Planta

, Volume 223, Issue 5, pp 1068–1080 | Cite as

How does glutamine synthetase activity determine plant tolerance to ammonium?

  • C. Cruz
  • A. F. M. Bio
  • M. D. Domínguez-Valdivia
  • P. M. Aparicio-Tejo
  • C. Lamsfus
  • M. A. Martins-Loução
Original Article

Abstract

The wide range of plant responses to ammonium nutrition can be used to study the way ammonium interferes with plant metabolism and to assess some characteristics related with ammonium tolerance by plants. In this work we investigated the hypothesis of plant tolerance to ammonium being related with the plants’ capacity to maintain high levels of inorganic nitrogen assimilation in the roots. Plants of several species (Spinacia oleracea L., Lycopersicon esculentum L., Lactuca sativa L., Pisum sativum L. and Lupinus albus L.) were grown in the presence of distinct concentrations (0.5, 1.5, 3 and 6 mM) of nitrate and ammonium. The relative contributions of the activity of the key enzymes glutamine synthetase (GS; under light and dark conditions) and glutamate dehydrogenase (GDH) were determined. The main plant organs of nitrogen assimilation (root or shoot) to plant tolerance to ammonium were assessed. The results show that only plants that are able to maintain high levels of GS activity in the dark (either in leaves or in roots) and high root GDH activities accumulate equal amounts of biomass independently of the nitrogen source available to the root medium and thus are ammonium tolerant. Plant species with high GS activities in the dark coincide with those displaying a high capacity for nitrogen metabolism in the roots. Therefore, the main location of nitrogen metabolism (shoots or roots) and the levels of GS activity in the dark are an important strategy for plant ammonium tolerance. The relative contribution of each of these parameters to species tolerance to ammonium is assessed. The efficient sequestration of ammonium in roots, presumably in the vacuoles, is considered as an additional mechanism contributing to plant tolerance to ammonium nutrition.

Keywords

Ammonium tolerance Glutamate dehydrogenase Glutamine synthetase Lactuca Lupinus Lycopersicon Pisum Spinacia 

Abbreviations

GDH

Glutamate dehydrogenase

GS

Glutamine synthetase

GS/GOGAT

Glutamine synthetase/glutamate synthase

NR

Nitrate reductase

Notes

Acknowledgements

This work was supported by the joint action HP2001-0020, the Fundação para a Ciência e Tecnologia (FCT) FEDER through the project POCTI 39230/BSE/2001, by the Ministerio de Ciencia y Tecnología through the project AGL2000-0934-C02-01, and Gobierno de Navarra through the project of Resolución 96/2000.

References

  1. Ameziane R, Bernhard K, Lightfoot D (2000) Expression of bacterial gdh A gene encoding a NADPH glutamate dehydrogenase in tobacco affects plant growth and development. Plant Soil 221:47–57CrossRefGoogle Scholar
  2. Bijlsma RL, Lambers H, Kooijman SALM (2000) A dynamic whole plant model of integrated metabolism of nitrogen and carbon. 1. Comparative ecological implications of ammonium-nitrate interactions. Plant Soil 220:49–69CrossRefGoogle Scholar
  3. Britto DT, Kronzucker HJ (2002) NH4+ toxicity in higher plants: a critical review. J Plant Physiol 159:567–584CrossRefGoogle Scholar
  4. Buchanan-Wollaston V (1997) The molecular biology of leaf senescense. J Exp Bot 48:181–199CrossRefGoogle Scholar
  5. Cárdenas-Navarro R, Adamowicz S, Robin P (1999) Nitrate accumulation in plants: a role for water. J Exp Bot 50:613–624CrossRefGoogle Scholar
  6. Claussen W, Lenz F (1999) Effect of ammonium or nitrate nutrition on net photosynthesis, growth, and activity of the enzymes nitrate reductase and glutamine synthetase in blueberry, raspberry and strawberry. Plant Soil 208:95–102CrossRefGoogle Scholar
  7. Cruz C, Lips SH, Martins-Loução MA (2003) Nitrogen use efficiency by a slow growing species as affected by CO2 levels, root temperature, N source and availability. J Plant Physiol 160:1421–1428PubMedCrossRefGoogle Scholar
  8. Elia A, Santamaria P, Serio F (1998) Nitrogen nutrition, yields and quality of spinach. J. Sci Food Agric 76:341–346CrossRefGoogle Scholar
  9. Falkengren-Grerup U (1995) Interspecies differences in the response of ammonium and nitrate in vascular plants. Oecologia 102:305–311CrossRefGoogle Scholar
  10. Ford BG, Woodall J (1995) Glutamine syntase in higher plants: molecular biology meets plant physiology. In: Wallsgrove RM (ed) Amino acids and their derivatives in higher plants. Cambridge University Press, Cambridge, pp 1–18Google Scholar
  11. Frechilla S, Lasa B, Aleu M, Juanarena N, Lamsfus C, Aparicio-Tejo PM (2002) Short-term ammonium supply stimulates glutamate dehydrogenase activity and alternative pathway respiration in roots of pea plants. J Plant Physiol 159:811–818CrossRefGoogle Scholar
  12. Gerendas J, Zhu Z, Bendixen R, Ratcliffe RG, Sattelmacher B (1997) Physiological and biochemical processes related to ammonium toxicity in higher plants. Z Pflanzenernaehr Bodenkd 160:239–251CrossRefGoogle Scholar
  13. Given CV (1978) Metabolic detoxification of ammonium in tissues of higher plants. Phytochemistry 18:375–382CrossRefGoogle Scholar
  14. Glevarec G, Bouton S, Jaspard E, Riou M-T, Cliquet J-B, Suzuki A, Limami AM (2004) Respective roles of glutamine synthetase/ glutamate synthase cycle and glutamate dhydrogenase in ammonium and amino acid metabolism during germination and post-germinative growth in the model legume Medicago truncatula. Planta 219:286–297CrossRefPubMedGoogle Scholar
  15. Groat RG, Vance CP (1981) Root nodules enzymes of ammonium assimilation in alfalfa (Medicago sativa L.). Plant Physiol 67:1198–1203PubMedCrossRefGoogle Scholar
  16. Harrison J, Brugiere N, Philipson B, Ferrario-Mery S, Becker T, Limami A, Hirel B (2000) Manipulating the pathway of ammonia assimilation through genetic engineering and breeding. Consequences of plant physiology and plant development. In: Martins-Loução MA, Lips SH (eds) Nitrogen in a sustainable ecosystem. Backhuys Publishers, Leiden, pp 89–101Google Scholar
  17. Hirel B, Lea PJ (2002) The biochemistry, molecular biology, and genetic manipulation of primary ammonium assimilation. In: Foyer CH, Noctor G (eds) Photosynthetic nitrogen assimilation and associated carbon and respiratory metabolism. Kluwer, London, pp 71–92Google Scholar
  18. Hosmer DW jr, Lemeshow S (1989) Applied logistic regression. Wiley, New YorkGoogle Scholar
  19. Kaiser WM, Man H-M, Kandlbinder A, Glaab J, Weiner H (2000) Nitrate reductase in higher plants. Post-translational regulation and comparison of rates of nitrate reduction in vivo and of nitrate reductase activity in vitro. In: Martins-Loução MA, Lips SH (eds) Nitrogen in a sustainable ecosystem. Backhuys Publishers, Leiden, pp 103–110Google Scholar
  20. Lancien M, Gadal P, Hodges M (2000) Enzyme redundancy and the importance the importance of 2-oxoglutarate in higher plant ammonium assimilation. Plant Physiol 123:817–824CrossRefPubMedGoogle Scholar
  21. Lasa B, Frechilla S, Aparicio-Tejo PM, Lamsfus C (2002) Role of glutamate dehydrogenase and phosphoenolpyruvate carboxilase activity in ammonium nutrition tolerance in roots. Plant Physiol Biochem 40:969–976CrossRefGoogle Scholar
  22. Lasa B, Frechilla S, Lamsfus C, Aparicio-Tejo PM (2001) The sensitivity to ammonium nutrition is related to nitrogen accumulation. Sci Horticult 91:143–152CrossRefGoogle Scholar
  23. Lee RB, Ratcliffe RG (1991) Observations on the subcellular distribution of the ammonium ion in maize root tissue using in vivo 14N-nuclear magnetic resonance spectroscopy. Planta 183:359–367CrossRefGoogle Scholar
  24. Loqué D, von Wirén N (2004) Regulatory levels for the transport of ammonium in plant roots. J Exp Bot 55:1293–1305CrossRefPubMedGoogle Scholar
  25. Loulakakis KA, Roubelakis-Angelakis KA, Kanellis AK (1994) Regulation of glutamate dehydrogenase and glutamine synthetase in avocado fruit during development and ripening. Plant Physiol 106:217–222PubMedGoogle Scholar
  26. Mäck G (1995) Organ specific changes in the activity and subunit composition of glutamine synthetase isoforms of barley (Hordeum vulgare L.) after growth on different level of NH4+. Planta 196:231–238CrossRefGoogle Scholar
  27. Magalhães JR, Ju GC, Rich PJ, Rhodes D (1991) Kinetics of 15NH4+ assimilation in Zea mays. Plant Physiol 94:647–656Google Scholar
  28. Magalhães JR, Huber DM (1989) Ammonium assimilation in different plant species as affected by nitrogen form and pH control in solution culture. Fert Res 21:1–6CrossRefGoogle Scholar
  29. Magalhães JR, Huber DM, Tsai CY (1992) Evidence of increased 15N-ammonium assimilation in tomato plants with exogenous α-ketoglutarate. Plant Sci 85:135–141CrossRefGoogle Scholar
  30. Martins-Loução MA, Cruz C (1999) Role of nitrogen source in carbon balance. In: Srivastava (ed) Nitrogen nutrition and plant growth. New Delhi, pp 231–282Google Scholar
  31. Matt P, Geiger M, Walch-Liu P, Engels C, Krapp A, Sttit M (2001a) The immediate cause of the diurnal changes of nitrogen metabolism in leaves of nitrate replete tobacco: a major imbalance between the rate of nitrate reduction and the rates of nitrate uptake and ammonium metabolism during the first part of the light period. Plant Cell Environ 24:177–190CrossRefGoogle Scholar
  32. Matt P, Geiger M, Walch-Liu P, Engels C, Krapp A, Sttit M (2001b) Elevated carbon dioxide increases nitrate uptake and nitrate reductase activity when tobacco is growing on nitrate, but increases ammonium uptake and inhibits nitrate reductase activity when tobacco is growing in ammonium nitrate. Plant Cell Environ 24:1119–1137CrossRefGoogle Scholar
  33. McCullagh P, Nelder JA (1989) Generalized linear models, 2nd edn. Chapman and Hall, LondonGoogle Scholar
  34. McNamara AL, Meeker GB, Shaw PD, Hageman RH (1971) Use of a dissimilatory nitrate reductase from Escherichia coli and formate as a reductive system for nitrate. J Agric Food Chem 19:229–231CrossRefGoogle Scholar
  35. Melo-Oliveira R, Cunha Oliveira I, Coruzzi GM (1996) Arabidopsis mutant analysis and gene regulation define a nonredundant role for glutamate dehydrogenase in nitrogen assimilation. Proc Natl Acad Sci USA 96:4718–4723CrossRefGoogle Scholar
  36. Miflin BJ, Habash DZ (2002) The role of glutamine synthetase and glutamate dehydrogenase in nitrogen assimilation and possibilities for improvement in the nitrogen utilization of crops. J Exp Bot 53:979–987CrossRefPubMedGoogle Scholar
  37. Miflin BJ, Lea PJ (1980) Ammonium assimilation. In: Miflin BJ (ed) The biochemistry of plants. Academic, New York, pp 169–202Google Scholar
  38. Migge A, Carrayol E, Hirel B, Becker TW (2000) Leaf specific over-expression of plastidic glutamine synthetase stimulates the growth of transgenic tobacco seedlings. Planta 210:252–260CrossRefPubMedGoogle Scholar
  39. Miller AJ, Cookson SJ, Smith SJ, Wells DM (2001) The use of microelectrodes to investigate compartmentation and the transport of metabolized inorganic ions in plants. J Exp Bot 52:541–549CrossRefPubMedGoogle Scholar
  40. Robinson D (2001) Root proliferation, nitrate inflow and their carbon costs during nitrogen capture by competing plants. Plant Soil 232:41–50CrossRefGoogle Scholar
  41. Salsac L, Chaillou S, Morot-Gaudry JF, Lessaint C, Jolivoe E (1987) Nitrate and ammonium nutrition in plants. Plant Physiol Biochem 25:805–812Google Scholar
  42. Schjoerring JK, Husted S, Mack G, Mattsson M (2002) The regulation of ammonium translocation in plants. J Exp Bot 53:883–890CrossRefPubMedGoogle Scholar
  43. Smil V (2001) Enriching the Earth. MIT Press, Cambridge, pp 1–17Google Scholar
  44. Smirnoff N, Stewart GR (1986) Nitrate assimilation and translocation by higher plants: comparative physiology and ecological consequences. Physiol Plant 64:133–140CrossRefGoogle Scholar
  45. Snell FD, Snell CT (1949) Colorimetric methods of analysis. Van Nostrand, New YorkGoogle Scholar
  46. Solozano L (1969) Determination of ammonium in natural waters by the phenolhypochlorite method. Limnol Oceanogr 14:799–801CrossRefGoogle Scholar
  47. Srivastava HS, Singh RP (1987) Role and regulation of 1-glutamate dehydrogenase activity in higher plants. Phytochemistry 26:597–610CrossRefGoogle Scholar
  48. Tercé-Laforgue T, Hirel B (2000) Is glutamine synthetase ammonium-regulated? In: Martins-Loução MA, Lips SH (eds) Nitrogen in a sustainable ecosystem. Backhuys Publishers, Leiden, pp 335–338Google Scholar

Copyright information

© Springer-Verlag 2005

Authors and Affiliations

  • C. Cruz
    • 1
  • A. F. M. Bio
    • 3
  • M. D. Domínguez-Valdivia
    • 2
  • P. M. Aparicio-Tejo
    • 2
  • C. Lamsfus
    • 2
  • M. A. Martins-Loução
    • 1
  1. 1.Departamento de Biologia Vegetal, Faculdade de Ciências de LisboaCentro de Ecologia e Biologia Vegetal – CEBVLisboaPortugal
  2. 2.Departamento de Ciencias del Medio NaturalUniversidad Publica de NavarraPamplona, NavarraSpain
  3. 3.Grupo de Ambiente do CMRPInstituto Superior TécnicoLisboaPortugal

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