番茄再生体系优化以及ProDH干扰载体的遗传转化

【目的】为了进一步探究番茄再生体系,对番茄再生体系进行优化,并利用筛选出的最优体系,将脯氨酸脱氢酶的干扰表达载体进行遗传转化,抗生素筛选获得再生植株。【方法】以44号和65号番茄品种为试验材料,外植体的选择有下胚轴和子叶,以MS为基本培养基,加入不同浓度比的6-BA和IAA,筛选合适的浓度比,来提高遗传转化率。【结果】结果表明：最适的愈伤培养基为MS + 2 mg?L-1 6-BA + 0.2 mg?L-1 IAA,最适分化培养基为MS + 2.0 mg?L-1 6-BA + 0.1 mg?L-1 IAA,最适生根培养基为MS + 0.2 mg?L-1 IAA；最适培养基浓度下,子叶的出愈率、分化率和生根率较下胚轴分别高25%、42%和20%,所以外植体选择子叶为宜,而在两个品种间也有差异,65号番茄总体比44号番茄再生率高。【结论】利用筛选出的最优再生体系,将脯氨酸脱氢酶的干扰表达载体对65号番茄遗传转化,获得抗性植株,以期为番茄抗寒性分子育种提供更多基础材料。

Optimization of Tomato Regeneration System and Genetic Transformation of ProDH Interference Vectors

To optimize the tomato regeneration system and screen out the optimal one, and at the same time, the ProDH interfering expression vector was genetically transformed to tomato, and regenerated plants were obtain by antibiotics screening. The test materials were hypocotyls and cotyledons of tomato variety‘Naiyun 2000? and‘O-33-1?, the basic medium was MS which respectively contained 1.0, 2.0, 3.0 mg/L 6-BA and 0.1, 0.2, 0.3 mg/L IAA. The appropriate concentration combinations were selected. The results showed that the optimal callus culture medium was MS + 2 mg/L 6-BA + 0.2 mg/L IAA, and the optimal differentiation medium was MS + 2.0 mg/L 6-BA + 0.1 mg/L IAA. The optimal rooting medium was MS + 0.2 mg/L IAA; in the optimum medium, the emergence rate, differentiation rate and rooting rate of cotyledon were higher than that of hypocotyls by 25%, 42% and 20% respectively, so the cotyledon was appropriate for explants. Meanwhile, there was also a difference between the two varieties, and‘O-33-1? had a higher regeneration rate than that of ‘Naiyun2000?. Using the optimized regeneration system, the ProDH interference expression vector was genetically transformed to‘O-33-1? to obtain resistant plants.