To reduce nitrate concentrations in vegetables grown under artificial lighting, we determined nitrate accumulation under various conditions of nitrate availability. Butterhead lettuce plants were grown with nutrient solutions of various concentrations, which were maintained according to electrical conductivity (EC) throughout the cultivation period. Under these conditions, growth in nutrient solutions with lower EC led to slight decreases in nitrate concentrations in leaves, but also decreased fresh and dry weights of shoots, leading to the risk of a yield loss under EC control management (ECM). By contrast, when total nitrate requirement was supplied only at the start of cultivation, nitrate concentrations in leaves were decreased significantly with only slightly reduced plant growth. Hence, marketable-sized butterhead lettuces with lower nitrate concentrations can be produced by supplying total nitrate requirements in the nutrient solution at the start of cultivation.