农杆菌介导大豆子叶节影响因素的研究  

The Factors Affecting Genetic Transformation of Soybean Cotyledon Node Mediated by Agrobacterium tumefacions

作　　者：杨晓倩[1] 李桂兰[1] 刘晨光[1] 董秋平[1] 张锴[1] 乔亚科[1] 

机构地区：[1]河北科技师范学院生命科技学院,昌黎066600

出　　处：《生物技术通报》2015年第12期91-96,共6页Biotechnology Bulletin

基　　金：转基因重大专项(2014ZX0800404B)

摘　　要：为提高大豆遗传转化效率,采用农杆菌介导遗传转化的方法,以大豆子叶节为外植体,研究共培养阶段浸染液浓度及外界培养条件(共培养温度和天数)和培养基添加物对转化效率和芽诱导率的影响。结果显示,浸染液OD600=0.5的诱导率最高,共培养温度为24℃,共培养天数为10 d具有较高转化效率,共培养基中加入一定浓度的单壁碳纳米管(Single-wall carbon nanotube,NM)对诱导卡那霉素抗性不定芽有促进作用。PCR检测表明PHR1基因已整合到T1代大豆基因组中,初步证明可在大豆基因组中稳定遗传。It was to improve the efficiency of soybean genetic transformation, we used the soybean cotyledon node as explants via Agrobactium-mediated transformation method, and investigated the effect of bacterial concentration, co-culture temperature and time on the transformation efficiency and the rate of bud induction. The results showed that under the OD600 = 0.5 of infection solution concentration, and induction rate was the highest. The conversion efficiency was higher under the environment of co-culture temperature being 24℃ for 10 days. The single-wall carbon nanotubes added to culture medium improved the induction of the adventitious bud with kanamycin-resistance. Moreover, the PCR results indicated that PHR1 gene was integrated into T1 genome generation, preliminarily proving that the heredity of gene in the soybean genome was stable.

关 键 词：农杆菌介导 大豆子叶节 共培养 单壁碳纳米管 

分 类 号：S565.1[农业科学—作物学]