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Light is an abiotic factor, and its quality, quantity, and photoperiod can be modulated to work as eustress inductors to regulate plant processes. It is known that red (R), blue (B), far-red (FR), and ultraviolet-A wavelengths can promote photomorphogenesis and secondary metabolite production in plants. Several ratios of R:B and the addition of end of-day FR, separately, have beneficial effects on plant development, whereas adding ultraviolet-A enhances the production of secondary metabolites such as phenols. However, the effects of extended photoperiods with a mixture of these four wavelengths and extra end-of-day FR have not been evaluated for plants of commercial interest. The objective of this study was to determine the effects on tomato seedlings (‘Saladette’, CORDOBA F1) of different overnight photoperiods using a fixed combination of R (625 nm), B (460 nm), FR (720 nm), and ultraviolet-A (410 nm). We expected increases in the production of specialized metabolites and the generation of beneficial changes in the seedling biomass and morphology. Four treatments involving overnight artificial light provided by a light-emitting diode (LED) module were established: TC (control), 0 h; T1, 4 hours; T2, 8 hours; and T3, 12 hours. All treatments were subjected to a 12-hour natural photoperiod and 12 hours of overnight artificial light. The experiment lasted 4 weeks, and plants were sampled every week for physical and phytochemical measurements. In general, seedlings subjected to 4-hour and 8-hour treatments presented better results than those subjected to the control and 12-hour overnight photoperiod treatments. Seedlings subjected to treatments with an 8-hour overnight photoperiod presented large accumulation of biomass in the stem rather than in the leaves because they had large stem dry weight, stem weight, and elongation and higher first, second, and third internode lengths; however, they had lower leaf area, leaf dry weight, and health index. Seedlings subjected to treatment with a 4-hour overnight photoperiod were visually bigger, with large leaf expansion, total length, stem weight, total weight, and specific leaf area; however, this treatment had a negative impact on the biomass accumulation, with lower leaf weight, stem dry weight, and health index. The 12-hour treatment had a negative impact on the leaf area, and thus the specific leaf area, of seedlings; however, the biomass accumulation was large, with higher leaf dry weight, total dry matter, and specific leaf area, but no difference in stem dry weight compared with the control. At the end of the experiment, the total phenolic content increased in all treatments compared with the control, but the flavonoid content decreased. Moreover, the antioxidant capacity was higher for T2 during the last 2 weeks of the experiment. Results are discussed according to the possibilities of using this light strategy for seedling production.