Abstract
Bacillus subtilis (B. subtilis), a versatile microorganism widely applied in agriculture and animal husbandry, has significant potential for supporting food security through crop protection and livestock productivity. On the basis of genomic and in vitro evidence, this study characterized a novel B. subtilis strain, G01, highlighting its potential as a candidate for biological control and food probiotics. Whole-genome sequencing revealed that G01 harbors nine secondary metabolite biosynthesis gene clusters, which play crucial roles in the biological control of microbial pathogens. Crucially, for potential food and feed-related uses, in vitro assays confirmed the potent broad-spectrum antimicrobial activity of G01 against phytopathogens and zoonotic bacteria, in addition to its ability to efficiently hydrolyze proteins and cellulose. Comparative genomics revealed unique gene clusters associated with antibacterial and probiotic-related functions in G01. Furthermore, genes involved in quorum sensing, biofilm formation, and spore production provide genomic support for its environmental resilience - a key characteristic for candidate biocontrol agents and probiotic precursors that target sustainable crop protection and livestock gut health management. This comprehensive genomic and in vitro functional analysis positions B. subtilis G01 as a promising candidate strain with theoretical potential for enhancing safety and efficiency in agricultural and livestock applications, laying a foundation for future in vivo validation of its practical efficacy.