山新杨遗传转化体系的优化及转MnSOD基因植株的鉴定

Prioritization of the system of genetic transformation of Populus davidiana × P. bollena and Identification of MnSOD Gene Transgenic Plants

为建立农杆菌介导的山新杨高效遗传转化体系,对MnSOD基因导入山新杨的遗传转化体系进行了优化,获得的较优转化体系如下：菌液浓度OD600=0.1,浸染2~5 min,共培养时加入200μmol/L乙酰丁香酮,脱菌时加入3 mg/L的AgNO3;对部分抗性植株经PCR及Northern杂交检测,证明MnSOD基因已经整合到山新杨基因组中并能够在转录水平上表达。

In order to establish the high-efficient genetic transformation system of Populus davidiana Dode × P. bollena Lauche by Agrobacterium tumefaciens,several key factors of Populus davidiana Dode× P. bollena Lauche transferring system were optimized during MnSOD transformation. The optimized conditions for transformation were obtained as follows： The explant was infected with Agrobacterium at the concentration of OD600= 0. 1 for 2 ~ 5 min,co-culture medium plus 200 μmol /L AS and put 3 mg /L AgNO3into the selective culture medium could increase the transformation ratio. PCR and Northern blot results showed that MnSOD gene was integrated into the genome and the transformed gene was expressed at the transcription level.