In the current study, biotic (T.harzianum, T. konignii, and B. subtilis) were used to examine the antifungal activity of these agents against maize root rot pathogens (F. oxysporum, F. solani or R. solani) in vitro. For these reasons, the experiments were done in the laboratory. Confrontation among biotic agents and the tested pathogenic fungi results shows that both Trichoderma spp and Bacillus subtilis prevent the linear growth of all the examined pathogenic fungi. T.harzianum appeared high level and significant effect of antagonism and diminuend the growth of F. oxysporum. While T. konignii was stronger in decreasing the linear growth of F. solani and R. solani. The antagonistic bacteria came an end as compared with antagonistic fungi in decreasing the linear growth of all examined pathogenic fungi. In screening of the bio-agent antagonist test, it was observed that the negative effect was estimated by using T. harzianum on IZ, L against F. oxysporum, and on OG as well as L on R. solani. While it gives of a potential antagonism on the three measures against F. solani as well as OG when grows rapidly over the mycelium of F. oxysporum and occurs IZ on R. solani. Moreover, T. koningii grows considerably faster than three tested pathogenic fungi and occurs lysis in hypha of both Fusarium spp viz, F. oxysporum, and F. solani. On the other side, the antagonistic B. subtilis lead to presence of an inhibition zone in all tested.