Seeds of four lentil genotypes (Castelluccio, Eston, Pantelleria, and Ustica) were subjected to five levels (0, 10, 15, 18, and 21%) of polyethylene glycol (PEG-6000). Germination percentage, root length, tissue water content (WC), a-and b-amylases, a-glucosidase activities, and osmolyte content were evaluated at 24, 48, and 72 h after starting the germination test. Water stress reduced seed germination percentage, root length, and seedling WC in all cultivars to different extent. The increase in proline content and total soluble sugars was greater for Eston and Castelluccio compared to the other genotypes. The activity of the enzymes involved in the germination process decreased in all cultivars; the activities of a-amylase and a-glucosidase were most negatively affected by osmotic stress, mainly in the drought sensitive Ustica and Pantelleria. Overall, Eston and Castelluccio were able to express greater drought tolerance and consequently could be used as a valuable resource for breeding programs.