Skip to main content
SpringerLink
Log in

Control of photosynthesis in barley leaves with reduced activities of glutamine synthetase or glutamate synthase

I. Plant characteristics and changes in nitrate, ammonium and amino acids

  • Published:
Planta Aims and scope Submit manuscript

Abstract

Wild-type and mutant plants of barley (Hordeum vulgare L. cv. Maris Mink) lacking activities of chloroplastic glutamine synthetase (GS) and of ferredox-in-dependent glutamate synthase (Fd-GOGAT) were crossed to generate heterozygous plants. Crosses of the F2 generation containing GS activities between 47‰ and 97‰ of the wild-type and Fd-GOGAT activities down to 63‰ of the wild-type have been selected to study the control of both enzymes on photorespiratory carbon and nitrogen metabolism. There were no major pleiotropic effects. Decreased GS had a small impact on leaf protein and the total activity of ribulose-1,5-bisphosphate carboxylase-oxygenase (Rubisco). The activation state of Rubisco was unaffected in air, but a decrease in GS influenced the activation state of Rubisco in low CO2. In illuminated leaves, the amino-acid content decreased with decreasing GS, while the content of ammonium rose, showing that even small reductions in GS limit ammonium re-assimilation and may bring about a loss of nitrogen from the plants, and hence a reduction in protein and Rubisco. Leaf amino-acid contents were restored, and ammonium and nitrate contents decreased, by leaving plants in the dark for 24 h. The ratios of serine to glycine decreased with a decrease in GS when plants were kept at moderate photon flux densities in air, suggesting a possible feedback on glycine decarboxylation. This effect was absent in high light and low CO2. Under these conditions ammonium contents exhibited an optimum and amino-acid contents a minimum at a GS activity of 65‰ of the wild-type, suggesting an inhibition of ammonium release in mutants with less than 65‰ GS. The leaf contents of glutamate, glutamine, aspartate, asparagine, and alanine largely followed changes in the total amino-acid contents determined under different environmental conditions. Decreased Fd-GOGAT resulted in a decrease in leaf protein, chlorophyll, Rubisco and nitrate contents. Chlorophyll a/b ratios and specific leaf fresh weight were lower than in the wild-type. Leaf ammonium contents were similar to the wild-type and total leaf amino-acid contents were only affected in low CO2 at high photon flux densities, but mutants with decreased Fd-GOGAT accumulated glutamine and contained less glutamate.

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.

Institutional subscriptions

Similar content being viewed by others

Abbreviations

Chl:

chlorophyll

FBPase:

fructose-1,6-bisphosphatase

Fd-GOGAT:

ferredoxin-dependent glutamine: 2-oxoglutarate aminotransferase

GS:

glutamine synthetase

PEP:

phosphoenolpyruvate

PFD:

photon flux density

Rubisco:

ribulose-1,5-bisphosphate carboxylase-oxygenase

References

  • Arnon, D.J. (1949) Copper enzymes in chloroplasts. Polypoxidase in Beta vulgaris. Plant Physiol. 24, 1–15

    Google Scholar 

  • Ashton, A.R., Burnell, J.N., Furbank, R.T., Jenkins, C.L.D., Hatch, M.D. (1990) Enzymes of C4 photosynthesis In: Methods in plant biochemistry, vol. 3, pp. 39–72, Lea, P.J., ed. Academic Press, London

    Google Scholar 

  • Aslam, M., Huffaker, R.C., Rains, D.W., Rao, K.P. (1979) Influence of light and ambient carbon dioxide concentration on nitrate assimilation by intact barley seedlings. Plant Physiol. 63, 1205–1209

    Google Scholar 

  • Betsche, T. (1983) Aminotransfer from alanine and glutamate glycine and serine during photorespiration in oat leaves. Plant Physiol. 71, 961–965

    Google Scholar 

  • Blackwell, R.D., Murray, A.J.S., Lea, P.J. (1987) Inhibition of photosynthesis in barley with decreased levels of chloroplastic glutamine synthetase activity. J. Exp. Bot. 38, 1799–1809

    Google Scholar 

  • Blackwell, R.D., Murray, A.J.S., Lea, P.J., Joy, K.W. (1988a) Photorespiratory amino donors, sucrose synthesis and the induction of CO2 fixation in barley deficient in glutamine synthetase and/ glutamate synthase. J. Exp. Bot. 39, 845–858

    Google Scholar 

  • Blackwell, R.D., Murray, A.J.S., Lea, P.J., Kendall, A.C., Hall, N.P., Turner, J.C., Wallsgrove, R.M. (1988b) The value of mutants unable to carry out photorespiration. Photosynth. Res. 16, 155–176

    Google Scholar 

  • Bowsher, C.G., Boulton, E.L., Rose, J., Nayagam, S., Emes, M.J. (1992) Reductant for glutamate synthase is generated by the oxidative pentose phosphate pathway in non-photosynthetic root plastids. Plant J. 2, 893–898

    Google Scholar 

  • Bradford, M. (1976) A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principles protein-dye-binding. Anal. Biochem. 72, 248–254

    Article  CAS  PubMed  Google Scholar 

  • Campbell, W.J., Ogren, W.L. (1990) Glyoxylate inhibition of ribulosebisphosphate carboxylase/oxygenase activation in intact, lysed, and reconstituted chloroplasts. Photosynth. Res. 23, 257–268

    Google Scholar 

  • Cataldo, D.S., Haroon, M., Schrader, L.E., Youngs, V.L. (1975) Rapid colorimetric determination of nitrate in plant tissue by nitration of salicylic acid. Commun. Soil Sci. Plant Anal. 72, 248–253

    Google Scholar 

  • Edwards, J.W., Walker, E.L., Coruzzi, G.M. (1990) Cell-specific expression in transgenic plants reveals nonoverlapping roles for chloroplast and cytosolic glutamine synthetase. Proc. Natl. Acad. Sci. USA 87, 3459–3463

    Google Scholar 

  • Farquhar, G.D., Firth, P.M., Wetslaar, R., Weir, B. (1980) On the gaseous exchange of ammonia between leaves and the environment: Determination of the ammonia compensation point. Plant Physiol. 66, 710–714

    Google Scholar 

  • Graan, T., Ort, D.R. (1984) Quantitation of the rapid electron donors to P700, the functional plastoquinone pool, and the ratio of the photosystems in spinach chloroplasts. J. Biol. Chem. 259, 14003–14010

    Google Scholar 

  • Häusler, R.E., Lea, P.J., Leegood, R.C. (1994) Control of photosynthesis in barley leaves with reduced activities of glutamate synthetase or glutamate synthase. II. Control of electron transport and CO2 assimilation. Planta 194, 418–435

    Google Scholar 

  • Heldt, H.W. (1979) Light-dependent changes in stromal H+ and Mg2+ concentrations controlling CO2 fixation. In: Encyclopedia of plant physiology N.S., vol. 6: Photosynthetic carbon metabolism and related processes, pp. 202–208, Gibbs, M., Latzko, E., eds. Springer Verlag, N.Y.

    Google Scholar 

  • Hoagland, D.R., Arnon, D.I. (1950) The water culture method for growing plants without soil. Calif. Agric. Exp. St. Circ. No. 397

  • Joy, K.W., Blackwell, R.D., Lea, P.J. (1992) Assimilation of nitrogen in mutants lacking enzymes of the glutamate synthase cycle. J. Exp. Bot. 43, 139–145

    Google Scholar 

  • 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 (Hordeum vulgare L.). Planta 159, 505–511

    Google Scholar 

  • Krause, G.H., Weis, E. (1991) Chlorophyll fluorescence and photosynthesis: the basics. Annu. Rev. Plant Physiol. Plant Mol. Biol. 42, 313–349

    Google Scholar 

  • Lea, P.J., Blackwell, R.D., Chen, F.L., Hecht, U. (1990) Enzymes of ammonia assimilation. In: Methods in plant biochemistry vol. 3, pp. 2–276, Lea, P.J., ed. Academic Press, London

    Google Scholar 

  • Lindroth, P., Mopper, K. (1979) High performance liquid chromatographie determination of subpicomole amounts of amino acids by precolumn fluorescence derivatization with o-phthaldialdehyde. Anal. Chem. 51, 1667–1674

    Google Scholar 

  • Matoh, T., Ida, S., Takahashi, E. (1980) A rapid and sensitive assay for ferredoxin-glutamate synthase. Bull. Res. Inst. Food Sci. Kyoto Univ. 43, 1–6

    Google Scholar 

  • Ort, D.R. (1986) Energy transduction in oxygenic photosynthesis: an overview of structure and mechanism. In: Encyclopedia of plant physiology, N.S., vol. 19: Photosynthesis III, pp. 179–196, Staehelin, L.A., Arntzen, C.J., eds. Springer Verlag, Berlin

    Google Scholar 

  • Peterson, L.W., Kleinkopf, G.E., Huffaker R.C. (1973) Evidence for a lack of turnover of ribulose 1,5-diphosphate carboxylase in barley leaves. Plant Physiol. 51, 1042–1045

    Google Scholar 

  • Plaxton W.C. (1990) Glycolysis. In: Methods in plant biochemistry vol. 3, pp. 145–173, Lea, P.J., ed. Academic Press, London

    Google Scholar 

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

    Google Scholar 

  • Stitt, M., Mieskes, G., Soling, H.D., Heldt, H.W. (1982) On a possible role of fructose 2,6-bisphosphate in regulating photosynthetic metabolism in leaves. FEBS Lett. 145, 217–222

    Google Scholar 

  • Ta, T.C., Joy, K.W. (1986) Metabolism of some amino acids in relation to the photorespiratory nitrogen cycle in pea leaves. Planta 169, 117–122

    Google Scholar 

  • Walker, K.A., Keys, A.J., Givan, C.V. (1984) Effect of l-methionine sulphoximine on the products of photosynthesis in wheat (Triticum aestivum) leaves. J. Exp. Bot. 35, 1800–1810

    Google Scholar 

  • Ward, D.A., Keys, A.J. (1989) A comparison between the coupled spectrophotometric and uncoupled radiometric assays for RuBP carboxylase. Photosynth. Res. 22, 167–171

    Google Scholar 

  • Weatherburn, M.W. (1967) Phenol-hypochlorite reaction for determination of ammonia. Anal. Chem. 39, 971–974

    Google Scholar 

  • Wendler, Ch., Barniske, M., Wild, A. (1990) Effect of phosphinothricin (glufosinate) on photosynthesis and photorespiration of C3 and C4 plants. Photosynth. Res. 24, 55–61

    Google Scholar 

  • Winter, H., Lohaus, G., Heldt, H.W. (1992) Phloem transport of amino acids in relation to their cytosolic levels in barley leaves. Plant Physiol. 99, 996–1004

    Google Scholar 

  • Portis, A.R. Jr., Salvucci, M.E., Ogren, W.L. (1986) Activation of ribulosebisphosphate carboxylase/oxygenase at physiological CO2 and ribulosebisphosphate concentrations by Rubisco activase. Plant Physiol. 82, 967–971

    Google Scholar 

  • Roberts, J.K.M., Pang, M.K.L. (1992) Estimation of ammonium ion distribution between cytoplasm and vacuole using nuclear magnetic resonance spectroscopy. Plant Physiol. 100, 1571–1574

    Google Scholar 

  • Schjoerring, J.K., Kyllingsbaek, A., Mortensen, J.V., Byskov-Nielsen, S. (1993) Field investigations of ammonia exchange between barley plants and the atmosphere. I. Concentration profiles and flux densities of ammonia. Plant Cell Environ. 16, 161–167

    Google Scholar 

  • Sharkey, T.D., Savitch, L.V., Butz, N.D. (1991) Photometric method for routine determination of kcat and carbamylation of rubisco. Photosynth. Res. 28, 41–48

    Google Scholar 

  • Somerville, C.R. (1986) Analysis of photosynthesis and photorespiration using mutants of higher plants and algae. Annu. Rev. Plant Physiol. 37, 467–507

    Google Scholar 

  • Somerville, C.R., Ogren, W.L. (1979) A phosphoglycolate phosphatase-deficient mutant of Arabidopsis. Nature 280, 833–836

    Google Scholar 

  • Somerville, C.R., Ogren, W.L. (1981) Photorespiration-deficient mutants of Arabidopsis thaliana lacking mitochondrial serine transhydroxymethylase activity. Plant Physiol. 67, 666–671

    Google Scholar 

Download references

Author information

Authors and Affiliations

  1. Robert Hill Institute, Department of Animal and Plant Sciences, University of Sheffield, S10 2UQ, Sheffield, UK

    Rainer E. Häusler & Richard C. Leegood

  2. Department of Biological Sciences, University of Lancaster, LA1 4YQ, Bailrigg, Lancaster, UK

    Ray D. Blackwell & Peter J. Lea

Authors
  1. Rainer E. Häusler
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Ray D. Blackwell
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Peter J. Lea
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Richard C. Leegood
    View author publications

    You can also search for this author in PubMed Google Scholar

Additional information

This research was jointly supported by the Agricultural and Food Research Council and the Science and Engineering Research Council, U.K. in the programme on Biochemistry of Metabolic Regulation in Plants (PG50/555).

Rights and permissions

Reprints and permissions

About this article

Cite this article

Häusler, R.E., Blackwell, R.D., Lea, P.J. et al. Control of photosynthesis in barley leaves with reduced activities of glutamine synthetase or glutamate synthase. Planta 194, 406–417 (1994). https://doi.org/10.1007/BF00197542

Download citation

  • Received:

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00197542

Key words

Search

Navigation