Six transgenic 'South Bay' lettuce lines (Lactuca sativa L.) with elevated levels of 5-enolpyruvyl shikimate-3-phosphate synthase (EPSPS) were evaluated for tolerance to the herbicide glyphosate, The six lines were selected from approximate to 150 independent transformation events using and Agrobacterium tumefaciens system. Three assay methods were used to identify gene expression with regard to glyphosate resistance. Leaf disks: of the transgenic lines were cultured on media containing 0 to 1280 muM glyphosate, Leaf disks of the control had lower dry weight (DW) at 40 muM and greater glyphosate than all the transgenic lines. The transgenic lines continued to grow even at 1280 muM. Plants 21 days old were sprayed in the greenhouse with rates of glyphosate at 0 to 35.84 kg.ha(-1). DW of all the lines were similar to the control, with a few exceptions, at glyphosate concentrations from 0 to 0.56 kg.ha(-1). At 2.24 to 8.96 kg.ha(-1) all of the transgenic lines had DW greater than the control, while at 17.92 and 35.84 kg.ha(-1) only B-32, B-33, C-3, and C-14 had DW greater than the control. The resistant line from the greenhouse experiment, B-32, grew normally in field trials at the highest glyphosate rate, 17.92 kg.ha(-1), while control plants died at 0.55 kg.ha(-1) glyphosate, Lines A-11 and C-3 had lower DW than B-32 at 2.24 kg ha(-1) glyphosate and greater, While Leaf disk assays can identify potential transformed lines expressing the EPSPS and glyphosate oxidase (GOX) gene, and greenhouse screening can evaluate seedling vigor after glyphosate application, field trials are necessary to evaluate plant growth and yield through the growing season. Chemical name used: N-(phosphono-methyl) glycine (glyphosate).