An efficient method to instantly assess drought-tolerant plants after germination is using osmoregulation in tissue culture media. In this study, the responses of three Iranian melon genotypes to sorbitol (0.1, 0.2, and 0.4 M) or polyethylene glycol (PEG) (0.009, 0.012, and 0.015 M) were evaluated as drought stress simulators in MS medium. ‘Girke’ (GIR), ‘Ghobadloo’ (GHO), and ‘Toghermezi’ (TOG) were the genotypes. GIR is reputed as a drought-tolerant genotype in Iran. The PEG or sorbitol decreased the coleoptile length, fresh weight, and photosynthetic pigments content while enhancing proline and malondialdehyde (MDA) contents. Protein content and antioxidant enzyme activity were utterly dependent on genotype, osmotic regulators, and their concentration. Coleoptile length, root and shoot fresh weight, root dry weight, proline and MDA content, and guaiacol peroxidase (GPX) activity can be used as indicators for in vitro screening of Cucumis melo L. genotypes. The results showed that sorbitol mimics drought stress better than PEG. Overall, our findings suggest that in vitro screening could be an accurate, rapid, and reliable methodology for evaluating and identifying drought-tolerant genotypes
