Using a binary tumor-inducing (Ti) plasmid vector system, several plant species were transformed with a kanamycin resistance marker (neomycin phosphotransferase gene). Four Nicotiana species, seven tomato cultivars, two potato cultivars, and Arabidopsis thaliana were transformed by the binary vector transformation method. In this method, various plant organ pieces were co-cultivated with Agrobacterium Transformation of Plants. Leaves were cut to about 0.1 to 0.5 cm2 and stems were cut to about 1 to 2 cm long. The plant sections were wounded with forceps and cultivated in 2 ml of MS liquid medium in 60 mm Petri dishes with about 108 A. Neomycin Phosphotransferase Assay. Plant materials used for the enzyme assay were from independently isolated tissues and free of the Agrobacterium. Plant tissues were lysed by homogenization in 1 ml/g of plant extraction buffer (0.5 M sucrose, 0.1% ascorbic acid, 0.1% cystine-HCl, and 0.1 M Tris-HCl [pH 7.5]). The lysate was centrifuged for 5 min in an Eppendorf centrifuge and supematant adjusted to 1 mg/ml of total protein. Twenty zl of the crude extract was used for the neomycin phosphotransferase assay as described previously (2,13

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