Studies On Callus Induction And Protoplast Isolation On Ziziphus Jujuba Mill

In this study, plantlets leaves of DongZao, plantlets twig and anther of yellow bud of DongZao were used as materials to treat the conditions of inducement callus, and plantlets leaves of DongZao, plantlets leaves of sour jujube tetraploid, suspension culture of anther of DongZao were used as materials to research the feasible conditions of protoplast isolation, including Cellulase "onozuka" R-10 and Macerozyme R-10 s' concentration, Mannital concentration, isolation time and isolation speed.1. Plantlet leaves of DongZao weren't feasible to induce callus. Used plantlet twig of DongZao to induce calls, the feasible culture medium was l/2MS+ZT3.0mg/L+NAA10.0mg/L or l/2MS+ZT0.5mg/L +NAA2.0mg/L and the state of callus were perfect. The feasible culture medium of anther to induce was l/2MS+TDZ0.4mg/L+PVP2.0mg/L.2.The multiplication culture medium of suspension culture of anther of DongZao was 1/2MS+TDZ0.4. After regeneration 7~9 days, suspension culture was used to isolation. Regeneration time was too short, the callus were too little to isolation;more than 10 days, the color of callus would become brown, vitality would reduce, the callus were not fit to isolate protoplast.3. Regeneration culture medium of anther callus was l/2MS+TDZ0.4mg/L, the callus' regeneration was quick and quantity was large. The callus could keep hearty vitality after two years.4.The favorable materials to isolate protoplast were plantlets leaves, suspension culture of anther. The optimal enzyme solution used for protoplasts isolation for plantlets leaves of DongZao was 0.7mol/L Mannital+1%MES +5g/L Cellulase "onozuka" R-10,5.The optimal enzyme solution used for protoplasts isolation for plantlets leaves of sour jujube tetraploid was 0.6mol/L Mannital +l%MES+10g/L Cellulase "onozuka" R-10+4— 5 g/L Macerozyme R-106.The optimal enzyme solution used for protoplasts isolation for suspension culture of anther was 0.6mol/L Mannital+1%MES+25g/L Cellulase "onozuka" R-10.7. The optimal centrifugal speed was 1010r/min during purification, 16 min was good. Grads purification's speed was 870 r/min, 7~8min. The speed which was too quick or too slow affected protoplasts' yield and viability.8. The optimal isolation time of Ziziphus jujube was 16~20h. The time was too short, the enzyme solution couldn't effect sufficiently;the time was too long, the protoplast would fracture to die.