影响根癌农杆菌介导香蕉胚性悬浮细胞遗传转化的因素

用含质粒载体pCAMBIA1301的根癌农杆菌EHA105转化贡蕉(Musa A A Pisang Mas cv. Mas)品种的胚性悬浮细胞,通过测定GUS基因瞬时表达率,对转化初期的主要影响因素进行了研究。结果表明,取处于对数期的胚性悬浮细胞和D600nm为0.2的菌液混合,在室温、8000r/min离心5min,然后在110r/min的旋转摇床上摇5min,在体细胞胚诱导培养基上共培养8d,瞬时表达率达到最高值25.30%。

Factors affecting A grobacterium-mediated genetic transformation of embryogenic cell suspension of banana

Embryogenic cell suspension (Musa AA Pisang Mas cv. Mas) was used as the receptors for transformation by Agrobacterium tumefaciens containing pCAMBIA1301. By detecting the transient expression of GUS gene, the factors affecting the early phase of Agrobacterium-mediated transformation of banana were investigated. The results showed that the transient expression of the highest rate was 25.30%, when mixing the bacterium D600 nm 0.2 concentration with the embryonic cell suspension,in the logarithmic phase then centrifugating at room temperature, 8 000 r/min for 5 min, and then shaking at 110 r/min in orbital shaker for 5 min, finally co-culturing for 8 d in somatic embryo induction medium.