Establishment of tissue culture and regeneration system for Xanthoceras sorbifolium Bunge
[Objective]To establish the embryogenesis and regeneration plant system of Xanthoceras sorbifolium Bunge,the high efficiency regeneration system established can lay a foundation for the genetic engineering of cultivating excellent strains and the genetic transformation of the fruit.[Method]The orthogonal experiments of different concentrations of auxin and cytokinin were designed with the young embryo,middle embryo and mature embryo of X.sorbifolium as explants.Through comparative experiments,the effects of different explants and different hormone concentration ratios on improving the callus induction rate of X.sorbifolium were determined,and the optimal basic medium and concentration ratio of each hormone for the callus induction proliferation,adventitious bud differentiation,elongation and rooting culture of X.sorbifolium were determined,so as to establish the tissue culture and regeneration system of X.sorbifolium.[Result]In the ratio of 1 mg/L 6-BA and 0.5 mg/L NAA,the callus with bright green and dense structure was induced,and the induction rate was 80.46% .The best variety to induce callus formation is'WF15'metaphase embryo.In the ratio of 1mg/L 6-BA and 0.3 mg/L IBA,a large number of callus showed bright green buds,some of them produced clumpy buds,and the highest induction rate of adventive buds was 75% .In the hormone ratio of 1.0 mg/L TDZ and 0.2mg/L IAA,the adventive buds could continue to grow well.After 30 days of subgeneration,each adventive bud differentiated into 3-4 branches with branch height of about 5cm.The best variety was'WF19'.The rooting rate of 0.3 mg/L IBA was the highest,the highest was 88.35%,and the length of the generated roots was about 9 cm.When the opening time was 36 h,the seedlings grow well and the survival rate of transplanting was 73.33%,which was significantly higher than other opening time.[Conclusion]The best medium for inducing callus formation is MS+1 mg/L 6-BA+0.5 mg/L NAA+30 g/L sucrose+7 g/L AGAR.The optimal medium for inducing adventitious bud formation is WPM+1 mg/L 6-BA+0.3 mg/L IBA+30 g/L sucrose+7 g/L AGAR.The medium for inducing adventive bud proliferation and differentiation is MS+1.0 mg/L TDZ+0.2 mg/L IAA+30 g/L sucrose+7 g/L AGAR.The optimal medium for rooting induction is 1/2 ms+0.3 mg/L IBA+30 g/L sucrose+7 g/L AGAR.
Xanthoceras sorbifolium Bungeexplanttissue cultureregeneration system
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