Agrobacterium tumefaciens is a Gram-negative bacterium. It causes plants to produce crown gall disease because of the transfer, integration and expression of oncogenes encoded by the T-DNA (transferred DNA) region of the tumour-inducing (Ti) plasmid. A set of transferred genes directs the production of bacterial nutrients, called opines, formed by condensation of an amino acid and a keto acid or a sugar. Transformed cells synthesize and secrete substantial quantities of particular opines, which A. tumefaciens then uses as a carbon and sometimes nitrogen source. Agrobacterium tumefaciens strains are usually classified on the basis of the opines they can catabolize. Because of the ability to transfer DNA between different kingdoms, A. tumefaciens is also frequently used to generate transgenic plants. This study analyses five poorly characterized wildtype Agrobacterium strains, 1D1108, 1D1460, 1D132, 1D1478 and 1D1487, isolated from Euonymus, cane, cherry, Salix and apple, respectively. Partial Ti-plasmid sequence analysis demonstrated that the five strains harbour the nopaline-type Ti plasmid. Tumorigenesis and transient transformation assays of the five analysed and six wildtype A. tumefaciens strains were performed with selected plant species, including two or three species of Brassicaceae, Asteraceae, Solanaceae, Apiaceae and Leguminosae. The A. tumefaciens strains 1D1108, 1D1460 and 1D1478 showed higher transformation efficiencies than the previously characterized A. tumefaciens strains with several economically important crops. These data suggest the potential use of these newly characterized wildtype A. tumefaciens strains in transient transformation assays with certain plant species.