选择新疆杨(Populus alba L.var.pyramidalis Bge.)为遗传转化受体材料,为建立根癌农杆菌介导新疆杨高效遗传转化系统,从预培养时间、侵染时间、共培养时间、添加乙酰丁香酮(AS)的时机、共培养培养基中添加乙酰丁香酮浓度、侵染菌液的制备方法、外植体继代方式等7个方面优化筛选。结果显示较合适的转化系统为:预培养8h,农杆菌菌液(OD600=0.4)侵染15min,共培养5d,侵染菌液的最优制备方法是液体培养活化农杆菌2次加离心收集菌体重悬,共培养培养基中添加乙酰丁香酮80μmol/L。新疆杨叶盘转化频率可达38.10%。

Several factors affecting transformation of Populus alba L. var. pyramidalis Bge. were studied by infecting with Agrobacterium tumefaciens LBA4404, such as pre-culture time, infection time, co-culture time, acetosyringone ( AS) appending method, AS concentration in the co-culture medium, bacterium liquid preparation method, explants state. An effective protocol in optimized conditions which were 8 h for preculture , 15 min for infection, 5 d for co-culture and dosage 80 μ mol/L AS in co-culture medium for transformation of leaf discs of P. alba var. pyramidalis was developed and the frequency of transformation reached to 38.10%.