Abstract: The Populus nigra × P. deltoides, the hybrid offspring of Aigeiros, is useful in the pathology of poplar against the infection of rust. The petiole, leaves and stem segments were cultured as explants to induce seeding. Optimal conditions of the regeneration system were determined for the primary culture, subculture, stem elongation culture and rooting culture. While the best primary culture medium (MS 0.5 mg/L 6-BA 0.05 mg/L NAA) was used, 86.7% of petioles transformed to seedlings. Further more, an adventitious bud induction rate of 96.7% was got when the subculture medium (MS 0.5 mg/L 6-BA 0.1 mg/L NAA) was adopted. A stem elongation rate of 91.7% was got when the stem culture medium (MS 0.1 mg/L 6-BA 0.05 mg/L NAA) was used. While the root media (1/2MS 0.25 mg/L NAA 20 g) was used, 85% of the seedlings were rooted. The survival rate of the regenerated seedlings was 91.5%. The Populus nigra × P. deltoides regeneration system can be rapidly propagated and used in gene inheritance and transformation. It also can provide material basis for the study of poplar susceptibility/resistance to the rust fungi.