Agrobacterium-mediated transformation of Eupatorium adenophorum

文献类型： 外文期刊

第一作者： Guo, Huiming

作者： Guo, Huiming;Zhang, Yongqiang;Cheng, Hongmei;Wan, Fanghao

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关键词： kanamycin: 8063-07-8;cefotaxime: 63527-52-6;naphthaleneacetic acid: 26445-01-2;6-benzyladenine: 1214-39-7;pBI121;callus formation

期刊名称：PLANT CELL TISSUE AND ORGAN CULTURE （ 影响因子：2.711； 五年影响因子：2.73 ）

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年卷期：

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收录情况： SCI

摘要： A simple and efficient protocol for Agrobacterium tumefaciens-mediated genetic transformation of Eupatorium adenophorum has been developed. Stem segments were pre-cultured on Murashige and Skoog (MS) medium for 24 h and then co-cultured with A. tumefaciens strain LBA4404 harboring the pBI121 plasmid on the same medium in darkness for 3 days. After co-cultivation, explants were transferred to MS medium supplemented with 1.0 mg l(-1) naphthaleneacetic acid (NAA), 1.0 mg l(-1) 6-benzyladenine (BA), 150 mg l(-1) kanamycin (Kan), and 200 mg l(-1) cefotaxime (Cef), and incubated at 25 +/- 1 degrees C under a 16-h photoperiod. Callus formation was induced for 3-4 weeks. Induced calluses were transferred to fresh selection medium and subcultured every 15 days. After 6-8 weeks, regenerated plantlets from embryogenic callus could be observed. Buds of 1 cm in length were transferred to half-strength MS medium containing 150 mg l(-1) Kan and 200 mg l-1 Cef, and rooted plantlets were obtained after 4 weeks. Putative transgenic plants were confirmed using GUS histochemical assay, polymerase chain reaction (PCR), and reverse transcriptase (RT)-PCR. A transformation efficiency of 2% was obtained.

分类号： Q942

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