This work was carried out in the Tissue Culture Laboratory, Horticulture Research Institute, Agricultural Research Center, Giza, Egypt from the period elapsed from 2014 to 2016 to study the effect of sterilization on propagation of Phytolacca dioica, L.. Results of this study could be briefed in the following outlines: Chlorox concentrations at 30 or 35 % gave the highest survival percentage of Phytolacca dioica, L.. In addition, soaking explants in the sterilant solution for 20 or 25 minutes resulted in the highest survival percentage. Similarly, the interaction treatment demonstrated that using chlorox at 35% for 25 minutes resulted in the highest survival%. In the multiplication stage it was found that using BAP at 10 ppm resulted in the highest number of shoots and leaves. BAP at 6 or 8 ppm induced the highest content of total chlorophyll. The longest shoots were a result of applying BAP at 3 ppm.  Inoculating explants on half strength medium resulted in the highest values of number of shoots and leaves, as well as highest contents of total chlorophyll and carotenoids. MS medium at 3 quarters strength shared in the first position of number of leaves. The longest shoots were a result of using MS medium at full strength. The interaction between BAP levels and MS medium strength, indicated that using MS medium at half strength supplemented with BAP at 10 ppm produced highest number of shoots. The same medium strength with BAP at 8 or 6 ppm gave the highest contents of total chlorophyll and carotenoids. MS medium at 3 quarters strength combined with PAP at 10 ppm gave highest number of shoots and leaves. MS medium at full strength plus BAP at either 3 or 10 ppm resulted in the highest shoot length and number of leaves.