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Transformation of multiple soybean cultivars by infecting cotyledonary-node with Agrobacterium tumefaciens
Abstract
Transformation of four soybean cultivars (Nannong88-1, Nannong18-6, Yu23 and Nannong 87C-38) by infecting cotyledonary-node with Agrobacterium tumefaciens strain EHA105 harboring pBI121
containing GFP reporter gene was conducted. The results indicated that the addition of thiol compounds (L-cysteine, dithiothreitol and sodium thiosulfate) in co-cultivation period increased the transformation efficiency of all four soybean cultivars, with Nannong 88-1 most increased up to 2.20%. Detection of GFP expression in the rooted plants was an effective selection system for the confirmation
of soybean transformation. And most GFP-positive plants were confirmed to be positive by Southern blot analysis, which showed that transformation of cotyledonary-node explants mediated by
Agrobacterium delivered T-DNA with one or two copies into soybean genome. In our study, the combination of Nannong88-1 with EHA105 is the optimum selection for explant and bacterial inoculum in soybean transformation, which could be applied in future functional study of soybean genes.
containing GFP reporter gene was conducted. The results indicated that the addition of thiol compounds (L-cysteine, dithiothreitol and sodium thiosulfate) in co-cultivation period increased the transformation efficiency of all four soybean cultivars, with Nannong 88-1 most increased up to 2.20%. Detection of GFP expression in the rooted plants was an effective selection system for the confirmation
of soybean transformation. And most GFP-positive plants were confirmed to be positive by Southern blot analysis, which showed that transformation of cotyledonary-node explants mediated by
Agrobacterium delivered T-DNA with one or two copies into soybean genome. In our study, the combination of Nannong88-1 with EHA105 is the optimum selection for explant and bacterial inoculum in soybean transformation, which could be applied in future functional study of soybean genes.