[1]李卉,武天龙.CaCl2诱导大豆花粉管通道农杆菌转基因研究[J].大豆科学,2007,26(01):55-59.[doi:10.3969/j.issn.1000-9841.2007.01.013]
　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]
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CaCl2诱导大豆花粉管通道农杆菌转基因研究

《大豆科学》[ISSN:1000-9841/CN:23-1227/S] 卷: 第26卷 期数: 2007年01期 页码: 55-59 栏目: 出版日期: 2007-02-25
Title:
TRANSFOMING AGROBACTERIUM INTO SOYBEAN BY MEANS OF POLLEN TUBE PATHWAY INDUCED BY CaCl2
文章编号:
1000-9841(2007)01-0055-05
作者:
李卉; 武天龙
上海交通大学农业与生物学院，上海201101
Author(s):
LI Hui; WU Tian-long
Agriculture & Biology College， Shanghai Jiaotong University， Shanghai, 201101
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
DOI:
10.3969/j.issn.1000-9841.2007.01.013
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.

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备注/Memo

基金项目： 863“大豆基因工程育种”（2004AA212150）

作者简介：李卉（1981-），女，在读硕士，从事作物遗传育种学研究。E-mail： syringali@sjtu.edu.cn

更新日期/Last Update: 2014-10-21