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Analysis of sources of oxolinic acid-resistant field strains of <Emphasis Type="Italic">Burkholderia glumae</Emphasis> based on rep-PCR analysis and nucleotide sequences of <Emphasis Type="Italic">gyrB</Emphasis> and <Emphasis Type="Italic">rpoD</Emphasis>
Authors:Yukiko Maeda  Mitsuo Horita  Hirosuke Shinohara  Akinori Kiba  Kouhei Ohnishi  Seiya Tsushima  Yasufumi Hikichi
Institution:(1) Laboratory of Plant Pathology and Biotechnology, Kochi University, Nankoku Kochi, 783-8502, Japan;(2) National Agricultural Research Center for Hokkaido Region, Sapporo, Japan;(3) National Agricultural Research Center for Tohoku Region, Fukushima, Japan;(4) Research Institute of Molecular Genetics, Kochi University, Nankoku, Japan;(5) National Institute for Agro-Environmental Science, Tsukuba, Japan
Abstract:Repetitive sequence-based polymerase chain reaction (rep-PCR) analysis using BOX and ERIC as primers showed a highly divergent phylogeny among field strains of Burkholderia glumae. To elucidate the sources of oxolinic acid (OA) resistance in field strains of B. glumae isolated from rice seedlings cultivated in Mie, Toyama, and Iwate prefectures, Japan, the amino acid at position 83 of GyrA (GyrA83), which is involved in OA resistance, and the DNA patterns from the rep-PCR and the partial nucleotide sequences of gyrB and rpoD from various strains were analyzed. The ten Mie strains, in which GyrA83 was isoleucine (Ile), were divided into two groups based on the band patterns in rep-PCR analysis, although the nucleotide sequences of gyrB and rpoD were identical among the strains. Based on the band patterns in the rep-PCR analysis and the gyrB and rpoD sequences, two highly OA-resistant Toyama strains, Pg-13 and Pg-14, for which GyrA83 was serine (Ser) and Ile, respectively, were in the same lineage. This suggests that the bacteria might acquire OA resistance faster than phylogenic diversity as determined with the repetitive sequences BOX and ERIC and with gyrB and rpoD. Furthermore, three Iwate strains (H95, H101, and H104), isolated from seedlings of different cultivars grown in different years and having Ile at GyrA83, are probably in the same lineage, suggesting that OA-resistant bacteria might be transferred among different cultivars.
Keywords:OA resistance            Burkholderia glumae                      gyrB                      rpoD            Repetitive sequence-based polymerase chain reaction (rep-PCR)
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