Abstract
Chickpea (Cicer arietinum L.) cv. C-235 inoculated with Rhizobium sp. (Cicer) strain cv4Az was raised in sand culture under natural conditions with nitrogen-free nutrient solution. 45-d-old plants were treated with 20 and 50 mM KNO3 and sampling made 2 and 6 d after treatment. KNO3 application induced premature nodule senescence. Light microscopic investigations showed that KNO3 treatments resulted in structural degradation of the central bacteroidal tissue. The mass of green nodules increased by 35 % under these treatments. This was accompanied by a rapid decline in leghemoglobin (Lb) content of the nodules being 51 - 67 % lower than in control. The total soluble nodule proteins showed relatively minor changes under KNO3 treatments thus suggesting preferential degradation of Lb. These changes were associated with a rapid decline in N2-fixing activity. However, the decline in total soluble sugars was relatively minor as compared to acetylene reducing activity, thus indicating that sugar deprivation is not the cause of decreased nitrogen fixation ability. Glutathione reductase and ascorbate peroxidase activity showed a 10 - 20 % decrease in comparison with the control. Accumulation of H2O2 and structural degradation of the nodular tissue are considered to be the factors leading to nodule senescence under nitrate treatments.
Similar content being viewed by others
References
Appleby, C.A.: Leghemoglobin and Rhizobium respiration.-Annu. Rev. Plant Physiol. 35: 443-478, 1984.
Becana, M., Aparacio-Tejo K., Sánchez-Díaz, M.: Nitrate and hydrogen peroxide metabolism in Medicago sativa nodules and possible effect on leghaemoglobin function.-Physiol. Plant. 72: 755-761, 1988.
Becana, M., Klucas, R.V.: Transition metals in legume root nodules: iron-dependent free radical production increases during nodule senescence.-Proc. nat. Acad. Sci. USA 89: 8958-8962, 1992.
Becana, M., Rodríguez-Barrueco, C.: Protective mechanisms of nitrogenase against oxygen excess and partially reduced oxygen intermediates.-Physiol. Plant. 75: 429-438, 1989.
Becana, M., Salin, M.L., Ji, L., Klucas, R.V.: Flavin-mediated reduction of ferric leghemoglobin from soybean nodules.-Planta 183: 575-583, 1991.
Becana, M., Sprent, J.I.: Nitrogen fixation and nitrate reduction in root nodules of legumes.-Physiol. Plant. 70: 757-765, 1987.
Dalton, D.A.: Antioxidant defence of plants and fungi.-In: Ahmad, S. (ed.): Oxidative Stress and Antioxidant Defences in Biology. Pp. 298-355. Chapman and Hall, New York 1995.
Dalton, D.A., Post, C.J., Langeberg, L.: Effect of ambient oxygen and of fixed nitrogen on concentrations of glutathione, ascorbate and associated enzymes in soybean root nodules.-Plant Physiol. 96: 812-818, 1991.
Dalton, D.A., Russell, S.A., Hanus, F.J., Pascoe, G.A., Evans, H.J.: Enzymatic reactions of ascorbate and glutathione that prevent peroxide damage in soybean root nodules.-Proc. nat. Acad. Sci. USA 83: 3811-3815, 1986.
DeLorenzo, C.A., Fernández-Pascual, M.M., De Felipe, M.R.: Protective enzymes against active oxygen species during nitrate-induced senescence of Lupinus albus nodules.-J. Plant Physiol. 144: 633-640, 1994.
DeLorenzo, C.A., Lucas, M.M., Vivo, A., De Felipe, M.R.: Effect of nitrate on peroxisome ultrastructure and catalase activity in nodules of Lupinus albus L. cv. Multolupa.-J. exp. Bot. 41: 1573-1578, 1990.
Escuredo, P.R., Minchin, F.R., Gogorcena, Y., Iturbe-Ormaetxe, I., Klucas, R.V., Becana, M.: Involvement of activated oxygen in nitrate induced senescence of pea root nodules.-Plant Physiol. 110: 1187-1195, 1996.
Faurie, O., Soussana, J.F.: Oxygen-induced recovery from short term nitrate inhibition of N2 fixation in white clover plants from spaced and dense stands.-Physiol. Plant. 89: 467-475, 1993.
Hardy, R.W.F., Burns, R.C., Holsten, R.D.: Application of C2H2-C2H4 assay for measurement of nitrogen fixation.-Soil Biol. Biochem. 5: 47-81, 1973.
Hardy, R.W.F., Havelka, U.D.: Photosynthate as a major factor limiting nitrogen fxation by field grown legumes with emphasis on soybean.-In: Nutman, P.S. (ed.): Symbiotic Nitrogen Fixation in Plants. Pp. 421-439. Cambridge University Press, Cambridge 1976.
Jun, H.K., Sarath, G., Moran, J.F., Becana, M., Klucas, R.V., Wagner, F.W.: Characteristics of modified leghemoglobins isolated from soybean (Glycine max Merr.) root nodules.-Plant Physiol. 104: 1231-1236, 1994.
Lowry, O.H., Rosenbrough, N.J., Farr, A.L., Randall, R.J.: Protein measurement with the Folin phenol reagent.-J. biol. Chem. 193: 265-275, 1951.
Minchin, F.R., Becana, M., Sprent, J.I.: Short-term inhibition of legume N2 fixation by nitrate. II. Nitrate effects on nodule oxygen diffusion.-Planta 180: 46-52, 1989.
Minchin, F.R., Iannetta, P.P.M., Fernández-Pascual, M., DeLorenzo, C., Witty, J.F., Sprent, J.I.: A new procedure for the calculation of oxygen diffusion resistance in legume nodules from flow-through gas analysis data.-Ann. Bot. 70: 283-289, 1992.
Naishitt, T., Sprent, J.I.: The long term effect of nitrate on the growth and nodule structure of the Caesalpinioid herbaceous legume Chamaecrista fasciculata Michaux.-J. exp. Bot. 44: 829-836, 1993.
Nakano, Y., Asada, K.: Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts.-Plant Cell Physiol. 22: 1236-1241, 1981.
Oghohorie, C.G.O., Pate, J.S.: The nitrate syndrome of the nodulated field pea (Pisum arvense L.).-In: Lie, T.A., Mulder, E.G. (ed.): Biological Nitrogen Fixation in Natural and Agriculture Habitats. Pp. 185-202. Martinus Nijhoff, The Hague 1971.
Pladys, D., Barthe, P., Rigaud, J.: Changes in intracellular pH in French bean nodules induced by senescence and nitrate treatment.-Plant Sci. 56: 99-106, 1988.
Pladys, D., Rigaud, J.: Senescence in French bean nodules: occurrence of different proteolytic activities.-Physiol. Plant. 63: 43-48, 1985.
Streeter, J.G.: Seasonal distribution of carbohydrates in nodules and stem exudate from field-grown soybean plants.-Ann. Bot. 48: 441-450, 1981.
Streeter, J.G.: Nitrate inhibition of legume nodule growth and activity. II. Short term studies with high nitrate supply.-Plant Physiol. 77: 325-328, 1985.
Streeter, J.G.: Inhibition of legume nodule formation and N2 fixation by nitrate.-Crit. Rev. Plant Sci. 7: 1-23, 1988.
Swaraj, K., Kuhad, M.S., Garg, O.P.: Dark treatment effects on symbiotic nitrogen fixation and related processes in Cicer arietinum (chickpea).-Environ. exp. Bot. 26: 31-38, 1986.
Swaraj, K., Laura, J.S., Bishnoi, N.R.: Nitrate induced nodule senescence and changes in activities of enzymes scavenging H2O2 in cluster bean (Cyamopsis tetragonoloba Taub.).-J. Plant Physiol. 141: 202-205, 1993.
Swaraj, K., Laura, J.S., Bishnoi, N.R.: Dark treatment effects on nitrogen fixation and enzymes associated with scavenging hydrogen peroxide in cluster bean nodules.-Plant Physiol. Biochem. 32: 115-119, 1994.
Taylor, D.C., Shelp, B.J., Belson, L.M., Grodzinski, B.: Carbon and nitrogen partitioning in young nodulated pea (wild type and nitrate reductase-deficient) plants exposed to NH4NO3.-Physiol. Plant. 74: 593-601, 1988.
Wasfi, M., Prioul, J.L.: A comparison of inhibition of French bean and soybean nitrogen fixation by nitrate, 1 % oxygen or direct assimilate deprivation.-Physiol. Plant. 66: 481-490, 1986.
Wilson, D.O., Reisenauer, H.M.: Cobalt requirements of symbiotically grown alfalfa.-Plant Soil 19: 364-373, 1963.
Yemm, E.W., Willis, A.J.: The estimation of carbohydrates in plant extracts by anthrone.-Biochem. J. 57: 508-514, 1954.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Sheokand, S., Babber, S. & Swaraj, K. Nodule structure and functioning in Cicer arietinum as affected by nitrate. Biologia Plantarum 41, 435–443 (1998). https://doi.org/10.1023/A:1001810813047
Issue Date:
DOI: https://doi.org/10.1023/A:1001810813047