|Thomson, James - Jim|
Submitted to: BMC Research Notes
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 2/22/2011
Publication Date: 3/2/2011
Citation: Thomson, J.G., Cook, M.A., Guttman, M.E., Smith, J.D., Thilmony, R.L. 2011. Novel sull binary vectors enable an inexpensive foliar selection method in Arabidopsis. BMC Research Notes. 4(44) Available: http://www.biomedcentral.com/1756-0500/4/44.
Interpretive Summary: Selectable markers are integral components of plant biotechnology research that allow the identification of genetically modified plant cells. A set of molecular tools (plant transformation vectors) that facilitate the engineering of plants has been generated and characterized. In addition a new screening method that allows the selection of greenhouse grown plants with a spray treatment of the selectable agent was developed. These plant transformation tools and the novel screening method will facilitate the identification of transformed plants and enable research characterizing the role of candidate genes or studying regulatory sequences that control gene expression.
Technical Abstract: Sulfonamide resistance is conferred by the sulI gene found on many Enterobacteriaceae R plasmids and Tn21 type transposons. The sulI gene encodes a sulfonamide insensitive dihydropteroate synthase enzyme required for folate biosynthesis. Transformation of tobacco, potato or Arabidopsis using sulI as a selectable marker generates sulfadiazine-resistant plants. Typically sulI-based selection of transgenic plants is performed on tissue culture media under sterile conditions. A set of novel binary vectors containing a sulI selectable marker expression cassette were constructed and used to generate transgenic Arabidopsis. We demonstrate that the sulI selectable marker can be utilized for direct selection of plants grown in soil with a simple foliar spray application procedure. A highly effective and inexpensive high throughput screening strategy to identify transgenic Arabidopsis without use of tissue culture was developed. Novel sulI containing Agrobacterium binary vectors designed to over-express a gene of interest or to characterize a test promoter in transgenic plants have been constructed. These new vector tools combined with the various beneficial attributes of sulfonamide selection and the simple foliar screening strategy provide a desirable alternative for plant biotechnology researchers. This set of binary vectors is freely available upon request.