The main objective of this study was to investigate the antimicrobial activity of three putative antagonist bacterial strains of Bacillus spp. against Xanthomonas vasicola pv. vasculorum (Xvv) and their potential to control bacterial leaf streak (BLS) of corn. Additionally, the study included investigations on the genome of one of these antagonist bacteria, such as genome sequencing and mining of genes involved in biofilm formation, swarming motility, and synthesis of secondary metabolites. The growth of Xvv was inhibited by both cell suspensions and cell-free supernatants of the bacterial strains LAFUEL 01, LAFUEL 02, and LAFUEL 03 in agar diffusion tests. All three antagonist strains significantly reduced the severity of BLS in the 3rd and 4th leaves of corn plants that were artificially inoculated at the V3 growth stage under greenhouse conditions. The 16S rRNA sequencing confirmed that the antagonistic bacterial strains belong to the genus Bacillus, with LAFUEL 03 having approximately 97% similarity to B. velezensis. B. velezensis LAFUEL 03 harbors genes related to the biosynthesis of secondary metabolites, biofilm formation/regulation, and swarming motility that enhances its potential for controlling BLS in corn and suggests a promising candidate for the development of a commercial biocontrol agent.