Skip to main content
SpringerLink
Log in

Nitrogen assimilation and partitioning in two nitrogen-fixing cultivars of Phaseolus vulgaris L.

  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

Two cultivars of Phaseolus vulgaris L., one responsive (Mexico 309) and one less-responsive (Rio Tibagi) to nodulation with Rhizobium were grown in Leonard jars in a greenhouse. Bean plants were either inoculated with a strain of Rhizobium leguminosarum bv. phaseoli (UMR-1899), a vesicular-arbuscular mycorrhizal (VAM) fungus (Glomus etunicatum) or were left non-inoculated (controls). At two harvests (21 and 28 days post-emergence), extracts containing soluble proteins and free amino acids were prepared from leaves, roots and nodules of field beans. Nodulated plants contained a significantly higher concentration of protein and amino acids in all plant parts. Nitrogen-fixing beans invested a significantly greater proportion of total N as protein-N and amino acid-N as compared to VAM or control beans. Abundant nodule-specific proteins (nodulins) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), identified and quantified using scanning densitometry. Rio Tibagi nodules contained a significantly lower concentration of glutamine synthetase (GS) subunits than did Mexico 309 nodules. Glutamate synthase (GOGAT) and GS activities were low relative to other legumes. The transferase/synthetase ratio for GS was also low indicating that the synthetase activity was caturated and was operating at maximal level in these young N2-fixing associations. Specific nodule activity (SNA) and the level of GS were correlated (r=0.90, p<0.05) for both cultivars at both harvests. GS activity was only 8 or 24% higher than SNA in nodules of Mexico 309 or Rio Tibagi cultivars, respectively, under conditions where substrate was not limiting. This suggests that early in the functioning of this symbiosis N assimilation by GS is the rate-limiting step in N2 fixation by these two bean cultivars, each with a different symbiotic efficiency. Phaseolus breeding programs that attempt to improve N2 fixation in beans should identify germplasm that expresses elevated levels of nodule-specific GS or GOGAT, and this material should be used along with effective R. leguminosarum bv. phaseoli strains that have already been selected, to determine superior host-microsymciont associations.

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

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Ames R N, Reid C P P, Porter L K and Cambardella C 1983 Hyphal uptake and transport of nitrogen from two 15N-labelled sources by Glomus mosseae, a vesicular-arbuscular mycorrhizal fungus. New Phytol. 95, 381–396.

    Google Scholar 

  • Barratt D H P 1980 Method for the detection of glutamine synthetase activity on starch gels. Plant Sci. Lett. 18, 249–254.

    Google Scholar 

  • Bethlenfalvay G J, Pacovsky R S, Bayne H G and Stafford A E 1982 Interactions between nitrogen fixation, mycorrhizal colonization, and host plant growth in the Phaseolus-Rhizobium-Glomus symbiosis. Plant Physiol. 70, 446–450.

    Google Scholar 

  • Boland M J, Fordyce A M and Greenwood R M 1978 Enzymes of nitrogen metabolism in legume nodules: A comparative study. Aust. J. Plant Physiol. 5, 553–559.

    Google Scholar 

  • Boland M J and Benny A G 1977 Enzymes of nitrogen metabolism in legume nodules: Purification and properties of NADH-dependent glutamate synthase from lupin nodules. Eur. J. Biol. Chem. 79, 355–362.

    Google Scholar 

  • Bradford M M 1976 A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.

    Google Scholar 

  • Campos F, Padilla J E, Vázquez M, Ortega J L, Enríquez C and Sánchez F 1987 Expression of nodule specific genes in Phaseolus vulgaris L. Plant Molec. Biol. 9, 521–532.

    Google Scholar 

  • Carvalho M T V and Derbyshire E 1985 Seed lectin variation in cultivars of beans (Phaseolus vulgaris L.) Phaseolus Information Exchange 1, 14–16.

    Google Scholar 

  • Cowan M C 1979 The effect of nitrogen source on levels of amino acids in peas. Plant and Soil 51, 279–282.

    Google Scholar 

  • Cullimore J V, Gebhardt C, Saarelainen R, Miflin B J, Idler K B and Barker R F 1984 Glutamine synthetase of Phaseolus vulgaris L.: Organ-specific gene expression of a multigene family. J. Mol. Appl. Genet. 2, 589–599.

    Google Scholar 

  • Davis B J 1964 Disc electrophoresis. II. Method and application to human serum proteins. Ann. NY Acad. Sci. 121, 404–427.

    Google Scholar 

  • Delaney A and Verma D P S 1988 Cloned nodulin genes for symbiotic nitrogen fixation. Plant Mol. Biol. Rep. 6, 279–285.

    Google Scholar 

  • Derbyshire E, Müller H P, Carvalho M T V and Crocomo O J 1981 Protein profiles of Brazilian beans (Phaseolus vulgaris) obtained by electrophoresis in slabs of polyacrylamide gel. Energia Nuclear Agric. (Piracicaba) 3, 100–109.

    Google Scholar 

  • Duque F F, Neves M C P, Franco A A, Victoria R L and Boddey R M 1985 The response of field grown Phaseolus vulgaris to Rhizobium inoculation and the quantification of N2 fixation using 15N. Plant and Soil 88, 333–343.

    Google Scholar 

  • Felix J-F, Obaton M, Messiaen C-M and Salsac L 1981 Nitrate reductase and nitrogenase activities of common beans (Phaseolus vulgaris L.) from different geographic locations. Plant and Soil 63, 427–438.

    Google Scholar 

  • Ferguson A R and Sims A P 1974 The regulation of glutamine metabolism in Candida utilis: The role of glutamine in the control of glutamine synthetase. J. Gen. Microbiol. 80, 159–171.

    Google Scholar 

  • Freund R J and Littell R C 1981 SAS for linear Models: A Guide to the ANOVA and GLM Procedures. SAS Institute, Raleigh.

    Google Scholar 

  • Gatehouse J A, Evans I M, Croy R R D and Boulter D 1986 Differential expression of genes during legume seed development. Philo. Trans. Royal Soc. (London) B314, 367–384.

    Google Scholar 

  • Groat R G, Vance C P and Barnes D K 1984 Host plant nodule enzymes associated with selection for increased N2 fixation in alfalfa. Crop Sci. 24, 895–898.

    Google Scholar 

  • Groat R G and Vance C P 1982 Root and nodule enzymes of ammonia assimilation in two plant-conditioned symbiotically ineffective genotypes of alfalfa (Medicago sativa L.). Plant Physiol. 69, 614–618.

    Google Scholar 

  • Hungria M and Neves M C P 1987 Cultivar and Rhizobium strain effects on nitrogen fixation and transport in Phaseolus vulgaris L. Plant and Soil 103, 111–121.

    Google Scholar 

  • Hungria M, Barradas C A A and Wallsgrove R M 1990 Nitrogen fixation assimilation and transport during the initial growth stage of Phaseolus vulgaris L. Planta. (In press.)

  • Israel D W 1981 Cultivar and Rhizobium strain effects on nitrogen fixation and remobilization by soybeans. Agron. J. 73, 509–516.

    Google Scholar 

  • Krishna K R and Bagyaraj D J 1983 Changes in free amino nitrogen and protein fractions of ground nut caused by inoculation with VA mycorrhiza. Ann. Bot. 51, 399–401.

    Google Scholar 

  • Laemmli U K 1970 Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227, 680–689.

    Google Scholar 

  • Lara M, Cullimore J V, Lea P J, Miflin B J, Johnson A W B and Lamb J W 1983 Appearance of a novel form of plant glutamine synthetase during nodule development in Phaseolus vulgaris L. Planta 157, 254–258.

    Google Scholar 

  • Legocki R P and Verma D P S 1980 Identification of ‘nodule-specific’ host proteins (nodulins) involved in the development of Rhizobium-legume symbioses. Cell 20, 152–163.

    Google Scholar 

  • Long S R 1989 Rhizobium-legume nodulation: Life together in the underground. Cell 56, 203–214.

    Google Scholar 

  • Minchin F R, Witty J F, Sheehy J E and Muller M 1983 A major error in the acetylene reduction assay: Decreases in nodular nitrogenase activity under assay conditions. J. Exp. Bot. 34, 641–649.

    Google Scholar 

  • Neptune A M L and Muraoka T 1978 Aplicão de ureia- 15N em feijoeiro (Phaseolus vulgaris L.) cultivar carioca. Rev. bras. Ci. Solo 2, 51–55.

    Google Scholar 

  • Oakley B R, Kirsch D R and Morris R 1980 A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels. Anal. Biochem. 105, 361–363.

    Google Scholar 

  • Pacovsky R S 1989 Carbohydrate, protein and amino acid status of Glycine-Glomus-Bradyrhizobium symbioses. Physiol. Plant. 75, 346–354.

    Google Scholar 

  • Pacovsky R S, Bayne H G and Bethlenfalvay G J 1984 Symbiotic interactions between strains of Rhizobium phaseoli and cultivars of Phaseolus vulgaris L. Crop Sci. 24, 101–105.

    Google Scholar 

  • Pacovsky R S, daSilva P, Carvalho M T V and Tsai S M 1991 Growth and nutrient allocation in Phaseolus vulgaris L. colonized by endomycorrhizae or Rhizobium. Plant and Soil 132, 127–137.

    Google Scholar 

  • Padilla J E, Campos F, Conde V, Lara M and Sánchez F 1987 Nodule-specific glutamine synthetase is expressed before the onset of nitrogen fixation in Phaseolus vulgaris L. Plant Mol. Biol. 9, 65–74.

    Google Scholar 

  • Pereira P A A and Bliss F A 1987 Nitrogen fixation and plant growth of common bean (Phaseolus vulgaris L.) at different levels of phosphorus availability. Plant and Soil 104, 79–84.

    Google Scholar 

  • Rennie R J and Kemp G A 1984 15N-determined time course for N2 fixation in two cultivars of field bean. Agron. J. 76, 146–154.

    Google Scholar 

  • Robert F M and Wong P P 1986 Isozymes of glutamine synthetase in Phaseolus vulgaris L. and Phaseolus lunatus L. root nodules. Plant Physiol. 81, 142–148.

    Google Scholar 

  • Rolfe B and Gresshoff P 1988 Genetic analysis of legume nodule initiation. Ann. Rev. Plant Physiol. 39, 297–319.

    Google Scholar 

  • Rosen H 1957 A modified ninhydrin colormetic analysis for amino acids. Arch. Biochem. Biophys. 67, 10–15.

    Google Scholar 

  • Sánchez F, Campos F, Padilla J, Bonneville J-M, Enríques C and Caput D 1987 Purification, cDNA cloning and developmental expression of the nodule-specific uricase from Phaseolus vulgaris L. Plant Physiol. 84, 1143–1147.

    Google Scholar 

  • Sengupta-Gopalan C and Pitas J W 1986 Expression of nodule specific glutamine synthetase genes during nodule development in soybeans. Plant Mol. Biol. 7, 189–199.

    Google Scholar 

  • Smith S E, St John B J, Smith F A and Nicholas D J D 1985 Activity of glutamine synthetase and glutamate dehydrogenase in Trifolium subterraneum L. and Allium cepa: Effects of mycorrhizal infection and phosphate nutrition. New Phytol. 99, 211–227.

    Google Scholar 

  • Thummler F and Verma D P S 1987 Nodulin-100 of soybean is the subunit of sucrose synthetase regulated by the availability of free heme in nodules. J. Biol. Chem. 262, 14730–14736.

    Google Scholar 

  • Tingey S V, Walker E L and Coruzzi G M 1987 Glutamine synthetase genes of pea encode distinct polypeptides which are differentially expressed in leaves, roots and nodules. EMBO J. 6, 1–9.

    Google Scholar 

  • VanKammen A 1984 Suggested nomenclature for plant genes involved in nodulation and symbioses. Plant Mol. Biol. Rep. 2, 43–45.

    Google Scholar 

  • Weiser G C, Grafton K F and Berryhill D L 1985 Nodulation of dry beans by commercial and indigenous strains of Rhizobium phaseoli. Agron. J. 77, 856–859.

    Google Scholar 

Download references

Author information

Author notes

  1. R. S. Pacovsky

    Present address: Department of Botany and Plant Pathology, Michigan State University, 48824-1312, East Lansing, MI, USA

Authors and Affiliations

  1. US Department of Agriculture, Western Regional Research Center, 94710, Albany, CA, USA

    R. S. Pacovsky & G. Fuller

Authors
  1. R. S. Pacovsky
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. Fuller
    View author publications

    You can also search for this author in PubMed Google Scholar

Rights and permissions

Reprints and permissions

About this article

Cite this article

Pacovsky, R.S., Fuller, G. Nitrogen assimilation and partitioning in two nitrogen-fixing cultivars of Phaseolus vulgaris L.. Plant Soil 132, 139–148 (1991). https://doi.org/10.1007/BF00011020

Download citation

  • Received:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00011020

Key words

Search

Navigation