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1Marine Biotechnology Institute, Kamaishi Laboratories, 3-75-1 Heita, Kamaishi City, Iwate 026-0001, Japan
2Unité des Entérobactéries, Unité INSERM 389, Institut Pasteur, 75724 Paris Cedex 15, France
Author for correspondence: Satoshi Yamamoto. Tel: +81 43 203 0226. Fax: +81 43 248 2109. e-mail: yamamotost{at}nichirei.co.jp
ABSTRACT
The phylogenetic relationships of 49 Acinetobacter strains, 46 of which have previously been classified into 18 genomic species by DNA-DNA hybridization studies, were investigated using the nucleotide sequence of gyrB, the structural gene for the DNA gyrase B subunit. The phylogenetic tree showed linkages between genomic species 1 (Acinetobacter calcoaceticus), 2 (Acinetobacter baumannii), 3 and TU13; genomic species 6, BJ15, BJ16 and BJ17; genomic species 5, BJ13 (synonym of TU14) and BJ14; genomic species 7 (Acinetobacter johnsonii), 10 and 11; and genomic species 8 and 9. The phylogenetic grouping of Acinetobacter strains based on gyrB genes was almost congruent with that based on DNA-DNA hybridization studies. Consequently, gyrB sequence comparison can be used to resolve the taxonomic positions of bacterial strains at the level of genomic species. However, minor discrepancies existed in the grouping of strains of genomic species 8, 9 and BJ17. The phylogenetic tree for these strains was reconstructed from the sequence of rpoD, the structural gene for the RNA polymerase 
70 factor. The latter tree was 100% congruent with the grouping based on DNA-DNA hybridization. The reliability of DNA-DNA hybridization may be superior to that of sequence comparison of a single protein-encoding gene in resolving closely related organisms since the former method measures the homologies between the nucleotide sequences of total genomic DNAs. Three strains that have not been characterized previously by DNA-DNA hybridization seem to belong to two new genomic species, one including strain ATCC 33308 and the other including strains ATCC 31012 and MBIC 1332.
Key Words: gyrB • rpoD • Acinetobacter • genomic species • DNA-DNA hybridization
The GSDB, DDBJ, EMBL and NCBI accession numbers for the sequences reported in this paper are listed in Table 1.
Present address: Food Science Laboratory, Research and Development Center, Nichirei Corporation, Shiminato 9, Mihama-Ku, Chiba City, Chiba 261--8547, Japan.
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