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A new plant transformation vector, pGKB5, was designed for T-DNA tagging in Arabidopsis thaliana using in planta transformation procedures in which adult plants are decapitated and inoculated with an Agrobacterium strain. The T-DNA region contains two selectable markers conferring resistance to kanamycin and Basta (phosphinothricin [glufosinate]). A promoterless β-glucuronidase reporter gene is fused to the right border of the T-DNA in order to identify gene fusions upon insertion in the plant genome. The Basta resistance allows selection of transformants in the greenhouse at high density. Using this system, transformants were obtained by in planta transformation procedures. Most of the transformants showed Mendelian segregation for one or two unlinked genetic loci. Hybridization analysis showed that the structure of the inserted T-DNA is relatively simple, with one or a few tandem-copies inserted at …