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Development of nitrogen-assimilating enzymes in sunflower cotyledons during germination as affected by the exogenous nitrogen source

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Abstract

Activities of nitrate reductase (NR; EC 1.6.6.1), nitrite reductase (NiR; EC 1.7.7.1), glutamine synthetase (GS; EC 6.3.1.2) and glutamate dehydrogenase (GDH; EC 1.4.1.3) were measured in cotyledons of sunflower (Helianthus annuus L. cv Peredovic) seedlings during germination and early growth under various external nitrogen sources. The presence of NO -3 in the medium promoted a gradual increase in the levels of NR and NiR activities during the first 7 d of germination. Neither NR nor NiR activities were increased in a nitrogen-free medium or in media with either NH +4 or urea as nitrogen sources. Moreover, the presence of NH +4 did not abolish the NO -3 -dependent appearance of NR and NiR activities. The increase of NR activity was impaired both by cycloheximide and chloramphenicol, which indicates that both cytoplasmic 80S and plastidic 70S ribosomes are involved in the synthesis of the NR molecule. By contrast, the appearance of NiR activity was only inhibited by cycloheximide, indicating that NiR seems to be exclusively synthesized on the cytoplasmic 80S ribosomes. Glutamine-synthetase activity was also strongly increased by external NO -3 but not by NH +4 or urea. The appearance of GS activity was more efficiently suppressed by cycloheximide than chloramphenicol. This indicates that GS is mostly synthesized in the cytoplasm. The cotyledons of the dry seed contain high levels of GDH activity which decline during germination independently of the presence or absence of a nitrogen source. Cycloheximide, but not chloramphenicol, greatly prevented the decrease of GDH activity.

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Abbreviations

GDH:

glutamate dehydrogenase

GS:

glutamine synthetase

NiR:

nitrite reductase

NR:

nitrate reductase

References

  • Agüera, E., De la Haba, P., Maldonado, J.M. (1987) In vitro stabilization and tissue distribution of nitrogen-assimilating enzymes in sunflower. J. Plant Physiol. 128, 443–449

    Google Scholar 

  • Akazawa, T. (1979) Ribulose-1,5-bisphosphate carboxylase. In: Encyclopedia of plant physiology, N.S., vol. 6: Photosynthesis II, pp. 208–229, Gibbs, M., Latzko, E., eds. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Börner, T., Mendel, R.R., Schiemann, J. (1986) Nitrate reductase is not accumulated in chloroplast-ribosome-deficient mutants of higher plants. Planta 169, 202–207

    Google Scholar 

  • Cammarts, D., Jacobs, M. (1985) A study of the role of glutamate dehydrogenase in the nitrogen metabolism of Arabidopsis thaliana. Planta 163, 517–526

    Google Scholar 

  • Datta, N., Rao, L.V.M., Guha-Mukherjee, S., Sopory, S.K. (1981) Regulation of nitrate reductase activity by ammonium in wheat. Plant Sci. Lett. 20, 305–313

    Google Scholar 

  • Feierabend, J. (1986) Investigation of the site of synthesis of chloroplastic enzymes of nitrogen metabolismby the use of heat-treated 70S ribosome-deficient rye leaves. Physiol. Plant. 67, 145–150

    Google Scholar 

  • Guerrero, M.G., Vega, J.M., Losada, M. (1981) The assimilatory nitrate-reducing system and its regulation. Annu. Rev. Plant Physiol. 32, 169–204

    Google Scholar 

  • Heath-Pagliuso, S., Huffaker, R.C., Allard, R.W. (1984) Inheritance of nitrite reductase and regulation of nitrate reductase, nitrite reductase, and glutamine synthetase isoenzymes. Plant Physiol. 76, 353–358

    Google Scholar 

  • Hewitt, E.J. (1966) Sand and water culture methods used in the study of plant nutrition, 2nd rev. edn. Commonweath Bureau of Horticultural and Plantation Crops. East Malling. Tech. Commun. No. 22

    Google Scholar 

  • Hirel, B., Vidal, J., Gadal, P. (1982) Evidence for a cytosolic-dependent light induction of chloroplastic glutamine synthetase during greening of etiolated rice leaves. Planta 155, 17–23

    Google Scholar 

  • Lea, P.J., Miflin, B.J. (1979) Photosynthetic ammonia assimilation. In: Encyclopedia of plant physiology, N.S., vol. 6: Photosynthesis II, pp. 445–456, Gibbs, M., Latzko, E., eds. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Losada, M., Paneque, A. (1971) Nitrite reductase. Methods Enzymol. 23, 487–491

    Google Scholar 

  • Lowry, O.H., Rosebrough, N.J., Farr, A.L., Randall, R.J. (1951) Protein measurement with the Folin-phenol reagent. J. Biol. Chem. 193, 265–275

    Google Scholar 

  • Loyola-Vargas, M., Sanchez de Jimenez, E. (1986) Regulation of glutamine synthetase/glutamate synthase cycle in maize tissues. J. Plant Physiol. 124, 147–154

    Google Scholar 

  • Mauriño, S.G., Echevarria, C., Mejias, J.A., Vargas, M.A., Maldonado, J.M. (1986) Properties of the in vivo nitrate reductase assay in maize, soybean and spinach leaves. J. Plant Physiol. 124, 123–130

    Google Scholar 

  • Mohanty, B., Fletcher, J.S. (1976) Ammonium influence on the growth and nitrate reductase activity of Paul's Scarlet rose suspension cultures. Plant Physiol. 58, 152–155

    Google Scholar 

  • Ratajczak, L., Ratajczak, W., Mazurowa, H. (1981) The effect of different carbon and nitrogen sources on the activity of glutamine synthetase and glutamate dehydrogenase in lupine embryonic axes. Physiol. Plant. 51, 277–280

    Google Scholar 

  • Rhodes, D., Rendon, G.A., Stewart, G.R. (1975) The control of glutamine synthetase level in Lemna minor L. Planta 125, 201–211

    Google Scholar 

  • Sawhney, S.K., Naik, M.S. (1973) Effect of chloramphenicol and cycloheximide on the synthesis of nitrate reductase and nitrite reductase in rice leaves. Biochem. Biophys. Res. Commun. 51, 67–73

    Google Scholar 

  • Shapiro, B.M., Stadtman, E.R. (1970) Glutamine synthetase (Escherichia coli). Methods Enzymol. 17A, 910–922

    Google Scholar 

  • Shen, T.C. (1969) The induction of nitrate reductase and the preferential assimilation of ammonium in germinating rice seedlings. Plant Physiol. 44, 1650–1655

    Google Scholar 

  • Singh, R.P., Srivastava, H.S. (1982) Glutamate dehydrogenase activity and assimilation of inorganic nitrogen in maize seedlings. Biochem. Physiol. Pflanz. 177, 633–642

    Google Scholar 

  • Sluiters-Scholten, C.M.T. (1973) Effect of chloramphenicol and cycloheximide on the induction of nitrate reductase and nitrite reductase in bean leaves. Planta 113, 229–240

    Google Scholar 

  • Smith, F.W., Thompson, J.F. (1971) Regulation of nitrate reductase in excised barley roots. Plant Physiol. 48, 219–223

    Google Scholar 

  • Timpo, E.E., Neyra, C.A. (1983) Expression of nitrate and nitrite reductase activities under various forms of nitrogen nutrition in Phaseolus vulgaris L. Plant Physiol. 72, 71–75

    Google Scholar 

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de la Haba, P., Agüera, E. & Maldonado, J.M. Development of nitrogen-assimilating enzymes in sunflower cotyledons during germination as affected by the exogenous nitrogen source. Planta 173, 52–57 (1988). https://doi.org/10.1007/BF00394487

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  • DOI: https://doi.org/10.1007/BF00394487

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