In order to cultivate new varieties of Lilium lancifolium with strong resistance and excellent ornament, the pollen viability of three ornamental lily cultivars ‘Black stone’, ‘Tresor’ and ‘Sorbonne’ was measured by liquid medium hanging drop method, which were respectively used to conduct distant crosses with female parent Lilium lancifolium Thunb. using bud stage pollination, normal pollination, delayed pollination, and stigma-cutting pollination. Ovules of the obtained hybrid progeny were cultured and multiplied, and the authenticity of the hybrid was identified by the method of SSR molecular marker. The results showed that the pollen germination rate of ‘Sorbonne’ was generally higher than that of ‘Black stone’ and ‘Tresor’; the seed setting rate with pollination at the bud stage was the highest, which was 14.44%; the hybrid combination with the largest number of capsules is Lilium lancifolium Thunb. × ‘Black stone’, with 17 capsules. Ovules of the hybrid capsules were cultured, and 9 F1 generation hybrid seedlings were obtained. The optimum medium formula for hybrid seedling propagation is MS+30 g/L sucrose+0.1 mg/L NAA+1 mg/L 6-BA+8 g/L agar. The optimal medium formula for bulb expansion and rooting is MS+80 g/L sucrose+0.1 mg/L NAA+ 0.05 mg/L 6-BA+0.75 g/L activated carbon+6.5 g/L agar, and the optimal culture environment is dark nourish. Finally, the two hybrid lines of Lilium lancifolium Thunb. × ‘Tresor’ were confirmed to be true hybrids through SSR molecular mark identification using four pairs of primers.