Abstract: To study the effect of ethyl methane sulfonate (EMS) induced mutation technology in the germplasm resources creation of wax gourd and to explore a suitable wax gourd seed induced mutation system, wax gourd seeds were subjected to mutagenesis using EMS phosphate buffer solution, and the effects of the treatment concentration and treating time on the wax gourd seed germination and growth were investigated. Moreover, the biological characters of M1 and M2 mutants were studied and analyzed. The results showed that the optimized induction condition was 1.2% EMS for 12 h. Under this condition, the seed germination frequency and seedling frequency of wax gourd were 54.67% and 51.83%, respectively, close to the lethal dose of 50%. And then 10000 wax gourd seeds were treated with this condition, 4752 seeds were sowed and pregerminated and 4500 converted into plant. The plants had the typical characteristics such as true leaf malformation and yellowing after EMS treatment, it proved that EMS induction was successful and 586 self- crossed wax gourds were obtained by self-pollination. After planting M2 strains, related mutants such as leaves, flowers, fruits and plant types were obtained, and the mutation frequencies were 27.22%, 26.80%, 20.47% and 16.06%, respectively. The total mutation frequency of wax gourd was 11.53%. This study establishes the EMS mutagenesis system of wax gourd seed and constructs the mutant library of wax gourd seed, laying a material foundation for the functional gene research of wax gourd.