Abstract: This study established the program of a high efficient in vitro regeneration procedure of five pepper cultivars. The effect of genotypes, explants, hormones, and days of seedlings on the tissue culture in pepper was investigated systematically. Three cultivars of pepper (Capsicum annuum L.), ‘Nongchengjiao No.2’, ‘0127’, and ‘0171’, were screened out of five cultivars for their high capacity of regeneration. The experimental results showed that the cotyledon explants taken from 12-day-old seedlings displayed the highest shoot-buds regeneration capacity. As far as culture medium was concerned, the buds induced well on MS+3% sucrose＋0.6% agar＋5.0mg/L BA＋0.5~1.0mg/L IAA, which made the shoot buds differentiation frequency of 98%. Moreover, 45.7% of induced buds elongated well on the medium of MS+3% sucrose＋0.6% agar＋3.0mg/L BA＋1.0mg/L IAA＋2.0mg/L GA3＋10.0mg/L AgNO3. The regenerated plants were transplanted into nutrient soil and developed into normal plants. So, an efficient program for plant regeneration from explants of pepper had been developed.