ABSTRACT. We attempted to create a new germplasm of cucumber cultivar Chinese long (9930) using different doses of ethyl methyl sulfonate (EMS) to induce variability. We tested EMS concentration (0, 0.5, 1.0, 1.5, 2, 3% v/v) with post-treatment (0.1 M Na 2 S 2 O 3 and water), EMS concentration (0, 0.5, 1.0, 1.5% v/v) over different treatment times (8, 16, 24 h), and EMS concentration (0, 0.5, 1.0, 1.5% v/v) with different treatment temperatures (20 and 28°C). In all experiments with increasing EMS concentration, germination percent, index, and rate were decreased. After addition of stop solution (0.1 M Na 2 S 2 O 3 ), posttreatment mutated seeds showed higher germination (84.44%) and rate (37.5%) than seeds treated with water (80 and 34.07%, respectively), 2441 EMS dose optimization in cucumber ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 14 (1): 2440-2449 (2015) while the germination index was high in seeds treated with water. At 20°C, the germination index (4.13) and rate (56.25%) were affected to a greater extent than at 28°C (7.68 and 91.31%, respectively). Treatment times of 16 and 24 h showed similar results for germination percent and rate, while the germination index was decreased over time. There were significant differences in seedling height, fresh true leaf weight, seedling weight, and plant survival with increasing EMS concentration and time. Higher variations in the form of dwarf seedlings were recorded after treatment with 1.5% EMS for 24 h. Based on germination and morphological data, an EMS concentration of 1.5% for 24 h at 20°C and post-treatment with stop solution (0.1 M Na 2 S 2 O 3 ) efficiently caused mutation.