| Drought is a significant issue because of the worldwide effects of climate change and the negative effects it has on wheat output. For improving production in water-scarce locations, screening drought-tolerant wheat cultivars is crucial. At the Plant Physiology Division of NIA, Tando Jam, the current experiment was conducted during the Rabi year of 2021. At both the P0.01 and P0.05 probability levels, the growth measurements at the seedling stage revealed extremely significant variations for all characteristics among genotypes, treatments, and interactions. The results demonstrated a stronger osmotic impact when using -0.5 MPa PEG. At the seedling stage, the genotype SDW-3 had higher decreases in a number of growth indices, such as shoot length (-11.3), root length (-72.6), and shoot fresh weight (-4.4), with an increase in PEG (-0.5 MPa) level. According to the results of the current study, all six wheat genotypes’ seed incubation shoot, root length, and shoot fresh weight drastically decreased under laboratory conditions due to osmotic stress. The genotype SDW-3 showed the greatest decrease under increased osmotic stress brought on by PEG-6000 (-5.0 MPa). As a result, the genotypes AST-1(V1), SDW-1, and SDW-2 may be employed in future breeding programmes and are drought resistant.