Planta

, Volume 168, Issue 3, pp 316–323

Carbon and nitrogen metabolism in barley (Hordeum vulgare L.) mutants lacking ferredoxin-dependent glutamate synthase

  • A. C. Kendall
  • R. M. Wallsgrove
  • N. P. Hall
  • J. C. Turner
  • P. J. Lea
Article

Abstract

Five mutant lines of barley (Hordeum vulgare L.), which are only able to grow at elevated levels of CO2, contain less than 5% of the wild-type activity of ferredoxin-dependent glutamate synthase (EC 1.4.7.1). Two of these lines (RPr 82/1 and RPr 82/9) have been studied in detail. Leaves and roots of both lines contain normal activities of NADH-dependent glutamate synthase (EC 1.4.1.14) and the other enzymes of ammonia assimilation. Under conditions that minimise photorespiration, both mutants fix CO2 at normal rates; on transfer to air, the rates drop rapidly to 15% of the wild-type. Incorporation of 14CO2 into sugar phosphates and glycollate is increased under such conditions, whilst incorporation of radioactivity into serine, glycine, glycerate and sucrose is decreased; continuous exposure to air leads to an accumulation of 14C in malate. The concentrations of malate, glutamine, asparagine and ammonia are all high in air, whilst aspartate, alanine, glutamate, glycine and serine are low, by comparison with the wild-type parent line (cv. Maris Mink), under the same conditions. The metabolism of [14C]glutamate and [14C]glutamine by leaves of the mutants indicates a very much reduced ability to convert glutamine to glutamate. Genetic analysis has shown that the mutation in RPr 82/9 segregates as a single recessive nuclear gene.

Key words

Ammonia/ammonium metabolism Glutamate synthase (ferredoxin dependent) Hordeum (mutant) Mutant (barley) Photorespiration 

Abbreviations

GDH

glutamate dehydrogenase (EC 1.4.1.2)

GS

glutamine synthetase (EC 6.3.1.2)

RuBP

ribulose 1,5-bisphosphate

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References

  1. Archireddy, N.R., Vann, D.R., Fletcher, J.S., Beevers, L. (1983) The influence of methionine sulphoximine on photosynthesis and nitrogen metabolism in excised pepper (Capsicum annuum) leaves. Plant Sci. Lett. 32, 73–78Google Scholar
  2. Arrabaca, M.N.C. (1981) The effects of temperature on photosynthetic and photorespiratory metabolism. Ph.D. thesis, University of LondonGoogle Scholar
  3. Bergman, A., Gardestrom, P., Ericson, I. (1981) Release and refixation of ammonia during photorespiration. Physiol. Plant. 53, 528–532Google Scholar
  4. 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: Oxford surveys of plant molecular and cell biology, vol. 1, pp. 141–169, Miflin, B.J. ed. Oxford University PressGoogle Scholar
  5. Brunetti, N., Hageman, R.H. (1976) Comparison of in vivo and in vitro assays of nitrate reductase in wheat (Triticum aestivum L.) seedlings. Plant Physiol. 58, 583–587Google Scholar
  6. Hall, N.P., Keys, A.J. (1983) Temperature dependence of enzymic decarboxylation and oxygenation of RuBP in relation to effects of temperature on photosynthesis. Plant Physiol. 72, 945–948Google Scholar
  7. Ikeda, M., Ogren, W.L., Hageman, R.H. (1984) Effect of methionine sulphoximine on photosynthetic carbon metabolism in wheat leaves. Plant Cell Physiol. 25, 447–452Google Scholar
  8. Kendall, A.C., Keys, A.J., Turner, J.C., Lea, P.J., Miflin, B.J. (1983) The isolation and characterisation of a catalase deficient mutant of barley. Planta 159, 505–511Google Scholar
  9. 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, 741–743Google Scholar
  10. Larsen, R.O., Cornwall, K.L., Gee, S.L., Bassham, J.A. (1981) Amino acid synthesis in photosynthesising spinach cells: effects of ammonia on pool sizes and rates of labelling from 14CO2. Plant Physiol. 68, 292–299Google Scholar
  11. Lea, P.J., Hall, N.P., Kendall, A.C., Keys, A.J., Turner, J.C., Miflin, B.J. (1984) The isolation and characterisation of photorespiratory mutants of barley (Hordeum vulgare). In: Advances in photosynthesis research, vol. III, pp. 841–844, Sybesma, C. ed. Martinus Nijhoff/Dr. W. Junk, The HagueGoogle Scholar
  12. Martin, F., Winspear, M.S., Macfarlane, J.D., Oaks, A. (1983) Effect of methionine sulphoximine on the accumulation of ammonia in C3 and C4 leaves. Plant Physiol. 71, 177–181Google Scholar
  13. McCullough, H. (1067) The determination of ammonia in whole blood by a direct colorimetric method. Clin. Chim. Acta 17, 297–304Google Scholar
  14. Miziorko, H.M., Lorimer, G.H. (1983) Ribulose-1,5-bisphosphate carboxylase/oxygenase. Annu. Rev. Biochem. 52, 507–535Google Scholar
  15. Möllering, H. (1974) Determination with malate dehydrogenase and glutamate-oxaloacetate transaminase. In: Methods of enzymatic analysis, vol. 3, pp. 1589–1593. Bergmeyer, H.U., ed. Academic Press, New York, LondonGoogle Scholar
  16. Ogren, W.L., Chollet, R. (1982) Photorespiration. In: Photosynthesis: development, carbon metabolism and plant productivity. vol. II. pp. 191–230, Govindjee, ed. Academic Press, New York LondonGoogle Scholar
  17. Platt, S.G., Anthon, G.E. (1981) Ammonia accumulation and inhibition of photosynthesis in methionine sulphoximine treated spinach. Plant Physiol. 67, 509–513Google Scholar
  18. Redgwell, R.J. (1980) Fractionation of plant extracts using ion-exchange Sephadex. Anal. Biochem. 107, 44–50Google Scholar
  19. 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. I, pp. 103–131, Miflin, B.J. ed. Oxford University Press, UKGoogle Scholar
  20. Somerville, C.R., Ogren, W.L. (1979) A phosphoglycolate phosphatase-deficient mutant of Arabidopsis. Nature 280, 833–836Google Scholar
  21. Somerville, C.R., Ogren, W.L. (1980) Inhibition of photosynthesis in Arabidopsis mutants lacking in leaf glutamate synthase activity. Nature 286, 257–259Google Scholar
  22. Somerville, S.C., Ogren, W.L. (1983) An Arabidopsis thaliana mutant defective in chloroplast dicarboxylate transport. Proc. Natl. Acad. Sci. USA 80, 1290–1294Google Scholar
  23. Somerville, S.C., Somerville, C.R. (1985) A mutant of Arabidopsis deficient in chloroplast dicarboxylate transport is missing an envelope protein. Plant Sci. Lett. 37, 217–220Google Scholar
  24. Suzuki, A., Vidal, J., Gadal, P. (1982) Glutamate synthase isoforms in rice. Immunological studies of enzymes in green leaf, etiolated leaf and root tissues. Plant Physiol. 70, 827–832Google Scholar
  25. Thomas, S.M., Thorne, G.N. (1975) Effect of nitrogen fertiliser on photosynthesis and ribulose 1,5-diphosphate carboxylase activity in spring wheat in the field. J. Exp. Bot. 26, 43–51Google Scholar
  26. Tolbert, N.E. (1979) Glycollate metabolism by higher plants and algae. In: Encyclopedia of plant physiology, N.S., vol. 6: Photosynthesis II, pp. 338–353, Gibbs, M., Latzko, E. eds. Springer, Berlin Heidelberg New YorkGoogle Scholar
  27. Waidyanatha, U.P. de S., Keys, A.J., Whittingham, C.P. (1975) Effects of carbon dioxide on metabolism by the glycollate pathway in leaves. J. Exp. Bot. 26, 15–26Google Scholar
  28. Walker, K.A., Givan, C.V., Keys, A.J. (1984a) 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, 60–65Google Scholar
  29. Walker, K.A., Keys, A.J., Givan, C. (1984b) Effect of l-methionine sulphoximine on the products of photosynthesis in wheat (Triticum aestivum) leaves. J. Exp. Bot. 35, 1800–1810Google Scholar
  30. Wallsgrove, R.M., Keys, A.J., Bird, I.F., Cornelius, M.J., Lea, P.J., Mifling, B.J. (1980) The location of glutamine synthetase in leaf cells and its role in the reassimilation of ammonia released in photorespiration. J. Exp. Bot. 31, 1005–1017Google Scholar
  31. 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, 232–236Google Scholar
  32. Wallsgrove, R.M., Lea, P.J., Miflin, B.J. (1982) The development of NAD(P)H-dependent and ferredoxin-dependent glutamate synthase in greening barley leaves. Planta 154, 473–476Google Scholar
  33. Yamaya, T., Oaks, A., Matsumoto, H. (1984) Characteristics of glutamate dehydrogenase in mitochondria prepared from corn shoots. Plant Physiol. 76, 1009–1013Google Scholar

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • A. C. Kendall
    • 1
  • R. M. Wallsgrove
    • 1
  • N. P. Hall
    • 1
  • J. C. Turner
    • 1
  • P. J. Lea
    • 1
  1. 1.Department of BiochemistryRothamsted Experimental StationHarpendenUK
  2. 2.Department of Biological SciencesUniversity of LancasterUK

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