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A new time-saving transformation system for Brassica napus

文献类型： 外文期刊

作者： Kong, Fanming 1 ; Li, Juan 1 ; Tan, Xiaoli 1 ; Zhang, Lili 1 ; Zhang, Zhiyan 1 ; Qi, Cunkou 2 ; Ma, Xiaoke 1 ;

作者机构： 1.Jiangsu Univ, Inst Life Sci, Zhenjiang 212013, Jiangsu, Peoples R China

2.Jiangsu Acad Agr Sci, Inst Ind Crop, Nanjing 200014, Peoples R China

关键词： Agrobacterium tumefactiens;cotyledonary node;6-benzyladenine;naphthalene acetic acid;transformation;Silwet L 77

期刊名称：AFRICAN JOURNAL OF BIOTECHNOLOGY （ 影响因子：0.573； 五年影响因子：0.794 ）

ISSN： 1684-5315

年卷期： 2009 年 8 卷 11 期

页码：

收录情况： SCI

摘要： By optimizing different parameters for infection, an efficient Agrobacterium-mediated transformation system for Brassica napus was developed. A series of combinations of optical densities, infection durations, concentrations of acetosyringone and silwet L77 were evaluated. Maximum transformation efficiency reached 18.93% when cotyledonary nodes were infected for 10 min with an Agrobacterium suspension of 0.8 OD and 200 mu M acetosyingone and 0.02% silwet L77 in the culture medium. In addition, the concentrations of 6-benzyladenine and naphthalene acetic acid were also optimized. Meanwhile, maximum transformation efficiency for green plantlet was 94.38% when cotyledonary nodes were cultivated in the medium containing 4 mg/L 6-benzyladenine and 0.04 mg/L naphthalene acetic after co-cultivation. Transformation was confirmed by polymerase chain reaction analysis, southern blotting analysis and green fluorescent protein assay.

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