Genetic diversity underlies the improvement of crops by plant breeding. Landraces of tomato (Solanum lycopersicum L.) can contain valuable alleles not common in modern germplasms. The aim was to measure genetic diversity present in 47 most common tomato varieties grown in Italy, 35 were varieties used for processing and 12 were landraces considered “salad varieties”. Furthermore, we demonstrated the possibility that the variety traceability can be extended through the entire production chain. Diversity was measured using 11 microsatellite markers and 94 genotypes. Among the markers used, a total of 48 alleles were detected. A dendrogram based on total microsatellite polymorphism grouped 47 varieties into three major clusters at 0.75 similarity coefficient, differentiating the modern varieties from tomatoes landraces. The DNA markers developed confirmed the possibility to support the genotype identification all along the tomato production chain. The number of alleles and genotypes identified in the present work is the largest considering papers on food traceability.