火焰南天竹高效稳定遗传转化体系的建立

Establishment of High Efficiency Agrobacterium-mediated Transformation System of Nandina domestica‘Fire Power'

以火焰南天竹的叶片为受体,对影响火焰南天竹遗传转化的因子进行优化,获得高效转化体系。结果表明:农杆菌菌液浓度为0D600为0.4;侵染时间20 min;共培养时间3 d可以获得较高的抗性芽诱导率。选用350 mg/L的羧苄青霉素(Cb)作为抑菌抗生素;25 mg/L的卡那霉素(Kan)作为筛选抗生素;PCR以及RT-PCR检测和荧光定量检测证明,基因已经成功的导入和整合到火焰南天竹基因组中。

Using the Nandina domestica‘Fire Power’leaves as the affection factors of Nandina domestica ‘Fire Power’genetic transformation, the high- efficiency transformation system was established. The results showed that the highest transformation rate could be achieved with Agrobacterium tumefaciens concentration OD600 of 0.4 and incubation-time of 20 mins for bacterial culture of 3 days. Using carboxy benzyl penicillin (Cb) of 350 mg/L as antibacterial antibiotics, 25 mg/L Kan as indicator for system optimization in resistance screening, and the PCR and RT-PCR detection and testing, results showed that the genes had been successfully imported and integrated into the Nandina domestica‘Fire Power’genome.