Drought-induced reduction in crop growth and productivity can be compensated by increasing atmospheric carbon dioxide (CO₂), a significant contributor to climate change. Drought stress (DS) affects crops worldwide due to dwindling water resources and irregular rainfall patterns. The experiment was set up under a randomized complete block design within a three-by-two factorial arrangement. Six SPAR chambers represent three blocks (10 replications each), where each chamber has 30 pots in three rows. Each chamber was maintained with 30/22 (day/night) °C temperature, with either ambient (aCO₂; 420 ppm) or elevated CO₂ (eCO₂; 720 ppm) concentrations. This experiment was designed to address the impact of DS on the physiological and biochemical attributes and study how the eCO₂ helps alleviate the adversity of DS in basil. The study demonstrated that DS + eCO₂ application highly accelerated the decrease in all forms of carotene and xanthophylls. eCO₂ positively influenced and increased anthocyanin (Antho) and chlorophyll (LChl). eCO₂ supplementation increased LChl content in basil under DS. Furthermore, DS significantly impeded the photosynthetic system in plants by decreasing CO₂ availability and causing stomatal closure. Although eCO₂ did not increase net photosynthesis (Pn) activity, it decreased stomatal conductance (gs) and leaf transpiration rate (E) under DS, showing that eCO₂ can improve plant water use efficiency by lowering E and gs. Peroxidase and ascorbate activity were higher due to the eCO₂ supply to acclimate the basil under the DS condition. This study suggests that the combination of eCO₂ during DS positively impacts basil’s photosynthetic parameters and biochemical traits than aCO₂.

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