Characterisation of transgene insertions in wheat using Fluorescence In Situ hybridisation (FISH).

摘要

Crop improvement via genetic transformation is an important area of agricultural research. However; very little is known of the relationship between the physical location of transgenes and their integration; expression and inheritance. Fluorescencein situ hybridisation (FISH) technology in plants has developed significantly over the past several years. Conventional FISH is now capable of detecting target sequences as small as lOkb within a large; complex genome such as that of wheat. We present results from a study using FISH technology to characterise and compare transgene integration sites resulting from either Agrobacterium- or particle bombardment-mediated transformation of wheat. Included in the latter category are populations of stable andsilenced transformants and populations containing homologous and heterologous transgenes with a high frequency of multiple locus insertion. Several wheat background genotypes are included in these 35 independently transformed lines. Transgene loci in lines showing their stable expression appeared equally distributed between genomes and individual chromosomes; however; a tendency for distal integration was evident.