【摘要】 为了建立栓皮栎体胚遗传转化体系,以未成熟合子胚诱导的体胚为转化受体,采用带有pCAMBIA1301质粒的根癌农杆菌菌株EHA105,其中含有hptⅡ和GUS基因,研究不同因子对遗传转化的影响。结果表明:选择培养中适宜的抗生素浓度为300 mg/L头孢霉素和30 mg/L潮霉素。预培养时间为15 d时,转化率达91.7%;菌液浓度为OD₆₀₀=0.5,乙酰丁香酮浓度为200μmol/L时,转化率可达100%。在添加潮霉素的选择培养基中培养4周后,组织化学染色和PCR检测证实,GUS报告基因已经转入到体胚中并正常表达。更多还原

【Abstract】 This study was conducted to establish a transformation system by using somatic embryogenesis in Quercus variabilis somatic embryos induced by immature zygotic embryos were used as transformation recipients. A strain of Agrobacterium tumefaciens（EHA105） harbouring a pCAMBIA1301 plasmid containing hpt II and β-glucuronidase（GUS） genes was used in the study. Several factors influencing genetic transformation were assayed. The results showed that the suitable antibiotic concentrations were 300 mg/L cephalosporin and 30 mg/L hygromycin. Transformation efficiencies were up to 91.7% with 15 days pre-culture time, to 100% with the concentrations of OD₆₀₀=0.5 A. tumefaciens and 200 μmol/L acetosyringone. After 4 weeks of selective culture in hygromycin-added medium, histochemical and PCR molecular analysis confirmed the presence of GUS reporter gene in the somatic embryos.更多还原