IDENTIFICATION OF GENETIC MARKERS TO AID THE DISTINCTION OF Bacillus subtilis FROM OTHER CLOSELY RELATED SPECIES

Bacillus subtilis is widely recognized for its agricultural, industrial, and biotechnology applications. However, its high genetic similarity to other members within the B. subtilis group often hinders accurate identification, leading to potential misclassification and inappropriate strain usage. This study aims to address these challenges by characterizing the genetic diversity among B. subtilis strains, subspecies, and closely related species and identifying candidate molecular markers for precise taxonomic resolution. A pan-genome analysis of 87 Bacillus genomes using the Roary pipeline revealed 906 conserved core genes. Phylogenomic reconstruction based on these genes clarified evolutionary relationships within the group. Furthermore, the core genes were examined to find nucleotide variant positions, which are exclusively present in B. subtilis. These unique variants were further examined against the corresponding nucleotide positions of other species via Nucleotide BLAST to verify their potential as diagnostic markers for reliably distinguishing B. subtilis from its close relatives. Collectively, our findings provide valuable insights into the genetic diversity of the B. subtilis group and propose candidate markers that can be incorporated into improved detection protocols. These advancements will enhance the accuracy of B. subtilis identification and minimize strain misapplication, ensuring the safe and effective application of B. subtilis across diverse fields.