Abstract: The effects of minimal media, hormones, sugar, light and supports of media on shoot propagation and root formation were studied in tissue culture of pumpkin (Cucurbita moschata). The results showed that 1) the best medium for shoot propagation was MS+BA 0.5～1.0 mg/L+IAA 0.1～0.5mg/L + sugar 30 g/L; 2) the suitable medium for adventitious root induction was 1/2MS+sugar 20g/L, and defatted cotton was the best support in rooting media; 3)diffused natural light was better than artificial light in tissue culture.