Abstract: In this research,Lilium var. ‘Constanta’(oriental lily hybrid)was chosen as female parent, while the endemic wild lily in China. Lilium regale Wilson was regarded as the male parent. Direct stigma pollination, cut-style pollination, and cut-style pollination with smear stigma fluid were carried out for pollination. Ovaries with different pollination method were collected 20 d after pollination and sterilized, the ovaries were transversely cut into 0.3 cm slices, and were embryo rescue cultured on the MS medium supplemented with 0.01 mg/L 6-BA and 0.1 mg/L NAA. SPAP method was used to identify the strains after embryo rescue culture. The results showed that stigma pollination was the most appropriate pollination method. Through the sliced ovaries rescue culture, seven strains were obtained and all the strains exist typical two parents’ molecular band. In conclusions, the barriers would appear after fertilization in this distant hybridization between Lilium var. ‘Constanta’ and Lilium regale Wilson. And all the seven rescued strains are hybrids.