Abstract: In this study, Tunisia soft seed pomegranate was used as material, and the leaves and stems were selected as explants. The suitable explants and induction medium were selected, and the proliferation culture, rooting and domestication of test tube seedlings were carried out. The results showed that when stem segments and leaves were used as explants, the induction rate of treatment 8 medium with MS + 6-BA 0.6 mg/L + NAA 0.3 mg/L was the highest, 82.78% and 45.00%, respectively（p＜0.01）, higher than other combinations extremely significantly. Compared with leaves, stem segment was an ideal explant material. The most suitable medium for subculture callus differentiation was MS + 6-BA 1.0 mg/L + NAA 0.4 mg/L medium in treatment 7, and the differentiation rate was 75.00%, which was significantly higher than that of other combinations（p＜0.05）, followed by MS + 6-BA 1.0 mg/L + NAA 0.2 mg/L, the differentiation rate was 73.89%. The most suitable for the rooting of Tunisia soft seed pomegranate tissue culture seedlings was that the formula of treatment 8 with 1/2 MS + IBA 0.5 mg/L + NAA 0.3 mg/L and the formula of treatment 7 with 1/2 MS + IBA 0.5 mg/L + NAA 0.1 mg/L,of which the rooting rate was more than 90.00% and tissue culture seedlings had good growth. The rooting seedlings were transplanted into garden soil, peat soil and perlite with a ratio of 2∶2∶1, and the survival rate was as high as 96.83%.