Detection and analysis of T-DNA in crown gall tumors and kanamycin-resistant callus of Robiniapseudoacacia

Five Agrobacteriumtumefaciens strains, and one A. rhizogenes strain, incited tumors on Robiniapseudoacacia L. (black locust) tissues that were inoculated invitro. Four phytohormone-independent tumors, incited by A. tumefaciens strains A6, A348, A274, and A208, were examined using Southern analysis. T-DNA sequences were detected in, and appeared to be integrated into, the DNA from all the tumors. Some tumors tested negative for the presence of octopine or nopaline. All of these tumors had T-DNAs that were truncated on the right ends, i.e., missing the region in which those particular opine synthetic loci are present on the Ti plasmid. Kanamycin-resistant callus was also obtained after inoculation of cotyledons with A. tumefaciens strain A281 carrying the binary vector pGA472, and Southern analysis indicated that nptII sequences were integrated into the R. pseudoacacia genome.