农杆菌介导的芦笋遗传转化体系的建立

The Establishment of Agrobacterium-mediated Genetic Transformation System for Asparagus

以芦笋"井岗701"胚状体为试验材料,在构建p CAMBIA3300-35S-hevein-NOS植物表达载体基础上,采用农杆菌介导的转基因方法,探究菌液浓度、AS浓度、侵染时间和共培养时间等4个因素对芦笋转基因效率的影响,以期建立高效的芦笋转基因体系。结果表明:用菌液浓度OD_(600)=0.6,AS终浓度为200μmol/L的农杆菌菌液进行侵染,侵染10 min后,暗培养4 d的转基因效率最佳,经PCR检测,阳性转化率达21%,获得了转基因幼苗。本实验构建了完整的农杆菌介导的芦笋遗传转化体系。

Embryoids derived from asparagus ＂Jinggang 701＂ were chosen as the experimental materials. With construction of plant expression vector-pCAMBIA3300-35S-hevein-NOS, the effects of liquid bacterial concentration, AS concentration, infection time and co-culture time on asparagus transgenic efficiency were explored by the agrobacterium-mediated transgenic method. The high-efficiency transgenic system of asparagus was expected to be established in this study. The result showed that the optimal infection conditions were liquid bacterial concentration OD600=0.6, adding AS up to final concentration of 200 μmol/L, infecting for 10min and culturing under darkness for 4d, which made the transformation rate up to 21% and got transgenic plant.