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通过高钙离子浓度、高pH值方法结合PEG处理诱导红豆草羟脯氨酸抗性系和苜蓿根癌农杆菌转化系原生质体融合 .融合机制表现为两种形式 ,第一种为相连膜在中间部位形成一个大的小泡 ,第二种为随相连膜的解体产生很多较小的泡状结构 .融合过程中形成的小泡直接来自凝集膜 ,而不是亚显微小泡相互融合的产物 .温度明显影响原生质体融合的频率和速度 .甘油和二甲基亚砜可以防止膜破裂 ,从而促进原生质体融合 .培养 2 0d以后 ,在选择培养基得到含 10～ 2 0个细胞的细胞团 .图 3参 6
The combination of high calcium concentration and high pH with PEG treatment induced the hydroxyproline resistance of the hybrids and the protoplast fusion of the A. tumefaciens Agrobacterium tumefaciens transformation system.The fusion mechanism appeared in two forms, The middle part forms a large vesicle and the second one produces many smaller vesicular structures with the disintegration of the connected membrane.The vesicles formed during the fusion process come directly from the aggregated membrane rather than from the product of the fusion of the sub-vesicles Temperature significantly affects the frequency and speed of protoplast fusion Glycerol and dimethylsulfoxide prevent protoplast fusion by preventing membrane rupture.After 20 days of culture, cell clusters containing 10 to 20 cells are obtained in selection medium Figure 3 Reference 6