蓝浆果农杆菌介导法基因转化条件研究  

Agrobacterium-mediated Transformation Factors of Blueberry

作　　者：刘捷[1] 杨丽娜[1] 章镇[1] 蔡斌华[1] 陶建敏[1] 

机构地区：[1]南京农业大学园艺学院,江苏南京210095

出　　处：《江苏农业学报》2008年第5期624-628,共5页Jiangsu Journal of Agricultural Sciences

基　　金：国家农业综合开发科技示范项目[财发(2001)14]国家"863"项目(P200310)

摘　　要：以蓝浆果品种蓝丰的叶片为转化受体,通过对农杆菌侵染时间、共培养时间、外植体暗培养时间、卡那霉素(Km)选择压以及头孢霉素(Cef)浓度等因素研究,确定转化的最佳试验参数。试验结果:农杆菌侵染5 m in,共培养4 d,暗处理21 d时gus基因瞬时表达率最高;在筛选初期添加Km 20 mg/L、Cef 250 mg/L,既能得到不定芽,又达到筛选的目的。利用GUS组织化学染色、PCR扩增方法对蓝浆果的Km抗性植株进行检测,初步证实AtNHX1基因已经整合到2株蓝丰株系基因组中。Transgenic plants were generated from leaves of bluecrop blueberry by Agrobacterium-mediated transformation with AtNHX1 gene, and the following factors affecting transformation were investigated： dipping time, dark-culture time, co-culture time, concentration of kanamycin （Km）, and concentration of cefotaxine （Cef）. By analyzing the frequency of gus gene transient-expression and bud regeneration rate, the optimal transformation conditions were worked out： 5 min of leaf dipping, 4 d of co-culture and 21 d of dark-culture. Resistant buds could be regenerated and selected on primary selection medium supplemented with Km 20 mg,/L and Cef 250 mg/L. PCR and GUS analysis confirmed that AtNHX1 gene was integrated into the genome of two bluecrop plants.

关 键 词：蓝浆果 农杆菌介导 基因转化 GUS基因 PCR检测 

分 类 号：S663.903.53[农业科学—果树学]