The impact of chitosan, Trichoderma harzianum, thyme oil and jojoba extract against F. oxysporum, wilt disease of strawberry, was studied in laboratory and throughout the following growing seasons of (2018 and 2019) in greenhouse. The antagonistic biocontrol agent, Trichoderma harzianum, was isolated and identified in lab at the Faculty of agriculture Saba Basha, Alexandria, Egypt. In laboratory tests, the effect of the tested biocontrol agents on fungal mean growth in vivo was determined. In the greenhouse experiments, the treatments were performed in three application methods: foliar, soil drench and the mixed foliar/soil method. In lab tests, Chitosan ® (4g/l) followed by T. harzianum and Top perfect ® (4ml/l) reduced the fungal growth of Fusarium oxysporum pathogen by 86.3, 85.9% and 78.2 %, respectively. In green house experiments, the data showed that the mixed application treatment (soil drench and foliar spray) is important for controlling Fusarium oxysporum pathogen. Thyme oil was less effective than the other treatments as biocontrol agent for the pathogen. The Highest determined chlorophyll content was determined in the treatment of T. harzianum (48.23), Chitosan® (47.66) and Top perfect® (44.97), without significant differences between T. harzianum and chitosan, especially in the mixed application treatment. Moreover, the treatments of Chitosan ® , T. harzianum and Top perfect ® , significantly, showed the highest fresh, dry weights and root length of plants and the least disease incidence and severity percentages compared with the untreated control.