基因枪法转化大豆萌动种子影响因素的研究  

Factors Affecting Particle Bombardment-mediated Transformation of Soybean Germinating Seeds

作　　者：薛仁镐[1] 

机构地区：[1]青岛农业大学生命科学学院,山东青岛266109

出　　处：《大豆科学》2008年第2期194-198,共5页Soybean Science

基　　金：山东省教育厅基金资助项目(J05K04);引进国际先进农业科学技术计划"948"资助项目(2006-G26)

摘　　要：以萌动1d的半片种子为靶组织,植物表达载体质粒pCAMBIA1301为外源DNA,通过GUS瞬时表达检测,研究了轰击次数、轰击距离、培养时间及靶组织放置方式对基因枪转化效率的影响。结果表明:轰击次数对转化效率有很大影响,轰击2或3次时GUS瞬时表达效率显著高于1次;在所试的3、6、9、12cm4个轰击距离中,9cm轰击距离对GUS瞬时表达效率最高;靶组织培养时间长短对转化效率也有很大影响,培养2d时,转化效率最高,平均每个半片种子达到217.5个蓝色斑点,随着培养时间的延长,GUS基因瞬时表达呈下降趋势,至第28天以后趋于稳定;靶组织不同放置方式对转化效率无明显的差异。本研究优化了影响基因枪法转化大豆萌动种子的主要因素,从而为基因枪介导大豆遗传转化技术体系的建立提供了参数。Although germinating seed of soybean has been successfully used as a target tissue in Agrobaeterium-mediated transformation ,it has not been investigated in particle bombardment-mediated transformation. In this study,factors such as bombardment times, bombardment distance,culture time, and placing orientation that affect particle bombardment-mediated transformation efficiency were studied via particle bombardment with a plant expression vector pCAMBIA1301 using 1 daygerminating half seeds of soybean as target tissue. The result showed that transformation efficiency based on GUS transient expression was significantly affected by bombardment parameters. The highest GUS transient expression（217.5 blue foci/ half seed） was obtained when particle bombardment was carried out under the 2 times of bombardment,9 cm distance of bombardment,and tissue culture for 2 days after bombardment. However,the GUS transient expression was not affected by placing orientation of target tissue. The factors affecting the particle bombardment-mediated transformation of soybean germinating seeds were optimized by GUS transient expression assay. These results will provide useful information for establishment of a particle bombardment- mediated soybean transformation system.

关 键 词：大豆 萌动种子 GUS瞬时表达 基因枪 

分 类 号：S565.1[农业科学—作物学]