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Application of temperature-gradient gel electrophoresis in taxonomy of coryneform bacteria

Andreas Felske^1,, Marc Vancanneyt², Karel Kersters^2,3 and Antoon D. L. Akkermans¹

¹Laboratory of Microbiology, Department of Biomolecular Sciences, Wageningen Agricultural University, Hesselink van Suchtelenweg 4, 6703 CT Wageningen, The Netherlands
²BCCM/LMG Culture Collection, University of Ghent, K.-L. Ledeganckstraat 35, B-9000 Ghent, Belgium
³Laboratory of Microbiology, University of Ghent, K.-L. Ledeganckstraat 35, B-9000 Ghent, Belgium

Author for correspondence: Andreas Felske. Tel: +34 5 4624711 (Institute) extension 131. Fax: +34 5 4624002. e-mail: Andreas{at}cica.es

ABSTRACT

Strains belonging to the Gram-positive coryneform soil bacteria were screened genotypically by temperature-gradient gel electrophoresis (TGGE). This method allows the sequence-specific separation of amplified fragments of 16S rRNA genes. A total of 115 reference strains representing the majority of the species of the genera Aeromicrobium, Agromyces, Arthrobacter, Aureobacterium, Cellulomonas, Curtobacterium, Nocardioides and Terrabacter were characterized. Depending on the genus investigated, the resolution limit of the technique appeared to be at the species or genus level or intermediate between the two. Aberrant TGGE profiles of strains within particular taxa revealed genomic heterogeneity and generic misclassification of nine strains studied. Beyond that, indications of 16S rRNA gene heterogeneity were found within the genomes of three Curtobacterium strains. The misclassifications revealed by TGGE were confirmed using whole-cell fatty acid methyl ester analysis and subsequent comparison with a database. TGGE has been demonstrated to be a useful tool in bacterial taxonomy.

Key Words: temperature-gradient gel electrophoresis • coryneform bacteria • 16S rDNA gene heterogeneity • fatty acid methyl ester

Present address: Institute ce Recursos Naturales y Agrobiologia, CSIC, Apartado 1052, 41080 Seville, Spain.

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