影响农杆菌介导甜瓜子叶遗传转化的因素

The Factors Influencing on Transformation of Cucumis melon Mediated by Agrobacterium

以甜瓜子叶作为根癌农杆菌介导转化的受体,通过GUS基因瞬时表达率的分析,研究此转化体系的最佳实验参数。实验结果表明,预培养时间、预培养后对外植体进行处理、感染时间、共培养时间、农杆菌工程菌的浓度等对转化效率都有一定的影响,但是根癌农杆菌诱导物AS并不能大幅度提高转化效果。对外植体进行重新处理,预培养2d,用OD560值为0.3的农杆菌工程菌感染15～25min,共培养3～4d的理想条件下,GUS瞬时表达率可达85%。

The optimal conditions for Agrobacteraum-mediated transformation of Cucumis melon cotyledons were investigated based on the rate of gus gene transient expression. Histochemical GUS assay showed that the transient expression rate was effected significantly by pre-culture time, explants treatments, infection time, coculture time and engineering bacterium concentration. While the influence by adding acetosyringone was not evident. The transient expression rate of gus gene could be reached 85% under the ideal conditions, which the explants were re-treated, pre-cultured 2 d, infected 15-25 min by engineering bacterium of 0.30 OD560, and cocultured 3-4 d.