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To evaluate the responses of some important agronomic traits of 14 bread wheat cultivars, a split plot experiment was conducted based on a randomized complete block design under non-stress, moderate and severe salinity stress conditions. In this study, Backcross and Roshan were identified as the most salinity-tolerant cultivars, while Mihan and Shirudi were considered as the most salinity-sensitive cultivars. The proline content, as well as Δ1-pyrroline-5-carboxylate synthase (P5CS) and vacuolar-type H+-ATPase subunit E (W36) gene expression levels, were examined in these cultivars under normal, moderate and high salinity stress conditions. The proline content and P5CS gene expression level increased with a rise in NaCl concentration. Further, a direct relationship was observed between the proline content and P5CS gene expression in all samples. Our results showed that W36 gene expression in Backcross and Roshan cultivars, as the most resistant ones, strongly increased with elevation of the NaCl concentrations. On the other hand, in the sensitive cultivars such as Mihan and Shirudi, small changes were observed in the gene expression levels with rising salinity levels. Additionally, Backcross and Roshan cultivars had the highest proline content as well as P5CS and W36 genes expression levels under moderate and high salinity stress levels.