根癌农杆菌介导欧美杨108遗传转化体系的优化

Optimization of Agrobacterium-mediated Populus euramericana 108 Genetic Transformation System

[目的]优化根癌农杆菌介导的欧美杨108遗传转化体系。[方法]以欧美杨108无菌苗的叶片为外植体,通过农杆菌介导法对其遗传转化体系进行优化研究。[结果]试验获得的优化转化体系如下:农杆菌介导欧美杨108遗传转化的潮霉素最佳叶片分化浓度为2mg/L,生根浓度为1 mg/L。优化筛选的最适转化条件为:预培养48 h,菌液浓度OD600为0.4,侵染15 min,共培养48 h。试验共获得18个抗性愈伤和抗性芽,转化频率为4.3%;经PCR检测初步证明TCS基因已整合至欧美杨108再生植株中。[结论]该研究建立了高效欧美杨108叶片农杆菌介导的遗传转化体系。

[Objective] The aim was to optimize Populus euramericana 108 genetic transformation system mediated by Agrobacterium.[Method] The leaf of P.euramericana aseptic seedling was taken as explants to study the optimization of P.euramericana genetic transformation system via Agrobacterium.[Result] The optimized conditions for transformation of P.euramericana were obtained as follows： the Hyg selection concentration was 2 mg/L for leaf-explants and 1 mg/L for shoot.The optimal prioritization optimal system was pre-cultured for 48 hours,infected with Agrobacterium at the concentration of OD600= 0.4 for 15 min,co-cultured for 48 hours.Adapting to Agrobacterinm-mediated to transform the leaf explants,18 resistant shoot were obtained and transformation frequency was 4.3%;The PCR detection proved that TCS gene had been inte-grated into the regenerated plants.[Conclusion] A highly efficient genetic transformation system of P.euramericana leaf mediated by Agrobacterium was established.