Distinct differentiation of closely related species of Bacillus subtilis group with industrial importance

► Species of Bacillus subtilis group showed limited variation in 16S rRNA gene. ► We identified species specific variations in 16S rRNA gene by in silico analysis. ► HinfI, RsaI and CfoI cut differently in the identified 16S rRNA variable regions. ► A simple 16S rRNA-RFLP distinctly differentiated B. subtilis group related species.