Potatoes are underground stem- tubers of the plant Solanum tuberosum, a perennial in the nightshade family Solanaceae. Potatoes can be infected by several fungi, causing severe losses in potato production. This study was designed for isolation and identification of potato root rot fungal pathogens in Assiut governorate, Egypt. Testing some biocontrol agents like Bacillus subtilis, Saccharomyces cerevisiae, and Trichoderma species, and testing the antifungal properties of zinc and titanium nanoparticles against potato root rot diseases caused by Fusarium spp. The assessment was conducted by measuring disease severity and morphological and physiological changes such as total antioxidants, total phenols, total flavonoids and total terpenes in the plant roots. Fusarium incarnatum, F. solani, and F. moniliforme were the dominant pathogens in 30 infected samples, with disease severity range from 0% to 83.3%. The results showed that T3 (Trichoderma atroviride), T4 (B. subtilis), T8 (ZnNPs 150 ppm), and T11 (TiNPs 150 ppm) effectively reduced disease severity to 0% and enhanced plant morphological and physiological characteristics compared with the infected control plants. The research showed various effective solutions for controlling Fusarium pathogens in soil the ability to apply these treatments in controlling potato root rot and wilt diseases.