in vitro values. These data were inter oreted as evidence for a gas diffusion barrier in the vesicles but not vegetative filaments of Frankia sp. HFPArI3.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Akkermans ADL (1971) Nitrogen fixation and nodulation of Alnus and Hippophae under natural conditions. PhD Thesis, University of Leiden
Becking JH (1977) Endophyte and association establishmentin non-leguminous nitrogen fixing plants. In: Newton W, Postgate JR, Rodriguez-Barrueco C. (eds.) Recent developments in nitrogen fixation. Academic Press, London, pp 551–567
Benson DR, Arp DJ, Burris RH (1979) Cell-free nitrogenase and hydrogenase from actinorhizal root nodules. Science 205:688–689
Bergersen FJ, Turner GL (1973) Kinetic studies of nitrogenase from soya-bean root nodule bacteroids. Biochem J 131:61–75
Bergersen FJ, Turner GL (1980) Properties of terminal oxidase systems of bacteroids from root nodules of soybean and cowpea and of N2-fixing bacteria grown in continuous culture. J Gen Microbiol 118:235–252
Berry A, Torrey JG (1979) Isolation and characterization in vivo and in vitro of an actinomycetous endophyte from Alnus rubra Bong. In: Gordon JC, Wheeler CT, Perry DA (eds) Symbiotic nitrogen fixation in the management of temperate forest. Oregon State University, Corvallis, OR, pp 69–83
Bond G (1960) The oxygen relation of nitrogen fixation in root nodules. Allg Mikrobiol 1:93–99
Bradford MM (1960) A rapid and sensitive method, for quantification of microgram quantities of protein utilizing the principle of protein dye binding. Anal Biochem 72:248–254
Brill WJ (1980) Biochemical genetics of nitrogen fixation. Microbiol Rev 44:449–467
Burggraaf AJP, Shipton WA (1983) Studies on the growth of Frankia isolates in relation to infectivity and nitrogen fixation (acetylene reduction). Can J Bot 61:2774–2782
Davis LC, Wang YL (1980) In vivo and in vitro kinetics of nitrogenase. J Bacteriol 141:1230–1238
Denison RF, Weisz PR, Sinclair TR (1983) Analysis of acetylene reduction rates of soybean nodules at low acetylene concentrations. Plant Physiol 73:648–651
Fessenden RJ, Knowles R, Brouzes R (1973) Acetylene ethylene assay studies on excised root nodules of Myrica asplenifolia L. Soil Sci Soc Amer Proc 37:893–898
Gauthier D, Diem HG, Dommergues Y (1981) In vitro nitrogen fixation by two actinomycete strains isolated from Casuarina nodules. Appl Environ Microbiol 41:306–308
Hallenbeck PC, Kostel PJ, Benemann JR (1979) Purification and properties of nitrogenase from the cyanobacterium Anabaena cylindrica. Eur J Biochem 98:275–284
Hardy RWF, Burns RC, Holsten RD (1973) Application of the acetylene reduction assay for measurement of nitrogen fixation. Soil Biol Biochem 5:47–81
Jensen BB, Cox RP (1983) Effect of oxygen concentration on dark nitrogen fixation and respiration in cyanobacteria. Arch Microbiol 135:287–292
Lambein F, Wolk CP (1973) Structural studies on the glycolipids from the envelope of the heterocyst of Anabcena cylindrica. Biochem 12:791–798
Lang NJ, Fay P (1971) The heterocysts of blue-green algae. II. Details of ultrastructure. Proc Roy Soc London B 178:193–203
Lommen PW, Schwintzer CR, Yocum CS, Gates DM (1971) A model describing photosynthesis in terms of gas diffusion and enzyme kinetics. Planta 98:195–220
Mian S, Bond G (1978) The onset of nitrogen fixation in young alder plants and its relation to differentiation in the nodular endophyte. New Phytol 80:187–192
Murry MA, Fontaine MS, Torrey JG (1984) Kinetics of growth and nitrogenase induction in Frankia sp. ArI3 in batch culture. Plant and Soil 78:61–79
Roades KS (1981) Oxygen sensitivity of nitrogen fixation in the cyanobacterium Anabaenopsis arnoldii. PhD Thesis, University of Michigan
Robson RL, Postgate JR (1980) Oxygen and hydrogen in biological nitrogen fixation. Ann Rev Microbiol 34:183–207
Skeffington RA, Stewart WDP (1976) Evidence from inhibitor studies that the endophyte synthesizes nitrogenase in the root-nodules of Alnus glutinosa L. Gaertn. Planta 129:1–6
Sloger C (1968) Nitrogen fixation by tissues of leguminous and nonleguminous plants. PhD Thesis, University of Florida
Tjepkema JD, Ormerod W, Torrey JG (1980) Vesicle formation and acetylene reduction activity in Frankia sp. CpI1 cultured in defined nutrient media. Nature 287:633–635
Tjepkema JD, Ormerod W, Torrey JG (1981) Factors affecting vesicle formation and acetylene (nitrogenase activity) in Frankia sp. CpI1. J Microbiol 27:815–823
Torrey JG, Callaham D (1982) Structural features of the vesicle of Frankia sp. CpI1 in culture. Can J Microbiol 28:749–757
van Straten J, Akkermans ADL, Roelofsen W (1977) Nitrogenase activity of endophyte suspensions derived from root nodules of Alnus, Hippophae, Shepherdia and Myrica spp. Nature 266:257–258
Waughman GJ (1972) The effect of varying oxygen tension, temperature and sample size on acetylene reduction by nodules of Alnus and Hippophae. Plant Soil 57:521–528
Wheeler CT, Gordon JC, Ching TM (1979) Oxygen relations of the root nodules of Alnus rubra Bong. New Phytol 82:449–457
Winship LJ, Tjepkema JD (1983) The role of diffusion in oxygen protection of nitrogenase in nodules of Alnus rubra. Can J Bot 61:2930–2936
Wittenberg JB, Bergersen FJ, Appleby CA, Turner GL (1974) Facilitated diffusion of O2; the role of leghemoglobin fixation by bacteroids isolated from soybean root nodules. J Biol Chem 249:4057–4066
Yates MG, Jones CW (1974) Respiration and nitrogen-fixation in Azotobacter. Adv Microbial Physiol 11:97–135
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Murry, M.A., Fontaine, M.S. & Tjepkema, J.D. Oxygen protection of nitrogenase in Frankia sp. HFPArI3. Arch. Microbiol. 139, 162–166 (1984). https://doi.org/10.1007/BF00401993
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00401993