根癌农杆菌介导白细胞介素-4基因转化甘蓝(Brassica oleracea L.)研究

Genetic Transformation of Cabbage (Brassica oleracea) with Interleukin-4 Gene Mediated by Agrobacterium

利用根癌农杆菌介导法，以白细胞介素-4（interleukin-4，IL-4）基因转化中甘11号甘蓝。本研究对可能影响il-4转化率的外植体类型、外植体的预培养时间、农杆菌的侵染时间以及外植体与农杆菌的共培养时间进行了优化。试验结果表明：il-4基因对带1-2mm子叶柄的子叶的转化率显著高于下胚轴；带柄子叶在预培养1d、侵染3min、共培养1d时，转化效果最好，转化率达23．3%;下胚轴在预培养1-3d，侵染3～5min，共培养1-2d时，转化效果较好，转化率最高可达13．3％。经PCR检测及PCR-Southem杂交，初步证明目的基因il-4已经转入甘蓝的再生植株。转il-4基因甘蓝的PCR阳性再生植株已经开花并产生了后代。

Intedeukin-4 gene is transformed into Zhong Gan 11 cabbage （Brassica oleracea L.） via mediated by Agrobacterium. In this study, we optimized the factors which might affected genetic transformation efficiencies such as explant types, pre-cultured time of explant, infective time of Agrobacterium and co-cultured time of explant by Agrobacterium. The results showed as followings： the transformation rate of cotyledon with 1-2 mm stem as explants can reach 23.3% much higher than that of hypocotyl; it is obvious better procedures for precultured 1 d, infection 3 min and co-cultured 1 d. As well as the procedure for hypocotyls with pre-cultured 1-3 d, infection 3-5 min and co-cultured 1-2 d in turn can reach 13.3% transformation rate. The results of PCR amplification and PCR-Southern blotting indicated that Intedenkin-4 gene should be introduced into the regenerative cabbage. The transgenic cabbages already have been bloomed and harvested.