Technical Abstract: Cultivated tomato (Solanum lycopersicum L.) dispersed out of Latin America beginning in the 16th century but little is known about worldwide patterns of diversity. A sample of 30 accessions was assembled to represent five regions (Europe, USA/Canada, Mexico/Central America, Asia or South America) and three time ranges representing recent breeding history (1930s – late 1940s, late 1950s – 1960s or 1980s – 2000s). DNA sequences were generated for 49 gene fragments (22.9 kb) in two plants per accession. All accessions were grown in the field for evaluation of highly heritable horticultural traits and assayed for nutritional content. °Brix, vitamin C and lycopene ranged from 4.3 – 8.5, 33.80 ug/ml – 99.02 ug/ml and 5.81 ug/g – 13.76 ug/g, respectively. Overall estimated p = 0.00112 (95% CI = 0.0090, 0.00134). South America was the only region with a significantly higher mean p relative to the other four regions. When accessions were grouped according to date of collection or year of cultivar release, mean p values did not significantly differ from each other. Based on AMOVA, significant variation was partitioned into accessions within regions (35.87%, df = 10, P = 0.000), individuals within accessions (60.74%, df = 45, P = 0.000) and within individuals (3.30%, df = 60, P = 0.000) but not among regions. Population structure analysis defined two clusters that were not strictly associated with a priori groups. South American germplasm seems promising as a source of new alleles but no geographical region should be viewed as a poor source of tomato genetic variation.