Sink and source activity in two tomato (Solanum lycopersicum L.) genotypes that vary in fruit Brix were investigated to identify differences that potentially underscore this trait. Solara (Brix 9%) accumulated almost twice the glucose, fructose, and sucrose in ripe fruit and had a higher horticultural yield (25% greater) compared to Moneymaker (Brix 5%). 14C-glucose feeding suggested large disparities in sucrose metabolism in ripe fruit between genotypes. Biochemical pathways in the leaf adjacent to a fruiting truss at night were also analyzed since in many species, this is the period when leaf reserves are mobilized to feed the plant. Surprisingly, leaf metabolism, i.e., starch and sugar content, the levels of polar metabolites assayed by GC-TOF MS and 14CO2-pulse-chase fluxes in detached leaves, did not change between the day and night in either genotype. Solara has a higher morphological source-to-sink ratio, and this may contribute to higher Brix in that genotype.