Dendranthema x grandiflorum is an important species of the genus Dendranthema, and for the development of callus cultures. In this study, explants such as leaves and internodes were used to evaluate the effects of different types of explants, plant growth regulating agents in various concentrations on callus production. The results showed that, the leaf explant was significantly enhanced callus production than internode explant according to the percentage of callus formation, fresh and dry weight parameters. MS supplemented with 0.2 mg/l-1 BAP and 0.5 mg/l-1 2,4-D yielded the highest efficiency of callus formation after 2 month of culture. The BAP 0.2 mg/l-1 was significantly enhanced callus production in the both fresh and dry weight parameters than BAP 0.1 mg/l-1. The use of 0.5 mg/l-1 2,4-D excelled all other treatments for fresh weight although, there is no significant differences from the 1 mg/l-1 2,4-D treatment, the high 2,4-D concentrations 2 mg/l-1 also performed better than the lower concentrations for dry weight. Similarly, the least fresh and dry weight were obtained from the control which was devoid of 2,4-D. Using 0.5 mg/l-1 2,4-D leaf explant excelled all other treatments for fresh weight than internode explant, but using 2 mg/l-1 2,4-D leaf explant excelled all other treatments for dry weight than internode explant. Similarly, the control of leaf and internode explant in the both concentrations (0 and 0.5 mg/l-1) yielded the lowest fresh weight, but in dry weight the least was obtained from the control of leaf explant. Using 0.1 mg/l-1 BAP with 0.5 mg/l-1 2,4-D and 0.2 mg/l-1 BAP with (0.5, 1, and 1.5 mg/l-1) 2,4-D excelled all other treatments for fresh weight, but the best is using 0.1 mg/l-1 BAP with 0.5 mg/l-1 2,4-D. The least fresh weight was obtained from the control which devoids of plant growth regulators. Using 0.1 mg/l-1 BAP with 2 mg/l-1 2,4-D leaf explant excelled all other treatments with espect to dry weight. The least dry weight was obtained from the control which devoids of plant growth regulators.