Fruits and vegetables are rich sources of antioxidants in human diets and their intake is associated with chronic disease prevention. Lettuce (Lactuca sativa L.) is a common vegetable in diets worldwide, but its nutritional content is relatively low. To elucidate the genetic basis of antioxidant content in lettuce, we measured the oxygen radical absorbance capacity (ORAC) and chlorophyll (Chl) content as a proxy of β-carotene in an F(8) recombinant inbred line (RIL) in multiple production cycles at two different production sites. Plants were phenotyped at the open-leaf stage to measure genetic potential (GP) or at market maturity (MM) to measure the influence of head architecture ('head' or 'open'). Main effect quantitative trait loci (QTL) were identified at MM (three Chl and one ORAC QTL) and GP (two ORAC QTL). No main effect QTL for Chl was detected at GP, but epistatic interaction was identified in one pair of marker intervals for each trait at GP. Interactions with environment were also detected for both main and epistatic effects (two for main effect, and one for epistatic effect). Main effect QTL for plant architecture and nutritional traits at MM colocated to a single genomic region. Chlorophyll contents and ORAC values at MM were significantly higher and Chl a to Chl b ratios were lower in 'open' types compared to 'head' types. The nutritional traits assessed for GP showed a significant association with plant architecture suggesting pleiotropic effects or closely linked genes. Taken together, the antioxidant and chlorophyll content of lettuce is controlled by complex mechanisms and participating alleles change depending on growth stage and production environment.