Zamiifolia(Zamioculcas sp.) is a perennial plant in theAraceaefamily and one of the new apartment plants in the world. In this study, in order to increase the effectiveness of the breeding program, tissue culture technique and explants of leaf parts were used. The results indicated that 2,4-D (1 mg/l) and BA (2 mg/l) hormones affected positively and significantly callus formation and simultaneous application of NAA and BA (both in 0.5 mg/l) caused the best results regarding seedling production and number, leaves, complete tubers, and root in tissue culture ofZaamifolia. In the study, three cultivars ofZamiifolia(green, black and Dutch) and 12 genotypes resulted from callus formation stage, irradiated with different gamma rays (0 to 175 Gy, with LD50as 68 Gy) were selected and the presence of genetic diversity was investigated using 22 ISSR primers. Applying ISSR marker showed that the highest amount of PIC values related to the F19(0.47) and F20(0.38) primers, which persuasively isolated the studied genotypes. Moreover, the highest efficiency was detected for AK66 marker based on the MI parameter. The PCA and clustering categorization via UPGMA methodology based on molecular information and Dice index, differentiated the genotypes into 6 groups. Genotypes 1(callus), 2(100 Gy) and 3(cultivar from Holland) created separated groups. The 4th group included 6(callus), 8(0 Gy), 9(75 Gy), 11(90 Gy), 12(100 Gy) and 13(120 Gy) genotypes appearing as the largest group. The 5th group included 7(160 Gy), 10(80 Gy), 14(140 Gy) and 15(Zanziber gem black) genotypes. The last group included 4(mather plant) and 5(callus) genotypes. In this context, genotypes 1, 5, and 6 had probably somaclonal variation. Moreover, genotypes that received doses of 100 and 120 Gy, had a medium diversity. There is a high possibility of introducing a cultivar with a low dose and high genetic diversity in the whole group. Genotype 7 in this classification, received the highest dose of 160 Gy. In this population, the Dutch variety, was used as a new variety. As a result, the ISSR marker could correctly group the genotypes. This is an interesting finding, and it could be hypothesized that the ISSR marker could correctly differentiateZaamifoliagenotypes and probably other ornamental plants under the effect of gamma rays mutagenesis in order to achieve novel variants.