A new time-saving transformation system for Brassica napus

文献类型: 外文期刊

第一作者: Kong, Fanming

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

作者机构:

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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