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Stricture and characteristics of chitinases from mature grapes

Research Project

Project/Area Number 14360018
Research Category

Grant-in-Aid for Scientific Research (B)

Allocation TypeSingle-year Grants
Section一般
Research Field 園芸・造園学
Research InstitutionUniversity of Yamanashi

Principal Investigator

TAKAYANAGI Tsutomu  University of Yamanashi, Department of Research Interdisciplinary Graduate school of medicine and Engineering, Professor, 大学院・医学工学総合研究部, 助教授 (00252007)

Co-Investigator(Kenkyū-buntansha) OKUDA Tohru  University of Yamanashi, Department of Research Interdisciplinary Graduate school of medicine and Engineering, Associate Professor, 大学院・医学工学総合研究部, 助教授 (10252008)
YOKOTSUKA Koki  University of Yamanashi, Department of Research Interdisciplinary Graduate school of medicine and Engineering, Professor, 大学院・医学工学総合研究部, 教授 (60020450)
Project Period (FY) 2002 – 2004
Project Status Completed (Fiscal Year 2004)
Budget Amount *help
¥14,600,000 (Direct Cost: ¥14,600,000)
Fiscal Year 2004: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2003: ¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 2002: ¥6,600,000 (Direct Cost: ¥6,600,000)
KeywordsChitinase / PR-protein / Grape / Vitis / Disease resistance
Research Abstract

Plant chitinases are thought to be closely related to the plants' defense system against phytopathogens. The structure and characteristics of chitinases separated from Muscat Bailey A (Bailey x Muscat Hamburg) grape skins were investigated. The chitinase fraction was obtained from the extract of the grape skins by affinity chromatography with chitin beads (Chitopearl BL-01) and then separated by cation-exchange chromatography (SP-Sepharose HP). The cation-exchange chromatography yielded two protein peaks (CHI-1 and CHI-2) that exhibited chitinase activity. The molecular weights of CHI-1 and CHI-2 were estimated to be 29 kDa and 27 kDa, respectively, by SDS-PAGE. The optimum pHs for CHI-1 and CHI-2 activity were 3.5-4.5 and 4.5-5.0, respectively. The enzymes were stable over the pH range of 3.0 to 7.0. The optimum temperature for CHI-1 activity (70℃) was higher than that for CHI-2 activity (50℃). CHI-1 was further separated by chromatofocusing into four chitinase isozymes, CHI-1a, -1b, -1c, and -1d. The N-terminal amino acid sequences of the chitinases (CHI-1a, -1b, -1c, -1d and CHI-2) were highly homologous with that of the class IV chitinase of Vitis vinifera. CHI-1 and CHI-2 showed an inhibitory effect on the growth of Botrytis cinerea (gray mold) and Glomerella cingulata (ripe rot). Chitinase activity in the grape skins of Muscat Bailey A was higher than those in the three Vitis vinifera cultivars (Cabernet Sauvignon, Koshu and Chardonnay). The grape skins of Muscat Bailey A, Cabernet Sauvignon and Koshu contained CHI-1 and CHI-2, whereas those of Chardonnay contained only CHI-1. These results suggested that the quantity and number of chitinase isozymes in grape skins are related to the disease resistance level of the grapevines.

Report

(4 results)
  • 2004 Annual Research Report   Final Research Report Summary
  • 2003 Annual Research Report
  • 2002 Annual Research Report
  • Research Products

    (10 results)

All 2005 2004 2003 Other

All Journal Article (7 results) Publications (3 results)

  • [Journal Article] ブドウ果実キチナーゼの品種間比較2005

    • Author(s)
      高柳 勉, 相川大介, 奥田 徹, 横塚弘毅
    • Journal Title

      日本ブドウ・ワイン学会誌 16

      Pages: 9-13

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Comparison of Chitinases in Grape Skins of Four Grape Cultivars2005

    • Author(s)
      Takayanagi, T., D.Aikawa, T.Okuda, K.Yokotsuka
    • Journal Title

      Journal of ASEV Japan 16

      Pages: 9-13

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] ブドウ果実キチナーゼの品種間比較2005

    • Author(s)
      高柳 勉, 相川大介, 奥田 徹, 横塚弘毅
    • Journal Title

      日本ブドウ・ワイン学会誌 16(1)

      Pages: 9-13

    • Related Report
      2004 Annual Research Report
  • [Journal Article] ブドウクラスIIIキチナーゼアミノ酸配列の品種間比較2004

    • Author(s)
      高柳 勉, 阿野明彦, 奥田 徹, 横塚弘毅
    • Journal Title

      日本ブドウ・ワイン学会誌 15(1)

      Pages: 29-33

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Annual Research Report 2004 Final Research Report Summary
  • [Journal Article] Comparative Analysis of Amino Acid Sequences of Class III Chitinase from Vitis Species2004

    • Author(s)
      Takayanagi, T., A.Ano, T.Okuda, K.Yokotsuka
    • Journal Title

      Journal of ASEV Japan 15

      Pages: 29-33

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Characterization of a class III chitinase from Vitis vinifera cv. Koshu2003

    • Author(s)
      Ano, A., Takayanagi, T., Uchibori, T., Okuda, T., Yokotsuka, K.
    • Journal Title

      Journal of Bioscience and Bioengineering 95(6)

      Pages: 645-647

    • NAID

      110002665143

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Journal Article] Characterization of a class III chitinase from Vitis vinifera cv.Koshu2003

    • Author(s)
      Ano, A., Takayanagi, T., Uchibori, T., Okuda, T., Yokotsuka, K.
    • Journal Title

      Journal of Bioscience and Bioengineering 95

      Pages: 645-647

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      2004 Final Research Report Summary
  • [Publications] Ano, A., Takayanagi, T., Uchibori, T., Okuda, T., Yokotsuka, K.: "Characterization of a class III chitinase from Vitis vinifera cv.Koshu"Journal of Bioscience and Bioengineering. 95. 645-647 (2003)

    • Related Report
      2003 Annual Research Report
  • [Publications] 高柳 勉, 阿野明彦, 奥田 徹, 横塚弘毅: "ブドウクラスIIIキチナーゼアミノ酸配列の品種間比較"日本ブドウ・ワイン学会誌. 15(掲載予定). (2004)

    • Related Report
      2003 Annual Research Report
  • [Publications] Ano, A., Takayanagi, T., Uchibori, T., Okuda, T., Yokotsuka, K.: "Characterization of a class III chitinase from Vitis vinifera cv.Koshu"Journal of Bioscience and Bioengineering. (in press). (2003)

    • Related Report
      2002 Annual Research Report

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Published: 2002-04-01   Modified: 2016-04-21  

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