Abstract
A sensitive tritium exchange assay was applied to the Rhizobium system for measuring the expression of uptake hydrogenase in free-living cultures of Rhizobium japonicum. Hydrogenase was detected about 45 hours after inoculation of cultures maintained under microaerophilic conditions (about 0.1% O2). The tritium exchange assay was used to screen a variety of different strains of R. japonicum (including major production strains) with the findings that about 30% of the strains expressed hydrogenase activity with identical results being observed using an alternative assay based on uptake of H2. The relative efficiency of intact soybean nodules inoculated with 10 different rhizobial strains gave results identical to those obtained using free-living cultures. The tritium exchange assay provides an easy, quick, and accurate assessment of H2 uptake efficiency of intact nodules.
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- Anand S. R., Krasna A. I. Catalysis of the H2-HTO exchange by hydrogenase. A new assay for hydrogenase. Biochemistry. 1965 Dec;4(12):2747–2753. doi: 10.1021/bi00888a027. [DOI] [PubMed] [Google Scholar]
- GINGRAS G., GOLDSBY R. A., CALVIN M. Carbon dioxide metabolism in hydrogen-adapted Scenedesmus. Arch Biochem Biophys. 1963 Feb;100:178–184. [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- O'Gara F., Shanmugam K. T. Regulation of nitrogen fixation in Rhizobium spp. Isolation of mutants of Rhizobium trifolii which induce nitrogenase activity. Biochim Biophys Acta. 1977 Dec 22;500(2):277–290. doi: 10.1016/0304-4165(77)90020-4. [DOI] [PubMed] [Google Scholar]
- O'gara F., Shanmugam K. T. Mutant strains of clover rhizobium (Rhizobium trifolii) that form nodules on soybean (Glycine max). Proc Natl Acad Sci U S A. 1978 May;75(5):2343–2347. doi: 10.1073/pnas.75.5.2343. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schubert K. R., Evans H. J. Hydrogen evolution: A major factor affecting the efficiency of nitrogen fixation in nodulated symbionts. Proc Natl Acad Sci U S A. 1976 Apr;73(4):1207–1211. doi: 10.1073/pnas.73.4.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Schubert K. R., Jennings N. T., Evans H. J. Hydrogen Reactions of Nodulated Leguminous Plants: II. Effects on Dry Matter Accumulation and Nitrogen Fixation. Plant Physiol. 1978 Mar;61(3):398–401. doi: 10.1104/pp.61.3.398. [DOI] [PMC free article] [PubMed] [Google Scholar]
- VALENTINE R. C., WOLFE R. S. ROLE OF FERREDOXIN IN THE METABOLISM OF MOLECULAR HYDROGEN. J Bacteriol. 1963 May;85:1114–1120. doi: 10.1128/jb.85.5.1114-1120.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]