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影响根癌农杆菌介导的香蕉基因转化早期的主要因素
引用本文:黄霞,黄学林,李哲,陈云凤,李筱菊.影响根癌农杆菌介导的香蕉基因转化早期的主要因素[J].中山大学学报(自然科学版),2002,41(5):68-72.
作者姓名:黄霞  黄学林  李哲  陈云凤  李筱菊
作者单位:中山大学生命科学学院∥基因工程教育部重点实验室,广东,广州,510275
基金项目:广东省科技厅科技攻关资助项目 (2KM0 3310N),广东省自然科学基金资助项目 (70 112 6 )
摘    要:用含质粒载体pCAMBIA2301的根癌农杆菌(Agrobacterium inoculation)转化“641”香蕉(Musa AAA Cavendish subgroup cv.64-1)的薄片外植体,通过测定GUS基因瞬时表达率,对转化早期的主要影响因素进行了研究。结果表明:农杆菌EHA105较适合于介导转化,将薄片置于固体高渗培养基上培养4h后,通过真空减压法使农杆菌接种于其上,获得了较高的瞬时表达率(41.67%),是对照(8.33%)的5倍,农杆菌悬液稀释5倍用于转化较好,共培养3d为宜,薄片外植体越靠近根部,瞬时表达率越高。而外植体预培养会降低瞬时表达率。

关 键 词:根癌农杆菌  香蕉  基因转化  瞬时表达  转基因植物  影响因素
文章编号:0529-6579(2002)05-0068-05
修稿时间:2002年2月1日

Factors Affecting the Early Phase of Agrobacterium- mediated Genetic Transformation of Banana
HUANG Xia,HUANG Xue_lin,LI Zhe,CHEN Yun_feng,LI Xiao_ju.Factors Affecting the Early Phase of Agrobacterium- mediated Genetic Transformation of Banana[J].Acta Scientiarum Naturalium Universitatis Sunyatseni,2002,41(5):68-72.
Authors:HUANG Xia  HUANG Xue_lin  LI Zhe  CHEN Yun_feng  LI Xiao_ju
Abstract:Micro_cross sections of '641' banana (\%Musa\% AAA Cavendish subgroup cv\^64_1) were used as the receptors for transformation by \%Agrobacterium tumefaciens \% containing pCAMBIA2301. By detecting the transient expression of GUS gene, the factors affecting the early phase of \%Agrobacterium\%_mediated transformation of banana (\%Musa\% spp\^) were investigated. The results indicated that in comparison with LBA4404, EHA105 was the more suitable strain for the transformation. The transient expression rate of GUS gene was increased 5 folds when the explant was pretreated with 0\^2 mol/L mannitol and the \%Agrobacterium\% inoculation was disposed by negative pressure produced by vacuum pump. Also, 3 day_coculture and 5 times dilution of the bacterium suspension were more suitable for the transformation. In addition, the explant, micro_cross section, obtained from closer to the root showed higher transient expression rate. But the preculture of explant was unfavourable for the banana transformation.
Keywords:banana (\%Musa\% spp\^)  \%Agrobacterium tumefaciens  \%  transformation  transient expression
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