影响基因枪轰击转化棉花胚性愈伤的因素初探

Elementary Exploration on Factors of Influencing Transformation via Particle Bombardment Embryonic Calluses of Cotton

本研究就基因枪法转化棉花胚性愈伤过程中的影响因素进行了探讨。结果表明,2,4 D在愈伤启动阶段很重要,而在胚性愈伤诱导阶段要尽量少加或不加,IAA的浓度比KT低时,有利于胚性愈伤的产生。胚性愈伤的表观状态、轰击次数和轰击距离是基因枪转化成功的关键因素。通过GUS基因在棉花胚性愈伤中的瞬间表达证明,在轰击前4h,轰击后16h采用0.3mol·L 1的甘露醇做渗透处理,轰击距离9cm,轰击次数2次,效果最好。

In the article,we studied the different factors influencing the tissue culture and biolistic bombardment in cotton transfer.The results showed 2,4-D is one of the key factors of callus initiation. In the process of the embryogenic calluses induction, it is benefit to production of embryogenic calluses that the concentration of 2,4-D was low or zero and the concentration of IAA was lower than KT.The embryogenic calluses physiology state, the distances and the times of bombardment were very important for success of transfermation. By the transient expression of GUS gene,the results suggested that the optimums of target distance and times of bombardment were 9 cm and 2, respectively.The osmotic treatment of embryogenic calluses with (0.3) mol·L^(-1)mannitol for 4 h before and 16 h after bombardments was the best combination.