农杆菌介导转录因子CBF转化甘蓝型油菜

文档编号:SW108  文档字数:10758,页数:24

摘 要：油菜是我国最主要的油料作物，有甘蓝型油菜、白菜型油菜和芥菜型油菜等9个种，其中许多是具有重要经济价值的油料与蔬菜作物。但是当环境温度低于10℃～20℃时油菜就会受到伤害，影响它的正常生长发育，严重的甚至造成死亡。目前很多作物在不同程度上都受到低温的影响，低温这一重要的环境因子正严重影响油菜的生长、发展、生存及分布。因此，针对耐逆性（抗寒性）相关的转基因油菜的研究，本试验以成熟油菜“超油2号”种子培养出的下胚轴愈伤组织为受体材料，运用农杆菌介导法将转录因子CBF基因导入供试品种的基因组中，得到的油菜植株经PCR检测，Southern杂交验证分析，获得转基因植株60株，转化频率为1.2%。已转入目的基因植株的耐逆性的鉴定正在准备之中。
关键词：CBF；根瘤农杆菌；PCR；Southern杂交

Agrobacterium tumefaciens-Mediated Transformation of Brassica napus L. with CBF Gene
Abstract: Rape is the most important oilseed crop, there are 9 kinds of rapes, Brassica napus, he cabbage rape, the leaf mustard rape and so on. Many of them have the important economic values. But when the environment temperature is lower than 10℃~ 20℃ rape may be injured, its normal growth would be affected, even worse, it would be died. At present, many crops affected by low temperature in the varying degree. But the low temperature of this important environmental factor that is seriously affecting rape's growth, development, survival and distribution. In this study, a gene for CBF was incorporated into genome of "super-oil No.2", a new Brassica napus L. strain with super-high oil content, by Agrobacterium tumefaciens-mediated transformation with hypocotyl segments as recipient materials. Based on PCR assay and Southern hybridization analysis, 60 transgenic plants were obtained and the transformation frequency was 1.2%. The identification of transgenic plants is being prepared.
Keywords : CBF; Agrobacterium tumefaciens ; PCR; Southern hybridization

 目  录
中文摘要 I
英文摘要 II
目录 III
1. 绪论 1
 1.1 冷驯化及CBF家族概述 1
 1.2 CBF家族与植物抗冻性 3
2. 材料与方法 7
 2.1 材料 7
 2.2 方法 8
3. 结果与分析 12
 3.1 转基因油菜植株的获得 12
 3.2 转化植株PCR检测 13
 3.3 转化植株Southern杂交分析 13
4. 总结与展望 15
致谢 16
参考文献 17

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