Abstract
AbstractTomato (Solanum lycopersicum), a model system for ripening of fleshy fruits, has ∼40,000 genes predicted byin silicohomology-based annotation. However, functional validation is lacking for most annotated tomato genes. Among the strategies for functional annotations, transposon-tagged mutagenesis is the most powerful approach. Transposon- tagged genes can be functionally validated by phenotyping and activation tagging. However, the lack of a robustin plantatransformation system precludes large-scale transposon- mutagenesis of tomato. To overcome this limitation, we developed two sets of starter lines in tomato, each carrying maize transposon element (Ds) and transposase (Ac), respectively. TheDsandAclines were crossed to allow theAc-mediated transposition of theDsin the F1generation. In the F2generation, the location of excisedDsat new sites in the tomato genome was monitored. TheDstransposition was interspersed on different chromosomes of the tomato, indicating unlinked transposition of theDs. The analysis of DNA sequences flankingDsshowed random integration ofDsin intergenic, genes, and the promoter region of the genome. Our study paves the way for the generation of large-scale transposon-tagged tomato lines usingAc/Dsstarter lines and provides a potential tool for the functional validation of genes in tomato.Short summaryWe developed two sets of starter lines in tomato, carrying maize transposon element (Ds) and transposase (Ac), to enable large-scale transposon-mutagenesis, facilitating functional validation of tomato genes and for generating an insertional mutant resource in tomato.
Publisher
Cold Spring Harbor Laboratory