Abstract
Nitrate-selective microelectrodes have been made using a quaternary ammonium sensor, methyl-tridodecylammonium nitrate, in a Polyvinylchloride matrix. These electrodes showed a log-linear response from 0.1 to 100 mol · m−3 nitrate with a typical slope of 55.6 mV per decade change in nitrate concentration. The only physiologically significant interfering anion was chloride but the lower limit of nitrate detection was 0.5 mol · m−3 in the presence of 100 mol · m−3 chloride which means this interference will not be important in most physiological situations. These microelectrodes were used to measure nitrate concentrations in internodal cells of Chara corallina cultured under low nitrate and nitrate-replete conditions for 6 to 30 weeks. Cells maintained in low nitrate only showed measurements which were less than the detection limit of the electrodes, while cells grown under nitrate-replete conditions showed two populations of measurements having means of 1.6 and 6.2 mol · m−3. Chemical analysis of the high-nitrate cells indicated that they contained a mean nitrate concentration of 5.9 mol · m−3. As vacuolar nitrate concentration would dominate this whole-cell measurement, it is concluded that the higher concentration measured with the electrodes represents vacuolar nitrate concentration and the lower value represents the cytoplasmic concentration. This intracellular distribution of nitrate could only be achieved passively if the electrical potential difference across the tonoplast is between +25 and + 35 mV.
Article PDF
Similar content being viewed by others
Avoid common mistakes on your manuscript.
Abbreviations
- APW:
-
artificial pond water
- Hepes:
-
4-(2-hydroxyethyl-1-piperazineethanesulfonic) acid
- MTDDA.NO3 :
-
methyltridodecylammonium nitrate
References
Ammann, D. (1986) Ion-selective microelectrodes, principles, de sign and application. Springer, Berlin Heidelberg New York
Deane-Drummond, C.E. (1985) Regulation of nitrate uptake into Chara corallina cells via NH +4 stimulation of NO −3 efflux. Plant Cell Environ. 8, 105–110
Deane-Drummond, C.E., Glass, A.D.M. (1982) Nitrate uptake into barley (Hordeum vulgare) plants. A new approach using 36ClO −3 as an analogue for NO −3 . Plant Physiol. 70, 50–54
Felle, H. (1987) Proton transport and pH control in Sinapis alba root hairs: a study carried out using double-barreled pH microelectrodes. J. Exp. Bot. 38, 340–354
Felle, H. (1988) Auxin causes oscillations of cytosolic free calcium and pH in Zea mays coleoptiles. Planta 174, 495–499
Flowers, T.J., Troke, P.F., Yeo, A.R. (1977) The mechanism of salt tolerance in halophytes. Annu Rev. Plant Physiol. 28, 89–121
Henriksen, G.H., Bloom, A.J., Spanswick, R.M. (1990) Measurement of net fluxes of ammonium and nitrate at the surface of barley roots using ion-selective microelectrodes. Plant Physiol. 93, 271–280
Lee, R.B. (1979) The release of nitrite from barley roots in response to metabolic inhibitors, uncoupling agents and anoxia. J. Exp. Bot. 30, 119–133
Lee, R.B., Clarkson, D.T. (1986) Nitrogen-13 studies of nitrate fluxes in barley roots. I Compartmental analysis from measurements of 13N efflux. J. Exp. Bot. 37, 1753–1767
Leigh, R.A., Wyn Jones, R.G. (1986) Cellular compartmentation in plant nutrition: the selective cytoplasm and the promiscuous vacuole. Adv. Plant Nutrition 2, 249–279
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
MacRobbie, E.A.C. (1970) The active transport of ions in plant cells. Quart. Rev. Biophys. 3, 251–294
Martinoia, E., Schramm, M.J., Kaiser, G., Kaiser, W.M., Heber, U. (1986) Transport of anions in isolated barley vacuoles. I. Permeability to anions and evidence for a Cl− uptake system. Plant Physiol. 80, 895–901
Miller, A.J., Sanders, D. (1987) Depletion of cytosolic free calcium induced by photosynthesis. Nature 326, 397–400
Okihara, K., Kiyosawa, K. (1988) Ion composition of the Chara internode. Plant Cell Physiol. 29, 21–25
Penny, M.G., Bowling, D.J.F. (1974) A study of potassium gra-dients in the epidermis of intact leaves of Commelina communis in relation to stomatal opening. Planta 119, 17–25
Penny, M.G., Kelday, L.S., Bowling, D.J.F. (1976) Active chloride transport in the leaf epidermis of Commelina communis in relation to stomatal activity. Planta 130, 291–294
Sanders, D. (1981) Physiological control of chloride transport in Chara corallina. II. The role of chloride as a vacuolar osmoticum. Plant Physiol. 68, 401–406
Steingröver, E., Ratering, P., Siesling, J. (1986) Daily changes in uptake, reduction and storage of nitrate in spinach grown at low light intensity. Physiol. Plant. 66, 550–556
Tester, M., Beilby, M.J., Shimmen, T. (1987) Electrical characteristics of the tonoplast of Chara corallina: a study using permeabilised cells. Plant Cell Physiol. 28, 1555–1568
Tsien, R.Y., Rink, T.J. (1981) Ca2+-selective electrodes: a novel PVC-gelled neutral carrier mixture compared with other currently available sensors. J. Neurosci. Methods 4, 73–86
Tyerman, S.D., Findlay, G.P. (1989) Current-voltage curves of single Cl − channels which which coexist with two types of K+ channels in the tonoplast of Chara corallina. J. Exp. Bot. 40, 105–117
Underbill, E.W. (1980) Glucosinolates. In: Encyclopedia of plant physiology, N. S. vol. 8; Secondary plant products, pp. 493–511, Bell, E.A., Charlwood, B.V., eds. Springer, Berlin Heidelberg New York
Wegmann D., Weiss, H., Ammann, D., Morf, W.E., Pretsch, E., Sugahara, K., Simon, W. (1984) Anion-selective liquid membrane electrodes based on lipophilic quaternary ammonium compounds. Mikrochim. Acta III, 1–16
Walker, N.A., Pitman, M.G. (1976) Measurement of fluxes across membranes. In: Encyclopedia of plant physiology, N.S., vol. 2; Transport in plants, pp. 93–126, Lüttge, U., Pitman, M.G., eds. Springer, Berlin Heidelberg New York
West, P.W., Ramachandran, T.P. (1967) Spectrophotometric determination of nitrate using chromotropic acid. Anal. Chim. Acta 35, 317–324
Wray, J.L. (1989) Molecular and genetic aspects of nitrite reduction in higher plants. In: Molecular and genetic aspects of nitrate assimilation, pp. 244–262, Wray, J.L., Kinghorn, J.R., eds. Oxford University Press, Oxford
Author information
Authors and Affiliations
Additional information
R.G. Zhen was awarded a Sino-British Friendship Scholarship, sponsored by the British Council. We are grateful to Dr. G. Dawson (Insecticides and Fungicides Department) for his advice on the synthesis of MTDDA.NO3, Mr. I. Jennings (Department of Biology, University of York) for developing the software used in this work and Dr. R.A. Leigh for critically reading the manuscript.
Rights and permissions
About this article
Cite this article
Miller, A.J., Zhen, RG. Measurement of intracellular nitrate concentrations in Chara using nitrate-selective microelectrodes. Planta 184, 47–52 (1991). https://doi.org/10.1007/BF00208235
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00208235