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%。
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 Agrobacterium GV3101, 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 plantswere 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.
出处
《大豆科学》
CAS
CSCD
北大核心
2007年第1期55-59,共5页
Soybean Science
基金
863"大豆基因工程育种"(2004AA212150)
