Abstract: The vitrification phenomenon of culturing body is one of the main obstacles to regenerated plantlet formation in the process of in vitro culture of megaspore zucchini (Cucurbita pepo). The aim of the research is to guarantee normal production of the embryoid, minimize the impact of vitrification phenomenon on plantlet regeneration from megaspore and enhance the survival rate of test-tube plantlets. In the experiment, 4 kinds of media-induced somatic embryogenesis were selected to induce the formation of excellent induction medium. And the environmental temperature, humidity and concentration of GA were adjusted to reduce the occurrence of vitrification shoots of culturing body. The results showed that the four kinds of different levels of culture medium could induce the formation of embryoid bodies. One of the best was No. D: B5 40 g/L Suc 8 g/L Agar 0.5 mg/L NAA 0.05 mg/L TDZ 30 mg/L AgNO3. Meanwhile, embryo induction was up to 96% and 76% adventitious bud was generated directly. In the condition of 25℃ , 85% humidity, and 0.5% rooting medium GA concentration, the vitrification shoots rate of culturing body was the lowest. On the basis of previous researches, the experiment optimized and established the best zucchini plant regeneration unfertilized room technology system, provided a reference for further study on zucchini.