| 番茄无选择标记转化体系的研究 |
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| 引用本文： | 邹芳玲,国艳梅,杜永臣,祝光涛,高建昌,胡鸿.番茄无选择标记转化体系的研究[J].中国蔬菜,2013,1(8):19-25. |
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| 作者姓名： | 邹芳玲 国艳梅 杜永臣 祝光涛 高建昌 胡鸿 |
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| 作者单位： | 中国农业科学院蔬菜花卉研究所，北京 100081 |
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| 基金项目： | 国家自然科学基金项目(30900988/C110502);农业部园艺作物生物学与种质创制重点实验室资助项目 |
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| 摘 要： | 以含多毛番茄GGPS 基因的表达载体pBI121-GGPS 和无标记载体pBINMF-GUS 为基础，利用载体pBINMF-GUS 的T-DNA 区无选择标记基因的特点，将目的基因GGPS 连接于pBINMF-GUS 的T-DNA 中，用农杆菌菌株EHA105、C58 介导转化番茄品种中蔬6 号，利用PCR 直接筛选转化体。结果表明：MS+1.0 mg·L-1 ZT+0.5 mg·L-1 IAA 为子叶最佳芽诱导培养基，农杆菌菌株EHA105 更利于中蔬6号的转化，获得阳性植株4 株，转化率为3.6%。PCR 及RT-PCR 检测结果表明：目的基因已整合到中蔬6 号基因组中，且在转录水平上得到表达。
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| 关 键 词： | 番茄 根癌农杆菌 无选择标记 遗传转化 |
Studies on Tomato Marker-free Transformation System |
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| ZOU Fang-ling,GUO Yan-mei,DU Yong-chen,ZHU Guang-tao,GAO Jian-chang,HU Hong.Studies on Tomato Marker-free Transformation System[J].China Vegetables,2013,1(8):19-25. |
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| Authors: | ZOU Fang-ling GUO Yan-mei DU Yong-chen ZHU Guang-tao GAO Jian-chang HU Hong |
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| Institution: | Institute of Vegetables and Flowers， Chinese Academy of Agricultural Sciences， Beijing 100081， China |
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| Abstract: | In this paper，the aimed gene GGPS was cut from the binary plant expression vector pBI121-GGPS and ligated to the marker-free vector pBINMF-GUS，so a new marker-free vector was formed，in which T-DNA segment only harboured GGPS gene and a GUS gene. This vector was successfully transferred into tomato（Lycopersicum esculentum Mill. ） cultivar‘ Zhongshu No. 6’ via Agrobacterium-mediated transformation used strains EHA105 and C58. The results showed that MS+1.0 mg·L-1 ZT+0.5 mg·L-1 IAA was the best shooting medium for‘ Zhongshu No. 6’. The transformation rate of strain EHA105 was 3.6%，which was better than the rate of strain C58. The results of PCR analysis indicated that GGPS gene was integrated into the genome of‘Zhongshu No. 6’. RT-PCR detection indicated that GGPS gene was expressed at RNA level. |
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| Keywords: | Agrobacterium tumefaciens Agrobacterium tumefaciens Marker-free Genetic transformation |
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