Lighting from red and blue light-emitting diodes (LEDs) is common for crop production in controlled environments. Continuous application of red or blue light at night has been shown to suppress sporulation by Peronospora belbahrii, the causal organism of basil downy mildew (DM), but the suppressing effects of intermittent applications of red and blue LEDs have not been thoroughly researched. This study examined the effects of red (λmax = 670 nm) and blue (λmax = 458 nm) LED top lighting, at two photosynthetic photon flux densities (PPFD = ≈12 and ≈60 µmol·m−2·s−1), using continuous (10-hour) nighttime and two intermittent nighttime exposures, to suppress basil DM sporulation. The two intermittent treatments consisted of one 4-hour exposure and three 1.3-hour exposures spaced 3 hours apart. Continuous nighttime treatments with blue or red LED top lighting at ≈60 µmol·m−2·s−1 were able to suppress basil DM sporulation by more than 99%. At a given nighttime dose of light that did not completely suppress sporulation, continuous lighting was more effective than intermittent lighting, and for these partially suppressing doses, red LEDs were not significantly different from blue LEDs for suppressing sporulation. The present study showed that horticultural lighting systems using red and blue LEDs to grow crops during the day can also be used at night to suppress basil DM sporulation by up to 100%.