Water deficit is one of the most important environmental stresses affecting the establishment and early growth of medicinal plants, and identifying genotypes adapted to such conditions is a crucial step toward the development of sustainable cultivation systems. In this study, the responses of four Glycyrrhiza glabra genotypes to drought stress were evaluated under two controlled environments, including laboratory and greenhouse conditions. In the laboratory experiment, drought stress was induced using polyethylene glycol (PEG 3000) at different osmotic potentials, while in the greenhouse experiment different soil moisture regimes based on percentages of field capacity were applied using a randomized complete block design with four replications. To characterize the response patterns of the genotypes, several traits were measured, including germination percentage, radicle length, plumule length, seed vigor index, chlorophyll content, proline concentration, and soluble sugar content. The results indicated that increasing drought stress significantly reduced germination parameters and seedling growth traits, and chlorophyll content also declined under stress conditions. In contrast, the accumulation of proline and soluble sugars increased under drought stress, suggesting the activation of osmotic adjustment mechanisms and protective responses in the plant. Significant differences were also observed among the genotypes in their responses to drought stress, indicating the presence of variability in their adaptive capacity. Overall, the combined evaluation of growth and biochemical indicators at the seedling stage can serve as an effective approach for identifying drought‑tolerant G.glabra genotypes.