Skip to main content
SpringerLink
Log in

Compartmentation and flux characteristics of nitrate in spruce

  • Published:
Planta Aims and scope Submit manuscript

Abstract

The radiotracer 13N was used to undertake compartmental analyses for NO −3 in intact non-mycorrhizal roots of Picea glauca (Moench) Voss. seedlings. Three compartments were defined, with half-lives of exchange of 2.5 s, 20 s, and 7 min. These were identified as representing surface adsorption, apparent free space, and cytoplasm, respectively. Influx, efflux, and net flux as well as cytoplasmic and apparent-free-space nitrate concentrations were estimated for three different concentration regimes of external nitrate. After exposure to external NO −3 for 3 d, influx was calculated to be 0.09 μmol·g−1·h−1 (at 10 μM [NO −3 ]o), 0.5μmol·g−1·h−1 (at 100 μM [NO sup−inf3 ]o), and 1.2 μmol · g−1· h−1 (at 1.5 mM [NO −3 ]o). Efflux increased with increasing [NO −3 ]o, constituting 4% of influx at 10 μM, 6% at 100 μM, and 21% at 1.5 mM. Cytoplasmic [NO −3 ] was estimated to be 0.3 mM at 10 uM [NO −3 ]o, 2mM at 100 μM [NO −3 ]o, and 4mM at 1.5 mM [NO −3 ]o, while free-space [NO −3 ] was 16 μM, 173 μM, and 2.2 mM, respectively. A series of experiments was carried out to confirm the identity of the compartments resolved by efflux analysis. Pretreatment at high temperature or application of 2-chloro-ethanol, sodium dodecyl sulphate or hydrogen peroxide made it possible to distinguish the metabolic (cytoplasmic) phase from the remaining two (physical) phases. Likewise, varying [Pi] of the medium altered efflux and thereby [NO −3 ]cyt, but did not affect [NO −3 ]free space.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Similar content being viewed by others

Abbreviations

[NO −3 ]cyt :

cytoplasmic NO −3 concentration

[NO −3 ]free space :

apparent-free-space NO −3 concentration

[NO −3 ]o :

concentration of NO −3 in the external solution

φ:

NO −3 flux

φco :

efflux from the cytoplasm

φoc :

influx to the cytoplasm

φnet :

net flux

φxylem :

flux to the xylem

φred/vac :

combined flux to reduction and the vacuole

References

  • Aguera, E., de la Haba, P., Fontes, A.G., Maldonado, J.M. (1990) Nitrate and nitrite uptake and reduction by intact sunflower plants. Planta 182, 149–154

    Google Scholar 

  • 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

    Google Scholar 

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

    Article  CAS  Google Scholar 

  • Chapin, F.S. III, van Cleve, K., Tyron, P.R. (1986) Relationship of ion absorption to growth rate in taiga trees. Oecologia 69, 238–242

    Google Scholar 

  • Clarkson, D.T. (1986) Regulation of the absorption and release of nitrate by plant cells. In: Fundamental, ecological and agricultural aspects of nitrogen metabolism in higher plants, pp. 3–27, Lambers, H., Neeteson, J.J., Stulen, I., eds. Martinus Nijhoff, Boston

    Google Scholar 

  • Cram, W.J. (1973) Chloride fluxes in cells of the isolated root cortex of Zea mays. Aust. J. Biol. Sci. 26, 757–779

    Google Scholar 

  • Devienne, F., Mary, B., Lamaze, T. (1994) Nitrate transport in intact wheat roots. I. Estimation of cellular fluxes and NO −3 distribution using compartmental analysis from data of 15NO −3 efflux. J. Exp. Bot. 45, 667–676

    Google Scholar 

  • Ferrari, T.E., Yoder, O.C., Filner, P. (1973) Anaerobic nitrite production by plants cells and tissues: evidence for two nitrate pools. Plant Physiol. 51, 423–431

    Google Scholar 

  • Flaig, H., Mohr, H. (1992) Assimilation of nitrate and ammonium by the Scots pine (Pinus sylvestris) seedling under conditions of high nitrogen supply. Physiol. Plant. 84, 568–576

    Google Scholar 

  • Glass, A.D.M. (1988) Nitrogen uptake by plant roots. Atlas Sci. Anim. Plant Sci. 1, 151–156

    Google Scholar 

  • Kamminga-van Wijk, C., Prins, H.B.A. (1993) The kinetics of NH +4 and NO −3 uptake by Douglas fir from single N-solutions and from solutions containing both NH +4 and NO −3 . Plant Soil 151, 91–96

    Google Scholar 

  • King, B.J., Siddiqi, M.Y., Glass, A.D.M. (1992) Studies of the uptake of nitrate in barley. V. Estimation of root cytoplasmic nitrate concentration using nitrate reductase activity — implications for nitrate flux. Plant Physiol. 99, 1582–1589

    Google Scholar 

  • Knoepp, J.D., Turner, D.F., Tingey, D.T. (1993) Effects of ammonium and nitrate on nutrient uptake and activity of nitrogen assimilating enzymes in western hemlock. For. Ecol. Manage. 59, 179–191

    Google Scholar 

  • Kronzucker, H.J., Glass, A.D.M., Siddiqi, M.Y. (1995a) Nitrate induction in spruce: an approach using compartmental analysis. Planta 196, 683–690

    Google Scholar 

  • Kronzucker, H.J., Siddiqi, M.Y., Glass, A.D.M. (1995b) Compartmentation and flux characteristics of ammonium in spruce. Planta 196, 691–698

    CAS  Google Scholar 

  • Lavoie, N., Vézina, L.-P., Margolis, H.A. (1992) Absorption and assimilation of nitrate and ammonium ions by Jack pine seedlings. Tree Physiol. 11, 171–183

    Google Scholar 

  • 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, 1753–1756

    Google Scholar 

  • Likens, G.E., Borman, F.H., Johnson, N.M. (1969) Nitrification: importance to nutrient losses for a cutover forest ecosystem. Science 163, 1205–1206

    Google Scholar 

  • Littke, W.R., Bledsoe, C.S., Edmonds, R.L. (1984) Nitrogen uptake and growth in vitro by Hebeloma crustuliniforme and other Pacific Northwest mycorrhizal fungi. Can. J. Bot. 62, 647–652

    Google Scholar 

  • Lodhi, M.A.K. (1978) Inhibition of nitrifying bacteria, nitrification and mineralization of spoil soils as related to their successional stages. Bull. Torrey Bot. Club 106, 284–289

    Google Scholar 

  • Mackion, 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, 359–3

    Google Scholar 

  • Marschner, H., Häussling, M., George, E. (1991) Ammonium and nitrate uptake rates and rhizosphere pH in non-mycorrhizal roots of Norway spruce (Picea abies L. Karst.). Trees 5, 14–21

    Google Scholar 

  • McNaughton, G.S., Presland, M.R. (1983) Whole plant studies using radioactive 13-nitrogen. I. Techniques for measuring the uptake and transport of nitrate and ammonium ions in hydroponically grown Zea mays. J. Exp. Bot. 34, 880–892

    Google Scholar 

  • Meeks, J.C. (1993) 13N techniques. In: Nitrogen isotope techniques, pp. 273–303, Knowles, R., Blackburn, T.H., eds. Academic Press, Inc., San Diego, Calif.

    Google Scholar 

  • Miller, A.J., Smith, S.J. (1992) The mechanism of nitrate transport across the tonoplast of barley root cells. Planta 187, 554–557

    Google Scholar 

  • Plassard, C., Barry, D., Eltrop, L., Mousin, D. (1993) Nitrate uptake in maritime pine (Pinus pinaster) and the ectomycorrhizal fungus Hebeloma cylindrosporum: effect of ectomycorrhizal symbiosis. Can. J. Bot. 72, 189–197

    Google Scholar 

  • Presland, M.R., McNaughton, G.S. (1984) Whole plant studies using radioactive 13-nitrogen. II. A compartmental model for the uptake and transport of nitrate ions by Zea mays. J. Exp. Bot. 35, 1277–1288

    Google Scholar 

  • Redinbaugh, M.G., Campbell, W.H. (1993) Glutamine synthetase and ferredoxin-dependent glutamate synthase in the maize (Zea mays) root primary response to nitrate. Plant Physiol. 101, 1249–1255

    Google Scholar 

  • Rice, E.L., Pancholy, S.K. (1972) Inhibition of nitrification by climax ecosystems. Am. J. Bot. 59, 1033–1040

    Google Scholar 

  • Robin, P., Conejero, G., Passama, L., Salsac, L. (1983) Evaluation de la fraction metabolisable du nitrate par la mesure in situ de sa réduction. Physiol. Vég. 21, 115–122

    Google Scholar 

  • Rygiewicz, P.T., Bledsoe, C., Zasoski, R.J. (1984a) Effect of ectomycorrhizae and solution pH on 15N-ammonium uptake by coniferous seedlings. Can J. Bot. 14, 885–892

    Google Scholar 

  • Rygiewicz, P.T., Bledsoe, C., Zasoski, R.J. (1984b) Effect of ectomycorrhizae and solution pH on 15N-nitrate uptake by coniferous seedlings. Can. J. For. Res. 14, 893–899

    Google Scholar 

  • Siddiqi, M.Y., Glass, A.D.M., Ruth, T.J., Rufty, T.W. (1990) Studies of the uptake of nitrate in barley. I. Kinetics of 13NO −3 influx. Plant Physiol. 93, 1426–1432

    Google Scholar 

  • Siddiqi, M.Y., Glass, A.D.M., Ruth, T.J. (1991) Studies of the uptake of nitrate in barley. III. Compartmentation of NO −3 . J. Exp. Bot. 42, 1455–1463

    Google Scholar 

  • Siddiqi, M.Y., King, B.J., Glass, A.D.M. (1992) Effects of nitrite, chlorate, and chlorite on nitrate uptake and nitrate reductase activity. Plant Physiol. 100, 644–650

    Google Scholar 

  • Smirnoff, N., Stewart, G.R. (1985) Nitrate assimilation and translocation by higher plants: comparative physiology and ecological consequences. Physiol. Plant. 64, 133–140

    Google Scholar 

  • Solomonson, L.P., Barber, M.J. (1990) Assimilatory nitrate reductase: functional properties and regulation. Annu. Rev. Plant Physiol. Plant Mol. Biol. 41, 225–253

    Google Scholar 

  • Vogt, M., Edmonds, R.L. (1982) NO −3 and NH +4 levels in relation to site quality in Douglas-fir soil and litter. Northwest Sci. 56, 83–89

    Google Scholar 

  • Walker, N.A., Pitman, M.G. (1976) Measurement of fluxes across membranes. In: Encyclopedia of plant physiology, vol. 2, part A, pp. 93–126, Lüttge, U., Pitman, M.G., eds., Springer Verlag, Berlin

    Google Scholar 

  • Wang, M.Y., Siddiqi, M.Y., Ruth, T.J., Glass, A.D.M. (1993) Ammonium uptake by rice roots. I. Fluxes and subcellular distribution of 13NH +4 . Plant Physiol. 103, 1249–1258

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Department of Botany, University of British Columbia, V6T 1Z4, Vancouver, B.C., Canada

    Herbert J. Kronzucker, M. Yaeesh Siddiqi & Anthony D. M. Glass

Authors
  1. Herbert J. Kronzucker
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. M. Yaeesh Siddiqi
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Anthony D. M. Glass
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Anthony D. M. Glass.

Additional information

The research was supported by a Natural Sciences and Engineering Research Council, Canada, grant to Dr. A.D.M. Glass and by a University of British Columbia Graduate Fellowship to Herbert J. Kronzucker. Our thanks go to Dr. M. Adam and Mr. P. Culbert at the particle accelerator facility TRIUMF on the University of British Columbia Campus for providing 13NO −3 , Drs. R.D. Guy and S. Silim for providing plant material, and Dr. M.Y. Wang, Mr. J. Mehroke and Mr. P. Poon for assistance in experiments and for helpful discussions.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kronzucker, H.J., Siddiqi, M.Y. & Glass, A.D.M. Compartmentation and flux characteristics of nitrate in spruce. Planta 196, 674–682 (1995). https://doi.org/10.1007/BF00197331

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00197331

Key words

Search

Navigation