农杆菌介导bar基因转化玉米幼胚的研究

通过农杆菌介导法,采用抗除草剂基因(bar)转化3个玉米自交系的幼胚,并对遗传转化影响因素进行优化。结果表明,2,4-D浓度为2 mg·L-1时,胚性愈伤组织诱导率较高;农杆菌最佳的侵染时间为20 min;乙酰丁香酮(AS)明显提高了幼胚的GUS瞬时表达频率,适宜浓度为200μmol·L-1;温和选择方法适于幼胚转化,不定芽诱导培养基、芽伸长培养基和生根培养基中潮霉素的浓度分别为8、6、2 mg.L-1;获得7株PCR检测以及除草剂抗性试验阳性植株。

Study on transformation of bar gene into maize(Zea mays L.)by Agrobacterium tumefacien mediated

bar gene was introduced into 3 maize inbred lines by Agrobacterium tumefacien mediated.The results showed that 2 mg·L-1 2,4-D was good to embryogenic callus.The suitable duration of infection was 20 min.The addition of AS(200 μmol·L-1) in infection and co-culture medium enhanced the instant expression frequency of immature embryo.The herbicide concentration should be lower than lethal concentration,8 mg·L-1 was better.Moderate selection fitted for transformation,the best screening mode was to use hygromycin in the concentration of 8,6 and 2 mg·L-1 in induction medium,buds elongation medium and rooting medium respectively.Seven plants showed positive reaction by PCR analysis and were confirmed to resist basta.