Transformation of four soybean cultivars (Nannong88-1, Nannong18-6, Yu23 and Nannong 87C-38) by infecting cotyledonary-node with Agrobacterium tumefaciensstrain 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.
Key words: Soybean, cotyledonary–node, Agrobacterium, transformation, GFP.
GFP, green fluorescent protein; BA, 6:benzylaminopurine; DTT,dithiolthreitol; IBA, indole-3-butyric acid
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