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1996 Fiscal Year Final Research Report Summary

Molecular biological study on bule light signaling system in stomatal guard cells.

Research Project

Project/Area Number 06454015
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物生理
Research InstitutionKyushu University

Principal Investigator

SHIMAZAKI Ken-ichiro  Kyushu Univ., Fac.Science, Professor, 理学部, 教授 (00124347)

Co-Investigator(Kenkyū-buntansha) KINOSHITA Toshinori  Kyushu Univ., Fac.Science, Assistant, 理学部, 教務員 (50271101)
DOI Michio  Kyushu Univ., Fac.Science, Assistant Professor, 理学部, 助手 (00167537)
WADA Hajime  Kyushu Univ., Fac.Science, Associate Professor, 理学部, 助教授 (60167202)
Project Period (FY) 1994 – 1996
Keywordsstomata / blue light effect / proton pump / signal transduction / phosphorylation / dephosphorylation / protein kinase / protein phosphatas
Research Abstract

Molecular mechanisms in blue light response of stomata were investigated regarding protein phosphorylation and dephosphorylation in guard cell protoplasts from Vicia faba. Blue light-dependent proton pumping has been shown to be inhibited by myosin light chain kinase (MLCK) inhibitor ML-7, suggesting that MLCk or MLCK-like protein may be involved in this stomatal response. We found that the antibody against MLCK from chicken gizzard reacted with guard cells proteins with molecular masses of 32 and 17kD.cDNA library of Vicia guard cells was immunoscreened using this polyclonal antibody. Two cDNA clones, VFPK1 and VFPK2, encoding putative protein kinases, were isolated from the library, and their nucleotide sequences were determined. The deduced amino acid sequences of VFPK1 and VFPK2 contain 271 and 219 amino acid residues, respectively, and are 88% identical at overlapping amino acid sequence level. The amino acid sequence revealed that they conserve invariant amino acid residues of kinase domain, and exhibited slight similarity to known protein kinases (<17%). Fusion protein of VFPK1 with glutathione-S-transferase was expressed in E.coli and the protein had protein kinase activity. These results suggest that VFPK1 cDNA encodes new protein kinase. However, the function and role in stomatal responses should be elucidated.
Furthermore, we found protein phosphorylation of 80 kD protein in guard cells in response to blue light. The blue light-dependent protein phosphorylation was inhibited by MLCK inhibitor, supporting the notion that MLCK-like protein might be involved in blue light response of stomatal guard cells.

  • Research Products

    (14 results)

All Other

All Publications (14 results)

  • [Publications] Toshinori Kinoshita: "Cytosolic concentration of Ca^<2+> Regulates the Plasma Membrane H^+-ATPase in Guard Cells of Fave Bean" Plant Cell. 7. 1333-1342 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshinori Kinoshita: "Evidence for Ca^<2+>-dependent protein phosphorylation in vitro in guard cells from Vicia faba L." Plant Science. 110. 173-180 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chang-Hyo Goh: "Properties of Proton Pumping in Response to Blue Light and Fusicoccin in Guard Cell Protoplasts Isolated from Adaxial Epidermis of Vicia Leaves" Plant Physiology. 109. 187-194 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Takeshi Sakaki: "Lipids and fatty acids in guard-cell protoplasts from Vicia faba leaves" Phytochemistry. 40. 1065-1070 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Chang-Hyo Goh: "Inhibition of Blue Light-Dependent H^+ Pumping by Abscisic Acid in Vicia Guard-Cell Protoplasts." Plant Physiology. 111. 433-440 (1996)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ken-ichiro Shimazaki: "Inhibition of the stomatal blue light response by verapamil at high concentration." Plant & Cell Physiology. (in press).

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Ken-ichiro Shimazaki: "Methods in Cell Biology" Academic Press,Orland USA, 13 (1995)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Toshinori Kinoshita: "Cytosolic concentration of Ca^<2+> Regulates the Plasma Membrane H^+-ATPase in Guard Cells of Fava Bean" Plant Cell. 7. 1333-1342 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Toshinori Kinoshita: "Evidence for Ca^<2+>-dependent protein phosphorylation in vitro in guard cells from Vicia faba L." Plant Science. 110. 173-180 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chang-Hyo Goh: "Properties of Proton Pumping in Response to Blue Light and Fusicoccin in Guard Cell Protoplasts Isolated from Adaxial Epidermis of Vicia Leaves" Plant Physiology. 109. 187-194 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takeshi Sakaki: "Lipids and fatty acids in guard-cell protoplasts from Vicia faba leaves" Phytochemistry. 40. 1065-1070 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ken-ichiro Shimazaki: "Analysis of the light signaling pathway in stomatal guard cells" Academic press, Inc.Methods in Cell Biology. 49. 501-513 (1995)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Chang-Hyo Goh: "Inhibition of Blue Light-Dependent H^+ Pumping by Abscisic Acid in Vicia Guard-Cell Protoplasts." Plant Physiology. 111. 433-440 (1996)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Ken-ichiro Shimazaki: "Inhibition of the stomatal blue light response by verapamil at high concentration." Plant & Cell Physiology. (in press).

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 1999-03-09  

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