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An approach to the biochemical mechanisms of heartwood formation using cell cultures

Research Project

Project/Area Number 03454081
Research Category

Grant-in-Aid for General Scientific Research (B)

Allocation TypeSingle-year Grants
Research Field 林産学
Research InstitutionKYUDHU UNIVERSITY

Principal Investigator

SAKAI Kokki  Kyushu University, Department of Forest Products, Professor, 農学部, 教授 (30015656)

Co-Investigator(Kenkyū-buntansha) TSUTSUMI Yuji  Shizuoka University, Department of Forest Resources Science, Research Associate, 農学部, 助手 (30236921)
SHIRAISHI Susumu  Kyushu University, Department of Forest Products, Associate Professor, 農学部, 助教授 (70226314)
KONDO Ryuichiro  Kyushu University, Department of Forest Products, Associate Professor, 農学部, 助教授 (80091370)
Project Period (FY) 1991 – 1993
Project Status Completed (Fiscal Year 1993)
Budget Amount *help
¥6,500,000 (Direct Cost: ¥6,500,000)
Fiscal Year 1993: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 1992: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 1991: ¥4,700,000 (Direct Cost: ¥4,700,000)
KeywordsCupressaceae / Heart wood / Thujaplicin / Hinokitiol / Biosynthesis / Elicitor / Mevaronate pathway / Cupressus lusitanica / トレーサー / 樹木 / トロポロン / フィトアレキシン / 木本植物 / エリシタ-
Research Abstract

(1). Biochimistry of lignification
Lignification process has been investigated in this project since it occurs prior to the heartwood formation in forest tree cells. A peroxidase isoenzyme was found to catalyze substrate specifically the dehydrogenetive polymerization of sinapyl alcohol. This enzyme was isolated and characterized from cell cultures of Populus alba.
(2). Establishment of cell cultures that produce heartwood constituents
Callus cultures of Cupressus lusitanica (Cupressanceae) and Pinus spp. produced their heartwood constituents, beta-thujaplicin and pinosylvins, respectively, although no heartwood constituents were detected in calluses of Cryptomeria japonica and Chamaecyparis obtusa. Thus, the culture of C.lusitanica cells is regarded as a good experimental material for studying phenomena of heartwood formation.
(3). Elicitor of beta-thujaplicin accumulation in callus cultures
The addition of yeast extract to callus cultures of C.lusitanica leads to a large increase in the p … More roduction of beta-thujaplicin that act as a phyoalexin of the treated cultures. The extract was fractionated by means of ethanol precipitation, ribonucleasetreatment, gel permiation choromatography and Cu-complexation. The most effective fraction was a polysaccharide composed of maily mannose and glucose.
(4). Biosythetic pathway of beta-thujaplicin
Callus cultures of C.lusitanica was fed with ^<14>C-mevaronate, ^<14>C-glucose or ^<14>C-malonate by the shot-gun method. Elicitor was added to the callus at the same time. The results proved that beta-thujaplicins synthsized via the mevaronate pathway.
(5). An attempt to isolation of the key enzyme(s) for beta-thujaplicin biosynthesis
We tried to isolate any enzyme(s) that catalyze biosynthesis of beta-thujaplicin from isopentenyl pyrophosphate. However, such enzyme(s) have not been detected in cell-free extract of callus cultures of C.lusitanica. Isoltion of the key enzyme(s), its characterization and proof of its presence in transition zone of C.lustanica trees should be subjects to be solved in the near future. Less

Report

(4 results)
  • 1993 Annual Research Report   Final Research Report Summary
  • 1992 Annual Research Report
  • 1991 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] 堤祐司・坂井克己: "Lignin Biosynthesis in Woody Angiosperm Tissues I." Mokuzai Gakkaishi. 39. 214-220 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 山口聡康・坂井克己: "Cupressus lusitanicaの培養細胞におけるヒノキチオール生合成経路" 日本木材学会大会研究発表要旨集. 43. 104 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 稲田聡子・堤祐司・坂井克己: "Cupressus lusitanica培養細胞のβ-ツヤプリシン生産を誘起するエリシター" 日本木材学会大会研究発表要旨集. 43. 105 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 坂井克己・山口聡康・稲田聡子: "Biosynthesis of Heartwood Constituent β-Thujaplicin in Cupressus lusitanica callus" Pro.7th Intl.Symp.Wood Pulp.Chem.3. 438-441 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 稲田聡子・堤祐司・坂井克己: "Elicitor of the β-Thujaplicin Accumulation in Callus Cultures of Cupressus lusitanica" J.Fac.Agric.,Kyushu Univ.38. 119-126 (1993)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 坂井克己・草場薫・堤祐司・白石忠夫: "Secondary Metabolites in Cell Culture of Woody Plants III." Mokuzai Gakkaishi. 40. 1-5 (1994)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 坂井克己: "樹木抽出成分の利用「樹木の組織培養による有用物質の生産」の項分担執筆" 日本木材学会研究分科会, 124 (1991)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] Y.Tsutsumi, K.Sakai: "Lignin Biosynthesis in Woody Angiosperm Tissues" Mokuzai Gakkaishi. 39. 214-220 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] T.Yamaguchi, K.Sakai: "Biosynthetic Pathway of Hinokitiol in Cell Cultures of Cupressus lusitanica" Nihonn Mokuzai Gakkai. 43. 104 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] S.Inada, Y.Tsutsumi, K.Sakai: "Elicitor that induces beta-Thujaplicin Formation in Cupressus lusitanica" Nihonn Mokuzai Gakkai. 43. 105 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Sakai, T.Yamaguchi, S.Inada: "Biosynthesis of Heartwood Constituent beta-Thujaplicin in Cupressus lusitanica callus" Pro. 7th Intl. Symp. Wood Pulp Chem. 3. 438-441 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] S.Inada, Y.Tsutsumi, K.Sakai: "Elicitor of the beta-Thujaplicin Accumulation in Callus Cultures of Cupressus lusitanica" J.Fac. Agric. Kyushu Univ.38. 119-126 (1993)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Sakai, K.Kusaba, Y.Tsutsumi, T.Shiraishi: "Secondary Metabolites in Cell Culture of Woody Plants" Mokuzai Gakkaishi. 40. 1-5 (1994)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] K.Sakai: in "Utilization of Wood Extractives" ed. by Japan Wood Res. Soc.Production of Valuable Metabolites by Tissue Culture of Woody Plants, 124 (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1993 Final Research Report Summary
  • [Publications] 堤祐司・坂井克己: "Lignin Biosynthesis in Woody Angiosperm Tissues I" Mokuzai Gakkaishi. 39. 214-220 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 山口聡康・坂井克己: "Cupressus lusitanicaの培養細胞におけるヒノキチオール生合成経路" 日本木材学会大会研究発表要旨集. 43. 104 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 稲田聡子・堤祐司・坂井克己: "Cupressus lusitanica培養細胞のbeta-ツヤプリシン生産を誘起するエリシター" 日本木材学会大会研究発表要旨集. 43. 105 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 坂井克己・山口聡康・稲田聡子: "Biosynthesis of Heartwood Constituant beta-Thujaplicin in Cupressus lusitanica callus" Pro.7th Intl.Symp.Wood Pulp.Chem,. 3. 438-441 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 稲田聡子・堤祐司・坂井克己: "Elicitor of the beta-Thujaplicin Accumulation in Callus Cultures of Cupressus lusitanica" J.Fac.Agric.,Kyushu Univ.38. 119-126 (1993)

    • Related Report
      1993 Annual Research Report
  • [Publications] 坂井克己・草場薫・堤祐司・白石忠生: "Secondary Metabolites in Cell Culture of Woody Plants III." Mokuzai Gakkaishi. 40. 1-5 (1994)

    • Related Report
      1993 Annual Research Report
  • [Publications] 草場 薫・坂井 克己 外2名: "Cupressus lusitanica培養細胞によるツヤプリシンの生産〜エリシターの効果" 第42回日本木材学会大会研究発表要旨集. 12- (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 稲田 聡子.堤 祐司.坂井 克己.: "樹木培養細胞のβ-ツナプリシン生産に対するエリシターの効果" 第2回樹木分子・細胞生物等シンポジウム講演要旨集. 52-55 (1992)

    • Related Report
      1992 Annual Research Report
  • [Publications] 草場 薫,坂井 克己 外: "Cupressus lusitanica培養細胞によるツヤプリシンの生産" 日本木材学会大会講演要旨集. 42. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 川畑 智晃,小坂 尚弘,坂井 克己: "針葉樹培養細胞に含まれる抽出成分の抗酸化性について" 日本木材学会大会講演要旨集. 42. (1992)

    • Related Report
      1991 Annual Research Report
  • [Publications] 坂井 克己(分担執筆): "木材の科学と利用技術 3.樹木抽出成分の利用" 日本木材学会研究分科会, 122 (1991)

    • Related Report
      1991 Annual Research Report

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Published: 1991-04-01   Modified: 2016-04-21  

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