Study On Tissue Culture And Plant Regeneration Of Punica Granatum Var.flavescens SW.

The study was mainly to probe an efficient way through tissue culture and rapid propagation of Punica granatum var.flavescens SW..Stems with axillary buds,leaves and calyx of young branches were taken as explants.Through using orthogonal experimental design to establish the technique system of tissue culture and plant regeneration,the affected factors of propagation,existing problems in this technique and the corresponding solutions,the best medium and culture progress are also described, which provide the theoretical base and technical measures for industrial producing of plantlet.The main results as follows:1.The most suitable season of taking explants:The most suitable season of taking explants on Punica granatum var.flavescens SW.is spring,the starting rate,the rate of forming callus are the highest, also the browning rate and contaminated rate are the lowest,especially on April.2.Anti-browning research:Every part of Punica granatum var.flavescens SW.distribute the antioxidant which can induce different browning levels.Then it can reduce the browning rate through putting the additions into the medium,so the treatment with Vc100mg/L and the treatment with AC1g/L are better,the browning rate individually contain 18.89%and 20.74%.3.Initial culture of stems with axillary:Through totally analyzing,the optional culture medium of initial culture was MS+0.05mg/L+1.0mg/L,as the highest initial rate was 76.67%,the highest callus induction rate was 43.33%,the highest shoot forming index was 3.3.4.Callus induction of leaves and calyx:The optional culture medium for leaves was MS+NAA0.2mg/L+BA4.0 mg/L,the highest initial rate was 59.39%;The optional culture medium for calyx was MS+NAA0.1mg/L+BA2.0 mg/L,the highest initial rate was 60.00%;5.Proliferating and differentiating cultivation:The optional proliferating shoots culture medium from stems with axillaries was MS+NAA0.1mg/L+BA2.0mg/L,on which it could multiply 4.8 times after 30d,and multiplication of efficiency shoot can contain 3.5;The optional proliferating callus culture medium for leaves was MS+2,4-D0.1mg/L+NAA0.2mg/L+BA2.0mg/L,proliferating rate was 76.67%;The optional proliferating callus culture medium from calyx was MS+2,4-D0.1mg/L+NAA0.1mg/L+BA2.0mg/L,proliferating rate was 56.67%.6.Rooting culture:Above three standards including the rooting rate,the root number,the root length,totally analysis from statistic show that the optional rooting culture medium from regenerated shoots was 1/2MS+è”—ç³–20g/L +NAA0.1mg/L +IBA 1.0mg/L+AC0.5g/L,rooting rate was 78.33%, root number was 38.00,root length was 2.47cm.7.Transplanting:Plantlets were transferred to the medium with vermiculite,ruby mica and sands which containing ratio of 1:2:1.The survival rate was up to 66.67%,and plantlets grow well.