Simple SummaryBiological control of plant diseases caused by fungal pathogens using antagonistic microorganisms including Bacilli has been considered to be an effective and safe alternative to chemical fungicides. Fusarium crown rot of wheat is a serious fungal disease affecting yield and grain quality. In this study, a newly isolated strain of Bacillus subtilis YB-15 from soil of wheat rhizosphere significantly inhibited Fusarium crown rot as well as improved growth of wheat seedlings. Multiple potential biocontrol and growth-promoting attributes of Bacillus subtilis YB-15 were determined in vitro and according to the whole genome sequencing analysis. Overall, the results demonstrated that Bacillus subtilis YB-15 has great potential for practical application in controlling plant fungal diseases and improving plant growth.Fusarium crown rot caused by Fusarium pseudograminearum is one of the most devastating diseases of wheat worldwide causing major yield and economic losses. In this study, strain YB-15 was isolated from soil of wheat rhizosphere and classified as Bacillus subtilis by average nucleotide identity analysis. It significantly reduced Fusarium crown rot with a control efficacy of 81.50% and significantly improved the growth of wheat seedlings by increasing root and shoot fresh weight by 11.4% and 4.2%, respectively. Reduced Fusarium crown rot may have been due to direct antagonism by the production of β-1, 3-glucanase, amylase, protease and cellulase, or by the ability of B. subtilis YB-15 to induce defense-related enzyme activities of wheat seedlings, both alone and in seedlings infected with F. pseudograminearum. Improved plant growth may be related to the ability of B. subtilis YB-15 to secrete indole acetic acid and siderophores, as well as to solubilize phosphorus. In addition, the genome of strain YB-15 was determined, resulting in a complete assembled circular genome of 4,233,040 bp with GC content of 43.52% consisting of 4207 protein-encoding genes. Sequencing the B. subtilis YB-15 genome further revealed genes for encoding carbohydrate-active enzymes, biosynthesis of various secondary metabolites, nutrient acquisition, phytohormone production, chemotaxis and motility, which could explain the potential of strain YB-15 to be plant growth-promoting bacteria and biological control agent. B. subtilis YB-15 appears to be a promising biocontrol agent against Fusarium crown rot as well as for wheat growth promotion.