We analyzed the effects of foliar spraying with amino acids, chitosan (CHS) and nanocomposites (NCs) of chitosan with the amino acids proline, l-cysteine and glycine betaine (CHS-Pro NCs; CHSCys NCs, CHS-GB NCs, respectively) on the changes in the physiological and biochemical parameters of iceberg lettuce grown at the control temperature (20 °C) and under chilling conditions (4 °C). The physicochemical parameters of the phospholipid monolayers (PLs) extracted from plants showed the effects of the treatments on the properties of the monolayers, namely, the packing density and flexibility. We observed increased accumulation of proline at 4 °C, and differences in the concentrations of sugars in most of the analyzed variants were a consequence of the lowered temperature and/or the use of organic compounds. A temperature of 4 °C caused a significant increase in the l-ascorbic acid level compared with that at 20 °C. Differences were also found in glutathione (GSH) content depending on the temperature and treatment with the tested organic compounds. CHS NCs loaded with Pro and GB were effective at increasing the amount of phenols under stress temperature conditions. We noted that a significant increase in the antioxidant activity of plants at 4 °C occurred after priming with Cys, CHS-Cys NCs, Pro and CHS-Pro NCs, and the CHS nanocomposites were more effective in this respect. Both low-temperature stress and foliar spraying of lettuce with various organic compounds caused changes in the activity of antioxidant enzymes. Two forms of dismutase (SOD), iron superoxide dismutase (FeSOD) and copper/zinc superoxide dismutase (Cu/ZnSOD), were identified in extracts from the leaves of iceberg lettuce seedlings. The application of the tested organic compounds, alone or in combination with CHS, increased the amount of malondialdehyde (MDA) in plants grown under controlled temperature conditions. Chilling caused an increase in the content of MDA, but some organic compounds mitigated t