Sesame (Sesamum indicum L.) is cultivated in Ethiopia as a cash crop, the second major source of foreign currency next to coffee, and a raw material for oil industries. Water deficit stress was a major constraint limiting sesame crop production worldwide. In the context of climate change and water scarcity, there is a need to develop and use drought-tolerant sesame cultivars. This study was conducted to evaluate the growth, physiological, biochemical, and yield responses of three sesame varieties (Gondar-1, Setit-1, and Adiarkay) under two water treatments (WW and WDS) with the objective of identifying water deficit-tolerant varieties. A field experiment consisted of variety of sesame grown under two water regimes arranged in a randomized complete block design was used for data collection. The two-way ANOVA results showed significant differences (p < 0.05) in plant height, number of branches, number of leaves, leaf area, relative water content, leaf membrane stability index, proline concentration, total soluble sugar, height of capsule stem length, number of capsules per plant, capsule length, number of seed per capsule, seed weight per capsule, seed weight per plant, seed yield per plot of sample but not chlorophyll content. Also, the results of mean productivity (66.61g), yield potential stress index, stress susceptibility index, stress tolerance index, water use efficiency, rank sum of studied varieties and due to less % reduction (29.33), yielding the highest seed weight(55.2 g pp) and overall productivity of Adiarkay (99.32 g pp), compared to other varieties confirmed that it was the most tolerant variety under water deficit stress, while Gondar-1 was found to be the highest susceptible to water deficit (1.19) and showed a significant reduction in growth and yield (42.33%). Thus, Adiarkay, followed by Setit-1, was found to be water deficit tolerant and high yielding, while Gondar-1 was the most susceptible and low yielding variety. The findings could assist farmers in choosing sesame varieties suited for their growing, particularly in regions prone to water scarcity. While researchers recommend additional sesame varieties to study the genetic mechanisms of water deficit tolerance for enhanced crop resilience.