Vernalization genes (Vrn), in addition to controlling the growth habit of wheat (spring and winter), play a key role in flowering time and early maturity of wheat. They are also of great interest in drought tolerance researches. To investigate the effect of Vrn genes on drought tolerance, four isogenic lines were developed. Isogenic lines were resulted from the crosses of the early heading Australian variety, Excalibur, with the late heading Iranian variety, Roshan, and followed by backcrossing with Roshan. Two experiments were conducted based on randomized complete block design with four replications under rainfed conditions of Sepidan during 2019-2020 and under well-watered conditions of Kerman during 2020-2021 cropping years. In the present research, seven important agronomic traits, including days to heading, days to ripening, grain filling period, spike number per meter square, grain number per spike, 1000-grain weight, and grain yield were assessed. Isogenic lines were assessed for drought tolerance using eight indices, including mean of productivity (MP), yield index (YI), stress tolerance index (STI ), geometric mean of productivity (GMP), stress susceptibility index (SSI ), yield stability index (YSI), and stress tolerance score (STS). The results of stress tolerance score (STS) showed that isogenic lines vrn-B1/Vrn-D1a and Vrn-B1a/vrn-D1 had the highest and the lowest stress tolerance score, respectively. In addition, the vrn-B1 and Vrn-D1a alleles, which cause early flowering, improve drought tolerance by utilizing an escape mechanism from dry conditions.