Abstract: Plant tissue culture setting up is one of the most important stages for modern plant breeding. Availability of a tissue culture protocol for genetic transformation toward resistance to diseases would be an important step for the establishment of a genetic transformation system in most plants. In current research, callus induction was investigated in Cuminum cyminum. The experiment was arranged in a factorial experiment with two factors. The first factor was 2,4-Dichlorophenoxyacetic acid (2,4-D) plant growth regulator in four levels (0, 0.5, 1 and 2 mg L-1 plus 0.1 mg L-1 kinetin) in the MS basal medium. The second factor was photoperiodic status in two levels (dark and 16/8 h light/dark). The experiment was layed out based on completely randomized design with three replications. Statistical analysis results showed that the highest percentage of callus induction was resulted at 1.0 mg L-1 2,4-D in both photoperiod conditions. Interaction effect of photoperiodx2,4-D concentrations showed significant (p≤0.05) difference of callus induction of cumin. It is concluded that 1 mg L-1 2,4-D treatment showed the most efficient influence on leaf explants callus induction.