大豆转化体系的优化和Dof 4基因转入大豆的研究  

Optimization of Soybean Transformation System and Transferring Dof 4 Gene into Soybean

作　　者：杜升伟[1,2] 刘业丽 姚丙晨[1,2] 白晨 苗兴芬[1] 刘春燕 陈庆山 胡国华[1,2] 

机构地区：[1]东北农业大学农学院,黑龙江哈尔滨150030 [2]黑龙江农垦科研育种中心,黑龙江哈尔滨150090

出　　处：《大豆科学》2010年第3期398-402,共5页Soybean Science

基　　金：国家高技术研究发展计划资助项目(2006AA100104-3);国家转基因专项资助项目(2008ZX08004-001-3)

摘　　要：研究优化了包括种子消毒方法、生长调节剂用量和抗生素种类在内的影响农杆菌介导大豆子叶节遗传转化效率的多个因素,并将Dof4基因转入绥农14中。结果表明:氯气熏蒸消毒方法对大豆伤害小,子叶节丛生芽分化率高;菌液侵染时间和共培养时间控制直接影响长菌和丛生芽状态。芽诱导培养阶段,当6-BA浓度为1.6mg·L-1,IBA浓度为0.1mg·L-1,分化率较高,畸形率和愈伤状况都较轻。最优的抗生素组合为头孢噻肟钠(Cefo-taxime Sodium)200mg·mL-1加羧苄青霉素(Carbenicillin)250mg·mL-1。In this paper,some major factors,which played important role in improving the transformation efficiency of soybean,such as the method of sterilizing the seeds,plant hormones,and antibiotics,were optimized.The Dof 4 was transferred into the soybean cultivar Suinong 14 by Agrobacterium-mediated cotyledonary node transformation.The result showed that,the chlorine suffocating was chosen to seeds sterilize,which did not harm to the seeds and have a higher differentiation rate for multiple shoot.However,duration of Agrobacterium infection and co-culture influenced the contamination and the state of multiple shoot.As for shoot induction,a higher regenerated rate,better callus induction rate,and lower deformity rate were obtained with 1.6 mg·L^-1 6-BA and 0.1 mg·mL^-1 IBA in this experiment.And the optimum combination of antibiotic and concentration in medium was 200 mg·mL^-1 Cefotaxime Sodium and 250 mg·mL^-1 Carbenicillin.

关 键 词：大豆 子叶节 Dof4 农杆菌介导转化 

分 类 号：S565.1[农业科学—作物学]