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以青花菜“绿秀”为研究材料,以带柄子叶为外植体,采用根癌农杆菌LBA4404菌株进行农杆菌遗传转化,研究影响青花菜遗传转化的主要因素,结果表明:乙酰丁香酮浓度、OD_(600)值、侵染时间等对瞬间表达都有一定的影响。因素中OD_(600)值影响最大,AS浓度影响次之,再次是侵染时间。最适宜的试验条件为:乙酰丁香酮浓度为100μmol·L~(-1)、OD_(600)值为0.3、侵染时间为5 min。抗性植株经PCR检测、Southern杂交和GUS组织化学染色法,表明目的基因已整合到青花菜基因组中并得到了很好的表达。
Taking Broccoli “Green Sauss ” as the research material, taking the cotyledons as explants, Agrobacterium tumefaciens strain LBA4404 was used to transform Agrobacterium tumefaciens, and the main factors affecting the genetic transformation of Broccoli were studied. The results showed that: Ketone concentration, OD_ (600) value, infection time and so on have a certain impact on the transient expression. Among the factors, OD_ (600) value had the greatest impact, followed by AS concentration, followed by infection time. The most suitable experimental conditions were as follows: acetosyringone concentration 100μmol·L -1, OD 600 value 0.3 and infection time 5 min. The resistant plants were detected by PCR, Southern blot and GUS histochemical staining, which showed that the target gene was integrated into the genome of broccoli and was well expressed.