LI Hui,WU Tian-long.TRANSFOMING AGROBACTERIUM INTO SOYBEAN BY MEANS OF POLLEN TUBE PATHWAY INDUCED BY CaCl2[J].Soybean Science,2007,26(01):55-59.[doi:10.3969/j.issn.1000-9841.2007.01.013]
CaCl2诱导大豆花粉管通道农杆菌转基因研究
- Title:
- TRANSFOMING AGROBACTERIUM INTO SOYBEAN BY MEANS OF POLLEN TUBE PATHWAY INDUCED BY CaCl2
- 文章编号:
- 1000-9841(2007)01-0055-05
- 关键词:
- 花粉管通道; 大豆; CaCl2; 质粒pCAMBIA3301; 农杆菌
- Keywords:
- Pollen tube pathway; CaCl2; Soybean; pCAMBIA3301; Agrobacterium
- 分类号:
- S565.103.52
- 文献标志码:
- A
- 摘要:
- 为了提高大豆花粉管通道法转基因效率,研究直接用农杆菌转化方法的可行性,以秋大豆品种上海本地青为受体材料,用花粉管通道法在三种CaCl2浓度水平下进行质粒pCAMBIA3301 转化,又以GV3101、LBA4404、EHA105三种农杆菌菌株直接导入花粉管通道为处理进行了遗传转化。以质粒为外源基因平均结实率为36.83%,T0代种子平均成活率为60.89%,T1植株喷施除草剂后平均存活率为8.24%。以农杆菌直接转化平均结实率为15.92%。T0代种子平均成活率为20.40%,T1植株喷施除草剂后平均存活率为7.79%。对存活的植株提取DNA进行PCR检测,以质粒为外源基因的处理共获得20株转化苗,三种CaCl2浓度水平的转化效率分别为0.56%、1.64%、1.40%,有两种水平的转化效率约为传统转化效率的三倍。以农杆菌直接转化的处理共获得5株转化苗,转化效率分别为0.17%、0.33%、0.29%。
- Abstract:
- In order to improve transgene efficiency of pollen tube pathway, and study the possibility of direct transformation of Agrobacterium, pCAMBIA3301 at 3 concentration levels of CaCl2 was transformed into Shanghai Bendiqing Soybean receptors by means of pollen tube pathway, meanwhile, 3 different varieties of Agrobacterium GV3101、LBA4404、EHA105 were introduced into soybean as another treatment. When pCAMBIA3301 was served as foreign gene transformed by pollen tube pathway, an average seed-set percentage of 36.83% and an average seedling percentage of 60.89% were achieved. An average seedling survival percentage was 8.24% when 0.2% Besta was conducted with T1 plants. When AgrobacteriumGV3101,LBA4404,EHA105 were served as foreign gene transformed by pollen tube pathway, an average seed-set percentage of 15.92% and an average seedling percentage of 20.40% were achieved. An average seedling survival percentage was 7.79% when 0.2% Besta was conducted with T1 plants. Leaf genome DNA from survived T1 plant of different treatments were analyzed by PCR, 20 positive plants were obtained with pCAMBIA3301 as foreign gene, 3 concentration levels of CaCl2 gained 0.56%, 1.64%,1.40% efficiency respectively, 2 of which were about 3 fold higher than that of traditional way. 5 positive plants were obtained which 3 different varieties of Agrobacterium were directly introduced into soybean, gained 0.17%, 0.33%, 0.29% efficiency respectively.
参考文献/References:
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备注/Memo
基金项目: 863“大豆基因工程育种”(2004AA212150)