In the present study, ameliorative abilities of plant growth promoting fungi (PGPF) under fusarium infection have been examined through stimulation of biochemical defense and Physio-biochemical performance in tomato plants. A total of 25 fungal isolates were isolated from the Beta vulgaris Rhizosphere cultivated soil (Tamiya, Fayoum Governorate, Egypt). These fungal isolates have been characterized in terms of the production of certain plant growth promotion active metabolites that enhance plant growth and suppress diseases. Four fungal isolates were selected as the most for plant growth promotion. The four fungal isolates were identified morphologically as Aspergillus niger, A. flavus, Mucor sp., and Penicillium sp. Under greenhouse conditions, tomato plant treated with these fungi separately showed significant reduction in wilt disease. Biochemical defense such as osmolytes, oxidative stress, and antioxidant enzymes activity were assessed at 60 days after planting. The results revealed that Fusarium oxysporum strain was highly destructive effect on tomato plants by PDI 87.5 %. Moreover, PGPF filtrates which applied to infected tomato improved osmolytes, total phenol and ascorbic acid. Interestingly, the deleterious impact of wilt disease on tomato plants were significantly reduced and it can be evident from reduced MDA and H2O2 levels. Therefore, these results highlighted that the soil harbors antagonistic fungi offering several plant growth-promoting fungi (PGPF), which can be exploited as a powerful biological control agent in tomato plant against Fusarium wilt.