In vitro cultivation and callus production are important tools for trait improvement, secondary metabolite production in medicinal plants, and ornamental propagation Chrysanthemum indicum var. indicum, due to its high medicinal and ornamental value, has attracted considerable attention from researchers in the field of tissue culture. The use of elicitors such as NaCl can influence callogenesis and the production of defensive compounds by inducing osmotic stress. The aim of this study was to investigate the process of callus induction and the effect of different NaCl concentrations on various morphophysiological, metabolic, biochemical, and antioxidant traits of callus in in vitro culture of Chrysanthemum indicum var. indicum. Seeds of Chrysanthemum indicum var. indicum were cultured in vitro, and explants were prepared from the leaves and stems of the plant. In the present study, the first experiment was conducted factorially in a completely randomized design with three factors: explant type, growth regulator combination, and light conditions. The highest callus induction percentage, the shortest time to callus initiation, and the greatest fresh and dry weight of callus were related to leaf explants treated with 2 mg BAP + 2 mg 2,4-D under light conditions. The second experiment was conducted as a completely randomized design to investigate the effect of different NaCl concentrations (0, 50, 100, and 150 mM) on callus traits. According to the results, NaCl treatment had a significant effect on increasing the metabolic, biochemical, and antioxidant traits of callus. T