农杆菌介导的旱稻遗传转化主要影响因素  

Main Factors Affecting Upland Rice Transformation Mediated by Agrobacterium tumefacien

作　　者：李朝炜[1] 冯丹[1] 翟会青 刘梅[1] 魏景芳[1] 

机构地区：[1]河北科技大学生物科学与工程学院,石家庄050018

出　　处：《湖北农业科学》2014年第15期3482-3487,共6页Hubei Agricultural Sciences

基　　金：国家转基因生物新品种培育科技重大专项(2011ZX08001-003)

摘　　要：以旱稻品种鲲旱1号为材料,对农杆菌介导的遗传转化中愈伤组织的诱导、筛选、分化等的影响因素进行了研究.将PPA抗虫基因和bar抗性基因成功导入旱稻愈伤组织,经过筛选和分化最终获得了抗性植株.结果表明,改良的NB培养基适合于旱稻愈伤组织的诱导;当农杆菌浓度在OD600nm为0.2～0.3,侵染时间为20 min时,转化效果最好;用40 mg/L的PPT筛选30d后,可以获得抗性愈伤组织;将已筛选的抗性愈伤组织在预分化培养基上培养5～7 d能有效提高分化率.Using the seeds of upland rice Kunhan No.1 as the experimental materials, the effects of main factors including induction of ealli, selection, and regeneration on the upland rice transformation frequency were investigated. The PPA gene and bar resistance gene were successfully transformed into upland rice calli. The resistant plants were obtained by screening and differentiation. The results showed that the improved NB medium was the optimal medium for inducing upland rice calli. The suitable concentration of the Agrobacterium was between 0.2 and 0.3. The incursion time was 20 min. Resistant calli can be obtained after being screened in the selective medium with 40 mg/L PPT for 30 days. The resistant calli were pre-differ- entiated 5-7 days before differentiation. The differentiation frequency of calli could be increased.

关 键 词：旱稻 农杆菌 遗传转化 

分 类 号：Q789[生物学—分子生物学]