Fusarium  oxysporum  Schlechtend : Fr   f.sp .  cubense (FOC) (E  F. Smith).  Snyder & Hansen  were the most causal organism  causing  wilt  disease of  banana plant  at Beheira , Gharbea  and  Assuit Governorates, Egypt. Fusarium  wilt disease  of banana showed  to be  similar to  those of  other  plants. Discoloration  brown  to dark  of  pseudostem  and  rhizomes  was  the most  indicator of Fusarium wilt  of  banana plants. In vitro four microbial  isolates  from the  banana  root and rhizosphere  showed  highly  antagonistic  activity  to  F.oxysporum  f. sp. cubense. These  isolates  were  identified  as Bacillus  subtilis (Ehrenberg) Cohn (No.1),  Pseudomonas  fluorescens Migula(No.3), Trichoderma  harzainum Rifai (No.6)  and Laetisaria arvalis Burds (4). Bacillus  subtilis (Ehrenberg) Cohn (No.1);  Pseudomonas  fluorescens Migula (No.3)  and T. harzianum isolates  were the most antagonistic isolates inhibiting  mycelial  growth of F. oxysporum f.sp. cubense.These antagonistic strains were excreted antifungal lytic enzymes on liquid medium containing dead mycelium of F. oxysporum  f. sp. cubense i.e. chitinase, β1-3,β1-4 glucanases and protease. Chitinase and protease enzymes were the most lytic produced by all antagonistic strains.while β1-3and β1-4 glucanases were produced at a moderate rate. Bacillus  subtilis (Ehrenberg) Cohn (No.1) was the most antagonistic strain producing chitinase and protease followed by L. arvalis and T. harzianum. On the other hand, Bacillus  subtilis and L. arvalis were the most antagonistic producing difusible and volatile antibiotics followed by P. fluorescens. while, T. harzianum was the least antagonistic phase of antibiotics. Pot trails conducted during a summer season, revealed that dipping   banana roots transplanting root in the suspension of B.subtilis,  P.fluorescens Migula, T.harzianum Rifai and Laetisaria arvalis Burds 3x10⁸ (CFU/ ml ) for one and two hours or soil drench with 100 and  200 ml / pot were effectively reduced wilt disease incidence of banana plants, respectively. Under field naturally infestation with  F.oxysporum f.sp. cubense  soil  dranch  of banana  around rhizome region by suspension of   B.subtilis,  P.fluorescens Migula, T.harzianum Rifai,  and fungicide (Topsin)  reduced the wilt  incidence  than control . Soil drenched  with  300 ml of  B. subtilis significantly  reduced  the  wilt incidence by  50% compared with control. Whereas the P. fluorescens Migula failed to control Fusarium  wilt.  B. subtilis  was the most promising  biocontrol agent  against  wilt  disease of  banana.