Abstract
We show that the pH dependence of the base-catalysed exchange rate of the ammonium ion provides a basis for discriminating between the cytoplasmic and vacuolar pools of ammonium in plant tissues. In vivo, 14N-nuclear magnetic resonance spectra were recorded with and without 1H-decoupling and information on the subcellular distribution of NH +4 was obtained from a lineshape analysis of the 1H-coupled spectrum. We applied this method to maize (Zea mays L.) root tissues and found that: (i), the cytoplasmic ammonium concentration was low, which was in accord with the large activity of glutamine synthetase present in the roots; and (ii), inhibition of glutamine synthetase with methionine sulphoximine increased the cytoplasmic ammonium concentration, and led to the appearance of ammonium in the xylem sap.
Similar content being viewed by others
Abbreviations
- GS:
-
glutamine synthetase
- MSO:
-
l-methionine sulphoximine
- NMR:
-
nuclear magnetic resonance
- Pi:
-
inorganic phosphate
References
Arima, Y., Kumazawa, K. (1977) Evidence of ammonium assimilation via the glutamine synthetase-glutamate synthase system in rice seedling roots. Plant Cell Physiol. 18, 1121–1129
Beck, E., Renner, U. (1990) Net fluxes and pools of nitrogenous compounds during suspension culture of photoautotrophic Chenopodium rubrum cells. Plant Cell Environ. 13, 111–122
Belton, P.S., Ratcliffe, R.G. (1985) NMR and compartmentation in biological tissues. Prog. NMR Spectrosc. 17, 241–279
Belton, P.S., Lee, R.B., Ratcliffe, R.G. (1985) A 14N nuclear magnetic resonance study of inorganic nitrogen metabolism in barley, maize and pea roots. J. Exp. Bot. 36, 190–210
Benoit, H., Ottavi, H., Pommier, J. (1963) Mesure du temps de corrélation caractérisant un échange chimique. C. R. Acad. Sci. 256, 399–402
Collander, R. (1959) Cell membranes: their resistance to penetration and their capacity for transport. In: Plant physiology — a treatise, vol. 2, pp. 3–102, Steward, F.C., ed. Academic Press, New York
Cooper, H.D., Ford, H.D., Mosquera-Pardo, A.C. (1989) Fluxes of nitrate and ammonium in wheat root cells. Agricultural and Food Research Council Meeting on Plant and Soil Nitrogen Metabolism, Lancaster University, abstract no. 38
Deignan, M.T., Lewis, O.A.M. (1988) The inhibition of ammonium uptake by nitrate in wheat. New Phytol. 110, 1–3
Emerson, M.T., Grunwald, E., Kromhout, R.A. (1960) Proton transfer studies by nuclear magnetic resonance. I. Diffusion control in the reaction of ammonium ion in aqueous acid. J. Chem. Phys. 33, 547–555
Fentem, P.A., Lea, P.J., Stewart, G.R. (1983a) Ammonia assimilation in the roots of nitrate- and ammonia-grown Hordeum vulgare (cv. Golden Promise). Plant Physiol. 71, 496–501
Fentem, P.A., Lea, P.J., Stewart, G.R. (1983b) Action of inhibitors of ammonia assimilation on amino acid metabolism in Hordeum vulgare L. (cv. Golden Promise). Plant Physiol. 71, 502–506
Handa, S., Huber, D.M., Warren, H.L., Tsai, C.Y. (1985) Nitrogen nutrition and N-assimilation in maize seedlings. Can. J. Plant Sci. 65, 87–93
Harvey, D.M.R. (1989) Effects of different external potassium concentrations on potassium concentrations in the cytoplasm of maize root cortical cells. J. Plant Physiol. 133, 768–769
Haynes, R.J., Goh, K.M. (1978) Ammonium and nitrate nutrition of plants. Biol. Rev. 53, 465–510
Joy, K.W. (1988) Ammonia, glutamine, and asparagine: a carbon-nitrogen interface. Can. J. Bot. 66, 2103–2109
Kaiser, J.J., Lewis, O.A.M. (1980) Nitrate-nitrogen assimilation in the leaves of Helianthus annuus L. New Phytol. 85, 235–241
Kleiner, D. (1981) The transport of NH3 and NH4/+ across biologcal membranes. Biochim. Biophys. Acta 639, 41–52
Lee, R.B., Drew, M.C. (1989) Rapid, reversible inhibition of nitrate influx in barley by ammonium. J. Exp. Bot. 40, 741–752.
Lee, R.B., Ratcliffe, R.G. (1983a) Phosphorus nutrition and the intracellular distribution of inorganic phosphate in pea root tips: a quantitative study using 31P-NMR. J. Exp. Bot. 34, 1222–1244
Lee, R.B., Ratcliffe, R.G. (1983b) Development of an aeration system for use in plant tissue NMR experiments. J. Exp. Bot. 34, 1213–1221
Lee, R.B., Ratcliffe, R.G., Southon, T.E. (1990) 31P-NMR measurements of the cytoplasmic and vacuolar Pi content of mature maize roots: relationships with phosphorus status and phosphate fluxes. J. Exp. Bot. 41, 1063–1078
Leigh, R.A., Wyn Jones, R.G. (1984) A hypothesis relating critical potassium concentrations for growth to the distribution and functions of this ion in the plant cell. New Phytol. 97, 1–13
Lewis, O.A.M., Chadwick, S. (1983) An 15N investigation into nitrogen assimilation in hydroponically-grown barley (Hordeum vulgare L. cv. Clipper) in response to nitrate, ammonium and mixed nitrate and ammonium nutrition. New Phytol. 95, 635–646
Lewis, O.A.M., James, D.M., Hewitt, E.J. (1982) Nitrogen assimilation in barley (Hordeum vulgare L. cv. Mazurka) in response to nitrate and ammonium nutrition. Ann. Bot. 49, 39–49
Lewis, O.A.M., Chadwick, S., Withers, J. (1983) The assimilation of ammonium by barley roots. Planta 159, 483–486
Loughman, B.C., Ratcliffe, R.G., Schwabe, J.W.R. (1989) Galactose metabolism in Zea mays root tissues observed by 31P-NMR spectroscopy. Plant Sci. 59, 11–23
Macklon, A.E.S. (1975) Cortical cell fluxes and transport to the stele in excised root segments of Allium cepa L. Planta 122, 109–130
Macklon, A.E.S., Ron, M.M., Sim, A. (1990) Cortical cell fluxes of ammonium and nitrate in excised root segments of Allium cepa L.; studies using 15N. J. Exp. Bot. 41, 359–370
MacMillan, A. (1956) The entry of ammonia into fungal cells. J. Exp. Bot. 7, 113–126.
Martin, J.B., Bligny, R., Rebeille, F., Douce, R., Leguay, J.J., Mathieu, Y., Guern, J. (1982) A 31P nuclear magnetic resonance study of intracellular pH of plant cells cultivated in liquid medium. Plant Physiol. 70, 1156–1161
Miflin, B.J., Lea, P.J. (1976) The pathway of nitrogen assimilation in plants. Phytochemistry 15, 873–885
Morgan, M.A., Jackson, W.A. (1988a) Inward and outward movement of ammonium in root systems: transient responses during recovery from nitrogen deprivation in presence of ammonium. J. Exp. Bot. 39, 179–191
Morgan, M.A., Jackson, W.A. (1988b) Suppression of ammonium uptake by nitrogen supply and its relief during nitrogen limitation. Physiol. Plant. 73, 38–45
Ogg, R.A. (1954) Study of ultrarapid reactions by nuclear magnetic resonance spectra. Discuss. Faraday Soc. 17, 215–234
Ogg, R.A., Ray, J.D. (1957) Quadrupole relaxation and structures in nitrogen magnetic resonances of ammonia and ammonium salts. J. Chem. Phys. 26, 1339–1340
Patel, D.D., Barlow, P.W., Lee, R.B. (1990) Development of vacuolar volume in the root tips of pea. Ann. Bot. 65, 159–169
Presland, M.R., McNaughton, G.S. (1986) Whole plant studies using radioactive 13-nitrogen. IV. A compartmental model for the uptake and transport of ammonium ions by Zea mays. J. Exp. Bot. 37, 1619–1632
Probyn, T.A., Lewis, O.A.M. (1979) The route of nitrate-nitrogen assimilation in the root of Datura stramonium L. J. Exp. Bot. 30, 299–305
Reisenauer, H.M. (1978) Absorption and utilization of ammonium nitrogen by plants. In: Nitrogen in the environment, vol. 2, pp. 157–170, Nielsen, D.R., MacDonald, J.G. eds. Academic Press, New York
Rhodes, D., Rendon, G.A., Stewart, G.R. (1975) The control of glutamine synthetase level in Lemna minor L. Planta 125, 201–211
Salsac, L., Chaillou, S., Morot-Gaudry, J.F., Lesainte, C., Jolivet, E. (1987) Nitrate and ammonium nutrition in plants. Plant Physiol. Biochem. 25, 805–812
Scherer, H.W., MacKown, C.T., Leggett, J.E. (1984) Potassium-ammonium uptake interactions in tobacco seedlings. J. Exp. Bot. 35, 1060–1070
Smith, F.A., Walker, N.A. (1978) Entry of methylammonium and ammonium ions into Chara internodal cells. J. Exp. Bot. 29, 107–120
Stewart, G.R., Mann, A.F., Fentem, P.A. (1980) Enzymes of glutamate formation: glutamate dehydrogenase, glutamine synthetase, and glutamate synthase. In: The biochemistry of plants, vol. 5, pp. 271–327, Miflin, B.J., ed. Academic Press, New York
Stulen, I., Lanting, L., Lambers, H., Posthumus, F., van de Dijk, S.J., Hofstra, R. (1981) Nitrogen metabolism of Plantago lanceolata as dependent on the supply of mineral nutrients. Physiol. Plant. 51, 93–98
Ullrich, W.R., Larsson, M., Larsson, C.M., Lesch, S., Novacky, A. (1984) Ammonium uptake in Lemna gibba G1, related membrane potential changes, and inhibition of anion uptake. Physiol. Plant. 61, 369–376
Vézina, L.P., Hope, H.J., Joy, K.W. (1987) Isoenzymes of glutamine synthetase in roots of pea (Pisum sativum L. cv. Little Marvel) and alfalfa (Medicago media Pers. cv. Saranac). Plant Physiol. 83, 58–62
Walker, N.A., Smith, F.A., Beilby, M.J. (1979) Amine uniport at the plasmalemma of Charophyte cells. II. Ratio of matter to charge transported and permeability of free base. J. Membr. Biol. 49, 283–296
Yoneyama, T., Kumazawa, K. (1974) A kinetic study of the assimilation of 15N-labelled ammonium in rice seedling roots. Plant Cell Physiol. 15, 655–661
Yoneyama, T., Kumazawa, K. (1975) A kinetic study of the assimilation of 15N-labelled nitrate in rice seedlings. Plant Cell Physiol. 16, 21–26
Author information
Authors and Affiliations
Additional information
On secondment to the Department of Plant Sciences, University of Oxford.
We acknowledge the financial support of the Agricultural and Food Research Council. R.B. Lee also thanks the Department of Plant Sciences, University of Oxford, for hospitality.
Rights and permissions
About this article
Cite this article
Lee, R.B., Ratcliffe, R.G. Observations on the subcellular distribution of the ammonium ion in maize root tissue using in-vivo 14N-nuclear magnetic resonance spectroscopy. Planta 183, 359–367 (1991). https://doi.org/10.1007/BF00197734
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00197734