Skip to main content
Log in

Carbon and nitrogen metabolism in a barley (Hordeum vulgare L.) mutant with impaired chloroplast dicarboxylate transport

  • Published:
Planta Aims and scope Submit manuscript

Abstract

A mutant line, RPr79/2, of barley (Hordeum vulgare L. cv. Maris Mink) has been isolated that has an apparent defect in photorespiratory nitrogen metabolism. The metabolism of 14C-labelled glutamine, glutamate and 2-oxoglutarate indicates that the mutant has a greatly reduced ability to synthesise glutamate, especially in air, although in-vitro enzyme analysis indicates the presence of wild-type activities of glutamine synthetase (EC 6.3.1.2) glutamate synthase (EC 1.4.7.1 and EC 1.4.1.14) and glutamate dehydrogenase (EC 1.4.1.2). Several characteristics of RPr79/2 are very similar to those described for glutamate-synthase-deficient barley and Arabidopsis thaliana mutants, including the pattern of labelling following fixation of 14CO2, and the rapid rise in glutamine content and fall in glutamate in leaves on transfer to air. The CO2-fixation rate in RPr79/2 declines much more slowly on transfer from 1% O2 to air than do the rates in glutamate-synthase-deficient plants, and RPr79/2 plants do not die in air unless the temperature and irradiance are high. Analysis of (glutamine+NH3+2-oxoglutarate)-dependent O2 evolution by isolated chloroplasts shows that chloroplasts from RPr79/2 require a fivefold greater concentration of 2-oxoglutarate than does the wild-type for maximum activity. The levels of 2-oxoglutarate in illuminated leaves of RPr79/2 in air are sevenfold higher than in Maris Mink. It is suggested that RPr79/2 is defective in chloroplast dicarboxylate transport.

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

Access this article

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

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Anderson, J.W., Done, J. (1977a) A polarographic study of glutamate synthase activity in isolated chloroplasts. Plant Physiol. 60, 354–359

    Google Scholar 

  • Anderson, J.W., Done, J. (1977b) Polarographic study of ammonia assimilation by isolated chloroplasts. Plant Physiol. 60, 504–508

    Google Scholar 

  • Bright, S.W.J., Lea, P.J., Arruda, P., Hall, N.P., Kendall, A.C., Keys, A.J., Kueh, J.S.H., Parker, M.L., Rognes, S.E., Turner, J.C., Wallsgrove, R.M., Miflin, B.J. (1984) Manipulation of the pathways of photorespiration and amino acid metabolism by mutation and selection. In: The genetic manipulation of plants and its application to agriculture, pp. 141–169, Lea, P.J., Stewart, G.P., eds. Oxford University Press, Oxford

    Google Scholar 

  • Dry, I.B., Wiskich, J.T. (1983) Characterization of dicarboxylate stimulation of ammonia, glutamine, and 2-oxoglutarate-dependent O2 evolution in isolated pea chloroplasts. Plant Physiol. 72, 291–296

    Google Scholar 

  • Kendall, A.C., Wallsgrove, R.M., Hall, N.P., Turner, J.C., Lea, P.J. (1986) Carbon and nitrogen metabolism in barley (Hordeum vulgare L.) mutants lacking ferredoxin-dependent glutamate synthase. Planta 168, 316–323

    Google Scholar 

  • Keys, A.J., Bird, I.F., Cornelius, M.J., Lea, P.J., Wallsgrove, R.M., Miflin, B.J., (1978) The photorespiratory nitrogen cycle. Nature 275, 741–743

    Google Scholar 

  • Larsen, R.O., Cornwall, K.L., Gee, S.L., Bassham, J.A. (1981) Amino acid synthesis in photosynthesising spinach cells: effect of ammonia on pool sizes and rates of labelling from [14C]O2. Plant Physiol. 68, 292–299

    Google Scholar 

  • Lilley, R.M., Fitzgerald, M.P., Rientis, K.G., Walker, D.A. (1975) Criteria of intactness and the photosynthetic activity of spinach chloroplast preparations. New Phytol. 75, 1–10

    Google Scholar 

  • Mann, A.F., Fentem, P.A., Stewart, G.R. (1980) Tissue localization of barley (Hordeum vulgare L.) glutamine synthetase isozymes. FEBS Lett. 110, 265–267

    Google Scholar 

  • Somerville, C.R. (1984) The analysis of photosynthetic carbon dioxide fixation and photorespiration by mutant selection. In: Oxford Surveys of Plant Molecular and Cell Biology, vol. 1, pp. 103–131, Miflin, B.J., ed. Oxford University Press, Oxford

    Google Scholar 

  • Somerville, C.R., Ogren, W.L. (1980) Inhibition of photosynthesis in Arabidopsis mutants lacking in leaf glutamate synthase activity. Nature 286, 257–259

    Google Scholar 

  • Somerville, S.C., Ogren, W.L. (1983) An Arabidopsis thaliana mutant defective in chloroplast dicarboxylate transport. Proc. Natl. Acad. Sci. USA 80, 1290–1294

    Google Scholar 

  • Walker, K.A., Givan, C.V., Keys, A.J. (1984) Glutamic acid metabolism and the photorespiratory nitrogen cycle in wheat leaves: metabolic consequences of elevated ammonia concentrations and of blocking ammonia assimilation. Plant Physiol. 75, 60–65

    Google Scholar 

  • Wallsgrove, R.M., Harel, E., Lea, P.J., Miflin, B.J. (1977) Studies on glutamate synthase from the leaves of higher plants. J. Exp. Bot. 28, 588–596

    Google Scholar 

  • Wallsgrove, R.M., Lea, P.J., Miflin, B.J. (1983) Intracellular localization of aspartate kinase and the enzymes of threonine and methionine biosynthesis in green leaves. Plant Physiol. 71, 780–784

    Google Scholar 

  • Wallsgrove, R.M., Lea, P.J., Miflin, B.J. (1979) Distribution of the enzymes of nitrogen assimilation within the pea leaf cell. Plant Physiol. 63, 232–236

    Google Scholar 

  • Wintermans, J.F.G.M., Demots, A. (1965) Spectrophotometric characteristics of chlorophylls a and b and their pheophytins in ethanol. Biochim. Biophys. Acta 109, 448–453

    Google Scholar 

  • Woo, K.C., Osmond, C.B. (1983) Characterization of dicarboxylate stimulation of ammonia, glutamine and 2-oxoglutarate-dependent O2 evolution in isolated pea chloroplasts. Plant Physiol. 72, 291–296

    Google Scholar 

  • Williamson, J.R., Corkey, B.E. (1979) Assay of citric acid cycle intermediates and related compounds — update with tissue metabolite levels and intracellular distribution. Methods Enzymol. 55, 200–222

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wallsgrove, R.M., Kendall, A.C., Hall, N.P. et al. Carbon and nitrogen metabolism in a barley (Hordeum vulgare L.) mutant with impaired chloroplast dicarboxylate transport. Planta 168, 324–329 (1986). https://doi.org/10.1007/BF00392356

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00392356

Key words

Navigation