【Objective】The objective of this study is to establish an in vitro organ regeneration system for the Liriodendron hybrid elite cultivar L. × sino-americanum ‘Nanlin-Jinsen E1’. This will be achieved using stem segments and shoot apical meristems from the Liriodendron hybrid elite cultivar, which possesses independent intellectual property rights, as experimental materials through tissue culture.【Method】Using stem segments and stem tips as explants, we conducted bud induction, elongation culture, and rooting culture of L. × sino-americanum ‘Nanlin-Jinsen E1’, ultimately obtaining fully regenerated plants and establishing a stable and efficient organ regeneration system in vitro.【Result】The optimal basic medium for inducing adventitious buds is MS medium; the optimal culture medium for inducing adventitious buds from stem explants is MS + 0.50 mg/L 6-BA + 0.100 mg/L NAA + 0.005 mg/L TDZ; the optimal culture medium for inducing adventitious buds from stem tip explants is MS + 0.50 mg/L 6-BA + 0.100 mg/L NAA + 0.020 mg/L TDZ; the optimal medium for plant growth regulators for elongation culture is MS + 0.01 mg/L GA₃; the optimal medium for rooting culture is 1/2 MS + 0.300 mg/L NAA; the optimal sucrose concentration for rooting culture is 20.0 g/L.【Conclusion】Using stem segments and tips as explants, the optimal experimental conditions for inducing adventitious buds, elongation culture, and rooting culture of L. × sino-americanum ‘Nanlin-Jinsen E1’ were obtained. A system for in vitro organ regeneration of L. × sino-americanum ‘Nanlin-Jinsen E1’ was established, providing a new and effective way for the optimal and rapid reproduction of Liriodendron hybrids excellent variety.