摘要以4个辣椒(Capsicum annuum L. )品种为试材,利用根癌农杆菌介导的带柄子叶转化法将拟南芥CBF-4基因转入辣椒,通过调整预培养、共培养以及延迟培养时间等因素,优化并完善了甜椒品种B12的遗传转化体系,并对获得的60株卡那霉素抗性再生植株进行PCR和RT-PCR检测表明,46株为阳性转基因植株,转化率高达76.67%。经过低温处理后,对阳性植株的抗寒性生理、形态指标分析发现,转基因植株的SOD、POD和MDA活性都明显高于未转基因植株,转基因植株比未转基因植株的株型紧凑,叶片肥厚、平直,叶色浓绿,叶面积大。
Abstract:Four cultivars of pepper(Capsicum annuum L.)were successfully transformed with the CBF-4 gene by Agrobacterium mediated transformation. This study established a highly efficient regeneration system of pepper . The main factors influencing regeneration and transformation rate were discussed and the best conduction of regeneration and transformation were determined. The positive regeneraion was obtained.60 Km-resistant regenerated plants were obtained, 46 of which were positive by the analysis of PCR and RT-PCR.The rate of transgene is 76.67%.The morphological indexes of transgenetic plants showed that it was compact plant type with thicker, darker green and larger area leaves compared with the non-transgenetic plants. The enzyme activities of SOD, POD and MDA in transgenetic plants are higher than that of the non-transgenetic plant (control group) after the cold resistant treatment.