<p>Water deficit in soil during the formative growth stage adversely affects the sugarcane productivity. Plants have developed a series of adaptive mechanisms to cope with these stresses. Characterization of plant genotypes under water deficit conditions will provide the basis for breeding new germplasms for efficient utilization of water and nutrients and adaptation to water stress. To achieve this goal, two tolerant (Co 98014 and Co 0118) and two sensitive (CoJ 85 and Co 89003) sugarcane genotypes, categorized in previous study were assessed for antioxidant responses, followed by differential expression of three aquaporin genes (ShPIP2-1, ShPIP-5 and ShPIP2-6) under two water deficit conditions. The MDA content was significantly increased 1.4 to 1.9 folds in tolerant genotypes than sensitive ones (0.4 to 0.7 folds). The percent change in H₂O₂ content was significantly increased upto 100.2 to 160.1% in the sensitive genotypes as compared to the tolerant genotypes (39.4 to 61.4%). The percent changes in SOD activity found greater (<i>p &lt;</i> 0.05) in tolerant genotypes (136.9 to 233.5%) than the sensitive (66.3 to 137.2%). The transcript analysis of AQPs revealed upregulation of ShPIP2;5 but downregulation of ShPIP2;1 and ShPIP2;6 when plants were subjected to water deficit conditions. The findings suggested the role of antioxidant activity and PIP2 AQPs in mitigating oxidative stress under drought. This biological insight highlights that genotypes with stronger antioxidant defense and efficient aquaporin regulation are better equipped to maintain cellular homeostasis and water transport, thereby enhancing survival under water deficit. These traits can serve as valuable physiological markers for selecting drought‑tolerant sugarcane varieties, contributing to the development of resilient cultivars suited for semi‑arid regions.</p>