Skip to main content
Log in

Manipulating the pathway of ammonia assimilation through genetic engineering and breeding: consequences to plant physiology and plant development

  • Published:
Plant and Soil Aims and scope Submit manuscript

Abstract

In this article we discuss the ways in which our understanding of the nature of the molecular controls of nitrogen assimilation have been increased by the use of leguminous and non-leguminous plants with modified capacities for ammonium assimilation. These modifications have been achieved through genetic engineering and breeding. An improved understanding of nitrogen assimilation will be vital if improvements in crop nitrogen use efficiency are to be made to reduce the need for excessive input of fertilisers. In this review we present an overall view of past work and more recent studies on this topic. In our work, using tobacco and Lotus as model plants, glutamine synthetase and glutamate synthase activites have been altered by stimulating or inhibiting in an organ- or tissue-specific manner the expression of the corresponding genes. The physiological impact of these genetic manipulations has been studied on plants grown under different nitrogen regimes.

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.

Similar content being viewed by others

References

  • Ameziane R, Bernhard K, Bates R and Lightfoot D A 1988 Metabolic engineering of C and N metabolism with NADPH glutamate dehydrogenase. In Abstract from the Annual Meeting of the American Society of Plant Physiologists June 27–July 1 1988, p 15. Madison, WIS.

  • Bauer D, Biehler K, Fock H, Carrayol E, Hirel B, Migge A and Becker T W 1997 A role for cytosolic glutamine synthetase in the remobilization of leaf nitrogen during water stress in tomato. Physiol. Plant. 99, 241–248.

    Article  CAS  Google Scholar 

  • Becker T W, Caboche M, Carrayol E and Hirel B 1992 Nucleotide sequence of a tobacco cDNA encoding plastidic glutamine 92 synthetase and light inducibility, organ specificity and diurnal rhythmicity in the expression of the corresponding genes of tobacco and tomato. Plant Mol. Biol. 19, 367–379.

    Article  PubMed  CAS  Google Scholar 

  • Bertin P 1997 Bases genetiques et physiologiques de la valorisation de la fumure azotée chez le mais. Thèse de Doctorat de l'Universite de Paris-Sud.

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

    CAS  Google Scholar 

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

    Article  Google Scholar 

  • Bourque J E 1995 Antisense strategies for genetic manipulations in plants. Plant Sci. 105, 125–149.

    Article  CAS  Google Scholar 

  • Botella R, Verbelen J P and Valpuesta V 1988 Immunocytolocalization of glutamine synthetase in green leaves and cotyledons of Lycopersicon esculentum. Plant Physiol. 88, 943–946.

    PubMed  CAS  Google Scholar 

  • Brangeon J, Hirel B and Forchion A 1989 Immunogold localisation of glutamine synthetase in soybean leaves, roots and nodules. Protoplasma 151, 88–97.

    Article  Google Scholar 

  • Brears T, Lui C, Knight T J and Coruzzi G M 1993 Ectopic expression of asparagine synthetase in transgenic tobacco. Plant Physiol. 103, 1285–1290.

    PubMed  CAS  Google Scholar 

  • Brugière N, Dubois F, Limami A M, Lelandais M, Roux Y, Sangwan R S and Hirel B 1999 Glutamine synthetase in the phloem plays a major role in controlling proline production. Plant Cell 11, 1995–2011.

    Article  PubMed  Google Scholar 

  • Cammaerts D and Jacobs M 1983 A study of the polymorphism and the genetic control of the glutamate dehydrogenase isozymes in Arabidopsis thaliana. Plant Sci. Lett. 31, 65–73.

    Article  CAS  Google Scholar 

  • Cammaerts D and Jacobs M 1985 A study of the role of glutamate dehydrogenase in the nitrogen metabolism of Arabidopsis thaliana. Planta 163, 157–260.

    Article  Google Scholar 

  • Carvalho E, Pereira S, Sunkel C and Salema R 1992 Detection of a cytosolic glutamine synthetase in leaves of Nicotiana tabacum L. by immunocytochemical methods. Plant Physiol. 100, 1591–1594.

    PubMed  CAS  Google Scholar 

  • Chen F and Cullimore J V 1988 Two enzymes of NADH-dependent glutamate synthase in root nodules of Phaseolus vulgaris L. Plant Physiol. 88, 1411–1417.

    PubMed  CAS  Google Scholar 

  • Donn G, Tisher E, Smith J A and Goodman H M 1984 Herbicide resistant alfalfa cells: an example of gene amplification in plants. J. Mol. Appl. Genet. 2, 621–635.

    PubMed  CAS  Google Scholar 

  • Degenhart N R, Barnes D K and Vance C P 1992 Divergent selection for nodule aspartate aminotransferase and asparagine synthetase activities in alfalfa. Crop Sci. 32, 313–317.

    Article  CAS  Google Scholar 

  • Dubois F, Brugière N, Sangwan R S and Hirel B 1996 Localisation of tobacco cytosolic glutamine synthetase enzymes and the corresponding transcripts show organ-and cell-specific pattern of protein synthesis and gene expression. Plant Mol. Biol. 31, 803–817.

    Article  PubMed  CAS  Google Scholar 

  • Eckes P, Schmidtt P, Daub W and Wegenmayer F 1989 Overproduction of alfalfa glutamine synthetase in transgenic tobacco plants. Mol. Gen. Genet. 217, 263–268.

    PubMed  CAS  Google Scholar 

  • Faure J-D, Vincentz M, Kronenberger J and Caboche M 1991 Coregulated expression of nitrate and nitrite reductases. Plant J. 1, 107–113.

    Article  CAS  Google Scholar 

  • Ferrario-Méry S, Suzuki A, Kunz C, Valadier M H, Roux Y, Hirel B and Foyer C H 1999 Modulation of amino acid metabolism in transformed tobacco plants deficient in Fd-GOGAT. Plant Soil 221, 67–79.

    Article  Google Scholar 

  • Forde B J and Cullimore J V 1989 The molecular biology of glutamine synthetase in higher plants. Oxf. Surv. Plant Mol. Biol. 6, 247–296.

    CAS  Google Scholar 

  • Groat R C, Vance C P and Barnes D K 1984 Host plant nodule enzymes associated with selection for increased N2 fixation in alfalfa. Crop Sci. 24, 895–898.

    Article  Google Scholar 

  • Hayakawa T, Nakamura T, Hattori F, Mae T, Ojima K and Yamaya T 1994 Cellular localisation of NADH-dependant glutamate synthase protein in vascular bundles of unexpanded leaf blades and young grains of rice plants. Planta 193, 455–460.

    Article  CAS  Google Scholar 

  • Hirel B, Marsolier M C, Hoarau J, Brangeon J, Scafer R and Verma D P S 1992 Forcing expression of a soybean root glutamine synthetase gene in tobacco leaves induces a native gene encoding cytosolic enzyme. Plant Mol. Biol. 20, 207–218.

    Article  PubMed  CAS  Google Scholar 

  • Hirel B, Miao G H and Verma D P S 1993 Metabolic and developmental control of glutamine synthetase genes in legume and non legume plants. In Control of plant gene expression. Ed. D P S Verma. pp 443–458. CRC Press, Boca Raton, FL.

    Google Scholar 

  • Hirel B, Phillipson B, Murchie E, Suzuki A, Kunz C, Ferrario S, Limami A, Chaillou S, Deléens E, Brugière N, Chaumont-Bonnet M, Foyer C H and Morot-Gaudry J F 1997 Manipulating the pathway of ammonia assimilation in transgenic non-legumes and legumes. Z. Pflanzenernähr. Bodenk. 160, 283–290.

    CAS  Google Scholar 

  • Jessen D L, Barnes D K, Vance C P and Heichel G H 1987 Variation for activity of nodule nitrogen and carbon assimilating enzymes in alfalfa. Crop Sci. 27, 627–631.

    Article  CAS  Google Scholar 

  • Jessen D L, Barnes D K and Vance C P 1988 Bidirectional selection in alfalfa for activity of nodule nitrogen and carbon assimilating enzymes. Crop Sci. 28, 18–22.

    Article  CAS  Google Scholar 

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

    CAS  Google Scholar 

  • Kamachi K, Yamaya T, Mae T and Ojima K 1991 A role for glutamine synthetase in the remobilisation of leaf nitrogen during natural senescence in rice leaves. Plant Physiol. 96, 411–417.

    PubMed  CAS  Google Scholar 

  • Keys A F, Bird I F, Cornelius M J, Lea P J, Wallsgrove R M and Miflin B J 1978 Photorespiratory nitrogen cycle. Nature 275, 741–742.

    Article  Google Scholar 

  • Knight T J and Langston-Unkefer P J 1988 Enhancement of symbiotic dinitrogen fixation by a toxin-releasing plant pathogen. Science 241, 951–954.

    CAS  PubMed  Google Scholar 

  • Kozaki A and Takeba G 1996 Photorespiration protects C3 plants from photooxidation. Nature 384, 557–560.

    Article  CAS  Google Scholar 

  • Lam H M, Coschigano K T, Oliveira I C, Melo-Oliveira R and Coruzzi G M 1996 The molecular genetics of nitrogen assimilation into amino acids in higher plants. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47, 569–593.

    Article  PubMed  CAS  Google Scholar 

  • Lara M, Cullimore J V, Lea P J, Miflin B J, Johnston A W B and Lamb J W 1983 Appearance of a novel form of glutamine synthetase during nodule development in Phaseolus vulgaris. Planta 157, 254–258.

    Article  CAS  Google Scholar 

  • Lea P J and Ireland R J 1999 The enzymes of glutamine, glutamate, asparagine and aspartate metabolism. In Plant Amino Acids. Ed. B K Singh. pp 1–47. Marcel Dekker, New York.

    Google Scholar 

  • Lea P J and Miflin B J 1980 Transport and metabolism of asparagine and other nitrogen compounds within the plant. In The Biochemistry of Plants. Eds P K Stumpf and E E Conn. pp 569–607. Academic Press, New York.

    Google Scholar 

  • Limami A, Phillipson B, Ameziane R, Pernollet N, Jiang Q, Roy R, Deleens E, Chaumont-Bonnet M, Gresshof PM and Hirel B 1999 Does root glutamine synthetase control plant biomass production in Lotus japonicus L.? Planta 209, 495–502.

    Article  PubMed  CAS  Google Scholar 

  • Loulakakis K A, Roubelakis-Angelakis K A and Kanellis A K 1994 Regulation of glutamatedehydrogenase and glutamine synthetase in avocado fruit during development and ripening. Plant Physiol. 106, 217–222.

    PubMed  CAS  Google Scholar 

  • Magalhaes J R, Ju G C, Rich P J and Rhodes D 1990 Kinetics of 15NH4 assimilation in Zea mays. Plant Physiol. 94, 647–656.

    Article  PubMed  CAS  Google Scholar 

  • McNally S F and Hirel B 1983 Glutamine synthetase isoforms in higher plants. Physiol. Vég. 21, 761–774.

    CAS  Google Scholar 

  • McNally S F, Hirel B, Gadal P, Mann A F and Stewart G R 1983 Glutamine synthetase of higher plants: evidence for a specific isoform content related to their possible physiologicalrole and their compartmentation within the leaf. Plant Physiol. 72, 22–25.

    PubMed  CAS  Google Scholar 

  • Melo-Oliveria R, Oliveria I C and Coruzzi G M 1996 Arabidopsis mutant analysis and gene regulation define a nonredundant role for glutamate dehydrogenase in nitrogen assimilation. Proc. Natl. Acad. Sci. USA 96, 4718–4723.

    Article  Google Scholar 

  • Miao G H, Hirel B, Marsolier M C, Ridge R W and Verma D P S 1991 Ammonium regulated expression of a soybean encoding cytosolic glutamine synthetase in transgenic Lotus corniculatus. Plant Cell 3, 11–22.

    Article  PubMed  CAS  Google Scholar 

  • Miflin R D and Lea P J 1976 The pathway of nitrogen assimilation in plants. Phytochem. 15, 873–885.

    Article  CAS  Google Scholar 

  • Miflin R D and Lea P J 1980 Ammonium assimilation. In The Biochemistry of Plants, Vol 5. Amino Acids and Derivatives. Ed. B J Miflin. pp 169–202. Academic Press, New York.

    Google Scholar 

  • Migge A, Carrayol E, Hirel B and Becker T W 1999 Leaf specific overexpression of plastidic glutamine synthetase stimulates the growth of transgenic tobacco seedlings. Planta. (In press).

  • Morris P F, Layzell D B and Canvin D T 1989 Photorespiratory ammonium does not inhibit photosynthesis in glutamate synthase mutants of Arabidopsis. Plant Physiol. 89, 498–500.

    PubMed  CAS  Google Scholar 

  • Oaks A 1992 A re-evaluation of nitrogen assimilation in roots. Bioscience 42, 103–111.

    Article  Google Scholar 

  • Oaks A and Hirel B 1985 Nitrogen metabolism in roots. Annu. Rev. Plant Physiol. 63, 345–365.

    Article  Google Scholar 

  • Pahlich E 1996 Remarks concerning the dispute related to the function of plant glutamate dehydrogenase: commentary. Can. J. Bot. 74, 512–515.

    Article  Google Scholar 

  • Peat L J and Tobin A 1996 The effect of nitrogen nutrition on the cellular localization of glutamine synthetase isoforms in barley roots. Plant Physiol. 111, 1109–1117.

    PubMed  CAS  Google Scholar 

  • Peeters K M U and Van Laere A J 1994 Amino acid metabolism associated with N-metabolism from the flag leaf of wheat (Triticum aestivum L.) during grain development. Plant Cell Environ. 17, 131–141.

    Article  CAS  Google Scholar 

  • Pérez-Rodriquez J and Valpuesta V 1996 Expression of glutamine synthetase genes during natural senescence of tomato leaves. Physiol. Plant. 97, 576–582.

    Article  Google Scholar 

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

    PubMed  CAS  Google Scholar 

  • Robertson J G, Farnden K J F, Warburton M P and Banks J A M 1975 Induction of glutamine synthetase during nodule development in lupin. Aust. J. Plant Physiol. 2, 265–272.

    Article  CAS  Google Scholar 

  • Robinson S, Slade A P, Fox G G, Phillips R, Ratcliffe R G and Stewart G R 1991 The role of glutamate dehydrogenase in plant metabolism. Plant Physiol. 95, 509–516.

    PubMed  CAS  Google Scholar 

  • Sechley K A, Yamaya T and Oaks A 1992 Compartmentation of nitrogen assimilation in higher plants. Int. Rev. Cytol. 134, 85–163.

    Article  CAS  Google Scholar 

  • Sommerville C R and Ogren WL 1980 Inhibition of photosynthesis in Arabidopsis mutants lacking leaf glutamate synthase activity. Nature 286, 257–259.

    Article  Google Scholar 

  • Suzuki A and Gadal P 1984 Glutamate synthase: physiochemical and functional properties of different forms in higher plants and other organisms. Physiol. Vég. 22, 471–486.

    CAS  Google Scholar 

  • Suzuki A, Vidal J and Gadal P 1982 Glutamate synthase isoforms in rice: immunological studies of enzymes of green leaf, etiolated leaf and root tissues. Plant Physiol. 70, 827–832.

    PubMed  CAS  Google Scholar 

  • Srivastava H S and Singh Rana P 1987 Role and regulation of glutamate dehydrogenaseactivity in higher plants. Phytochemistry 26, 597–610.

    Article  CAS  Google Scholar 

  • Temple S J, Bagga S and Sengupta-Gopalan C 1994 Can glutamine synthetase activity bemodulated in transgenic plants by the use of recombinant DNA technology? Transgenic Plants and Plant Biochemistry. Biochem. Soc. Trans. 22, 915–920.

    CAS  Google Scholar 

  • Temple S J, Bagga S and Sengupta-Gopalan C 1998b Down regulation of specific members of the glutamine synthetase gene family in alfalfa by antisense RNA technology. Plant Mol. Biol. 37, 535–547.

    Article  PubMed  CAS  Google Scholar 

  • Temple S T, Knight T J, Unkefer P J and Sengupta-Gopalan C 1993 Modulation of glutamine synthetase gene expression in tobacco by the introduction of an alfalfa glutamine synthetasegene in sense and antisense orientation: molecular and biochemical analysis. Mol. Gen. Genet. 236, 315–325.

    Article  PubMed  CAS  Google Scholar 

  • Temple S J, Vance C P and Gantt J S 1998a Glutamate synthase and nitrogen assimilation. Trends Plant Sci. 3, 51–56.

    Article  Google Scholar 

  • Trepp G B, Plank D W, Gantt J S and Vance C P 1999a NADHglutamate synthase in alfalfa root nodules. Immunocytochemical localisation. Plant Physiol. 119, 829–837.

    Article  PubMed  CAS  Google Scholar 

  • Trepp G B, van de Mortel M, Yoshoika H, Miller S S, Samac D A, Gantt J S and Vance C P 1999b NADH-glutamate synthase in alfalfa roots. Genetic regulation and cellular expression. Plant Physiol. 119, 817–828.

    Article  PubMed  CAS  Google Scholar 

  • Vezina L P and Langlois J R 1989 Tissue and cellular distribution of glutamine synthetase in roots of pea (Pisum sativum) seedlings. Plant Physiol. 90, 1129–1133.

    PubMed  CAS  Google Scholar 

  • Vincent R, Fraiser V, Chaillou S, Limami M A, Deleens E, Phillipson B, Couat C, Boutin J and Hirel B 1997 Overexpression of a soybean gene encoding cytosolic glutamine synthetase in shoots of transgenic Lotus corniculatus L. plants triggers changes in ammonium assimilation and plant development. Planta 201, 424–433.

    Article  PubMed  CAS  Google Scholar 

  • Woodall J, Boxall J G, Forde B G and Pearson J 1996 Changing perspectives in plant nitrogen metabolism: the central role of glutamine synthetase. Sci. Prog. 79, 1–26.

    CAS  Google Scholar 

Download references

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Harrison, J., Brugière, N., Phillipson, B. et al. Manipulating the pathway of ammonia assimilation through genetic engineering and breeding: consequences to plant physiology and plant development. Plant and Soil 221, 81–93 (2000). https://doi.org/10.1023/A:1004715720043

Download citation

  • Issue Date:

  • DOI: https://doi.org/10.1023/A:1004715720043

Navigation