Vertical farming systems offer an efficient solution for sustainable food production in urban areas. However, managing nitrate (NO₃⁻) levels remains a significant challenge for improving crop yield, quality, and safety. This study evaluated the effects of nitrate availability on growth performance, nutrient uptake, and water use efficiency in a vertical hydroponic system that intercropped lettuce (Lactuca sativa) with alfalfa (Medicago sativa). The experiment was conducted in a controlled vertical hydroponic system using Nutrient Film Technique (NFT) channels, with nitrogen levels set at 0, 33, 66, 100, and 133% of the standard concentration. The results indicated that the intercropping treatment with 66% nitrate (IC-N_66%) improved water use efficiency by 38% and slightly increased leaf area compared to the other intercropping treatments. However, the control group, which consisted of a monoculture with full nitrate supply, achieved the highest overall biomass. Ion concentrations, including nitrate, calcium, magnesium, and micronutrients, were moderately affected by the intercropping strategy and nitrate levels. These findings suggest that moderate nitrate input, combined with nitrogen-fixing legumes, can enhance resource efficiency in hydroponic systems without significantly compromising yield. These findings offer a promising framework for incorporating legumes into hydroponic systems, minimizing the need for synthetic inputs while maintaining yield. These results support the use of agroecological intensification strategies in highly efficient soilless systems.