Abstract
Tissue and cellular localization of NADH-dependent glutamate synthase (NADH-GOGAT, EC 1.4.1.14) in the unexpanced leaf blades and young grains of rice (Oryza sativa L.) was investigated using tissue-print immunoblot and immunocytological methods with an affinity-purified anti-NADH-GOGAT immunoglobulin G. Tissue-print immunoblots showed that the NADH-GOGAT protein was mostly located in large and small vascular bundles of the unexpanded blades. When the cross-sections (10μ in thickness) prepared from the paraffin-embedded blades were stained with the antibody, the NADH-GOGAT protein was detected in vascular-parenchyma cells and mestome-sheath cells. In developing grains, the NADH-GOGAT protein was detected in both phloem- and xylem-parenchyma cells of dorsal and lateral vascular bundles, and in the nucellar projection, nucellar epidermis, and aleurone cells. On the other hand, ferredoxin (Fd)-dependent GOGAT (EC 1.4.7.1) was located mainly in mesophyll cells of the leaf blade and in chloroplast-containing cross-cells of the pericarp of the grains. The spatial expression of these GOGAT proteins indicates distinct and non-overlapping roles in rice plants. In the leaf blades and young grains, NADH-GOGAT could be involved in the synthesis of glutamate from the glutamine that is transported through the vascular system from roots and senescing tissues.
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Abbreviations
- Fd-GOGAT:
-
ferredoxin-dependent glutamate synthase
- GS:
-
glutamine synthetase
- IgG:
-
immunoglobulin G
- NADH-GOGAT:
-
NADH-dependent glutamate synthase
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We would like to thank Dr. Ann Oaks, University of Guelph, Canada, for her helpful comments and critical reading of the manuscript. This work was supported in part by a Grant-in-Aid for Scientific Research on Priority Areas (Nos. 04273202 and 05266202), in part by Grants-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan (Nos. 03304005, 03454061, 04660063, and 05660080), and by Fellowships of the Japan Society for the Promotion of Science for Junior Scientists to T. Hayakawa. (No. 0634).
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Hayakawa, T., Nakamura, T., Hattori, F. et al. 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 (1994). https://doi.org/10.1007/BF00201826
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DOI: https://doi.org/10.1007/BF00201826