Transgenic cotton plants have been produced that express a chimeric gene that confers tolerance to the active ingredient (glyphosate) in the nonselective, broad spectrum herbicide Roundup^®. This achievement was made possible by the development of a reliable regeneration protocol coupled with Agrobacterium-mediated transformation. Hypocotyl explants were inoculated with an Agrobacterium strain containing a chimeric coding sequence for NPT 11 (conferring kanamycin resistance) and for a glyphosate-tolerant EPSP synthase. Following a co-culture period, the explants were cultured on media containing kanamycin. Plants were obtained via somatic embryogenesis from kanamycin-resistant calli. Confirmation of transformation was done by a NPT II dot blot assay, assays for overexpression of the EPSP synthase enzyme, and the expression of extracted EPSP synthase in the presence of glyphosate. Progeny were grown from transformed plants and sprayed with various rates of glyphosate at the 2-4 leaf stage. After 2 weeks, the plants were evaluated for damage. No visible damage was observed in the transgenic plants while significant growth reduction was present in the controls with necrosis in the shoot apex. In the three progeny sets tested, all exhibited a three to one segregation ratio, indicative of a single insert.