Regeneration from VFNT Cherry tomato was optimized prior to transformation. Cotyledon and hypocotyl sections from 7-day-old in vitro grown VFNT Cherry tomato were cultured on medium containing MS salts, B5 vitamins and the following per liter: sucrose, 30g; zeatin, 1 mg; IAA, 0.1 mg; agar, 8.0g or GelriteR, 2.2g. Culture conditions investigated included cotyledon and upper hypocotyl explant lengths, light vs. dark germination, and agar vs. GelriteR. The conditions which resulted in the highest average number of shoots per explant were cotyledon basal explants 2 mm in length, 3.95; cotyledons from dark germination, 6.2; and hypocotyls from light germination, 8.6. An equal number of shoots regenerated on medium containing agar or GelriteR, however, shoots regenerated on medium containing agar were more vigorous. Cotyledon and hypocotyl sections were cocultivated with the Agrobacterium tumefaciens binary vector pBI121 containing the neomycin phosphotransferase II (NPTII) and B-glucuronidasc (GUS) genes. Transformants were selected by regeneration and rooting on medium containing kanamycin. Southern blot and PCR analysis indicated regeneranrs contain the NPTII and GUS genes. Mapping the chromosomal location of the NPTII gene is in progress.