Journal or Publishing Institution: Theoretical and Applied Genetics
Study: https://link.springer.com/article/10.1007/s001220051364
Author(s): Svitashev, S., Ananiev, E., Pawlowski, W.P. and Somers, D.A.
Article Type: Peer Reviewed Study
Record ID: 2241
Abstract: Transgene loci in 16 transgenic oat (Avena sativa L.) lines produced by microprojectile bombardment were characterized using phenotypic and genotypic segregation, Southern blot analysis, and fluorescence in situ hybridization (FISH). Twenty-five transgene loci were detected; 8 lines exhibited single transgene loci and 8 lines had 2 or 3 loci. Double FISH of the transgene and oat C- and A/D-genome-specific dispersed and clustered repeats showed no preferences in the distribution of transgene loci among the highly heterochromatic C genome and the A/D genomes of hexaploid oat, nor among chromosomes within the genomes. Transgene integration sites were detected at different locations along individual chromosomes, although the majority of transformants had transgenes integrated into subtelomeric and telomeric regions. Transgene integration sites exhibited different levels of structural complexity, ranging from simple integration structures of two apparently contiguous transgene copies to tightly linked clusters of multiple copies of transgenes interspersed with oat DNA. The size of the genomic interspersions observed in these transgene clusters was estimated from FISH results on prometaphase chromosomes to be megabases long, indicating that some transgene loci were significantly larger than previously determined by Southern blot analysis. Overall, 6 of the 25 transgene loci were associated with rearranged chromosomes. These results suggest that particle bombardment-mediated transgene integration may result from and cause chromosomal breakage and rearrangements.
Keywords: Genetic engineering, Oat (A. sativa L.), Microprojectile bombardment, FISH, Transgene DNA integration, Avena, Avena sativa, plants, biolistics, biotechnology, breakage, cereals, characterization, chromosome breakage, chromosomes, fluorescence in situ hybridization, genetic transformation, genetically engineered organisms, genomes, oats, southern blotting, transgenic plants, fluorescence in situ hybridisation, genetic manipulation, genetically engineered plants, genetically modified organisms, genetically modified plants, GEOs, GMOs, transgenic organisms
Citation: Svitashev, S., Ananiev, E., Pawlowski, W.P. and Somers, D.A., 2000. Association of transgene integration sites with chromosome rearrangements in hexaploid oat. Theoretical and Applied Genetics, 100(6), pp.872-880.