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2001 Fiscal Year Final Research Report Summary

Fusarium oxysporum genes expressed in planta

Research Project

Project/Area Number 11660047
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field 植物保護
Research InstitutionYamaguchi University

Principal Investigator

ITO Shin-ichi  Faculty of Agriculture, Yamaguchi University, Professor, 農学部, 教授 (30243629)

Project Period (FY) 1999 – 2001
KeywordsFusarium oxysporum f. sp. cucumerinum / Fusarium oxysporum f. sp. lycopersici / mRNA / gene / gene silencing
Research Abstract

1) Fusarium oxysporum f. sp. cucumerinum (Foc) genes expressed specifically in cucumber xylem sap were examined at mRNA and protein levels. Twenty-nine Foc cDNA clones isolated by suppression subtractive hybridization were sequenced and the resultant nucleotide sequences were subjected to homology searches. Two cDNA clones showing high homology with NIPSNAP gene and putative cation transporter gene were further examined for their expression in planta. RT-PCR analysis with specific primers designed from the two sequences revealed that their corresponding genes are actually expressed in host plants. Foc proteins accumulated in cucumber xylem sap in vitro were analyzed by two-dimensional SDS polyacrylamide gel electrophoresis. Twenty-two protein spots were extracted from the gels and subjected to peptide mass fingerprinting analysis. Sequence alignment searches showed that no proteins in databases were matched to the peptide mass fingerprintings obtained.
2) Tomatinase gene (Fotoml) of Fusarium oxysporum f. sp. lycopersici which has been known to be expressed in planta was chosen as a target gene for gene disruption. Eighteen mutants obtained showed increase in sensitivity to α-tomatine and decrease in tomatinase activity. Inoculation assay showed reduced pathogenicity of the mutants, suggesting an important role of tomatinase in pathogenicity of this fungus. Northem blot analysis showed that antisense RNA of Fotoml was expressed when one of the mutants was induced by α-tomatine, suggesting posttranscreptional gene silencing is involved in the suppression of tomatinase activity at least in this mutant strain.

  • Research Products

    (4 results)

All Other

All Publications (4 results)

  • [Publications] 松下 華子: "Fusatium oxysporum各分化型からのトマチナーゼ遺伝子の検出"日本植物病理学会報. 66巻3号. 138 (2000)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] 高原 浩之: "トマト萎ちょう病菌におけるトマチナーゼ遺伝子のコサプレッション"日本植物病理学会報. 67巻2号. 286 (2001)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Matsusita H: "Detection of tomatinase gene from formae speciales of Fusarium oxysporum"Jpn J. Phytopathol. 66-3. 286 (2000)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Takahara H: "Cosuppression of the tomatinase gene in Fusarium oxysporum f. sp. lycopersici"Jpn J. Phytopathol. 67-2. 138 (2001)

    • Description
      「研究成果報告書概要(欧文)」より

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Published: 2003-09-17  

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