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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。
The agrobacterium-mediated method was used to transform the immature embryos of three maize inbred lines with the bar-resistant gene and the factors affecting genetic transformation were optimized. The results showed that the induction rate of embryogenic callus was higher when the concentration of 2,4-D was 2 mg · L-1; the best time of Agrobacterium infection was 20 min; the content of acetosyringone (AS) Embryonic GUS transient expression frequency, the appropriate concentration of 200μmol·L-1; moderate selection method suitable for immature embryo transformation, adventitious bud induction medium, bud elongation medium and rooting medium hygromycin concentrations were 8,6 , 2 mg.L-1; obtained seven PCR test and herbicide resistance test positive plants.