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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。
Based on the plant expression vector p CAMBIA3300-35S-hevein-NOS, asparagus “Konggang701 ” embryoid body was used as experimental material. Agrobacterium-mediated transgenic method was used to investigate the effects of bacterial concentration, AS concentration, Dyeing time and co-culture time on the efficiency of transgenic asparagus, in order to establish efficient asparagus transgenic system. The results showed that Agrobacterium tumefaciens with OD600 (600) = 0.6 and AS (final concentration 200μmol / L) were used for infection. After 10 min of infection, the efficiency of transgene under dark culture for 4 d was the best. Positive conversion rate of 21%, obtained transgenic seedlings. In this experiment, a complete agrobacterium-mediated genetic transformation system of asparagus was constructed.