• Search Research Projects
  • Search Researchers
  • How to Use
  1. Back to previous page

Analysis of molecular mechanisms of defense systems by ascorbate peroxidase against oxidative damages caused by excess light energy

Research Project

Project/Area Number 12660090
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 応用微生物学・応用生物化学
Research InstitutionKinki University

Principal Investigator

SHIGEOKA Shigeru  Kinki University, Agriculture, professor, 農学部, 教授 (80140341)

Project Period (FY) 2000 – 2001
Project Status Completed (Fiscal Year 2001)
Budget Amount *help
¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
Keywordsascorbate peroxidase / ascorbate / active oxygen / photooxidative stress / alternative splicing / regulation of gene expression / post-transcriptional regulation / transgenic plants / 選択的スプライシング / アルコルビン酸ペルオキシダーゼ
Research Abstract

Alternative splicing events in the 3'-terminal region of chloroplast ascorbate peroxidase (chlAPX) pre-mRNA in spinach and tobacco, which produced four types of mRNA variants, one form (tAPX-I) encoding thylakoid-bound APX (tAPX) and three forms (sAPX-I, -II, -III) encoding stromal APX (sAPX), were regulated in a tissue-specific manner. The ratio of the level of SAPX mRNAs (sAPX-I, -II, and -III) to tAPX-I mRNA was close to 1 in leaf, whereas the ratio in root was greatly elevated due to an increase in sAPX-III and a decrease in tAPX-I resulting from the alternative excision of intron 11 and intron 12, respectively. A putative splicing regulatory cis-element (SRE), which is highly conserved in the sequences of chlAPX genes of higher plants, was identified upstream of the acceptor site in intron 12. Gel-shift analysis showed that SRE interacts strongly with a nuclear protein from leaves, but not those from the roots of spinach and tobacco. These results indicate that the tissue-specific alternative splicing of chlAPX pre-mRNA is regulated by the splicing enhancer SRE.
Transgenic tobacco plants (TpTAP-12) overexpressing thylakoid membrane-bound ascorbate peroixdase (tAPX) targeted to chloroplasts were generated. The tAPX activity in TbTAP-12 plants was approximately 37-fold higher than that of the wild-type. The abilities of CO_2 fixation and PSII activity in between wild-type and TpTAP-12 plants were not changed.
The activities of thiol-modulated enzymes in the Calvin cycle, antioxidative enzymes, and levels of antioxidants were also same values in both plants. Transgenic tobacco plants -suppressing chloroplastic ascorbate peroixdase isoenzymes are generating using antisense technology.
Five lines of those plants at T_1 generation were isolated.

Report

(3 results)
  • 2001 Annual Research Report   Final Research Report Summary
  • 2000 Annual Research Report
  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] 重岡 成: "Regulation and function of ascorbate peroxidase isozyrnes"Journal of Experimental Botany. (印刷中). (2002)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 吉村和也: "Characterization of monoclonal antibodies against ascorbate peroxidase isoenzymes : purification and epitope-mapping using im munoaffinity colu mn chromatography"Biochimica et Biophysica Acta. 1526. 168-174 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 重岡 成: "光合成生物め活性酸素代謝の応答機構-特集によせて-"生物工学会誌. 79. 303-304 (2001)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] S. Shigeoka et al.: "Regulation and function of ascorbate peroxidase isozymes"Journal of Experimental Botany. (in press). (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] K. Yoshimura et al.: "Characterization of monoclonal antibodies against ascorbate peroxidase isoenzymes : purification andepitope-mapping using immunoaffinity column chromatography"Biochimica et Biophysica Acta. 1526. 168-174 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] S. Shigeoka et al: "Response of active oxygen metabolisms in photosynthetic organisms"Biological engineering (in Japanese). 79. 303-304 (2001)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2001 Final Research Report Summary
  • [Publications] 重岡 成: "Regulation and function of ascorbate peroxidase isozymes"Journal of Experimental Botany. (印刷中). (2002)

    • Related Report
      2001 Annual Research Report
  • [Publications] 吉村和也: "Characterization of monoclonal antibodies against ascorbate peroxidase isoenzymes : purification and epitope-mapping using immunoaffinity colunn chromatography"Biochimica et Biophysica. 1526. 168-174 (2001)

    • Related Report
      2001 Annual Research Report
  • [Publications] 重岡 成: "光合成生物の活性酸素代謝の応答機構-特集によせて-"生物工学会誌. 79. 303-304 (2001)

    • Related Report
      2001 Annual Research Report

URL: 

Published: 2001-04-01   Modified: 2016-04-21  

Information User Guide FAQ News Terms of Use Attribution of KAKENHI

Powered by NII kakenhi