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Applied and Environmental Microbiology, September 2004, p. 5613-5620, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5613-5620.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.

Implications of Amino Acid Substitutions in GyrA at Position 83 in Terms of Oxolinic Acid Resistance in Field Isolates of Burkholderia glumae, a Causal Agent of Bacterial Seedling Rot and Grain Rot of Rice

Yukiko Maeda, Akinori Kiba, Kouhei Ohnishi, and Yasufumi Hikichi*

Laboratory of Plant Pathology and Biotechnology,1 Research Institute of Molecular Genetics, Kochi University, Nankoku, Kochi, Japan2

Received 13 February 2004/ Accepted 8 April 2004

Oxolinic acid (OA), a quinolone, inhibits the activity of DNA gyrase composed of GyrA and GyrB and shows antibacterial activity against Burkholderia glumae. Since B. glumae causes bacterial seedling rot and grain rot of rice, both of which are devastating diseases, the emergence of OA-resistant bacteria has important implications on rice cultivation in Japan. Based on the MIC of OA, 35 B. glumae field isolates isolated from rice seedlings grown from OA-treated seeds in Japan were divided into sensitive isolates (OSs; 0.5 µg/ml), moderately resistant isolates (MRs; 50 µg/ml), and highly resistant isolates (HRs; ≥100 µg/ml). Recombination with gyrA of an OS, Pg-10, led MRs and HRs to become OA susceptible, suggesting that gyrA mutations are involved in the OA resistance of field isolates. The amino acid at position 83 in the GyrA of all OSs was Ser, but in all MRs and HRs it was Arg and Ile, respectively. Ser83Arg and Ser83Ile substitutions in the GyrA of an OS, Pg-10, resulted in moderate and high OA resistance, respectively. Moreover, Arg83Ser and Ile83Ser substitutions in the GyrA of MRs and HRs, respectively, resulted in susceptibility to OA. These results suggest that Ser83Arg and Ser83Ile substitutions in GyrA are commonly responsible for resistance to OA in B. glumae field isolates.

* Corresponding author. Mailing address: Laboratory of Plant Pathology and Biotechnology, Kochi University, 200 Monobe, Nankoku, Kochi 783-8502, Japan. Phone: 81-88-864-5218. Fax: 81-88-864-5219. E-mail: yhikichi{at}cc.kochi-u.ac.jp.

Applied and Environmental Microbiology, September 2004, p. 5613-5620, Vol. 70, No. 9
0099-2240/04/$08.00+0     DOI: 10.1128/AEM.70.9.5613-5620.2004
Copyright © 2004, American Society for Microbiology. All Rights Reserved.



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