The tomato (Solanum esculentum Mill.) is one of the most widely cultivated vegetable crops worldwide. It serves as a favorable host for plant-parasitic nematodes, particularly the root-knot nematode (Meloidogyne javanica). Investigations were carried out to determine the effectiveness of bio-agents against root-knot nematodes in pot conditions. The bio-agents included Metarhizium anisopliae, Bacillus subtilis, Verticillium lecanii, Trichoderma harzianum and, Trichoderma asperellum) against root-knot nematode in pot condition. Effect of bio-agents was also estimated for accumulation of PPO and PAL and Root-knot nematode-infected tomato roots containing phenol. The results of the experiment showed that all the bio-agents significantly increased the levels of PPO, PAL, and phenol in tomato roots, improved plant growth parameters, and reduced nematode reproduction compared to the untreated control. The best treatment with the highest PPO, PAL, and phenol activity among the investigated bio-agents was determined to be Trichoderma harzianum at 3 gm/kg soil, followed by Bacillus subtilis and Metarhizium anisopliae. It improves the plant growth parameter and lowers the number of nematodes in the pot.