基于草甘膦筛选的菘蓝转化体系的建立

摘要

The transformation system of Isatis indigotica by using Glyphosate as the selectable marker was established in this study.Tetraploid Isatis indigotica was transformed via Agrobacterium tumefaciens strain LBA4404 carrying the plasmid pASM12.The transgenie explants were directly selected on the media containing 8 mg/L Glyphosate, and the transgenic Glyphosate -resistant plantlets were confirmed by genomic PCR techniques and Glyphosate resistance test.PCR analysis showed the transformation frequency of the cotyledons of lsatis indigotica was 30％, and that of hypocotyles was 40％.The results of Glyphosate resistance test indicated that most of the transgenic plantlets could resist to Glyphosate at different levels.%介绍了以菘蓝草甘膦筛选的遗传转化体系,以携带植物表达载体pASM12的根癌农杆菌LBA4404对四倍体菘蓝进行叶盘法转化,在选择培养基中加入8 mg/L的草甘膦进行筛选,获得草甘膦抗性的再生植株,对再生植株进行PCR检测和草甘膦抗性试验,结果表明:PCR检测表明子叶的转化频率达30%,下胚轴的转化频率达40%;对PCR阳性植株的草甘膦抗性试验表明,多数植株在不同程度上表现出对草甘膦的抗性.