Log in

Co-suppressed glutamine synthetase2 gene modifies nitrogen metabolism and plant growth in rice

  • Articles
  • Molecular Biology
  • Published:
Chinese Science Bulletin

Abstract

A full-length cDNA that encodes the rice chloroplastic glutamine synthetase 2 gene was isolated from a Minghui 63-normalized cDNA library; and GS2 rice transformants were obtained by an Agrobacterium tumefaciens-mediated transformation method. Transcripts of the GS2 gene were shown to accumulate at higher levels in the primary transgenic plants in the T0 generation; whereas plants in the T1 generation exhibited a co-suppressed chlorosis phenotype (yellow leaves) accompanied by decreased plant height, few tillers and decreased dry weight. The plants with yellow leaves also displayed a significant decline in GS2 messenger RNA (mRNA) transcriptional level and chlorophyll content; a decrease in total GS activities of ˜50% was also found. Although there was no decrease in the concentration of total free amino acids, a change in the concentration of individual amino acids was observed. Our result also indicates a decreased metabolic level (soluble protein content and ammonium concentration) in GS2 co-suppressed plants. A correlation between chlorophyll content and GS2 mRNA expression level was also observed. The GS2 co-suppressed plants showed better performance when complemented with exogenous glutamine, indicating that the lack of an organic nitrogen pool inside the cell is the possible reason for the chlorosis phenotype of the transformants.

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

Access this article

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (France)

Instant access to the full article PDF.

Similar content being viewed by others

References

  1. Hirel B, Lea P J. Ammonium Assimilation. In: Lea P J, Morof Gaudry J F, eds. Plant Nitrogen. Berlin: Springer-Verlag, 2001. 79–99

    Google Scholar 

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

    Article  Google Scholar 

  3. Ireland R J, Lea P J. The enzymes of glutamine, glutamate, asparagines and aspirate metabolism. In: Singh B K, ed. Plant Amino Acids: Biochemistry And Biotechnology. New York: Marcel Dekker, 1999. 49–109

    Google Scholar 

  4. Stewart G R, Mann A F, Fentem P A. Enzymes of glutamate formation: glutamate dehydrogenase, glutamine synthetase and glutamate synthase. In: Miflin B J, ed. The Biochemistry Of Plants: Amino Acids And Derivatives. New York: Academic Press, 1980. 271–327

    Google Scholar 

  5. Zozaya-Garza M, Sengupta-Gopalan C. Glutamine synthetase gene isolation from an alfalfa leaf cDNA library. Plant Physiol, 1999, 119: 1568

    Google Scholar 

  6. Lightfoot D A, Green N K, Cullimore J V. The chloroplast-located glutamine synthetase of Phaseolus vulgaris L.: nucleotide sequence, expression in different organs and uptake into isolated chloroplast. Plant Mol Biol, 1988, 11: 191–202

    Article  Google Scholar 

  7. Tingey S V, Tsai F-Y, Edwards J W, et al. Chloroplast and cytosolic glutamine synthetase are encoded by homologous nuclear genes which are differentially expressed in vivo. J Biol Chem, 1988, 163: 9651–9657

    Google Scholar 

  8. Keegstra K, Cline K. Protein import and routing system of chloroplasts. Plant Cell, 1999, 11: 557–570

    Article  Google Scholar 

  9. May T, Soll J. Chloroplast precursor protein translocon. FEBS Lett, 1999, 452: 53–56

    Article  Google Scholar 

  10. Lam H-M, Coschigano K, Shultz C, et al. Use of Arabidopsis mutants and genes to study amide amino acid biosynthesis. Plant Cell, 1995, 7: 887–898

    Article  Google Scholar 

  11. Kozaki A, Tabeka G. Photorespiration protects C3 plants from photooxidation. Nature, 1996, 384: 557–560

    Article  Google Scholar 

  12. Hoshida H, Tanaka Y, Hibino T, et al. Enhanced tolerance to salt stress in transgenic rice that overexpresses chloroplast glutamine synthetase. Plant Mol Biol, 2000, 43: 103–111

    Article  Google Scholar 

  13. Husted S, Mattsson M, Mollers C, et al. Photorespiratory NH4 + production in leaves of wild-type and glutamine synthetase 2 antisense oilseed rape. Plant Physiol, 2002, 130: 989–998

    Article  Google Scholar 

  14. Chu Z, Peng K, Zhang L, et al. Construction and characterization of a normalized whole-life-cycle cDNA library of rice. Chinese Sci Bull, 2003, 48: 229–235

    Google Scholar 

  15. Maniatis T A, Fritsch E F, Sambrook J. Molecular Cloning: A Laboratory Manual. Cold Spring Harbor: Cold Spring Harbor Laboratory Press, 1992

    Google Scholar 

  16. Sambrook J, Fritsch E F, Maniatis T. Molecular cloning: A laboratory manual, 2nd ed. Cold Spring Harbor: Cold Spring Harbor Laboratory Press, 1989

    Google Scholar 

  17. Hiei Y, Ohta S, Komari T, et al. Efficient transformation of rice (Oryza sativa L.) mediated by Agrobacterium and sequence analysis of the boundaries of the T-DNA. Plant J, 1994, 6: 271–282

    Article  Google Scholar 

  18. Yoshida S, Forno D A, Cook J H, et al. Laboratory manual for physiological studies of rice. 3rd ed. Manila: International Rice Research Institute, 1976

    Google Scholar 

  19. Arnon D I. Copper enzymes in isolated chloroplasts. Polyphenoloxidase in Beta vulgaris. Plant Physiol, 1949, 24: 1–15

    Article  Google Scholar 

  20. Melo P M, Lima L M, Santos I M, et al. Expression of the plastid-located glutamine synthetase of Medicago truncatula. Accumulation of the precursor in root nodules reveals an in vivo control at the level of protein import into plastids. Plant Physiol, 2003, 132: 390–399

    Article  Google Scholar 

  21. Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein binding. Anal Biochem, 1976, 72: 248–254

    Article  Google Scholar 

  22. O’Neal D, Joy K W. Glutamine synthetase of pea leaves: Purification, stabilization and pH optima. Arch Biochem Biophys, 1973, 159: 113–122

    Article  Google Scholar 

  23. Gordon S A, Fleck A, Bell J. Optimal conditions for the estimation of ammonium by the Berthelot reaction. Ann Clin Biochem, 1978, 15: 270–275

    Google Scholar 

  24. Lancien M, Gadal P, Hodges M. Enzyme redundancy and the importance of 2-oxoglutarate in higher plant ammonium assimilation. Plant Physiol, 2000, 123: 817–824

    Article  Google Scholar 

  25. Wallsgrove R M, Turner J C, Hall N P, et al. Barley mutants lacking chloroplast glutamine synthetase-biochemical and genetic analysis. Plant Physiol, 1987, 83: 155–158

    Article  Google Scholar 

  26. Edwards J W, Walker E L, Coruzzi G M. Cell specific expression in transgenic plants reveals nonoverlap** roles for chloroplast and cytosolic glutamine synthetase. Proc Natl Acad Sci USA, 1990, 87: 3459–3463

    Article  Google Scholar 

  27. Zhao X Q, Shi W M. Expression analysis of the glutamine synthetase and glutamate synthase gene families in young rice (Oryza sativa) seedlings. Plant Sci, 2006, 170: 748–754

    Article  Google Scholar 

  28. Blackwell R D, Murray A J S, Lea P J, et al. Photorespiratory amino donors, sucrose synthesis and the induction of CO2 fixation in barley deficient in glutamine synthetase and/or glutamate synthase. J Exp Bot, 1988, 39: 845–858

    Article  Google Scholar 

  29. Somerville S R, Ogren W L. Inhibition of photosynthesis in Arabidopsis mutants lacking leaf glutamate synthase activity. Nature, 1980, 286: 257–259

    Article  Google Scholar 

  30. Coschigano K T, Melo-Oliveira R, Lim J, et al. Arabidopsis gls mutants and distinct Fd-GOGAT genes: Implications for photorespiration and primary nitrogen metabolism. Plant Cell, 1998, 10: 741–752

    Article  Google Scholar 

  31. Ferrario-Mery S, Suzuki A, Kunz C, et al. Modulation of amino acid metabolism in transformed tobacco plants deficient in Fd-GOGAT. Plant Soil, 2000, 221: 67–79

    Article  Google Scholar 

  32. Hausler R E, Blackwell R D, Lea P J, et al. Control of photosynthesis in barley leaves with reduced activities of glutamine synthetase and glutamate synthase: Plant characteristics and changes in nitrate, ammonium and amino acids. Planta, 1994, 194: 406–417

    Article  Google Scholar 

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

    Article  Google Scholar 

  34. Carvalho H G, Lopes-Cardoso I A, Lima L M, et al. Nodule-specific modulation of glutamine synthetase in transgenic Medicago truncatula leads to inverse alterations in asparagines synthetase expression. Plant Physiol, 2003, 133: 243–252

    Article  Google Scholar 

  35. Harrison J, Pou M A, Sene O, et al. Does lowering glutamine synthetase activity in nodules modifying nitrogen metabolism and growth of Lotus japonicus? Plant Physiol, 2003, 133: 253–262

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to **ngMing Lian.

Additional information

This work was supported in part by grants from the National Basic Research Program of China (Grant No. 2005CB120905), the National Special Key Project of China on Functional Genomics of Major Plants and Animals, the National Natural Science Foundation of China and the Cultivation Fund of the Key Scientific and Technical Innovation Project, Ministry of Education of China (Grant No. 707045).

Electronic supplementary material

About this article

Cite this article

Cai, H., **ao, J., Zhang, Q. et al. Co-suppressed glutamine synthetase2 gene modifies nitrogen metabolism and plant growth in rice. Chin. Sci. Bull. 55, 823–833 (2010). https://doi.org/10.1007/s11434-010-0075-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11434-010-0075-9

Keywords

Navigation