Tomato affected by several pathogenic factors, including root-knot nematodes (Meloidogyne spp.). This research was conducted to evaluate the efficacy of three amino acid Arginine, Serine, and Leucine, in their normal and Nano forms, at three different concentrations (10, 20, 30 ppm). The evaluation was performed under laboratory conditions to assess their effectiveness in inhibiting egg hatching and the viability Second stage Juveniles (J2) of the targeted nematode species. The laboratory results demonstrated the efficiency of all treatments in reducing the percentage of unhatched eggs, particularly at higher concentrations. The Nano-form of the three amino acids at a concentration of 30 ppm exhibited the highest efficacy compared to the normal form and showed significant superiority over the control treatment. After 72 hours, the percentage of unhatched eggs for Arginine, Serine, and Leucine Nano-amino acids was (21.00, 31.33, and 30.67), respectively, with a kill rate of (79%, 68.88%, and 69.33%) compared to the control treatment, which recorded 75% unhatched eggs and a kill rate of 25%. The impact of these treatments on the viability Second stage juveniles(J2) also varied significantly with different concentrations, showing a dose-dependent effect over time and significant differences compared to the control group. The highest concentration of 3000 ppm was the most effective, resulting in the highest kill rate for Arginine, Serine, and Leucine Nano-amino acids, which reached (95.27%, 90.27%, 88.89%), respectively, compared to the control treatment, which had a kill rate of 48.60%.