Abstract: By using the seed and cotyledon of loofah as experimental materials, the effects of different disinfection treatments, different seedling ages, different types and concentrations of plant growth regulator on the regeneration system of loofah cotyledon were studied. The results showed that the combination of disinfection with alcohol for 40 s and disinfection with 0.1% mercuric chloride for 10 min was the best combination for seed disinfection, with the seed contamination rate of 5.56% and seed bud ratio of 94.44%. The green cotyledons slightly spread at about 7 days were selected as the best explants, and the most appropriate bud inducing medium was MS+2.0 mg·L^-1 6-BA+30 mg·L^-1 sucrose and 6 g·L^-1 agar powder, with an induction rate of over 64%. The optimal culture medium for adventitious bud proliferation was MS+2.0 mg·L^-1 6-BA+4 mg·L^-1 AgNO₃, and the proliferation coefficient was 3.27. The proliferating plants were healthy and growing well. During the root induction of loofah explants, 1/2 MS was used as the rooting medium, and the rooting rate was up to 100%. The root was cultured for 3-4 weeks until the main root grew to more than 5 cm, and the melon seedlings grew well. In the later transplanting, the suitable culture medium was the ratio between peat soil and perlite of 5:1, and the success rate of cultivation could reach above 95%.