双价表达载体转化辣椒的体系优化

Transformation system optimization of binary expression vector with resistance to virus and insect on pepper

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4～6 d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/L NAA＋0.1mg/L IAA.经过优化的甘露糖体系筛选共获得28株To代转基因植株,其中阳性率为21.4％,转化率为2.5％.

To establish the efficient Agrobacterium-mediated transformation system of pepper （Capsicum annuum L.） Flamingo-bill explant, the binary expression vector containing Cry2A α2 and PamPA P was transferred into pepper mediated by Agrobacterium tumefaciens and the transformation system was optimized. The results showed that adventitious differential 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 cocuhivation. 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.