Use of 16S rRNA, 23S rRNA, and <i>gyrB</i> Gene Sequence Analysis To Determine Phylogenetic Relationships of <i>Bacillus cereus</i> Group Microorganisms

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  • Sergei G. Bavykin
    BioChip Technology Center, Argonne National Laboratory, Argonne, Illinois, 60439
  • Yuri P. Lysov
    BioChip Technology Center, Argonne National Laboratory, Argonne, Illinois, 60439
  • Vladimir Zakhariev
    BioChip Technology Center, Argonne National Laboratory, Argonne, Illinois, 60439
  • John J. Kelly
    BioChip Technology Center, Argonne National Laboratory, Argonne, Illinois, 60439
  • Joany Jackman
    Applied Physics Laboratory, Johns Hopkins University, Laurel, Maryland 20723
  • David A. Stahl
    Department of Civil and Environmental Engineering, University of Washington, Seattle, Washington 98195
  • Alexey Cherni
    BioChip Technology Center, Argonne National Laboratory, Argonne, Illinois, 60439

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<jats:title>ABSTRACT</jats:title> <jats:p> In order to determine if variations in rRNA sequence could be used for discrimination of the members of the <jats:italic>Bacillus cereus</jats:italic> group, we analyzed 183 16S rRNA and 74 23S rRNA sequences for all species in the <jats:italic>B. cereus</jats:italic> group. We also analyzed 30 <jats:italic>gyrB</jats:italic> sequences for <jats:italic>B. cereus</jats:italic> group strains with published 16S rRNA sequences. Our findings indicated that the three most common species of the <jats:italic>B. cereus</jats:italic> group, <jats:italic>B. cereus, Bacillus thuringiensis</jats:italic> , and <jats:italic>Bacillus mycoides</jats:italic> , were each heterogeneous in all three gene sequences, while all analyzed strains of <jats:italic>Bacillus anthracis</jats:italic> were found to be homogeneous. Based on analysis of 16S and 23S rRNA sequence variations, the microorganisms within the <jats:italic>B. cereus</jats:italic> group were divided into seven subgroups, Anthracis, Cereus A and B, Thuringiensis A and B, and Mycoides A and B, and these seven subgroups were further organized into two distinct clusters. This classification of the <jats:italic>B. cereus</jats:italic> group conflicts with current taxonomic groupings, which are based on phenotypic traits. The presence of <jats:italic>B. cereus</jats:italic> strains in six of the seven subgroups and the presence of <jats:italic>B. thuringiensis</jats:italic> strains in three of the subgroups do not support the proposed unification of <jats:italic>B. cereus</jats:italic> and <jats:italic>B. thuringiensis</jats:italic> into one species. Analysis of the available phenotypic data for the strains included in this study revealed phenotypic traits that may be characteristic of several of the subgroups. Finally, our results demonstrated that rRNA and <jats:italic>gyrB</jats:italic> sequences may be used for discriminating <jats:italic>B. anthracis</jats:italic> from other microorganisms in the <jats:italic>B. cereus</jats:italic> group. </jats:p>

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