In order to optimize the tissue culture and rapid propagation system for Lilium, L. oriental ‘Sorbonne’, L. sulphureum Baker and L. formolongi ‘Raizan 3’ were studied, using tissue culture technology, to explore the optimum conditions of three different explants’ induction and differentiation, plantlets proliferation and bulblets enlargement. The results showed that the suitable medium for seeds germination of L. sulphureum Baker was MS medium supplemented with 0.01 mg/L NAA and 60 g/L sugar, while MS medium supplemented with 2.0 mg/L 6–BA, 0.3 mg/L NAA and 0.5 mg/L 6–BA, 0.2 mg/L NAA were the best to promote the induction of adventitious shoots for ‘Sorbonne’ and ‘Raizan 3’ bulb scales respectively. The optimal medium for plantlets proliferation from three different genotypes was MS medium containing 2.0 mg/L 6–BA and 0.2 mg/L NAA. In addition, 3MS medium with 0.5 g/L AC, 3.0 mg/L PP333 and 80 g/L sugar was fitted for ‘Sorbonne’ bulblets enlargement.