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Development of Gene Introduction and Vector System in Plant Pathogenic Fungus

Research Project

Project/Area Number 01560057
Research Category

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

Allocation TypeSingle-year Grants
Research Field 植物保護
Research InstitutionKinki University

Principal Investigator

TOYODA Hideyoshi  Kinki University, Agriculture, Associate Professor, 農学部 (00150805)

Co-Investigator(Kenkyū-buntansha) YOSHIDA Motonobu  Kinki University, Research Insti, Food Science,, 講師 (80192425)
松田 克礼  近畿大学, 農学部, 研究生
松田 克のり  近畿大学, 農学部, 研究生
Project Period (FY) 1989 – 1990
Project Status Completed (Fiscal Year 1990)
Budget Amount *help
¥2,700,000 (Direct Cost: ¥2,700,000)
Fiscal Year 1990: ¥1,100,000 (Direct Cost: ¥1,100,000)
Fiscal Year 1989: ¥1,600,000 (Direct Cost: ¥1,600,000)
Keywordselectroporation / rRNA gene / homologous recombination / in situ hybridization / microinjection / トマト萎凋病菌 / リボゾ-ムRNA遺伝子 / in situ検出 / 細胞壁構造 / 固型NMR / キチンーキトサン複合系 / キケン分解性放線菌 / プロモ-タ- / デンプン / アミラ-ゼ生産遺伝子 / mRNA / 遺伝子バンク / 水銀無毒化遺伝子
Research Abstract

In a first year, we established an efficient system for direct gene transfer to macrospores of Fusarium oxysporum f. sp. lycopersici (race I), which causes the wilting in tomato plant, by electroporation. This work has been published in Journal of Phytopathology (1989). This method enabled us to introduce foreign genes into plant pathogic fungus without any special treatments such as protoplast formation and its regeneration. In a last year, therefore, we attempted to construct the vector system functional in this fungus. For this purpose. We first isolated the genes encoding rRNA in fungal chromosome by synthesizing cDNAs from rRNAs extracted from mycelia of this fungus. These cDNAs were hybridized with genomic DNAs which had been prepared from chromosomal DNA, and the cDNA specific to 28S rRNA was isolated. The DNA sequence of this clone was determined by dideoxy method and cleaved into two parts of the fragment. In order to construct the vector, the marker gene (hygromycin B) was flanked between these two fractions. This plasmid vector allowed us to ensure the homologous recombination of DNA sequences between the vector and chromosome. With this vector, macroconidia were successfully transformed by electroporation. In addition, we developed the method for in situ hybridization of mRNA with the probe clarified in this study. The probe DNA was introduced into mycelia of this fungus by microinjection in order to ensure cytoplasmic hybridization in mycelial cytoplasm. These results were submitted to the Journal, Agricultural and Biological Chemistry.

Report

(3 results)
  • 1990 Annual Research Report   Final Research Report Summary
  • 1989 Annual Research Report
  • Research Products

    (25 results)

All Other

All Publications (25 results)

  • [Publications] 松田 克礼,他: "Direct introduction of fluorescent isothiocyanaTeーcajngated albumin into intact macroconibia of F・oxysporum by electroporation" Journal of Phytopathology. 125. 89-96 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 豊田 秀吉,他: "Transient expression of the βーGlucuronidase gene introduced into borley coleoptile cells by microinjection" Plant Cell Reports. 9. 299-302 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 深溝 慶,他: "Solid state^<13>cーNMR analysis of cell wall components of Fusarium oxysporum" Agricultural and Biological Chemistry. 54. 2761-2762 (1990)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 豊田 秀吉,他: "In situ hybridization of cytoplasmic mRNA in filamentous plant pathsgenic Fmgus,Fusarium oxysporum f sp <lycopersici>___ー" Agriwltural and Biolojical Chemistry.

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 豊田 秀吉 他: "植物病害と遺伝子工学" 朝日出版社, 225 (1989)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] Y. Matsuda: "Direct introduction of fluorescent isothiocyanate-conjugated albumin into intact macroconidia of Fusarium oxysporum f. sp. lycopersici by microinjection" Journal of Phytopathology. 125. 89-96 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] H. Toyoda: "Transient expression of the beta-glucuronidase gene introduce into barley coleoptile cells by microinjection." Plant Cell Reports. 9. 299-302 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] T. Fukamizo: "Solid state ^<13>C-NMR analysis of cell wal components of Fusarium oxysporum." Agricultural and Biological Chemistry. 54. 299-302 (1990)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] H. Toyoda: "In situ hybridization of cytoplasmic mRNA in filamentous plant pathogenic fungus, Fusarium oxysporum f. sp. lycopersici." Agricultural and Biological Chemistry. (1991)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] H. Toyoda: Asahi Publishing. Plant Diseases and Genetic Engineering., 225 (1989)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1990 Final Research Report Summary
  • [Publications] 豊田 秀吉: "培養カルス系による耐病性植物の選抜" 化学と生物. 28. 12-19 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 豊田 秀吉・堀越 浩二・稲葉 幸司・大内 成志: "Plant Regeneration of callus tissues mduced from leaf explants of strawberry" 植物組織培養. 7. 38-41 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 深溝 慶・園田 浩二・豊田 秀吉・大内 成志・後藤 幸男: "Solid state ^<13>CーNMR analysis of cell wall components of Fusarium oxysporum" Agric.Biol.Chem.54. 2761-2762 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 豊田 秀吉: "野菜の培養変異と育種への利用" バイオホルティ-. 4. 9-14 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 豊田 秀吉・山家 高子・松田 克礼・大内 成志: "Transient expression of the βーgluculonidase gene introduced into barley coleoptile cells by microinjection" Plant Cell Reports. 9. 299-302 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 豊田 秀吉他9名: "Callus induction and plant regeneration from barley coleoptile tissues" 植物組織培養. 7. 202-204 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 豊田 秀吉: "最新バイオテクノロジ-全書3 野菜の組織・細胞培養と育種" 農業図書, 250 (1990)

    • Related Report
      1990 Annual Research Report
  • [Publications] 豊田 秀吉: "植物病害と遺伝子工学" 朝日出版社, 225 (1989)

    • Related Report
      1990 Annual Research Report
  • [Publications] Y.Matsuda,H.Toyoda,T.Nishiguchi,S.Ouchi: "Direct introduction of fluorescein isothio cyanate-conjugated albumin into in tact macroconidia of Fusarium oxysporum f.sp.lyoopersici by electroporation." Journal of Phytopathology. 125. 89-96 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] Y.Matsuda,H.Toyoda S.Ouchi: "Application of microinjection to appressoria and haustoria to Erysiphe graminis f.sp.hordei" Ann.Phytopath.Soc.Japan. 55. 67-68 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] H.Toyoda,T.Oki,Y.Matsuda,K.Katsuragi,T.Nishigudi,S.Ochi: "Transformation of contituent cells of tomato callus ajgregates by intranuclear mieroinjection" Ploant Tissue Culture Letters. 6. 95-97 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] H.Toyoda,K.Shimizu.K.Chatani,N.Kita,Y.Matsuda,S.Ouchi: "Selection of bacterial-wilt resistant tomato through tissue cultare" Plant Cell Reports. 8. 317-320 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] H.Toyoda,K.Chatani,Y.Matsuda,S.Ouchi: "Multiplication of tobacco mosaic virus in tobacco callus tissues and in ritro selection for vival disease resistance" Plant Cell Reports. 8. 433-436 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 豊田秀吉(分担執筆): "植物病理学実験マニュアル" 養賢堂, 160 (1989)

    • Related Report
      1989 Annual Research Report
  • [Publications] 豊田秀吉(分担執筆): "植物病害と遺伝子工学" 朝日出版社, 225 (1989)

    • Related Report
      1989 Annual Research Report

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Published: 1989-04-01   Modified: 2016-04-21  

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