Specific Oligonucleotide Primers Based on Sequences of the 16S-23S rDNA Spacer Region for the Detection of Burkholderia gladioli by PCR :

  • FURUYA,Naruto
    Laboratory of Plant Pathology, Division of Plant Pathology and Pesticide Science, Department of Applied Genetics and Pest Management, Faculty of Agriculture, Kyushu University
  • URA,Hiroyuki
    Yame Regional Agriculture Extension Center
  • IIYAMA,Kazuhiro
    Laboratory of Plant Pathology, Division of Plant Pathology and Pesticide Science, Department of Applied Genetics and Pest Management, Faculty of Agriculture, Kyushu University
  • MATSUMOTO,Masaru
    Laboratory of Plant Pathology, Division of Plant Pathology and Pesticide Science, Department of Applied Genetics and Pest Management, Faculty of Agriculture, Kyushu University
  • TAKESHITA,Minoru
    Laboratory of Plant Pathology, Division of Plant Pathology and Pesticide Science, Department of Applied Genetics and Pest Management, Faculty of Agriculture, Kyushu University
  • TAKANAMI,Yoichi
    Laboratory of Plant Pathology, Division of Plant Pathology and Pesticide Science, Department of Applied Genetics and Pest Management, Faculty of Agriculture, Kyushu University

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Specific primers were designed based on the sequences of the spacer region between the 16S and 23S ribosomal DNA (rDNA) for direct, rapid and specific detection of Burkholderia gladioli. These primers were named GLA-f and GLA-r. PCR performed on boiled bacterial suspensions yielded an amplification product of approximately 300 bp. No products from other bacterial species, including B. glumae were amplified, even after complete DNA extraction by the cetyltrimethyl-ammonium bromide (CTAB) method. Using the specific primers designed in this study, the PCR method can detect B. gladioli in plant samples within 6 hr. These data demonstrate the potential of specific PCR for the detection of B. gladioli.

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