摘要
以MYB转录因子AtPHR1表达载体为转化对象,采用农杆菌介导子叶节转化技术将其转入栽培大豆,研究不同萌发时间、预培养时间、菌液和侵染液浓度、大豆基因型以及筛选剂浓度对转化效率的影响。结果表明:以萌发5 d大豆幼苗制备子叶节外植体,预培养1 d,农杆菌菌液OD600为0.7,侵染液OD600为0.5或0.7进行转化,100 mg·L-1卡那霉素进行筛选的抗性芽诱导率最高;5个供试品种中,以五星2号的转化率最高,约为2.0%;经PCR检测转化后的大豆植株,获得了含有转录因子AtPHR1的5个大豆新材料。
In this study, a MYB transcription factor AtPHR1 was introduced into soybean varieties by Agrobacterium-mediated eotyledonary-node transformation method to study the influence factors on transformation efficiency. The studied factors includ- ed seedling age,pre-incubation period,strain density,infectious concentration, kanamycin concentration and soybean genotype. The results showed that a much higher induction rate of resistant shoots was obtained when the seedling age was 5 d from ger- mination with 1 d pre-culture, the strain density （ OD60o ）was 0.7 and infectious concentration was 0.5 or 0.7, and also the ka- namyein concentration was 100mg· L-1 The soybean variety, Wuxing 2, with the highest transformation efficiency was about 2.0%. Finally, five new PCR positive transgenic materials with transcription factor AtPHR1 were gained.
出处
《大豆科学》
CAS
CSCD
北大核心
2013年第2期154-159,共6页
Soybean Science
基金
农业科技成果转化资金(2012GB2A200032)
国家转基因重大专项(2009ZX08004-004B)
河北省自然科学基金(C2010000749)
关键词
MYB转录因子
农杆菌介导
大豆
遗传转化
MYB transcription factor
Agrobacterium-mediated method
Soybean
Genetic transformation
