The induction of variation in chrysanthemums using gamma radiation under in vitro conditions is an effective technique in ornamental plants. The purpose of this study is to obtain new mutants by isolating desirable properties from the three-colored single chrysanthemum mutant using in vitro cultures. Bud explants were cultured four times, the plantlets were acclimatized, and 520 plants were planted in outdoor conditions. Observations of the mutants were collected during flowering time, and 97 of the mutants were compared to the control group. Plants with pink, white, and chimeric flowers were obtained. Mutant plants with white flowers constituted the majority of the population followed by plants with variegated flower colors. The population is divided into six clusters, based on the plant height, plant diameter, flower number, flower diameter, number of flower colors, ray flowers’ number, leaves’ number, stem weight, and lengths and widths of leaves. The population decreased in plant height, flower number, and stem weight, while other features increased compared to the control group. Thus, a new population with similar characteristics to the parent plant was obtained from a single mutant. Chrysanthemum plants exposed to mutagens showed major changes in flower parts as well as other parts of the plant.
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