Zeng P, Vadnais D A, Zhang Z, Polacco J C
Department of Biochemistry, University of Missouri-Columbia, 65211, USA.
Plant Cell Rep. 2004 Feb;22(7):478-82. doi: 10.1007/s00299-003-0712-8. Epub 2003 Sep 30.
Modern genetic analysis and manipulation of soybean ( Glycine max) depend heavily on an efficient and dependable transformation process, especially in public genotypes from which expressed sequence tag (EST), bacterial artificial chromosome and microarray data have been derived. Williams 82 is the subject of EST and functional genomics analyses. However, it has not previously been transformed successfully using either somatic embryogenesis-based or cotyledonary-node transformation methods, the two predominant soybean transformation systems. An advance has recently been made in using antioxidants to enhance Agrobacterium infection of soybean. Nonetheless, an undesirable effect of using these antioxidants is the compromised recovery of transgenic soybean when combined with the use of the herbicide glufosinate as a selective agent. Therefore, we optimized both Agrobacterium infection and glufosinate selection in the presence of L-cysteine for Williams 82. We have recovered transgenic lines of this genotype with an enhanced transformation efficiency using this herbicide selection system.
现代大豆(Glycine max)的遗传分析与操作在很大程度上依赖于高效且可靠的转化过程,尤其是对于那些已获得表达序列标签(EST)、细菌人工染色体和微阵列数据的公共基因型而言。Williams 82是EST和功能基因组学分析的对象。然而,此前无论是基于体细胞胚胎发生的转化方法还是子叶节转化方法(两种主要的大豆转化系统)都未能成功转化该品种。最近在利用抗氧化剂增强农杆菌对大豆的感染方面取得了进展。尽管如此,使用这些抗氧化剂的一个不良影响是,当与除草剂草铵膦作为选择剂一起使用时,转基因大豆的恢复受到影响。因此,我们针对Williams 82在L-半胱氨酸存在的情况下优化了农杆菌感染和草铵膦选择。我们利用这种除草剂选择系统获得了该基因型的转基因株系,其转化效率得到了提高。