Abstract
Introduction of T-DNAs with foreign genes by Agrobacterium-mediated transformation is widely used in plants. Multiple-introduced complex T-DNA loci, however, are difficult to clarify by conventional DNA gel blot analysis. We performed fluorescence in situ hybridization on extended DNA fibers (fiber FISH) of transgenic tobacco plants harboring multiple 37-kb T-DNA constructs. Five and seven types of integrations were successfully visualized in two transgenic lines. Most of the loci suffered duplication, deletion and/or translocation, indicating the complex integration events of the medium-size T-DNA. We concluded that fiber-FISH analysis is a powerful tool to analyze organization of multiple T-DNA loci in detail.
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