Rosa roxburghiif.eseiosaKu is a variety ofRosa roxburghii, with two known genotypes: Wuci 1 and Wuci 2. The lack of prickle on the peel ofR. roxburghiif.eseiosamakes it easy to pick and process, but its fruit size is small. Therefore, we aim to induce polyploidy in order to obtain a larger fruit variety ofR. roxburghiif.eseiosa. In this study, current-year stems of Wuci 1 and Wuci 2 were used as materials for polyploid induction, which was carried out through colchicine treatment coupled with tissue culture and rapid propagation technology. Impregnation and smearing methods were effectively used to produce polyploids. Using flow cytometry and a chromosome counting method, it was found that one autotetraploid of Wuci 1 (2n= 4x= 28) was obtained by the impregnation method before primary culture, with a variation rate of 1.11%. Meanwhile, seven Wuci 2 bud mutation tetraploids (2n= 4x= 28) were produced by smearing methods during the training seedling stage. When tissue-culture seedlings were treated with 20 mg/L colchicine for 15 days, the highest polyploidy rate was up to 60%. Morphological differences between different ploidys were observed. The side leaflet shape index, guard cell length, and stomatal length of the Wuci 1 tetraploid were significantly different from those of the Wuci 1 diploid. The terminal leaflet width, terminal leaflet shape index, side leaflet length, side leaflet width, guard cell length, guard cell width, stomatal length, and stomatal width of the Wuci 2 tetraploid were significantly different from those of the Wuci 2 diploid. Additionally, the leaf color of the Wuci 1 and Wuci 2 tetraploids changed from light to dark, with an initial decrease in chlorophyll content followed by an increase. In summary, this study established an effective method for inducing polyploids inR. roxburghiif.eseiosa, which could provide a foundation for the breeding and development of new genetic resources forR. roxburghiif.eseiosaand otherR. roxburghiivarieties in the future.