红豆草与苜蓿原生质体融合过程中小泡的形成(英文)

VESICLE FORMATION IN FUSION PROCESS OF SAINFOIN AND ALFALFA PROTOPLASTS

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

Protoplast fusion between sainfoin ( Onobrychis viciaefolia Scop) hydroxyproline resistance cell line (Hyp r) and alfalfa ( Medicago sativa L.) cell line transformed with Agrobacterium tumefaciens 702 (M 7) was induced via polyethylene glycol (PEG) solution with high pH and high Ca 2+ concentration. The fusion kinetics exhibited in two paths, the first was that a big vesicle formed near the center of the linked membrane, and the second was that many small vesicles formed along with the disintegration of the linked membrane. The vesicles formed in the fusion process were derived directly from the agglutinated membrane, but not resulted from the fusion of submicroscopic vesicles. Temperature remarkably affected the frequency and the speed of protoplast fusion. Glycerol and dimethylsulfoxide could protect the membrane from bursting, and had a positive effect on fusion. After 20 d by culture, cell colonies consisting of 10～20 cells were formed in selection medium. Fig 3, Ref 6