Abstract: In order to advance the bulblet in Vitro bolt and shorten the time of the bulb formation, the bulblet quality and the bolting rate were studied through different cultivation methods of the bulblet in Vitro of oriental hybrid lily. Results showed that: there was no difference in the physiological change rule between cold treatment in substrate and that in bottles, indicating that the contents of starch and soluble protein both decreased during the cold treatment, while the contents of soluble sugar and reducing sugar increased continuously. The bolting rate in dark was better than that in light whether the cold treatment was in substrate or in bottles. The bolting rate with the cold treatment in substrate was better than that in bottles. Factory production of lily seedlings should adopt dark culture cultivation in combination with cold treatment in substrate for bulblet before planting.