Establishment Of Callus System Of Populus Tremula ×P.tremuloides And Analysis Of The Somaclonal Variation

In this paper, the leaves, petioles and stems of 20 clones of Populus tremula ×P tremuloides were cultured on WPM, NT, IS, and MS media supplemented with different growth regulators, respectively. A rapid regeneration system was developed via the establisment of callus, the ploidy of regenerated plantlets from calli was surveyed, and the AFLP polymorphism of genome DNA of calli and the regenerated plantlets from calli were performed. The conclusions were as follows:1. The medium WPM+ 1.0 mg/L 6-BA +(0.1-0.5) mg/L NAA+2% sucrose was the most suitable medium for the induction of calli of Populus tremula X P. tremuloides. Adventitious buds could form from calli cultured on this medium, with a differentiation rate of 98.33%.2.The medium WPM+6-BA (1.0 mg/L) +2% sucrose was the most suitable medium for the induction of adventitious buds of Populus tremula X P. tremuloides. Explants of this old hybride aspen, cultured under this condition, were easy to differentiate and form adventitious buds.3. Effects of different growth regulators on regenerated plantlet were different. The variation of chromonsome number of regenerated plantlets induced by 6-BA was smaller than that induced by 2,4-D. The shape of chromonsome was changed in the regenerated plantlets induced by 2,4-D.4. 12 pairs of primer wer screened for AFLP of Populus tremula ×P. tremuloides genomic DNA. AFLP technique was improved to make it suitable for polymorphism analysis of Populus tremula X P tremuloides genomic DNA.5.The AFLP polymorphisms between the regenerated plantlets induced by 6-BA and 2,4-D were different. Among the plantlets induced by 2,4-D, total 3457 fragments were detected, and 895 of them were polymorphism, accounting for 25.89%. While 2867, 565 and 19.71% induced by 6-BA.6. The AFLP polymorphism was related to the culture time. With the increase of culture time, the AFLP polymorphism was increased significantly.7. Medium WPM supplemented with 1.0 mg/L 6-BA was the right medium for rapid propagation of Populus tremula ×R tremuloides with small variation in tissue culture.