In Japan, red perilla leaves are used in the food and coloring industries, as well as in crude medicine. Perilla leaves contain a high concentration of phytochemicals such as perillaldehyde (PA) and rosmarinic acid (RA). The effects of UV-B radiation intensity (0.05–0.2 W m−2, UV-BBE: 0.041–0.083 W m−2), duration (3 or 6 h), and irradiation method (continuous or intermittent) for artificial nocturnal lighting using UV-B fluorescent lamps were evaluated on growth, flowering, and leaf phytochemical concentration in greenhouse-grown perilla. Under continuous UV-B irradiation at 0.1 W m−2 for 3 or 6 h, leaf color changed from red to green and leaf fresh weight decreased, compared with the control treatment. No leaf color change was observed under the 3-h treatment with UV-B radiation at 0.05 W m−2, wherein leaf fresh weight was similar to that of the control. Furthermore, RA concentration under continuous UV-B irradiation at 0.05 W m−2 for 3 h increased two-fold compared to that under control treatment, while PA concentration was not affected by UV-B irradiation. Thus, our data showed that continuous UV-B irradiation at 0.05 W m−2 for 3 h could effectively produce RA-rich perilla leaves without reducing in phenotypic quality or productivity. However, a 6-h intermittent illumination inhibited flowering without altering phytochemical concentration.