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A novel dominant selectable system for the selection of transgenic plants under in vitro and greenhouse conditions based on phosphite metabolism

机译:基于亚磷酸盐代谢的体外和温室条件下选择转基因植物的新型主导选择系统

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Antibiotic and herbicide resistance genes are currently the most frequently used selectable marker genes for plant research and crop development. However, the use of antibiotics and herbicides must be carefully controlled because the degree of susceptibility to these compounds varies widely among plant species and because they can also affect plant regeneration. Therefore, new selectable marker systems that are effective for a broad range of plant species are still needed. Here, we report a simple and inexpensive system based on providing transgenic plant cells the capacity to convert a nonmetabolizable compound (phosphite, Phi) into an essential nutrient for cell growth (phosphate) trough the expression of a bacterial gene encoding a phosphite oxidoreductase (PTXD). This system is effective for the selection of Arabidopsis transgenic plants by germinating T0 seeds directly on media supplemented with Phi and to select transgenic tobacco shoots from cocultivated leaf disc explants using nutrient media supplemented with Phi as both a source of phosphorus and selective agent. Because the ptxD/Phi system also allows the establishment of large-scale screening systems under greenhouse conditions completely eliminating false transformation events, it should facilitate the development of novel plant transformation methods.
机译:抗生素和除草剂抗性基因目前是植物研究和作物发展的最常用的选择标志物基因。然而,必须仔细控制抗生素和除草剂的使用,因为对这些化合物的易感性在植物物种中的易感性范围内各种各样地变化,并且因为它们也可以影响植物再生。因此,仍然需要新的可选标记系统,这些标记系统有效地用于广泛的植物物种。在这里,我们报告了一种基于提供转基因植物细胞的简单且廉价的系统,该系统将不可再生化合物(亚磷酸盐,PHI)转化为细胞生长(磷酸盐)的基本营养素(磷酸盐)槽中编码亚磷酸盐氧化酶的细菌基因的表达(PTXD) )。该系统是通过直接发芽在补充有PHI的培养基上的培养基中的T0种子来选择拟南芥转基因植物,并使用补充有PHI作为磷和选择性药物来源的营养培养物选择转基因烟草芽的转基因烟草芽。因为PTXD / PHI系统还允许在温室条件下建立大规模筛选系统,完全消除虚假转型事件,它应该促进新型植物转化方法的发展。

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