An experiment was conducted to investigate the physiological and biochemical responses of two hexaploids viz., C 306 (water stress tolerant) and Hira (water stress susceptible), and two tetraploids, HW 24 (Triticum dicoccum) and A 9-30-1 (Triticum durum) wheat genotypes to water stress under pot culture condition. Water stress was imposed for a uniform period of 10 days at 50, 60 and 70 days after sowing (DAS) and observations were recorded at 60, 70 and 80 DAS. Total dry matter and plant height were recorded at harvest. Water stress caused a decline in relative water content (RWC), chlorophyll and carotenoid content, membrane stability and nitrate reductase activity and increased accumulation of proline at all stages and abscisic acid (ABA) at 80 DAS in all the genotypes. Both the tetraploids showed a lower reduction in RWC and highest ABA accumulation under water stress. Among the hexaploids Hira showed the most decline in RWC and the lowest ABA accumulation. The tetraploids also showed comparatively higher carotenoid content and membrane stability, closely followed by C 306, while Hira showed the minimum response under water stress. Nitrate reductase activity and chlorophyll content under irrigated conditions were highest in Hira but under water stress the lowest per cent decline was observed in C 306, followed by HW 24, A 9-30-1, and Hira. Proline accumulation under water stress conditions was highest in hexaploids C 306 and Hira and lowest in tetraploids HW 24 and A 9-30-1. Tetraploids HW 24, followed by A 9-30-1 maintained higher plant height and total dry matter (TDM) under water stress and also showed a lower per cent decline under stress than hexaploids C 306 and Hira. From the results it is clear that proline accumulation did not contribute to better drought tolerance of tetraploids than hexaploids. It is also apparent that water stress tolerance is the result of the cumulative action of various physiological processes, and all the parameters/processes may not be positively associated with the drought tolerance of a particular tolerant genotype.