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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。
Using MYB transcription factor AtPHR1 expression vector as transformation target, Agrobacterium tumefaciens-mediated cotyledonary node transformation was used to transform it into cultivated soybean. The effects of different germination time, pre-culture time, concentration of bacteria and inoculum, soybean genotype and screening agent Effect of concentration on conversion efficiency. The results showed that the cotyledonary node explants were prepared from 5-day germination soybean seedlings, pre-cultured for 1 day, Agrobacterium tumefaciens OD600 was 0.7 and the inoculum OD600 was 0.5 or 0.7 for transformation. 100 mg · L -1 kanamycin The highest induction rate of the resistant buds was found in the five varieties tested. The transformation rate of Wuxing 2 was the highest among the five tested cultivars, which was about 2.0%. Five transgenic soybean plants containing the AtPHR1 transcription factor were obtained by PCR. new material.