Springer Nature is making SARS-CoV-2 and COVID-19 research free. View research | View latest news | Sign up for updates

Hydrogen uptake by Robinia pseudoacacia nodules

  • 28 Accesses

  • 3 Citations


Hydrogen uptake is thought to increase the efficiency of nitrogen fixation by recycling H2 produced by nitrogenase that would otherwise be lost by diffusion. Here we demonstrate the capacity of eight Rhizobium strains to take up molecular hydrogen. Uptake by nodule homogenates from Robinia pseudoacacia was measured amperometrically under nitrogenase repression. Markedly lower activities were found than in soybean nodules. In addition hydrogenase activity was detected by the ability of bacteroids to reduce methylene blue in the presence of hydrogen. It was demonstrated that hydrogenase structural genes are present in the black locust symbiont, Rhizobium sp. strain R1, using hybridization with a plasmid, which contained hydrogenase genes from R. leguminosarum bv. viceae.

This is a preview of subscription content, log in to check access.


  1. Arp JD (1992) Hydrogen cycling in symbiontic bacteria. In: Stacey G, Burris RH, Evans HJ (eds) Biological nitrogen fixation. Chapman and Hall, New York, pp 432–460

  2. Basit HA, Angle JS, Salem S, Gewaily EM, Kotob SI, van Berkum P (1991) Phenotypic diversity among strains of Bradyrhizobium japonicum belonging to serogroup 110. Appl Environ Microbiol 57: 1570–1572

  3. Batzli JM (1991) Indigenous rhizobial diversity of Robinia pseudoacacia L., and nodulation studies of Maackia amurensis maxim, & rupr. and Sophora japonica L. M. Sc. Thesis, Department of Horticulture, University of Maryland, College Park, Maryland

  4. Batzli JM, Graves WR, van Berkum P (1992) Diversity among rhizobia effective with Robiniap seudoacacia L. Appl Environ Microbiol 58: 2137–2143

  5. van Berkum P (1987) Expression of uptake hydrogenase and hydrogen oxidation during heterotrophic growth of Bradyrhizobium japonicum. J Bacteriol 169: 4565–4569

  6. Dixon ROD (1967) Hydrogen uptake and exchange by pea root nodules. Ann Bot 31: 179–188

  7. Dixon ROD (1968) Hydrogenase in pea root nodule bacteroids. Arch Microbiol 62: 272–283

  8. Drevon JJ, Kalia VC, Heckmann MO, Salsac L (1987) Influence of the Bradyrhizobium japonicum hydrogenase on the growth of Glycine and Vigna Species. Appl Environ Microbiol 53: 610–612

  9. Evans HJ, Burris RH (1992) Highlights in biological nitrogen fixation during the last 50 years. In: Stacy G, Burris RH, Evans HJ (eds) Biological nitrogen fixation. Chapman and Hall, New York, pp 1–42

  10. Evans HJ, Hanus FJ, Haugland RA, Cantrell MA, XU LS, Russell SA, Lambert GR, Harker AR (1985) Hydrogen recycling in nodules affects nitrogen fixation and growth of soybeans. In: Shibles R (ed) Proceedings of the world soybean research conference. III. Westview Press, Boulder, pp 935–942

  11. Fuhrmann J (1990) Symbiotic effectiveness of indigenous soybean bradyrhizobia as related to serological, morphological, rhizobia toxine and hydrogenase phenotypes. Appl Environ Microbiol 56: 224–229

  12. Hanus FJ, Carter KR, Evans HJ (1980) Techniques for measurement of hydrogen evolution by nodules. Methods Enzymol 69: 731–739

  13. Haugland RA, Hanus FJ, Cantrell MA, Evans HJ (1983). Rapid colony screening method for identifying hydrogenase activity in Rhizobium japonicum. Appl Environ Microbiol 45: 892–897

  14. Herbert D, Philipps PJ, Strange RE (1971) Chemical analysis of microbial cells. In: Norris JR, Ribbons DW (eds) Methods in microbiology, vol. 5 b. Academic Press, New York, pp 249–252

  15. van Kessel C, Roskoski JP, Wood T, Montano J (1983) 15N2 fixation and H2 evolution by six species of tropical leguminous trees. Plant Physiol 72: 909–910

  16. Lambert GR, Hanus FJ, Rüssel SA, Evans HJ (1985) Determination of the hydrogenase status of individual legume nodules by a methylene blue reduction assay. Appl Environ Microbiol 50: 537–539

  17. Martinez-Romero E, Segovia L, Mercante FM, Franco AA, Graham P, Pardo MA (1991) Rhizobium tropici, a novel species nodulating Phaseolus vulgaris L. beans and Leucaena sp. trees. Int J Sys Bacteriol 41: 417–426

  18. Röhm M, Werner D (1991) Nitrate levels affect the development of the black locust-Rhizobium symbiosis. Trees 5: 227–231

  19. Röhm M, Werner D (1992) Robiniapseudoacacia-Rhizobium symbiosis: isolation and characterization of a fast-nodulation and efficiently nitrogen fixing Rhizobium strain. Nitrogen Fix Tree Res Rep 10: 193–197

  20. Schubert, Evans (1976) Hydrogen evolution: a major factor affecting the efficiency of nitrogen fixation in nodulated symbionts. Proc Nat Acad Sci USA 73: 1207–1211

  21. Simpson FB, Burris RH (1984) A nitrogen pressure of 50 atmospheres does not prevent evolution of hydrogen by nitrogenase. Science 224: 1095–1097

  22. Streit W, Kosch K, Werner D (1992) Nodulation competitivenes of Rhizobium leguminosarum bv. phaseoli and Rhizobium tropici strains measured by use of the glucuronidase (gus) gene fusion. Biol Fert Soils 14: 140–144

  23. Wang R, Healey FP, Myers J (1971) Amperometric measurement of hydrogen evolution in Chlamydomonas. Plant Physiol 48: 108–110

  24. Werner D, Stripf R (1978) Differentiation of Rhizobium japonicum. I. Enzymatic comparison of nitrogenase repressed and derepressed free living cells and of bacteroids. Z Naturforsch 33: 245–252

  25. Werner D, Wilcockson J, Kalkowsky B (1975 a) Nitrogen fixing activity in Rhizobium japonicum separated from plant cell cultures. Z Naturforsch 30: 687–688

  26. Werner D, Wilcockson J, Zimmermann E (1975 b) Absorption and selection of rhizobia by ion exchange papers. Arch Microbiol 105: 27–32

  27. Zablotowicz RM, Russell SA, Evans HJ (1980) Effect of the hydrogenase system in Bradyrhizobium japonicum on the nitrogen fixation and growth of soybeans at different stages of development. Agron J 72: 555–559

Download references

Author information

Rights and permissions

Reprints and Permissions

About this article

Cite this article

Röhm, M., Streit, W., Evans, H.J. et al. Hydrogen uptake by Robinia pseudoacacia nodules. Trees 8, 99–103 (1993). https://doi.org/10.1007/BF00197687

Download citation

Key words

  • Robinia pseudoacacia L.
  • Hydrogen uptake
  • Hydrogenase
  • Nitrogen fixation