Abstract
Transformed Nicotiana plumbaginifolia plants with constitutive expression of nitrate reductase (NR) activity were grown at different levels of nitrogen nutrition. The gradients in foliar NO −3 content and maximum extractable NR activity observed with leaf order on the shoot, from base to apex, were much decreased as a result of N-deficiency in both the transformed plants and wild type controls grown under identical conditions. Constitutive expression of NR did not influence the foliar protein and chlorophyll contents under any circumstances. A reciprocal relationship between the observed maximal extractable NR activity of the leaves and their NO −3 content was observed in plants grown in nitrogen replete conditions at low irradiance (170 μmol photons·m−2 ·s−1). This relationship disappeared at higher irradiance (450 μmol photons·m−2·S−1) because the maximal extractable NR activity in the leaves of the wild type plants in these conditions increased to a level that was similar to, or greater than that found in constitutive NR-expressors. Much more NO −3 accumulated in the leaves of plants grown at 450 μmol photons·m−2·s−1 than in those grown at 170 μmol photons·m−2·s−1 in N-replete conditions. The foliar NO −3 level and maximal NR activity decreased with the imposition of N-deficiency in all plant types such that after prolonged exposure to nitrogen depletion very little NO −3 was found in the leaves and NR activity had decreased to almost zero. The activity of NR decreased under conditions of nitrogen deficiency. This regulation is multifactoral since there is no regulation of NR gene expression by NO −3 in the constitutive NR-expressors. We conclude that the NR protein is specifically targetted for destruction under nitrogen deficiency. Consequently, constitutive expression of NR activity does not benefit the plant in terms of increased biomass production in conditions of limiting nitrogen.
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Abbreviations
- Chl:
-
chlorophyll
- N:
-
nitrogen
- NR:
-
NADH-nitrate reductase
- WT:
-
wild type
References
Arnon, D.I. (1949) Copper enzymes in isolated chloroplasts. Polyphenoloxidases in Beta vulgaris. Plant Physiol. 34, 1–15
Benamar, S., Pizelle, G., Thiery, G. (1989) Activité nitrate réductase non induite par le nitrate dans les feuilles d'Alnus glutinosa. Plant Physiol. Biochem. 27, 107–112
Bradford, M.M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254
Cataldo, D.A., Haroon, M., Schrader, L.E., Youngs, V.L. (1975) Rapid colorimetric determination of nitrate in plant tissue by nitration of salicylic acid. Commun. Soil Sci. Plant Anal. 6, 71–80
Coic, Y., Lesaint, C. (1975) La nutrition minérale et en eau des plantes en horticulture avancée. Document Technique de SCPA 23, 1–22
Cooper, H.D., Clarkson, D.T. (1989) Cycling of amino-nitrogen and other nutrients between shoots and roots in cereals. A possible mechanism integrating shoot and root in the regulation of nutrient uptake. J Expt. Bot. 40, 753–762
Deng, M.D., Moureaux, T., Leydecker, M.T., Caboche, C. (1990) Nitrate reductase expression is under the control of a circadian rhythm and is light inducible in Nicotiana tabacum leaves. Planta 180, 257–261
Duke, S.H., Duke, S.O. (1984) Light control of extractable nitrate reductase activity in higher plants. Physiol. Plant. 62, 485–493
Foyer, C.H., Ferrario, S. (1994) Modulation of carbon and nitrogen metabolism in transgenic plants with a view to improved biomass production. Biochem. Soc. Trans. (Special issue), in press
Foyer, C.H., Lefebvre, C., Provot, M., Vincentz, M., Vaucheret, H. (1993) Modulation of nitrogen and carbon metabolism in transformed Nicotiana plumbaginifolia mutant E23 lines expressing either increased or decreased nitrate reductase activity. In: Aspects of applied biology, vol. 34: Physiology of varieties, pp. 137–145, White, E., Kettlewell, P.S., Parry, M.A., Ellis, R.P., eds. Association of Applied Biologists, Wellesbourne, Warwick, UK
Foyer, C.H., Lescure, J.C., Lefebvre, C., Morot-Gaudry, J.F., Vincentz, M., Vaucheret, H. (1994) Adaptations of photosynthetic electron transport, carbon assimilation and carbon partitioning in transgenic Nicotiana plumbaginifolia plants to changes in nitrate reductase activity. Plant Physiol. 104, 171–178
Galangau, F., Daniel-Vedele, F., Moureaux, T., Dorbe, M.F., Leydecker, M.T., Caboche, M. (1988) Expression of leaf nitrate reductase genes from tomato and tobacco in relation to light dark regimes and nitrate supply. Plant Physiol. 88, 383–388
Gastal, F., Saugier, B. (1989) Relationships between nitrogen uptake and carbon assimilation in whole plants of tall fescue. Plant. Cell Environ 12, 407–418
Gojon, A., Wakrim, R., Passama, L., Robin, P. (1991) Regulation of NO −3 assimilation by anion availability in excised soybean leaves. Plant Physiol. 96, 398–405
Huber, J.L., Huber, S.C., Campbell, W.H., Redinbaugh, M.G. (1992) Comparative studies of the light modulation of nitrate reductase and sucrose-phosphate synthase activities in spinach leaves. Plant Physiol. 100, 706–712
Kaiser, W.M., Spill, D., Glaab, J. (1993) Rapid modulation of nitrate reductase in leaves and roots: indirect evidence for the involvement of protein phosphorylation/dephosphorylation. Physiol. Plant. 89, 557–562
Kenis, J.D., Silvente, S.T., Luna, C.M., Campbell, W.H. (1992) Induction of nitrate reductase in detached corn leaves: the effect of the age of the leaves. Physiol. Plant. 85, 49–56
Khamis, S., Lamaze, T., Lemoine, Y., Foyer, C.H. (1990) Adaptation of the photosynthetic apparatus in maize leaves as a result of nitrogen limitation. Plant Physiol. 94, 1436–1443
Kleinhofs, A., Warner, R.L. (1990) Advances in nitrate assimilation. In: The biochemistry of plants, vol. 16: Intermediary nitrogen metabolism, pp. 89–120, Miflin, B.J., Lea, P.J. eds. Academic Press, San Diego
Lillo, C., Henriksen, A. (1984) Comparative studies of diurnal variations of nitrate reductase activity in wheat, oat and barley. Physiol. Plant. 62, 89–94
Moureaux, T., Leydecker, M.T., Meyer, C. (1989) Purification of nitrate reductase from Nicotiana plumbaginifolia by affinity chromatography using 5′AMP-Sepharose and monoclonal antibodies. Eur. J. Biochem. 179, 617–620
Quilleré, I., Dufossé, C., Roux, Y., Foyer, C.H., Caboche, M., Morot Gaudry, J.F. (1994) The effects of deregulation of NR gene expression on growth and nitrogen metabolism of Nicotiana plumbaginifolia plant. J. Expt. Bot. 45, 1205–1211
Rufty, T.W. Jr., Volk, R.J., MacKown, C.T. (1987) Endogenous NO −3 in the root as a source of substrate for reduction in the light. Plant Physiol. 84, 1421–1426
Rufty, T.W. Jr., Huber, S.C., Volk, R.J. (1988) Alteration in leaf carbohydrate metabolism in response to nitrogen stress. Plant Physiol. 88, 725–730
Shaner, D.L., Boyer, J.S. (1976) Nitrate reductase activity in maize (Zea mays L.) leaves. I. Regulation by nitrate flux. Plant Physiol. 58, 499–504
Srivastava, H.S. (1980) Regulation of nitrate reductase activity in higher plants. Phytochemistry 19, 725–733
Streit, L., Martin, B.A., Harper, J.E. (1987) A method for the separation and partial purification of the three forms of nitrate reductase present in wild-type soybean leaves. Plant Physiol. 84, 654–657
Vincentz, M., Caboche, M. (1991) Constitutive expression of nitrate reductase allows normal growth and development of Nicotiana plumbaginifolia plants. EMBO J. 10, 1027–1035
Vincentz, M., Moureaux, T., Leydecker, M.T., Vaucheret, H., Caboche, M. (1993) Regulation of nitrate and nitrite reductase expression in Nicotiana plumbaginifolia leaves by nitrogen and carbon metabolites. Plant J. 3, 315–324
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Ferrario, S., Valadier, MH., Morot-Gaudry, JF. et al. Effects of constitutive expression of nitrate reductase in transgenic Nicotiana plumbaginifolia L. in response to varying nitrogen supply. Planta 196, 288–294 (1995). https://doi.org/10.1007/BF00201387
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DOI: https://doi.org/10.1007/BF00201387