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UV-B radiation has been previously reported to induce protective or deleterious effects on plants depending on the UV-B irradiation doses. To elucidate how these contrasting events are physiologically coordinated, we exposed sweet basil plants to two UV-B doses: low (8.5 kJ m^−2 day^−1, 30 min exposure) and high (68 kJ m^−2 day^−1, 4 h exposure), with the plants given both doses once continuously in a single day. Physiological tests during and after both UV-B exposures were performed by comparing the stress-induced damage and adverse effects on photosynthetic activity, the concentration and composition of photosynthetic and non-photosynthetic pigments, and stress-related hormones biosynthesis in basil plants. Our results showed that upon receiving a high UV-B dose, a severe inactivation of oxygen evolving complex (OEC) activity at the PSII donor side and irreversible PSII photodamage caused primarily by limitation of the acceptor side occurred, which overloaded protective mechanisms and finally led to the death of the plants. In contrast, low UV-B levels did not induce any signs of UV-B stress injuries. The OEC partial limitation and the inactivation of the electron transport chain allowed the activation of photoprotective mechanisms, avoiding irreversible damage to PSII. Overall results indicate the importance of a specific response mechanisms regulating photoprotection vs irreversible photoinhibition in basil that were modulated depending on the UV-B doses.