The Establishment And Optimization Of The Apple (Malus Pumila Mill.) Leaf-explants Regeneration System

In this experiment , as the experimental materials genotype three kind of the apple (Malus pumila Mill.)cultivars of Royal Gala,Yanfu No.10 and Changfu No.2,the apple asepsis quick numerous system were establishmented by using for initially generation of culture medium for neen't increase any hormone in MS; subculture stem generation of raise to increase the best hormone combination on the MS culture medium is: Imperial golmo to lasa: 6-BA 0.5mg/L+NAA 0.05 mg/L; Yanfu No.10, Changfu No.2: 6-BA 0. 5~1.0mg/L+NAA 0.05~0.10 mg/L.In the apple leaf blade regeneration adventitious bud experiment, studied the test tube seedling leaf blade regeneration adventitious bud separately to plant the body genotype, the nitrogen shape and the proportion with outside(NH4+:NO3-), the plant hormone combination, the dark raise time, the leaf blade seedling age and the different raise method various factorss relations,Discovered the most suitable leaf blade regeneration adventitious bud raise condition, established optimizes stably, the highly effective exsomatize leaf blade regeneration adventitious bud system.The experiment screens the imperial golmo to lasa highly effective adventitious bud induction culture medium is: The increase nitrogen shape and proportion(NH4+:NO3-)=2:0, the additional hormone combination and the density are the 6-BA 2.0mg/L+ NAA 0.15mg/L MS culture medium; Screens Fuji is Yanfu No.10, Changfu No.2highly effective adventitious bud induction culture medium is: Attaches 6-BA 4.0mg/L+ NAA 0.15mg/L the MS culture medium.Choose the 20~25d seedling age middle color deeply in the test tube seedling leaf blade green, the growing trend strong leaf blade outside took plants the body to be advantageous to the leaf blade regeneration; The dark raise adventitious bud to the apple leaf blade regeneration by no means essentially, after the vaccination the complete illumination raise leaf blade also can regenerate, but may obviously enhance the leaf blade after the 14d dark raise the regeneration bud number and the regeneration frequency; Yanfu No.10 in after the melon seed leaf, the imperial golmo to lasa leaf blade altogether raise, the leaf blade color deepens, expands the curl to be obvious, injuries the organization condition to be advantageous in adventitious bud regeneration; 1μmol/L AgNO3 to adventitious bud the leaf blade regeneration to have the remarkable promoter action, the leaf blade expands rapidly, injuries the organization to form quickly, the leaf blade regeneration percentage achieves 100%, average each regeneration bud number 5.86. The regeneration adventitious bud to take root the available non- phaseomannite 1/2MS culture medium, additional hormone IBA 0.5 mg/L, the average effective radical may reach 4.6, takes root rate to reach 82%.