Identifiable endophytic bacteria with plant growth-promoting traits promise to ensure sustainable agriculture. However, information on the versatility and exploration of sunflower-associated bacteria as bioinoculants is less studied. Here, we present the whole-genome sequence and annotation of Stenotrophomonas maltophilia JVB5 isolated from the sunflower root endosphere from the North West province, South Africa. The whole-genome analysis revealed a genome size of 4 771 305 bp, a sequence read count of 8 764 890, a 66% guanine-cytosine content, 57 tRNAs, 268 contigs, and 4 160 protein-coding genes with functions in various metabolic pathways. Pathways involved in the indole acetic acid production were found in the S. maltophilia JVB5 genome. The whole-genome annotation predicted notable genes involved in bacterial colonisation, antibiosis, and plant growth promotion. The predicted genes are involved in the sulfur metabolism, and the oxidative stress may enhance the plant growth promotion and boost plant the resistance to stress. Upon inoculation, S. maltophilia JVB5 efficiently colonised the sunflower root under greenhouse conditions with a significant improvement on the wet plant weight of 437.20 g compared to the uninoculated control with a 331.04 g wet weight. The genomic analysis revealing specific functional genes in the bacteria genome suggests their bioprospecting in agriculture. Hence, understanding the mechanisms employed by S. maltophilia JVB5 based on the predicted multifunctional genes will help harness their bioresource in sustainable plant health.