以冀无2031为材料，采用正交设计方法，研究了农杆菌介导的棉花下胚轴遗传转化过程中菌液浓度、侵染时间、共培养时间和共培养温度对抗性愈伤转化效率的影响。结果表明，4个因素均对抗性愈伤的诱导率有极显著影响，其诱导效应依次是共培养温度>菌液OD值>侵染时间>共培养时间。按照正交设计的原则，最终得到农杆菌介导转化棉花下胚轴的最佳条件是：菌液OD值为0.3，侵染15 min，24 ℃共培养24 h，下胚轴抗性愈伤的转化效率最高。用珂312、中521和农大94-7对所选出的最佳组合进行了进一步验证，结果3个品种在优选组合下的抗性愈伤组织诱导率都很高，分别为93.85%、86.72%和85.83%。
Abstract
The influences of Agrobacterium concentration,infection time,the time and the temperature of co-cultute on the induction ratios of kanamycin-resistant calli were studied by Agrobacterium-mediated transformation using the hypocotyls of Jiwu 2031 as explants.The factors were optimized using orthogonal design.The results showed that the transformation ratio was the highest under the regime of OD=0.3 of bacteria concentration,15 min of infection time,24 ℃ of co-culture temperature and 24 h of co-culture time.High frequency of kanamycin-resistant calli formation was observed when infecting hypocotyls of Coker 312,Zhong 521 and Nongda 94-7 using the optimized combination of factors,and the induction ratios reached 93.85%,86.72% and 85.83%,respectively.
关键词
棉花
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农杆菌介导 /
正交设计 /
抗性愈伤
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Key words
cotton
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Agrobacterium-mediated /
orthogonal design /
kanamycin-resistant callus
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