Submitted to: European Journal of Horticultural Science
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 8/20/2005
Publication Date: 12/1/2006
Citation: Ipek, A., Masson, P., Simon, P.W. 2006. Genetic transformation of an Ac/Ds - based transposon tagging system in carrot (Daucus carota). European Journal of Horticultural Science. 71:245-251. Interpretive Summary: The DNA of an organism is mostly localized and stable in chromosomes, but some small pieces of DNA move around from one region of a chromosome to another. We introduced two of these mobile DNA elements from corn, called Ac and Ds, into carrot. We found that these elements are mobile in carrot. This research is of interest to researchers and seed companies since it will help us find other carrot genes.
Technical Abstract: Maize transposable elements, Activator (Ac) and Ds have been transformed into several heterologous plant species for transposon tagging of genes and they have been successfully used to taf and to clone genes in Arabidopsis, flax, pentunia, tobacco, and tomato. To investigate the possibility of transposon tagging and subsequent cloning of carrot genes, carrot was transformed with modified maize transposable elements Acativator-transposase (Ac-transposase) gene and Ds using Agrobacterium tumefaciens strain LBA 4404. Callus initially transformed with Ac-transposase was then transformed with Ds. Transgenic carrot calli and plants carrying only Ds or both modified Ac and Ds were analyzed for transposition of Ds. Ds did not transpose in any of the transgenic plants carrying Ds only. On the other hand, excision of Ds was detected in all of tissue culture-regenerated plants carrying both Ac-transposase and Ds. Reinsertion of Ds into new chromosomal sites was detected in transformed plants based on Southern blotting and sequence analysis of Ds insertion sites using TAIL-PCR. Our results indicated that Ds will transpose in the carrot genome if Ac-transposase is present and could be used for transposon tagging of carrot genes.