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以中华猕猴桃伏牛95-2(Actinidia chinensis‘Funiu 95-2’)叶片为试验材料,对不同侵染方式、预培养时间、菌液浓度、共培养时间等影响β-葡萄糖苷酸酶基因(β-glucuronidase,GUS)瞬时表达率的因素进行了研究。结果表明,叶片预培养3天,菌液浓度A_(600)值为0.3,真空渗入方式侵染10 min,共培养4天条件下,GUS基因瞬时表达率最高,达到92.2%。对转基因抗性植株进行PCR检测和GUS组织化学染色,初步证明外源基因已整合到中华猕猴桃伏牛95-2的基因组中。
The leaves of Actinidia chinensis’Funiu 95-2 ’from China were used as experimental material to study the effects of different infection methods, pre-culture time, concentration of bacteria and co-culture time on the expression of β-glucuronidase gene β-glucuronidase, GUS) were investigated. The results showed that the rate of transient expression of GUS gene reached 92.2% under the conditions of 3 days of pre-culture, 0.3 of bacterial concentration and 10 minutes of infiltration by vacuum infiltration. The transgenic plants were detected by PCR and GUS histochemical staining, initially proving that the exogenous genes have been integrated into the genome of Actinidia chinensis Fuxin 95-2.