Journal Article

Organ and Cellular Localization of Asparagine Synthetase in Rice Plants

Kentaro Nakano, Tatsuro Suzuki, Toshihiko Hayakawa and Tomoyuki Yamaya

in Plant and Cell Physiology

Published on behalf of Japanese Society of Plant Physiologists

Volume 41, issue 7, pages 874-880
Published in print July 2000 | ISSN: 0032-0781
Published online July 2000 | e-ISSN: 1471-9053 | DOI: http://dx.doi.org/10.1093/pcp/pcd006
Organ and Cellular Localization of Asparagine Synthetase in Rice Plants

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  • Biochemistry
  • Molecular and Cell Biology
  • Plant Sciences and Forestry

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DNA gel blot analysis suggested that asparagine synthetase (AS; EC 6.3.5.4) occurred as a single gene in rice. A fusion protein consisting of 17 kDa tagged-region from pET32a(+) expression plasmid and 42 kDa N-terminal region of rice AS was first expressed in Escherichia coli. The resulting polypeptide was purified and a mono-specific antibody for rice AS was prepared after affinity-purification with the antigen. Immunoblotting revealed a high content of AS protein in the leaf sheath at the second position from the fully expanded top leaf and in grains at the middle stage of ripening. Accumulation of mRNA for AS was also observed in these organs. During the ripening of the spikelets, the AS protein contents increased during the first 21 days after flowering, then declined rapidly. Immunolocalization analysis revealed signals for AS protein in the companion cells of vascular bundles of leaf sheath and phloem-parenchyma cells, nucellar projection, and nucellar epidermis of dorsal vascular bundles of grains.

Keywords: Key words: Asparagine synthetase (EC 6.3.5.4) — Cellular localization — Nitrogen metabolism — Protein distribution — Rice (Oryza sativa L.); Abbreviations: AS, asparagine synthetase; DAF, days after flowering; DIG, digoxigenin; Fd, ferredoxin; GS1, cytosolic glutamine synthetase; GOGAT, glutamate synthase; IgG, immunoglobulin G; PBS, phosphate-buffered saline.

Journal Article.  5293 words.  Illustrated.

Subjects: Biochemistry ; Molecular and Cell Biology ; Plant Sciences and Forestry

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