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本研究以GUS基因在子叶节区的瞬时表达为依据,通过探讨影响农杆菌转化效率的因素,优化了大豆子叶节非组织培养遗传转化体系;利用该体系对冀豆16号进行Bar基因的遗传转化,并使用针刺法对转基因植株进行草铵膦筛选。结果表明,侵染液中附加3%蔗糖、OD6 00=0.6、以脱脂棉作为菌液附着介质同时不添加表面活性剂Silwet L-77、侵染1次的GUS阳性率最高达到62.13%。草铵膦抗性植株经PCR检测获得T0阳性植株10个,转化率为2.5%。经PCR和RT-PCR鉴定共获得3株T1阳性植株,初步证明目的基因已整合到大豆基因组中。
In this study, based on the transient expression of GUS gene in cotyledonary node region, the genetic transformation system of soybean cotyledonary node non-tissue culture was optimized by investigating the factors affecting the transformation efficiency of Agrobacterium tumefaciens. Using this system, Transformation and the use of acupuncture on transgenic plants glufosinate screening. The results showed that with the addition of 3% sucrose and OD6 00 = 0.6, the highest positive rate of GUS infection reached 62.13% with cotton wool as the medium for bacterial attachment without the addition of surfactant Silwet L-77. Glufosinate-resistant plants T0-positive plants were obtained by PCR detection 10, the conversion rate of 2.5%. A total of 3 T1 positive plants were obtained by PCR and RT-PCR. The results showed that the target gene had been integrated into soybean genome.