Capparis spinosa, belonging to the Capparaceae, was studied to understand the effects of different types of mutagenesis. The study explored gamma rays, ultraviolet rays, colchicine, and caffeine. Various aspects, including appearance, chemical composition, and genetic variation, were evaluated. Radiation mutagenesis of Capparis spinosa using gamma rays (0.5, 1, and 1.5 kGy) and ultraviolet rays (2, 4, and 6 hours) resulted in a decrease in shoot number and length but an increase in the number of leaves. Chemical mutagenesis using colchicine (50, 100, and 150 mg/l) and caffeine (50, 100, and 150 mg/l) led to an increase in shoot and leaf numbers. Genetic variation was assessed using ten primers and the SRAP-PCR technique. The samples treated with colchicine at a concentration of 50 mg/l showed the highest similarity to the control group (89%), while the lowest similarity (81%) was observed in the samples treated with gamma rays at a dose of 0.5 kGy. The highest rutin amount (4.483 mg/ml) was produced when the plant was exposed to ultraviolet rays for 4 hours and gave high inhibition zone (31mm) against Bacillus subtilis bacteria while the lowest amount (1.213 mg/ml) was obtained from plants treated with caffeine at a concentration of 150 mg/l. Overall, the best treatment for Capparis spinosa was radiation using ultraviolet rays for 4 hours. This treatment resulted in a high number of shoots (2.83 shoots per plant), shoot length (0.66 cm), leaf number (6 leaves per shoot), callus growth (1.849 g), and rutin content (4.483 mg/ml).