Persian violet flowers are considered esthetically attractive, leading to the high economic value of this plant. Plant breeding is fundamental to crop improvement, and the induction of mutation by tissue culture technology in combination with irradiation has been beneficially applied to generate plants with novel desirable characteristics. In this research, single or double rounds of UV-C irradiations were carried out on plant tissue cultures to initiate the in vitro rooting and mutation of Persian violets. It was found that single low-intensity UV-C exposure, when applied to Persian violet microshoots for 4 h, could induce the maximum number of roots and the highest root length without the use of a plant growth regulator. Overall, the single and double UV-C irradiation of Persian violet microshoots led to 44 different types of Persian violet flower mutations. Under single high-intensity UV-C irradiation for 6 h, up to nine petals were initiated, whereas single low-intensity UV-C irradiation did not influence the morphological variation of Persian violet flowers. Thus, Persian violet microshoots respond differently in terms of in vitro rooting and flowering depending on the UV-C intensity and exposure duration. These outcomes may be applied to micropropagation and in vitro plant breeding.