农杆菌培养方式和预培养基激素配比对甘蓝型油菜下胚轴转化效应分析  

Effects of Agrobacterium Culture Methods and Hormone Combinations in Pre-culture Medium on Hypocotyle Transformation in Brassica napus

作　　者：孙洁[1] 黄团[1] 李加纳[1] 殷家明[1] 柴友荣[1] 丁彩霞[1] 

机构地区：[1]西南大学农学与生命科技学院,重庆市油菜工程技术研究中心,重庆400716

出　　处：《西南大学学报（自然科学版）》2007年第12期49-53,共5页Journal of Southwest University(Natural Science Edition)

基　　金：国家自然科学基金重点资助项目(30330400);863计划资助项目(2006AA10Z110).

摘　　要：利用根癌农杆菌共培养法转化甘蓝型油菜"华双4号"下胚轴,通过组织化学染色法检测报告基因gus的表达情况,研究了农杆菌培养方式和预培养基激素配比对转化效果的影响.结果表明,用LB培养基活化后的农杆菌再用添加AS的MS培养液继续培养6～30h对转化有明显的促进作用;不同的农杆菌培养方式对转化的影响有明显的差异,在试验的6种农杆菌培养方式中s2-5的转化效果最好,即收集活化后的农杆菌用MS+AS+抗生素重悬培养24h,再用MS+AS培养6h;2,4-D与6BA之间的相互作用对转化有重要影响,在试验的预培养基激素组合中,2,4-D2.00mg/L6BA0.25mg/L的组合转化效率最高;抗性愈伤率与转化效率之间不一定具有相关关系.The hypocotyl of Brassica napus L. ＂Huashuang No. 4＂ was transformed mediated by Agrobacterurn turnefaciens and the expression of the reporter gene gus was detected by histochemical pigmentation. The effects of Agrobacteriurn culture method and hormone combination in the pre-culture medium on transformation were analyzed. The results showed that subculture of Agrobacterium in MS with AS for some time after activating in LB medium promoted transformation. Obvious differences were observed among the six Agrobacterium culture methods adopted, of which s2-5 gave the best transformation result. Combination of 2,4-D with 6-BA in pre-culture had better effect than others. The combination of 2.00 mg/L 2, 4-D and 0. 25 mg/L 6BA was the best in transformation efficiency. The experiments also showed that resistant callus percentage was not necessarily correlated with transformation efficiency.

关 键 词：甘蓝型油菜 下胚轴 农杆菌培养方式 预培养基 转化 

分 类 号：S634.3[农业科学—蔬菜学]