Drought is considered one of the major constraints of plant growth and productivity worldwide. Plants respond to drought through different mechanisms including physiological, biochemical, and gene expression modulation. Studying these mechanisms will provide better understanding of drought response mechanisms and will help breeders in developing new cultivars. In this study, growth, biochemical, and molecular responses of four wheat ( Desf.) landraces to drought stress (300 mM mannitol) were investigated at the seedling stage. Reverse transcription-polymerase chain reaction was used to assess gene expression level for a drought stress responsive gene (). Germination percentage, shoot length, root length, and root number for all landraces were decreased significantly under drought stress. However, drought stress caused an increase in proline content, lipid peroxidation level, and transcript level. According to the studied traits, the Karak landrace showed long shoots (48% relative to its control), the longest roots (45% relative to its control) and the highest proline content (483% relative to its control). The results indicate that from the landraces studied, Karak may be selected as the most tolerant wheat landrace and may help in wheat breeding programs for adaptation to drought-prone environments.