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Nitrate and nitrite uptake and reduction by intact sunflower plants

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Abstract

Nitrogen-starved sunflower plants (Helianthus annuus L. cv. Peredovic) cannot absorb NO 3 or NO 2 upon initial exposure to these anions. Ability of the plants to take up NO 3 and NO 2 at high rates from the beginning was induced by a pretreatment with NO 3 . Nitrite also acted as inducer of the NO 2 -uptake system. The presence of cycloheximide during NO 3 -pretreatment prevented the subsequent uptake of NO 3 and NO 2 , indicating that both uptake systems are synthesized de novo when plants are exposed to NO 3 . Cycloheximide also suppressed nitrate-reductase (EC 1.6.6.1) and nitrite-reductase (EC 1.7.7.1) activities in the roots. The sulfhydryl-group reagent N-ethylmaleimide greatly inhibited the uptake of NO 3 and NO 2 . Likewise, N-ethylmaleimide promoted in vivo the inactivation of nitrate reductase without affecting nitrite-reductase activity. Rates of NO 3 and NO 2 uptake as a function of external anion concentration exhibited saturation kinetics. The calculated Km values for NO 3 and NO 2 uptake were 45 and 23 μM, respectively. Rates of NO 3 uptake were four to six times higher than NO 3 -reduction rates in roots. In contrast, NO 2 -uptake rates, found to be very similar to NO 3 -uptake rates, were much lower (about 30 times) than NO 2 -reduction rates. Removal of oxygen from the external solution drastically suppressed NO 3 and NO 2 uptake without affecting their reduction. Uptake and reduction were also differentially affected by pH. The results demonstrate that uptake of NO 3 and NO 2 into sunflower plants is mediated by energy-dependent inducible-transport systems distinguishable from the respective enzymatic reducing systems.

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Abbreviations

CHI:

cycloheximide

NEM:

N-ethylmaleimide

NiR:

nitrite reductase

NR:

nitrate reductase

pHME:

p-hydroxymercuribenzoate

References

  • Agüera, E., de la Haba, P., Maldonado, J.M. (1987a) Induction of nitrate reductase, nitrite reductase and glutamine synthetase by nitrate and/or nitrite in germinating sunflower cotyledons Effect of plant hormones. Plant Physiol. (Life Sci. Adv.) 6, 255–258

    Google Scholar 

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

    Google Scholar 

  • Aslam, M., Huffaker, R.C. (1989) Role of nitrate and nitrite in the induction of nitrite reductase in leaves of barley seedlings. Plant Physiol. 91, 1152–1156

    Google Scholar 

  • Bagchi, S.N., Rai, U.N., Rai, A.N., Singh, H.N. (1985) Nitrate metabolism in the cyanobacterium Anabaena cycadeae: Regulation of nitrate uptake and reductase by ammonia. Physiol. Plant. 63, 322–326

    Google Scholar 

  • Beevers, L., Hageman, R.H. (1983) Uptake and reduction of nitrate: bacteria and higher plants. In: Encyclopedia of plant physiology, N.S., vol. 15A: Inorganic plant nutrition, pp. 351–375, Läuchli, A., Bieleski, R.L., eds. Springer, Berlin Heidelberg New York

    Google Scholar 

  • Behl, R., Tischner, R., Raschke, K. (1988) Induction of a highcapacity nitrate-uptake mechanism in barley roots prompted by nitrate uptake through a constitutive low-capacity mechanism. Planta 176, 235–240

    Google Scholar 

  • Breteler, H., Luczak, E. (1982) Utilization of nitrite and nitrate by dwarf bean. Planta 156, 226–232

    Google Scholar 

  • Cawse, P.A. (1967) The determination of nitrate in soil solutions by ultraviolet spectrophotometry. Analyst 92, 311–315

    Article  CAS  Google Scholar 

  • Córdoba, F., Cárdenas, J., Fernández, E. (1986) Kinetic characterization of nitrite uptake and reduction by Chlamydomonas reinhardtii. Plant Physiol. 82, 904–908

    Google Scholar 

  • Cresswell, R.C., Syrett, P.J. (1981) Uptake of nitrate by the diatom Phaeodactylum tricornutum. J. Exp. Bot. 32, 19–25

    Google Scholar 

  • De la Haba, P., Agüera, E., Maldonado, J.M. (1988) Development of nitrogen-assimilating enzymes in sunflower cotyledons during germination as affected by the exogenous nitrogen source. Planta 173, 52–57

    Google Scholar 

  • Deane-Drummond, C.E. (1984) The apparent induction of nitrate uptake by Chara corallina cells following pretreatment with or without nitrate and chlorate. J. Exp. Bot. 35, 1182–1193

    Google Scholar 

  • Dhugga, K.S., Waines, J.G., Leonard, R.T. (1988) Correlated induction of nitrate uptake and membrane polypeptides in corn roots. Plant Physiol. 87, 120–125

    Google Scholar 

  • Goyal, S.S., Huffaker, R.C. (1986) The uptake of NO 3 , NO 2 , and NH +4 by intact wheat (Triticum aestivum) seedlings. Plant Physiol. 82, 1051–1056

    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 

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

  • Ibarlucea, J.M., Llama, M.J., Serra, J.L., Macarulla, J.M. (1983) Mixed-transfer kinetics of nitrite uptake in barley (Hordeum vulgare L. cv. Miranda) seedlings. Plant Sci. Lett. 29, 339–347

    Google Scholar 

  • Ingemarsson, B., Oscarson, P., af Ugglas, M., Larsson, C.-M. (1987) Nitrogen utilization in Lemna. II. Studies of nitrate uptake using 13NO 3 . Plant Physiol. 85, 860–864

    Google Scholar 

  • Jackson, W.A., Johnson, R.E., Volk, R.J. (1974) Nitrite uptake patterns in wheat seedlings as influenced by nitrate and ammonium. Physiol. Plant. 32, 108–114

    Google Scholar 

  • Jackson, W.A., Kwik, K.D., Volk, R.J., Butz, R.G. (1976) Nitrate influx and efflux by intact wheat seedlings: effects of prior nitrate nutrition. Planta 132, 149–156

    Google Scholar 

  • Kochian, L.V., Lucas, W.J. (1982) Potassium transport in corn roots. I. Resolution of kinetics into a saturable and linear component. Plant Physiol. 70, 1723–1731

    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 

  • McClure, P.R., Omholt, T.E., Pace, G.M., Bouthyette, P.Y. (1987) Nitrate-induced changes in protein synthesis and translation of RNA in maize roots. Plant Physiol. 84, 52–57

    Google Scholar 

  • MacKown, C.T., McClure, P.R. (1988) Development of accelerated net nitrate uptake. Effects of nitrate concentration and exposure time. Plant Physiol. 87, 162–166

    Google Scholar 

  • Madueño, F., Vega-Palas, M.A., Flores, E., Herrero, A. (1988) A cytoplasmic-membrane protein repressible by ammonium in Synechococcus R2: altered expression in nitrate-assimilation mutants. FEBS Lett. 239, 289–291

    Google Scholar 

  • Mauriño, S.G., Echevarría, C., Mejías, 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 

  • Morgan, M.A., Jackson, W.A., Volk, R.J. (1985a) Uptake and assimilation of nitrate by corn roots during and after induction of the nitrate uptake system. J. Exp. Bot. 36, 859–869

    Google Scholar 

  • Morgan, M.A., Volk, R.J., Jackson, W.A. (1985b) p-Fluorophenylalanine-induced restriction of ion uptake and assimilation by maize roots. Plant Physiol. 77, 718–721

    Google Scholar 

  • Orsi, B.A., Tipton, K.F. (1979) Kinetic analysis of progress curves. Methods Enzymol. 63, 159–183

    Google Scholar 

  • Serra, J.L., Llama, M.J., Cadenas, E. (1978) Nitrate utilization by the diatom Skeletonema costatum. I. Kinetics of nitrate uptake. Plant Physiol. 62, 987–990

    Google Scholar 

  • Snell, F.D., Snell, C.T. (1949) Colorimetric methods of analysis, vol. 2. Van Nostrand, New York

    Google Scholar 

  • Tompkins, G.A., Jackson, W.A., Volk, R.J. (1978) Accelerated nitrate uptake in wheat seedlings: effects of ammonium and nitrite pretreatments and of 6-methylpurine and puromycin. Physiol. Plant. 43, 166–171

    Google Scholar 

  • Ullrich, W.R. (1987) Nitrate and ammonium uptake in green algae and higher plants: mechanism and relationship with nitrate metabolism. In: Inorganic nitrogen metabolism, pp. 32–38, Ullrich, W.R., Aparicio, P.J., Syrett, P.J., Castillo, F., eds. Springer, Berlin Heidelberg New York

    Google Scholar 

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This research was supported by grant PB86-0232 from the Dirección General de Investigatión Científica y Técnica (Spain). One of us (E.A.) thanks the Consejeria de Educación y Ciencia de la Junta de Andalucia for the tenure of a fellowship. We thank Miss G. Alcalá and Miss C. Santos for their valuable technical and secretarial assistance.

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Agüera, E., de la Haba, P., Fontes, A.G. et al. Nitrate and nitrite uptake and reduction by intact sunflower plants. Planta 182, 149–154 (1990). https://doi.org/10.1007/BF00239997

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