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Reverse genetics approach to characterize a function of NADH-glutamate synthase1 in rice plants

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Abstract

Rice plants grown in anaerobic paddy soil prefer to use ammonium ion as an inorganic nitrogen source for their growth. The ammonium ions are assimilated by the coupled reaction of glutamine synthetase (GS) and glutamate synthase (GOGAT). In rice, there is a small gene family for GOGAT: there are two NADH-dependent types and one ferredoxin (Fd)-dependent type. Fd-GOGAT is important in the re-assimilation of photorespiratorily generated ammonium ions in chloroplasts. Although cell-type and age-dependent expression of two NADH-GOGAT genes has been well characterized, metabolic function of individual gene product is not fully understood. Reverse genetics approach is a direct way to characterize functions of isoenzymes. We have isolated a knockout rice mutant lacking NADH-dependent glutamate synthase1 (NADH-GOGAT1) and our studies show that this isoenzyme is important for primary ammonium assimilation in roots at the seedling stage. NADH-GOGAT1 is also important in the development of active tiller number, when the mutant was grown in paddy field until the harvest. Expression of NADH-GOGAT2 and Fd-GOGAT in the mutant was identical with that in wild-type, suggesting that these GOGATs are not able to compensate for NADH-GOGAT1 function.

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References

  • Abiko T, Obara M, Ushioda A, Hayakawa T, Hodges M, Yamaya T (2005) Localization of NAD-isocitrate dehydrogenase and glutamate dehydrogenase in rice roots: candidates for providing carbon skeletons to NADH-glutamate synthase. Plant Cell Physiol 46:1724–1734

    Article  CAS  PubMed  Google Scholar 

  • Cañas RA, Quilleré I, Christ A, Hirel B (2009) Nitrogen metabolism in the developing ear of maize (Zea mays): analysis of two lines contrasting in their mode of nitrogen management. New Phytol 184:340–352

    Article  PubMed  Google Scholar 

  • Cao Y, Glass ADM, Crawford NM (1993) Ammonium inhibition of arabidopsis root growth can be reversed by potassium and by auxin resistance mutations aux1, axr1, and axr2. Plant Physiol 102:983–989

    Article  CAS  PubMed  Google Scholar 

  • Fukumorita T, Chino M (1982) Sugar, amino acid, and inorganic contents in rice phloem sap. Plant Cell Physiol 23:273–283

    CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Hayashi H, Chino M (1990) Chemical composition of phloem sap from the upper most internode of the rice plant. Plant Cell Physiol 31:247–251

    CAS  Google Scholar 

  • Hirochika H, Sugimoto K, Otsuki Y, Tsugawa H, Kanda M (1996) Retrotransposons of rice involved in mutations induced by tissue culture. Proc Natl Acad Sci USA 93:7783–7788

    Article  CAS  PubMed  Google Scholar 

  • Hirose N, Hayakawa T, Yamaya T (1997) Inducible accumulation of mRNA for NADH-dependent glutamate synthase in rice roots in response to ammonium ions. Plant Cell Physiol 38:1295–1297

    CAS  Google Scholar 

  • International Rice Genome Sequencing Project (2005) The map-based sequence of the rice genome. Nature 436:793–800 (http://rgp.dna.affrc.go.jp/IRGSP/)

    Google Scholar 

  • Ireland RJ, Lea PJ (1999) The enzymes of glutamine, glutamate, asparagine and aspartate metabolisms. In: Singh BK (ed) Plant amino acids. Biochemistry and biotechnology. Marcel Dekker Inc., New York, pp 49–109

    Google Scholar 

  • Ishiyama K, Hayakawa T, Yamaya T (1998) Expression of NADH-dependent glutamate synthase protein in the epidermis and exodermis of rice roots in response to the supply of nitrogen. Planta 204:288–294

    Article  CAS  PubMed  Google Scholar 

  • Ishiyama K, Inoue E, Tabuchi M, Yamaya T, Takahashi H (2004) Biochemical background and compartmentalized functions of cytosolic glutamine synthetase for active ammonium assimilation in rice roots. Plant Cell Physiol 45:1640–1647

    Article  CAS  PubMed  Google Scholar 

  • Kendall AC, Wallsgrove RM, Hall NP, Turner JC, Lea PJ (1986) Carbon and nitrogen metabolism in barley (Hordeum vulgare L.) mutants lacking ferredoxin-dependent glutamate synthase. Planta 168:316–323

    Article  CAS  Google Scholar 

  • Kiyomiya S, Nakanishi H, Uchida H, Tsuji A, Nishiyama S, Futatsubashi M, Tsukada H, Ishioka NS, Watanabe S, Ito T, Mizuniwa C, Osa A, Matsuhashi S, Hashimoto S, Sekine T, Mori S (2001) Real time visualization of 13N-translocation in rice under different environmental conditions using positron emitting tracer imaging system. Plant Physiol 125:1743–1754

    Article  CAS  PubMed  Google Scholar 

  • Kojima M, Kamada-Nobusada T, Komatsu H, Takei K, Kuroha T, Mizutani M, Ashikari M, Ueguchi-Tanaka M, Matsuoka M, Suzuki K, Sakakibara H (2009) High sensitive and high-throughput analysis of plant hormones using MS-probe modification and liquid chromatography-tandem mass spectrometry: an application for hormone profiling in Oryza sativa. Plant Cell Physiol 50:1201–1214

    Article  CAS  PubMed  Google Scholar 

  • Kronzucker HJ, Britto DT, Davenport RJ, Tester M (2001) Ammonium toxicity and the real cost of transport. Trends Plant Sci 6:335–337

    Article  CAS  PubMed  Google Scholar 

  • Lancien M, Martin M, Hsieh M-H, Leustek T, Goodman H, Coruzzi GM (2002) Arabidopsis glt1-T mutant defines a role for NADH-GOGAT in the nonphotorespiratory ammonium assimilation. Plant J 29:347–358

    Article  CAS  PubMed  Google Scholar 

  • Lea PJ, Miflin BJ (2003) Glutamate synthase and the synthesis of glutamate in plants. Plant Physiol Biochem 41:555–564

    Article  CAS  Google Scholar 

  • Lea PJ, Sodek L, Parry MAJ, Shewry PR, Halford NG (2007) Asparagine in plants. Ann Appl Biol 150:1–26

    Article  CAS  Google Scholar 

  • Mae T, Ohira K (1981) The remobilization of nitrogen related to leaf growth and senescence in rice plants (Oryza sativa L.). Plant Cell Physiol 22:1067–1074

    CAS  Google Scholar 

  • Martin A, Lee J, Kichey T, Gerentes D, Zivy M, Tatout C, Dubois F, Balliau T, Valot B, Davanture M, Tercé-Laforgue T, Quilleré I, Coque M, Gallais A, Gonzalez-Moro M-B, Bethencourt L, Habash DZ, Lea PJ, Charcosset A, Perez P, Murigneux A, Sakakibara H, Edwards KJ, Hirel B (2006) Two cytosolic glutamine synthetase isoforms of maize are specifically involved in the control of grain production. Plant Cell 18:3252–3274

    Article  CAS  PubMed  Google Scholar 

  • Obara M, Sato T, Sasaki S, Kashiba K, Nakamura I, Ebitani T, Yano M, Yamaya T (2004) Identification and characterization of QTL on chromosome 2 for cytosolic glutamine synthetase content and panicle number in rice (Oryza sativa L.). Theor Appl Genet 110:1–11

    Article  CAS  PubMed  Google Scholar 

  • Rahman A, Bannigan A, Sulaman W, Pechter P, Blancaflor BE, Baskin IT (2007) Auxin, actin and growth of Arabidopsis thaliana primary root. Plant J 50:514–528

    Article  CAS  PubMed  Google Scholar 

  • Sakamoto T, Matsuoka M (2008) Identifying and exploiting grain yield genes in rice. Curr Opin Plant Biol 11:209–214

    Article  CAS  PubMed  Google Scholar 

  • Sakurai N, Hayakawa T, Nakamura T, Yamaya T (1996) Changes in the cellular localization of cytosolic glutamine synthetase protein in vascular bundles of rice leaves at various stages of development. Planta 200:306–311

    Article  CAS  Google Scholar 

  • Sakurai N, Katayama Y, Yamaya T (2001) Overlapping expression of cytosolic glutamine synthetase and phenylalanine ammonia lyase in immature leaf blades of rice. Physiol Plant 113:400–408

    Article  CAS  PubMed  Google Scholar 

  • Shinozaki K, Sakakibara H (2009) Omics and bioinformatics: an essential toolbox for systems analyses of plant functions beyond 2010. Plant Cell Physiol 50:1177–1180

    Article  CAS  PubMed  Google Scholar 

  • Sonoda Y, Ikeda A, Saiki S, von Wirén N, Yamaya T, Yamaguchi J (2003) Distinct expression and function of three ammonium transporter genes (OsAMT1;1-1;3) in rice. Plant Cell Physiol 44:726–734

    Article  CAS  PubMed  Google Scholar 

  • Tabuchi M, Sugiyama K, Ishiyama K, Inoue E, Sato T, Takahashi H, Yamaya T (2005) Severe reduction in growth rate and grain filling of rice mutants lacking OsGS1;1, a cytosolic glutamine synthetase1;1. Plant J 42:641–651

    Article  CAS  PubMed  Google Scholar 

  • Tabuchi M, Abiko T, Yamaya T (2007) Assimilation of ammonium ions and reutilization of nitrogen in rice (Oryza sativa L.). J Exp Bot 58:2319–2327

    Article  CAS  PubMed  Google Scholar 

  • Tian Q, Chen F, Liu J, Zhang F, Mi G (2008) Inhibition of maize root growth by high nitrate supply is correlated with reduced IAA levels in roots. J Plant Physiol 165:942–951

    Article  CAS  PubMed  Google Scholar 

  • Tobin AK, Yamaya T (2001) Cellular compartmentation of ammonium assimilation in rice and barley. J Exp Bot 52:591–604

    Article  CAS  PubMed  Google Scholar 

  • Wallsgrove RM, Turner JC, Hall NP, Kendall AC, Bright SWJ (1987) Barley mutants lacking chloroplast glutamine synthetase—biochemical and genetic analysis. Plant Physiol 83:155–158

    Article  CAS  PubMed  Google Scholar 

  • Yamaya T, Oaks A (2004) Metabolic regulation of ammonium uptake and assimilation. In: Amâncio S, Stulen I (eds) Nitrogen acquisition and assimilation in higher plants. Kluwer academic, Dordrecht, pp 35–63

    Chapter  Google Scholar 

  • Yamaya T, Hayakawa T, Tanasawa K, Kamachi K, Mae T, Ojima K (1992) Tissue distribution of glutamate synthase and glutamine synthetase in rice leaves. Occurrence of NADH-dependent glutamate synthase protein and activity in the unexpanded, nongreen leaf blades. Plant Physiol 100:1427–1432

    Article  CAS  PubMed  Google Scholar 

  • Yamaya T, Obara M, Nakajima H, Sasaki S, Hayakawa T, Sato T (2002) Genetic manipulation and quantitative-trait loci mapping for nitrogen recycling in rice. J Exp Bot 53:917–925

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We are grateful to Dr. Autar K. Mattoo, USDA, ARS, Beltsville, MD, USA and Dr. Louis J. Irving, Tohoku University, Japan for helpful comments and critical reading of the manuscript. The Tos17 line used in this work was developed by Drs. A. Miyao and H. Hirochika (Rice Genome Project, National Institute of Agrobiological Sciences, Tsukuba, Japan). Seeds of the Tos17-inserted rice mutant were provided by the Rice Genome Resource Center (Tsukuba, Japan). This work was supported in part by a grant from the Ministry of Agriculture, Forestry and Fisheries of Japan (Genomics for Agricultural Innovation, IPG-0008 to TY) and in part by a Grant-in-Aids for Scientific Research (A) (19208007 to TY) and (B) (20380042 to TH) from the JSPS.

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Correspondence to Tomoyuki Yamaya.

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Tamura, W., Hidaka, Y., Tabuchi, M. et al. Reverse genetics approach to characterize a function of NADH-glutamate synthase1 in rice plants. Amino Acids 39, 1003–1012 (2010). https://doi.org/10.1007/s00726-010-0531-5

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  • DOI: https://doi.org/10.1007/s00726-010-0531-5

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