The present study aimed to establish an efficient in vitro propagation protocol for purple-leaf plum (Prunus cerasifera Ehrh). Clorox at 15% (v/v) or HgCl2 at 0.1 % (w/v) for 10 min each gave the best results for both decontamination (100 and 90.2) and survival (90 and 92) percentages, respectively. The effect of three cytokinin types (BAP, Kin and 2ip) at four concentrations (0.5, 1.0, 1.5 and 2.0) mg/L were studied during multiplication stage in comparing with hormonal-free medium. Applying BAP at 1.5 mg/L induced the initiated buds to develop into shoots (100% shooting) during the three successive subcultures. Adding BAP at 1.5 or 2.0 mg/L gave the highest significant shootlets multiplication rate (2.13 and 2.15 shootlet/ explant, respectively). While the longest shootlets (1.92 cm) were recorded on MS medium augmented with 1.5 mg/L of 2ip. Adding 0.4 mg/L of IBA to the culture medium lead to 100% rooting of shootlets. Using peat moss + perlite (1:1 v/v) as a putting medium for ex vitro acclimatization of plantlets resulted in the best results for survival, height, stem fresh and dry weights of acclimatized plantlets. The amplified DNA fragments using 844-A (for ISSR) and A2 (for RAPD) primers to compare between donor and micropropagated plants. Data showed that the maximum number of bands (11 and 9), respectively of DNA fragments with molecular size ranging between 226.98- 1007.92 and 215.16- 1619.62 bp. Data indicated that micropropagated plants and donor plant are much closed to each other and the micropropagation protocol genetically safe.