Drought stress is the most important factor leading to a decrease in both the quantity and quality of wheat yield, a strategic crop. This stress has a negative impact on various vital biochemical and physiological processes in wheat. This study aimed to assess the activity of glutathione reductase and ascorbate peroxidase enzymes, as well as to investigate the expression levels of genes that control these enzymes and their relationship with yield and some yield components in the field under three moisture conditions for two wheat cultivars. The experiment was conducted in a split-plot design within a randomized complete block design with four replications. The main factor consisted of three levels of moisture stress: -0.3 bar (field capacity as the control treatment), -2 bar, and -4 bar, applied in field conditions using a tensiometer. The secondary factor included two wheat cultivars, Kalateh and Ehsan. The results of the analysis of variance indicated that the effects of different moisture levels and the differences between the two cultivars were significant (p≤0.01) for all evaluated traits. Yield and its related traits were negatively impacted by drought stress, leading to a decrease. The activity of ascorbate peroxidase and glutathione reductase enzymes was influenced by the interaction between drought stress and cultivar. In general, their activity increased with the occurrence of stress, consistent with the increase in gene expression. The increase in gene expression under stress conditions was four times higher in the Kalate cultivar compared to the control, and two times higher than in the stress condition. The results suggest that drought stress resulted in a decrease in yield and related traits, likely due to oxidative stress and disruption of plant biological processes. Plants under drought stress aim to reduce oxidative damage by upregulating genes related to antioxidant enzymes, resulting in decreased yield but increased resistance through biological reactions. This pattern was more pronounced in the Kalateh cultivar, indicating its biological resilience to stress. Overall, the results indicated that the Kalateh cultivar performed better than the Ehsan cultivar.