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1998 Fiscal Year Final Research Report Summary

Study on the molecular breeding of transgenic plants resistant to bacterial diseases

Research Project

Project/Area Number 08456032
Research Category

Grant-in-Aid for Scientific Research (B)

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

Principal Investigator

YONEYAMA Katsuyoshi  Meiji University, School of Agriculture, Prof., 農学部, 教授 (50110060)

Project Period (FY) 1996 – 1998
Keywordsdisease-resistant transgenic plants / kiwifruit canker disease / phaseolotoxin / grain rot of rice / toxoflavin / IS2 like sequence
Research Abstract

1) Creation of transgenic plants resistant to kiwifruit canker disease
Pseudomonas syringae pv. actinidiae produces the toxin phaseolotoxin, a pathoge ic factor of kiwifruit canker. Phaseolotoxin inhibits ornithine carbamoyltransferase (OCTase) involved in arginine biosynthesis, but Ps. pv. phaseolicola is resistant to phaseolotox in by producing the toxin-insensitive OCTase. We clonedthe gene encoding the phaseolotoxin-insensitive OCTase gene (argK) , and the gene was fused to the chloroplast transit peptide of RuBisCO small subunit of tobacco because OCTase is located in the chioroplast of plants. The constructed plasmid pMY6 was introduced to kiwifruit plants by agro-infection method. After sele tion of kanamycin, fifty clones were obtained as the resistant plants. In all the clones, toxin resistance was examined, and 7 transgenic clones were obtained as phaseolotoxin-resistant plants. In OCTase enzyme assay, the transgenic clones showed that the transgene was expressed, and the prod … More uct was translocated into the chloroplasts of kiwifruit plants.
2) Mechanism of gene expression related to toxin biosynthesis in Burkholderia gumae
The wild type of B.glumae was mutagenized by transposon(Tn), and one of the mutants, B.glumae <overhead double bar>, was deficient in toxin biosynthesis and pathogenicity. Whentransformed with the genomic library of the wild strain, the toxin biosynthesis and pathogenicity of B.glumae <overhead double bar> was complemented with the cosmid pNP147. Thisshowed that toxoflavin was an essential factor for pathogenicity of B.glumae. The subcloning of pNP147 gave a complemented fragment of 2.7kb DNA.An ORF of the fragment showed a high similarity to IS2 of E.coil in the nucleotide sequence. On the other hand, the mutant <overhead double bar> had no the IS2-like sequence near at 3'-downstream of Tn insertion site, although the wild strains had the IS2 sequence at the corresponding site. These results suggest that IS2 may be important for the regulation of toxofl Less

  • Research Products

    (9 results)

All Other

All Publications (9 results)

  • [Publications] YONEYAMA, K.et al.: "Toxoflavin is an essential factor for virulence of Burkholderia glumae causing rice seedling rot disease," Ann phytopatlol.Soc.Jpn. 64. 91-96 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Arie, T.et al: "Gel penetrate-blotted assay, a novel method for selorogical detection of Fusarium spp.in soil," J.Pesticide Sci.22. 321-325 (1997)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Hayashi, Y.et al.: "Characterization of the antigenic determinant on Fusarium oxysporum recognized by a genus-specific monoclonal antibody," J.Gen.Microbiol. 44. 43-47 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Arie, T.et al.: "Immunological detection of end-polygalacturonase secretion by Fusarium oxysporum in plant tissue and sequencing of its encoding gene" Ann.Phytopathol.Soc.Jpn. 64. 7-15 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] K.Yoneyama: "Fungicidal activity : Chemical and biological approaches to plant protection" Jhon Wiley & Sons Ltd.New York., 254 (1998)

    • Description
      「研究成果報告書概要(和文)」より
  • [Publications] Yoneyama, K.et al.: "Toxoflavin is an essential factor for virulence of Burkholderia glumae causing rice seedling rot disease" Ann.Phytopathol.Soc.Jpn.64. 91-96 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Yoneyama, K.: "Novel approaches to disease control" John Wiley & Sons, in Fungicidal activity. 221-224 (1998)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Arie, T.et al.: "Gel penetrate-blotted assay, a novel method for selorogical detection of Fusarium spp.in soil" J.Pesticide Sci.22. 321-325 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
  • [Publications] Arie, T.et al.: "Immunological detection of end-polygalacturonase secretion by Fusarium oxysporum in plant tissue and sequencing of its encoding gene" Ann.Phytopathol.Soc.Jpn.64. 7-15 (1998)

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

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Published: 1999-12-08  

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