84K杨基因转化受体系统的建立

以84K杨为研究对象,通过建立叶片再生系统及卡那霉素敏感性试验,确立了稳定高效的基因转化受体系统。结果表明,以MS培养基为基本培养基,0.5～1.0mg/L6-BA+0.01～0.1mg/LNAA组合时,叶片出芽率不到50%,每叶平均生芽数5～6个;1.0～1.5mg/L6-BA+0.02mg/L2,4-D组合时,叶片出芽率达90%～100%,每叶平均生芽数约20个。用后者附加不同质量浓度梯度卡那霉素,确定叶片外植体芽诱导卡那霉素临界筛选质量浓度为20mg/L。利用对84K杨芽生根效果较好的培养基GMS+0.01mg/LNAA+0.25mg/LIBA,附加不同质量浓度梯度卡那霉素,筛选出芽生根卡那霉素临界质量浓度亦为20mg/L。

Establishment of gene transformation receptor system for poplar 84K

Through the establishment of high frequency regeneration system for leaf and kanamycin sensitivity tests,a stable and efficient gene transformation receptor system for poplar 84K was set up,which laid a sound foundation for Agrobacterium-mediated transformation of disease resistance gene by means of leaf disc method.The results indicated that budding percentage was less than 50% and average adventitious buds per leaf was 5-6 when basal MS medium supplemented with 0.5-1.0 mg/L 6-BA+(0.01-)0.1 mg/L NAA.While 1.0-1.5 mg/L 6-BA+0.02 mg/L 2,4-D were added to MS medium,(90%-100%) budding percentage and 20 average adventitious buds per leaf were obtained.Utilizing the latter medium,adding different concentrations of kanamycin,the optimal selecting pressure 20 mg/L of kanamycin for adventitious bud was confirmed.GMS+0.01 mg/L NAA+0.25 mg/L IBA was better to induce root,by which the optimal selecting pressure for root induction was also 20 mg/L of kanamycin for poplar 84K when adding kanamycin different concentrations.