金鱼藤(Asarina procumbens)‘维多利亚瀑布’遗传转化体系建立

Establishment of genetic transformation system of Asarina procumbens’Victoria Falls’

以金鱼藤‘维多利亚瀑布’品种为试验材料,建立根癌农杆菌介导金鱼藤遗传转化体系,以获得转LiSEP3基因金鱼藤植株。结果表明,金鱼藤愈伤组织最佳侵染时间为16 min,最佳菌液侵染浓度为OD600=0.6。菌液侵染后得到正常膨大生长抗性愈伤43个。抗性金鱼藤愈伤经PCR检测,呈阳性转基因愈伤20个。选取15株由阳性愈伤发育成的转基因金鱼藤植株幼苗,RT-PCR检测,最终获得阳性植株9株。

In this study,Asarina procumbens’Victoria Falls’variety was used as the experimental material,and Agrobacterium tumefaciens-mediated genetic transformation system of Asarina pro-cumbens was established to obtain Asarina procumbens plants transfected with LiSEP3 gene.The results showed that the infection time was 16 min and the optimum concentration of bacterial solution was OD600=0.6.The total 43 normal enlarged callus were obtained after bacterial infection.The callus of the resistant Asarina procumbens was detected by PCR,and there were 20 positive transgenic calluses.The total 15 transgenic Asarina procumbens seedlings developed from positive callus were selected and detected by RT-PCR.Among them,nine seedlings were positive.