根癌农杆菌介导反义fad2-1基因转化大豆的研究

Agrobacterium tumefaciens-mediated Antisense fad2-1 Gene Transfer to Cotyledonary Node Cells of Soybean(Glycine max L.)

通过根癌农杆菌介导法将反义fad2-1基因导入大豆子叶节,研究了农杆菌菌液浓度、预培养时间、共培养时间、恢复培养时间、乙酰丁香酮浓度和pH值对大豆子叶节形成丛生芽的影响,建立了有效的转化体系,获得了转基因大豆再生植株。PCR、PCR-Southern blotting检测结果表明,反义fad2-1基因成功转入并整合于大豆基因组。GC检测结果表明,转反义fad2-1基因大豆种子油酸含量比非转基因大豆种子的油酸含量提高了14.44%,而亚油酸含量降低了20.27%。

Soybean is the most important oil crop in the world.fad2-1 is a major gene related to soybean oil components.In the present study,an efficient gene transfer system was established to transfer Agrobacterium tumefaciens-mediated antisense fad2-1 gene into cotyledonary node cells of soybean.Experimental factors considering in the system included Agrobacterium tumefaciens concentration,pre-culture duration,co-culture time,recover culture time,pH value,acetosyringone（AS） on the cluster shoot formation from infected soybean cotyledonary node explants with Agrobacterium tumefaciens.With this system,kanamycin-resistant regenerated soybean plants were obtained.The integration of antisense fad2-1 gene into the soybean genome was confirmed by PCR,PCR-Southern blotting,and GC analysis indicated that the fad2-1 gene in the soybean genome was suppressed by the incorporated antisense one.The seed oleic content of transformed soybean was 14.44% higher than the receptor line,while the linoleic content was 20.27% lower than the receptor line.