| 双价表达载体转化辣椒的体系优化 |
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| 引用本文： | 高玉尧,许文天,黄皓,陈长明,曹必好,陈国菊,刘实忠,雷建军.双价表达载体转化辣椒的体系优化[J].广东农业科学,2014,41(13):132-135. |
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| 作者姓名： | 高玉尧 许文天 黄皓 陈长明 曹必好 陈国菊 刘实忠 雷建军 |
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| 作者单位： | 华南农业大学园艺学院，中国热带农业科学院湛江实验站 |
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| 基金项目： | 国家“863”计划项目（2009AA10Z104）；广东省重大科技专项（2010A020102001） |
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| 摘 要： | 将含有PamPAP和Cry2Aa2的双价表达载体通过农杆菌介导法转化辣椒,并对其遗传转化体系进行优化，从而建立辣椒Flamingo-bill农杆菌介导的高效遗传转化体系。结果表明：Flamingo-bill外植体的不定芽容易伸长,不定芽分化率明显高于带柄子叶;延迟筛选4~6d可显著提高Flamingo-bill外植体不定芽的分化率和抗性率；Flamingo-bill外植体最适筛选浓度为甘露糖18g/L和蔗糖12 g/L；抗性苗生根最佳培养基为MS+0.2 mg/LNAA+
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| 关 键 词： | 辣椒 遗传转化 Flamingo-bill外植体 PMI/甘露糖筛选 |
Transformation system optimization of binary expression vector with resistance to virus and insect on pepper |
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| Abstract: | To establish the efficient Agrobacterium-mediated transformation system of pepper (Capsicum annuum L.)Flamingo-bill explant, the binary expression vector containing Cry2Aa2 and PamPAP was transferred into pepper mediatedby Agrobacterium tumefaciens and the transformation system was optimized. The results showed that adventitiousdifferential rates of Flamingo-bill was significantly higher than that of cotyledon with petiole in three pepper cultivars, and the adventitious shoots of Flamingo-bill were easier to elongate. The highest adventitious differential rate and resistant rate were obtained when the Flamingo -bill explants were selected at 4 ~6 days later after cocultivation. The optimal concentration for screening the transformed Flamingo-bill explants was 18 g/L mannose and 12 g/L sucrose. The optimal
rooting medium for resistant seedlings was MS+NAA 0.2 mg/L+IAA 0.1 mg/L. 28 transgenic pepper plants were obtained with optimizated PMI/Man selection system, of which the positive and transformation rate were 21.4% and 2.5% , respectively. |
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| Keywords: | pepper genetic transformation Flamingo-bill explant PMI/Man selection system |
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