超声波辅助农杆菌介导的长春花(Catharanthus roseus)遗传转化

Sonication-Assisted Agrobacterium-Mediated Transformation of Catharanthus roseus

研究以根癌农杆菌介导结合超声波处理和乙酰丁香酮诱导,通过对GUS基因瞬时表达来研究影响长春花转化效率的因素。对转化方法包括超声波处理的功率和时间、乙酰丁香酮(3,5-methoxy-4-hydroxyacetophenone,AS)的浓度、共培养时间进行了系统优化。结果表明:在添加AS 100μmol·L^(-1)的1/2 MS培养液中,超声波处理功率80 W,处理时间10 min,共培养2 d,根癌农杆菌介导的长春花下胚轴遗传转化能获得最佳的GUS基因瞬间表达率。本研究为进一步开展根癌农杆菌介导稳定转化长春花奠定了基础。

The influencing factors of transformation of the hypocotyls of Catharanthus roseus using sonication-assisted, Agrobacterium-mediated method with Acetosyringone inducement were investigated. The results show that the optimal GUS transient expression conditions are that the hypocotyls were treated with 80 W power sonication for 10 minutes after immersed in 1/2 MS suspension containing 100 μmol·L^-1 Acetosyringone and then co-cuhured for 2 days. This study lays a foundation for the establishment of Agrobacterium-mediated stable transformation of C. roseus.