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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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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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