Water extract from spent mushroom substrate (WESMS) of Pleurotus eryngii suppressed bacterial wilt disease of tomato caused by Ralstonia solanacearum by 70% without any direct antibacterial activity against the pathogen. WESMS-treated tomato had increased contents of free phenolic compounds (increased by 3%) and total salicylic acid (increased by 75%), and significantly enhanced plant height, leaf number, and fresh weight compared to those of a water-treated tomato sample. These results suggest that the treatment of tomato with WESMS can suppress bacterial wilt disease by enhancing plant defense factors and overall plant health.