Discovery of marine Bacillus species by 16S rRNA and rpoB comparisons and their usefulness for species identification

Systematic studies of the Bacillus group have been biased towards terrestrial and pathogenic isolates, and relatively few studies have examined Bacillus species from marine environments. Here we took twenty Bacillus strains from diverse marine environments and sequenced their 16S rRNA. Using molecular comparisons, we separated the strains into thirteen Bacillus genotypes and identified 9 species: B. aquaemaris. B. badius, B. cereus group, B. firmus, B. halmapalus, B. hwajinpoensis, B. litoralis, B. sporothermodurans, B. vietnamensis, and three indistinguishable Bacilli. In addition, we sequenced the DNA-directed RNA polymerase beta subunit (rpoB) gene and assessed its discriminative power in identifying Bacilli. Phylogenetic trees of Bacillus rpoB genes separated each Bacillus according to their taxonomic positions and were supported statistically. The resolution of Bacillus on the rpoB phylogenetic tree was approximately 4.5 times greater than on the 16S rRNA phylogenetic tree. These results demonstrate that the polymorphism of the Bacillus rpoB gene can be used to identify Bacillus species, providing an improved identification scheme for Bacillus species.