Tomato (cv.Peto-86) is one of the most important crops in the world. In Egypt and many other countries it is often attacked by several pathogens causing root rot. Trichoderma and Bacillus species are widely the most feasible biocontrol microorganisms that suppress several pathogens. The efficiency of both antagonistic organisms was evaluated in vitro. as well as in vivo against root rot of tomatoes. The inoculation with T. viride and or B. subtilis of tomato plant suppressed F. salani as indexed by survival rate. Field experiments were carried out at Senowras Fayoum governorate during 2017 and 2018 seasons,. The obtained results showed that, the tested fungicide Topsin-M70 and the two biocontrol agents significantly reduced the incidence of root rot of tomato. The fungicide was the most efficient treatment in this regard compared with the two tested bioagents. Two bioagent treatments favored greater proliferation of rhizophere microflora and dehydrogenase activity in the rhizosphere. It is recommended to use these strains as a common biocontrol application in agriculture.