Abstracts: In order to establish an efficient and stable genetic transformation system of Lycium ruthenicum, effectively reduce the vitrification rate of regenerated seedlings, promote the study of gene function and improve the efficiency of genetic improvement. In this paper, the leaves of Lycium ruthenicum were used as explants, and the genetic transformation method mediated by Agrobacterium (LBA4404, EHA105) was used, which adjusted the type of basic medium, added the corresponding concentration of plant hormones, screened out the optimal callus induction medium, differentiation and selection medium and rooting induction medium. The genetic transformation rate of Lycium ruthenicum increased to more than 65%, and the vitrification rate of seedlings decreased to less than 10%. In this combination culture system lays an important foundation for molecular breeding of Lycium ruthenicum. The results showed that: (1) The optimal Agrobacterium infection concentration ( OD600 ) was 0.6 and the infection time was 25 min in the efficient combination culture system of Lycium ruthenicum leaves. Under this condition, the infected leaves were placed in the callus induction medium, and the resistance callus induction rate was 78.2%-96%;(2) The optimum differentiation and selection culture medium for genetic transformation of Lycium ruthenicum was : MS + inositol 50 mg / L + nicotinic acid 0.25 mg / L + VB6 0.25 mg / L + Fe salt mother liquor 1 mL / L + glycine 1.0 mg / L + VB1 0.05 mg / L + 6-BA 0.25 mg / L + sucrose 30 g / L + agar 6 g / L + Kanamycin 30 mg / L + Timentin 300 mg / L ( pH = 6.0 ) ;The optimal rooting culture medium was WPM + IBA 0.25 mg / L + sucrose 30 g / L + agar 6 g / L + Kanamycin 30 mg / L + Timentin 300 mg / L ( pH = 6.0 ).(3) In the optimal differentiation and selection culture medium, the seedling vitrification rate infected by Agrobacterium LBA4404-pBI121 was about 65 %, while the seedling vitrification rate infected by Agrobacterium EHA105-pBI121 was less than 10 %.(4) The rooting efficiency of regenerated seedlings of Lycium ruthenicum could reach about 81.2 % by using low-salt WPM medium of woody plants. (5) The ratio of the number of positive callus to the total number of inoculated leaves was used as the evaluation index of genetic transformation efficiency. In the optimal genetic transformation system, the genetic transformation rates of Agrobacterium LBA4404-pBI121 and EHA105-pBI121 were 51% and 65.2%, respectively. The efficient combination culture system of Lycium ruthenicum leaves can significantly improve its genetic transformation rate and reduce the incidence of vitrified seedlings.