Background <p>A serious tomato disease that results in plant death and high crop losses is the bacterial wilt. Crop rotation and the utilization of resistant cultivars are examples of control approaches. Since management using other approaches produces inconsistent outcomes, biological control is preferred.</p> Results <p>The efficiency of microbial antagonists against <i>Ralstonia solanacearum</i> is discussed in this research. Four antagonists (<i>Bacillus subtilis, Bacillus pumilus, Bacillus chitinosporus</i> and <i>Trichoderma asperellum</i>) demonstrated the greatest potential for disease control (53–71% efficacy) and enhanced plant growth against <i>R. solanacearum</i> both in vitro and in vivo. There were increases in <i>PR</i>-2 and <i>PR</i>-3 expression levels relative to mock-inoculated plants. The elevated activation of protection correlated with increased up-regulations of <i>PR</i>-2 and <i>PR</i>-3, exhibiting relative expression levels of 4.7- and 5.4-folds for <i>T. asperellum</i>, respectively. The isolates resulted in increased plant height (cm), fresh and dry weights (g), number of leaves, number of flowers, yield/plant, and chlorophyll contents (SPAD). Additionally, tomato fruit quality was improved by total soluble solids (TSS) percentage, hardness, pH, diameter (mm), and vitamin C mg/100g.</p> Conclusion <p>The results suggest that <i>Bacillus subtilis, B. pumilus, B. chitinosporus</i> and<i> T. asperellum</i> might be useful in protecting tomato plants against bacterial wilt disease. These findings support further investigation in the biological regulation of bacterial wilt by the rhizosphere beneficial isolates.</p>