LED Light Intensity Regulates Nitrogen Assimilation Enzyme Activity and Metabolic Responses in Iceberg and Leaf Lettuce (Lactuca sativa L.)

Light availability is a key environmental factor regulating nitrogen assimilation, carbon metabolism, and nutritional quality in leafy vegetables grown in controlled environments. However, how practical lighting regimes used in plant factories with artificial lighting (PFALs) influence the coordination between nitrogen assimilation and central carbon metabolism across different lettuce cultivar types remains insufficiently understood. This study investigated how moderate differences in photosynthetic photon flux density (PPFD) influence nitrogen metabolism and metabolic coordination in hydroponically cultivated lettuce. Two cultivars representing contrasting morphological types, iceberg lettuce (‘Celebration’) and leaf lettuce (‘Sunny’), were grown under LED light intensities of 150 and 200 µmol·m−2·s−1. Nitrate, nitrite, and ammonium concentrations were measured together with the activities of nitrate reductase (NRA) and nitrite reductase (NiRA), as well as ascorbic acid content. Metabolomic profiling was additionally performed to characterize broader metabolic responses. Higher light intensity enhanced nitrate reduction capacity in both cultivars, but the resulting patterns of nitrogen accumulation were strongly genotype-dependent. The leaf lettuce cultivar ‘Sunny’ exhibited increased NRA and reduced nitrate accumulation under higher light intensity, whereas the iceberg lettuce cultivar ‘Celebration’ accumulated more nitrate under the same conditions. Ammonium responses further suggested differences in downstream nitrogen assimilation processes. Elevated light intensity also increased ascorbic acid levels in both cultivars. Metabolomic analysis revealed contrasting cultivar-specific shifts in central carbon metabolism, particularly involving soluble sugars and tricarboxylic acid cycle intermediates, indicating differential coordination between carbon metabolism and nitrogen utilization. Overall, these findings demonstrate that moderate changes in light intensity within the practical PFAL cultivation range can significantly influence the integration of carbon and nitrogen metabolism in lettuce. Importantly, cultivar-specific physiological traits determine how these metabolic responses translate into nitrate accumulation and nutritional quality in controlled-environment production systems.