Plant and Soil

, Volume 152, Issue 1, pp 1–17 | Cite as

Methods for enhancing symbiotic nitrogen fixation

  • Gudni Hardarson


Biological nitrogen fixation of leguminous crops is becoming increasingly important in attempts to develop sustainable agricultural production. However, these crops are quite variable in their effectiveness in fixing nitrogen. By the use of the 15N isotope dilution method some species have been found to fix large proportions of their nitrogen, while others like common bean have been considered rather inefficient. Methods for increasing N2 fixation are therefore of great importance in any legume work. Attempts to enhance nitrogen fixation of grain legumes has been mainly the domain of microbiologists who have selected rhizobial strains with superior effectiveness or competitive ability. Few projects have focused on the plant symbiont with the objective of improving N2 fixation as done in the FAO/IAEA Co-ordinated Research Programme which is being reported in this volume. The objective of the present paper is to discuss some possibilities available for scientists interested in enhancing symbiotic nitrogen fixation in grain legumes. Examples will be presented on work performed using agronomic methods, as well as work on the plant and microbial symbionts. There are several methods available to scientists working on enhancement of N2 fixation. No one approach is better than the others; rather work on the legume/Rhizobium symbiosis combining experience from various disciplines in inter-disciplinary research programmes should be pursued.

Key words

grain legumes N2 15N isotope dilution nitrogen fixation Rhizobium symbiosis 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Afza R, Hardarson G, Zapata F and Danso S K A 1987 Effects of delayed soil and foliar N fertilization on yield and N2 fixation of soybean. Plant and Soil 97, 361–368.Google Scholar
  2. Agboola A A and Fayemi A A A 1972 Fixation and excretion of nitrogen by tropical legumes. Agron. J. 64, 409–412.Google Scholar
  3. Barnes D K, Heichel G H, Vance C P, Viands D R and Hardarson G 1981 Successes and problems encountered while breeding for enhanced N2 fixation in alfalfa. In Genetic Engineering of Symbiotic Nitrogen Fixation and Conservation of Fixed Nitrogen. Eds. J MLyons, R CValentine, D APhillips, D WRains and R CHuffaker, pp. 233–248. Plenum Press, New York.Google Scholar
  4. Barnes D K, Heichel G H, Vance C P and Ellis W R 1984 A multiple-trait breeding program for improving the symbiosis for N2 fixation between Medicago sativa L and Rhizobium meliloti. Plant and Soil 82, 303–314.Google Scholar
  5. Bergersen F J (Ed.) 1980 Methods for Evaluating Biological Nitrogen Fixation. Wiley, Chichester 701 p.Google Scholar
  6. Beringer J E 1989 Biotechnology in relation to beneficial microorganisms. In Biotechnology Study Project Papers; Summaries of Commissioned Papers. World Bank, ISNAR, and ACIAR, Canberra, pp 30–31.Google Scholar
  7. Bliss F A 1993 Breeding common bean for improved biological nitrogen fixation. Plant and Soil 152, 71–79.Google Scholar
  8. Boote K J, Gallagher R N, Robertson W K, Hinson K and Hammond L C 1978 Effect of foliar fertilization on photosynthesis, leaf nutrition and yield of soybean. Agron. J. 70, 787–791.Google Scholar
  9. Bowen G D and Danso S K A 1987 Nitrogen research for perennial crops. IAEA Bulletin 29, 5–8 p.Google Scholar
  10. Brockwell J, Diatloff A, Roughly R J and Date R A 1982 Selection of rhizobia for inoculants. In Nitrogen Fixation in Legumes. Ed. J MVincent pp 173–191. Academic Press, Sydney.Google Scholar
  11. Carroll B J, McNeil D L and Gresshoff P M 1985 A supernodulation and nitrate-tolerant symbiotic (nts) soybean mutant. Plant Physiol. 78, 34–40.Google Scholar
  12. Craswell E T 1990 Biological Nitrogen Fixation: Investments and Expectations from a Donor's Perspective Working Paper No. 30. ACIAR, Canberra 8 p.Google Scholar
  13. Cregan P B, Keyser H H and Sadowsky 1988 Soybean genotypes which reduce the competitiveness of the Bradyrhizobium japonicum serotype strains comprising serocluster 123. In Nitrogen Fixation: Hundred Years After. Eds. HBothe, F JdeBruijn and W ENewton. Gustav Fischer, Stuttgart 782 p.Google Scholar
  14. Döbereiner J 1982 The Brazilian program in biological nitrogen fixation. In Biological Nitrogen Fixation Technology for Tropical Agriculture. Eds. P HGraham and S CHarris. pp 687–688. CIAT, Cali.Google Scholar
  15. FAO 1984 Legume Inoculants and Their Use. Food and Agriculture Organization of the United Nations, Rome 63 p.Google Scholar
  16. Danso S K A, Zapata F, Hardarson G and Fried M 1987 Nitrogen fixation in faba bean as affected by plant population density in sole or intercropped systems with barley. Soil Biol. Biochem. 19, 411–415.Google Scholar
  17. Eaglesham A R J, Ayanaba A, Rao V R and Eskew D L 1981 Improving the nitrogen nutrition of maize by intercropping with cowpea. Soil Biol. Biochem 13, 169–171.Google Scholar
  18. Gibson A H 1980 Methods for legumes in glasshouses and controlled environment cabinets. In Methods for Evaluating Biological Nitrogen Fixation. Ed. F JBergersen pp 139–184. Wiley, Chichester.Google Scholar
  19. Giller K E and Wilson K J 1991 Nitrogen Fixation in Tropical Cropping System CAB International 313 p.Google Scholar
  20. Halliday J 1984 Integrated approach to nitrogen fixing trees germplasm development. Braz. J Agric. Res. 19, 91–117.Google Scholar
  21. Hardarson G and Jones D G 1979a Effect of temperature on competition amongst strains of Rhizobium trifolii for nodulation of two white clover varieties. Ann. Appl. Biol. 92, 229–236.Google Scholar
  22. Hardarson G and Jones D G 1979b The inheritance of preference for strains of Rhizobium trifolii by white clover (Trifolium repens L.). Ann. Appl. Biol. 92, 329–333.Google Scholar
  23. Hardarson G and Lie T A 1984 Breeding Legumes for Enhanced Symbiotic Nitrogen Fixation. Martinus Nijhoff/Dr. W. Junk Publishers, Dordrecht 166 p.Google Scholar
  24. Hardarson G, Heichel G H, Vance C P and Barnes D K 1981 Evaluation of alfalfa and Rhizobium meliloti for compatibility in nodulation and nodule effectiveness. Crop Sci. 21, 562–567.Google Scholar
  25. Hardarson G, Zapata F and Danso S K A 1984a Field evaluation of symbiotic nitrogen fixation by rhizobial strains using 15N methodology. Plant and Soil 82, 369–375.Google Scholar
  26. Hardarson G, Zapata F and Danso S K A 1984b Effect of plant genotype and nitrogen fertilizer on symbiotic nitrogen fixation by soybean cultivars. Plant and Soil 82, 397–406.Google Scholar
  27. Hardarson G, Danso S K A and Zapata F 1987 Biological nitrogen fixation in field crops. In Handbook of Plant Science in Agriculture Ed. B RChristie pp 165–192. CRC Press Inc., Boca Raton, FL.Google Scholar
  28. Hardarson G, Danso S K A and Zapata F 1988 Dinitrogen fixation measurement in alfalfa-ryegrass swards using nitrogen-15 and influence of the reference crop. Crop Sci. 28, 101–105.Google Scholar
  29. Hardarson G, Golbs M and Danso S K A 1989 Nitrogen fixation in soybean (Glycine max L. Merrill) as affected by nodulation patterns. Soil Biol. Biochem. 21, 783–787.Google Scholar
  30. Hardarson G, Danso S K A, Zapata F and Reichardt K 1991 Measurements of nitrogen fixation in fababean at different N fertilizer rates using the 15N isotope dilution and ‘A-value’ methods. Plant and Soil 131, 161–168.Google Scholar
  31. Hardarson G, Bliss F A, Cigales-Rivera M R, Henson R A, Kipe-Nolt J A, Longeri L, Manrique A, Peña-Cabriales J J, Pereira P, Sanabria C A and Tsai S M 1993 Genotypic variation in biological nitrogen fixation by common bean. Plant and Soil 152, 59–70.Google Scholar
  32. Heichel G H, Barnes D K, Vance C P and Sheaffer C C 1989 Dinitrogen fixation technology for alfalfa improvement. J. Prod. Agric. 2, 24–32.Google Scholar
  33. Herridge D F and Betts J H 1988 Field evaluation of soybean genotypes selected for enhanced capacity to nodulate and fix nitrogen in the presence of nitrate. Plant and Soil 110, 129–135.Google Scholar
  34. IITA 1985 Grain Legume Improvement Program: Research Highlights 1981–1984. IITA, Nigeria. 83 p.Google Scholar
  35. Keyser H H, Cregan P B and Sadowsky M J 1988 Soybean genotypes which restrict nodulation of strains of Bradyrhizobium japonicum serocluster 123. In Nitrogen Fixation: Hundred Years After. Eds. HBothe, F JdeBruijn and W ENewton. Gustav Fischer, Stuttgart 784 p.Google Scholar
  36. Kuykendall L D and Weber D F 1978 Genetically marked Rhizobium identifiable as inoculum strain in nodules of soybean plants grown in field populated with Rhizobium japonicum. Appl. Environ. Microbiol. 36, 915–919.Google Scholar
  37. McDermott T R and Graham P H 1989 Bradyrhizobium japonicum inoculant mobility, nodule occupancy, and acetylene reduction in the soybean root system. Appl. Environ. Microb. 55, 2493–2498.Google Scholar
  38. Peoples M B, Faizah A W, Rerkasem B and Herridge D F 1989 Methods for Evaluating Nitrogen Fixation by Nodulated Legumes in the Field. ACIAR, Canberra 76 p.Google Scholar
  39. Poole W D, Randall G W and Ham G E 1983 Foliar fertilization of soybean. I. Effect of fertilizer source, rates and frequency of application. Agron. J. 75, 195–200.Google Scholar
  40. Ramos M L G, Ribeiro W OJr and Kipe-Nolt J A 1993 Effect of fungicides on survival of Rhizobium on seeds and the nodulation of bean (Phaseolus vulgaris L) Plant and Soil 152, 145–150.Google Scholar
  41. Rosas J C and Bliss F A 1986 Host plant traits associated with estimates of nodulation and nitrogen fixation in common bean. Hort Science 21, 287–289.Google Scholar
  42. Roughley R J 1980 Environmental and cultural aspects of the management of legumes and Rhizobium. In Advances in Legume Sciences Eds. R JSummerfield and A HBunting. pp. 97–103. Royal Botanic Gardens, Kew.Google Scholar
  43. Roughley R J, Sprent J I and Day J M 1983 Nitrogen fixation. In The Faba Bean (Vicia faba L.) Ed. D DHabblethwaite. pp 230–260. Butterworths, London.Google Scholar
  44. Senaratne R, Amornpimol C and Hardarson G 1987 Effect of combined nitrogen on nitrogen fixation of soybean (Glycine max L. Merrill) as affected by cultivar and rhizobial strain. Plant and Soil 103, 45–50.Google Scholar
  45. Silvester-Bradley R, Ayarza M A, Mendez J E and Moriones R 1983 Use of undisturbed soil cores for evaluation of Rhizobium strains and methods for inoculation of tropical forage legumes in a Colombian Oxisol. Plant and Soil 74, 237–247.Google Scholar
  46. Somasegaran P and Hoben H J 1985 Methods in Legume-Rhizobium Technology. NifTAL, MIRCEN, Hawaii 367 p.Google Scholar
  47. Tauer L W 1989 Economic impact of future biological nitrogen fixation technologies on United States agriculture. Plant and Soil, 119, 261–270.Google Scholar
  48. Viands D R, Barnes D K and Heichel G H 1981 Nitrogen Fixation in Alfalfa: Responses to Bidirectional Selection for Associated Characteristics. Technical Bulletin No. 1643. USDA-ARS 18 p.Google Scholar
  49. Vincent J M 1970 A Manual for the Practical Study of Root-Nodule Bacteria. Blackwell Scientific Publications, Oxford 164 p.Google Scholar
  50. Wolyn D J, Attewell J, Ludden P W and Bliss F A 1989 Indirect measure of N2 fixation in common bean (Phaseolus vulgaris L.) under field conditions. Role of lateral root nodules. Plant and Soil 113, 181–187.Google Scholar
  51. Zapata F, Danso S K A, Hardarson G and Fried M 1987 Time course of nitrogen fixation in field-grown soybean using nitrogen-15 methodology. Agron. J. 79, 172–176.Google Scholar

Copyright information

© Kluwer Academic Publishers 1993

Authors and Affiliations

  • Gudni Hardarson
    • 1
  1. 1.FAO/IAEA Programme, Soil Science UnitAgency's LaboratoriesSeibersdorfAustria

Personalised recommendations