Organ and cellular localization of asparagine synthetase in rice plants

Kentaro Nakano, Tatsuro Suzuki, Toshihiko Hayakawa, Tomoyuki Yamaya

Research output: Contribution to journalArticlepeer-review

43 Citations (Scopus)

Abstract

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 coil. 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.

Original languageEnglish
Pages (from-to)874-880
Number of pages7
JournalPlant and Cell Physiology
Volume41
Issue number7
DOIs
Publication statusPublished - 2000

Keywords

  • Asparagine synthetase (EC 6.3.5.4)
  • Cellular localization
  • Nitrogen metabolism
  • Protein distribution
  • Rice (Oryza sativa L.)

ASJC Scopus subject areas

  • Physiology
  • Plant Science
  • Cell Biology

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