Zinc (Zn) is an essential micronutrient for plant growth, serving as a co-factor in enzymatic processes and pigment biosynthesis. In horticultural crops such as lettuce, Zn fertilization is increasingly relevant for optimizing yield and nutritional quality. In this study, a greenhouse pot experiment was conducted using Lactuca sativa L. cv. Romana Verano (Ramiro Arnedo) to evaluate the effects of four Zn sources with contrasting physio-chemical properties—ZnSO₄, a synthetic chelate containing DTPA, EDTA, and HEDTA, a Zn–lignosulphonate complex, and ZnO nanoparticles—applied to soil at rates of 15, 30, 60, and 120 mg Zn·kg⁻¹. Morphometric traits, photosynthetic pigmentation, and photosystem performance were assessed to determine differences in plant response. Results showed that low to moderate Zn supply (15–60 mg Zn·kg⁻¹) maintained growth, leaf number, stem diameter, and biomass without significant changes compared to the control. In contrast, the highest dose (120 mg Zn·kg⁻¹), particularly in chelated forms, led to reductions in growth and yield exceeding 80%, reflecting supra-optimal effects. Although lignosulphonate and nanoparticles sources lowered soil Zn availability, they did not affect lettuce growth or yield, indicating their potential as safer agricultural alternatives to conventional Zn fertilizers. Photosynthetic efficiency, measured through chlorophyll fluorescence and electron transport activity, was positively modulated by adequate Zn levels but declined at excessive concentrations. These findings highlight that Zn efficiency strongly depends on its chemical form and applied dose, providing practical insights for optimizing Zn fertilization strategies in lettuce and other horticultural crops.