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本研究利用以双单倍体马铃薯DM的茎段为试验材料,对影响马铃薯遗传转化的条件进行了优化,主要从对通过抗生素浓度、预培养时间、侵染时间、农杆菌浓度和共培养时间等几个因素进行了的分析,对影响马铃薯遗传转化的条件进行了优化。研究表明:头孢霉素(cef)的浓度在600 mg/L时既不影响愈伤的正常生长还能有效的抑制农杆菌的生长。潮霉素(hyg)浓度在8 mg/L时对愈伤达到半致死状态,是因为较低的潮霉素浓度无法起到对抗性愈伤的筛选作用,所以合适的浓度应为愈伤达半致死状态浓度。得到更多抗性愈伤的最佳条件为:外植体预培养时间13天,侵染时间为8 min,农杆菌浓度以OD600=0.4左右,且共培养时间3天。上述的条件的优化大大的提高了马铃薯DM的遗传转化效率。
In this study, using the stem segments of double haploid potato DM as the experimental material, the conditions affecting potato genetic transformation were optimized, mainly from the analysis of the antibiotic concentration, pre-culture time, infection time, Agrobacterium concentration and co-culture time And other factors were analyzed, the conditions affecting potato genetic transformation were optimized. The results showed that cefotaxime (Cef) concentration of 600 mg / L did not affect the normal growth of callus but also effectively inhibited the growth of Agrobacterium. The hygromycin half-lethal condition at a hyg concentration of 8 mg / L is because the lower hygromycin concentration does not act as a screening for resistance callus, so a suitable concentration should be callus Semi-lethal concentration. The best conditions for more resistant callus were as follows: explant preincubation time 13 days, infection time 8 min, Agrobacterium concentration OD600 = 0.4, and co-culture time 3 days. Optimization of the above conditions greatly improves the genetic transformation efficiency of potato DM.