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Breeding of plants resistant to fungal infections by introduction of a chitinase gene from filamentous fungi.

Research Project

Project/Area Number 07556021
Research Category

Grant-in-Aid for Scientific Research (A)

Allocation TypeSingle-year Grants
Section試験
Research Field 応用微生物学・応用生物化学
Research InstitutionThe University of Tokyo

Principal Investigator

TAKAGI Masamichi  The University of Tokyo, Graduate School of Agricultural and Life Sciences, Faculty of Agriculture, Professor, 大学院・農学生命科学研究科, 教授 (50018339)

Co-Investigator(Kenkyū-buntansha) TERAKAWA Teruhiko  Hokko Chemical Ind. Co., Central Research Laboratories, Research Scientist, 開発研究所, 研究員
KOIKE Masaru  Hokko Chemical Ind. Co., Central Research Laboratories, Manager, 開発研究所, 主任研究員
HARADA Satoshi  KAGOME Co. Ltd., Research Institute, Research Scientist, 総合研究所, 研究員
TANAKA Hiroshi  KAGOME Co. Ltd., Research Institute, Manager, 総合研究所, 主任研究員
HORIUCHI Hiroyuki  The University of Tokyo, Graduate School of Agricultural and Life Sciences, Facu, 大学院・農学生命科学研究科, 助手 (00209280)
原田 聡  カゴメ(株), 総合研究所, 研究員
Project Period (FY) 1995 – 1996
Project Status Completed (Fiscal Year 1996)
Budget Amount *help
¥8,800,000 (Direct Cost: ¥8,800,000)
Fiscal Year 1996: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 1995: ¥6,200,000 (Direct Cost: ¥6,200,000)
KeywordsRhizopus / chitinase / glass / tomato / rice
Research Abstract

Chitinase I of Rhizopus, oligosporus, belonging to zygomycete filamentous fungi, is a pae-pro enzyme. We have cloned chil gene encoding chitinase I.To investigate the effects of expression of fungal chitinase on anti-fungal activity of the transgenic plant, we deleted introns and 3'-region encoding pro-wequence from chil gene and introduced it into some important plants and analyzed phenotypes of the plants. First, the modified chil gene described above was placed between the 35S promoter of Cauliflower mosaic virus and nopaline synthase (Nos) gene terminator and was introduced it into tobacco by the Agrobacterium tumefaciens leaf disc system. Total DNAs were isolated from two of the transformants. Integration of chil gene in these genome DNAs was confirmed by PCR and Southern blot analysis. The expressions of chitinase I in lysates from the young leaves of them were detected by Western blot analysis with anti-chitinase I antibody. Chitinase activities in these lysates were three- to f … More our-fold higher than that from the control plant. Then, the discomycete pathogens Sclerotinia sclerotiorum and Botrytis cirerea were infected on the leaves of them. The symptoms observed with these two transformants were remarkably suppressed as compared with leaves of the control plants. Next, the modified chil gene was placed between the maize ubiquitin gene promoter and Nos gene terminator and was cotransformed into rice with a plasmid containing hygromycin phosphotransferase gene as a selectable marker by the method of particle bombardment. 123 transformants were obtained and regenerated to plants. Total DNAs of them were prepared and used as substrates of PCR analysis and Southern blot analysis. Integration of chil gene was confirmed in 24 transformants. A weak signal of 43 kDa was detected in lysates of the leaves of 4 transformants by Western blot analysis. These transformants were regenerated to plants and infected by Magnaporthe grisea which could cause rice blast disease. The anti-fungal activity of these plants seemed to raise as compared with that of control plants. We also introduced the modified chil gene into tomato and glass and are now trying the regeneration of these transgenic tomato and glass calli. Less

Report

(3 results)
  • 1996 Annual Research Report   Final Research Report Summary
  • 1995 Annual Research Report
  • Research Products

    (7 results)

All Other

All Publications (7 results)

  • [Publications] 寺川輝彦,他: "A fungal chitinase gene from Rhizopus oligosporus confers the antifungal activity to the transgenic tobacco." Plant Cell Rep.(印刷中). (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] Terakawa T,Takaya N,Noriuchi H,Koike M,Takagi M: "gal chitinase gene from Rhizopus oligosporus confers the antifungal ty to the transgenic tobacco." Plant Cell Reports.(in press). (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1996 Final Research Report Summary
  • [Publications] 寺川輝彦 他: "A fungal chitinase gene from Rhizopus oligosporus confers the antifungal activity to the transgenic tobacco." Plant Cell Rep.(印刷中).

    • Related Report
      1996 Annual Research Report
  • [Publications] 本山高幸 他: "The Aspergillus nidulans chsD gene encodes a chitin synthase which has a redundunt function with the chsA gene product in conidiophore formation" Mol. Gen. Genet.(印刷中).

    • Related Report
      1995 Annual Research Report
  • [Publications] 堀内裕之 他: "Cloning of Rhizopus niveus pyr4 gene and its use for transformation of Rhizopus delemar" Curr. Genet.27. 472-478 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 高谷直樹 他: "Analysis of the 3-phosphoglycerate Kinase 2 promoter in Rhizopus niveus" Gene. 152. 121-125 (1995)

    • Related Report
      1995 Annual Research Report
  • [Publications] 高谷直樹 他: "Cloning and characterization the Rhizopus niveus leu1 gene and its use for homologous transformation" Biosci. Biotech. Biochem.60. 448-452 (1996)

    • Related Report
      1995 Annual Research Report

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Published: 1995-04-01   Modified: 2016-04-21  

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