The present study investigates the effect of plant growth regulators (PGRs) and fungicide treatments on the non-enzymatic antioxidant potential of Ocimum basilicum L. (basil), a medicinal herb known for its antioxidant properties. The research aimed to evaluate the influence of gibberellic acid (GA3), propiconazole (PPZ), and their combinations with beneficial microbes on key non-enzymatic antioxidants, namely total phenols, ascorbic acid, and reduced glutathione, across different growth stages of the plant. The experiment was carried out with treatments applied at three growth stages: 25, 50, and 75 days after planting (DAP). Results showed a significant increase in the total phenol, ascorbic acid, and reduced glutathione contents in treated plants compared to the control group. The application of PPZ demonstrated a stronger enhancement in these antioxidants than GA3, with the highest increase observed at 75 DAP in the propiconazole-treated plants. Specifically, propiconazole treatment resulted in a 121.35% increase in reduced glutathione content over the control. Additionally, a positive correlation was observed between the enhanced antioxidant levels and the overall growth parameters of the plants. These findings suggest that the application of PGRs like GA3 and fungicides such as propiconazole can significantly boost the non-enzymatic antioxidant defenses in Ocimum basilicum, potentially improving its medicinal value.