To investigate the relationship between the lighting direction-induced morphophysiological traits and post-harvest storage of lettuce, the effects of different lighting directions (top, T; top + side, TS; top + bottom, TB; side + bottom, SB; and top + side + bottom, TSB; the light from different directions for a sum of light intensity of 600 μmol·m·s photosynthetic photon flux density (PPFD)) on the growth morphology, root development, leaf thickness, stomatal density, chlorophyll concentration, photosynthesis, and chlorophyll fluorescence, as well as the content of nutrition such as carbohydrates and soluble proteins in lettuce were analyzed. Subsequently, the changes in water loss rate, membrane permeability (measured as relative conductivity and malondialdehyde (MDA) content), brittleness (assessed by both brittleness index and β-galactosidase (β-GAL) activity), and yellowing degree (evaluated based on chlorophyll content, and activities of chlorophyllase (CLH) and pheophytinase (PPH)) were investigated during the storage after harvest. The findings indicate that the TS treatment can effectively reduce shoot height, increase crown width, enhance leaves' length, width, number, and thickness, and improve chlorophyll fluorescence characteristics, photosynthetic capacity, and nutrient content in lettuce before harvest. Specifically, lettuce's leaf thickness and stomatal density showed a significant increase. Reasonable regulation of water loss in post-harvested lettuce is essential for delaying chlorophyll degradation. It was utilized to mitigate the increase in conductivity and hinder the accumulation of MDA in lettuce. The softening speed of leafy vegetables was delayed by effectively regulating the activity of the β-GAL. Chlorophyll degradation was alleviated by affecting CLH and PPH activities. This provides a theoretical basis for investigating the relationship between creating a favorable light environment and enhancing the post-harvest preservation of leafy vegetables, thus prolonging their post-harvest storage period through optimization of their morphophysiological phenotypes.