Abstract <p>Polyhydroxylated fullerenes are actively studied for their effects on plant growth, development, and tolerance to abiotic stress in various crops. The scientific interest in these carbon nanomaterials is due to their unique structure, good water solubility, and biocompatibility. However, the effects of fullerenol C₆₀(OH)₂₄ (FNPs) on physiological and biochemical traits and low-temperature tolerance of wheat seedlings (<i>Triticum aestivum</i> L., cv. Zlata) remain unclear. In this study, a FNPs concentration of 0.1 mg/L was selected for seed nanopriming, as it enhanced seed germination energy and germination percentage. Nanopriming promoted seedling growth, elevated chlorophyll <i>a</i> content in leaves, and enhanced both photosynthetic and dark respiration rates. Structural changes in mesophyll cells were also observed, including an increase in chloroplast area. A decrease in total SOD activity was detected in leaves of seedlings grown from fullerenol-primed seeds, while no differences were observed in the activity of H₂O₂-scavenging enzymes (POX, CAT, APX). At the same time, these seedlings accumulated higher levels of proline and ascorbic acid, whereas TBARS and H₂O₂ contents were lower compared with the control variant. The transcript levels of the key COR genes <i>Wcor15</i> and <i>Wcor726</i> were also increased after nanopriming with FNPs. The protective effect of FNPs under low-temperature stress (–2°C for 24 h) was confirmed by increased total antiradical activity (according to the degree of neutralization of the DPPH radical) and reduced electrolyte leakage. Thus, this study demonstrates for the first time that fullerenol acts as a stimulator of plant growth and development, enhances photosynthetic activity, and functions as an effective antioxidant that mitigates low-temperature-induced damage to cell membranes in wheat seedlings. An attempt was made to identify candidates to markers of nanopriming at different levels of the plant organism organization.</p>