To determine the role of microwaves in the stress resistance of plants to enhanced ultraviolet-B (UV-B) radiation, Isatis indigotica Fort. seeds were subjected to microwave radiation for 8 s (wavelength 125 mm, power density 1.26 mW mm⁻², 2450 MHz). Afterwards they were cultivated in plastic pots in an artificial-glass greenhouse maintained at 25°C, 70% relative humidity, and 400 μmol mol⁻¹ CO₂, under visible-light conditions of 1500 μmol m⁻² s⁻¹ for 8 h day⁻¹. When the seedlings were 10 days old, they were subjected to 10.08 kJ m⁻² UV-B (PAR: 220 μmol m⁻² s⁻¹) radiation for 8 days. Changes in a number of physiological and biochemical characteristics and in the thermal decomposition enthalpy of biomass were measured and used as indicators of the protective capacity of microwave radiation in this experiment. Our results revealed that microwave pretreatment of seeds enhanced UV-B stress resistance in the seedlings by decreasing the concentration of malondialdehyde (MDA) and increasing the concentration of ascorbic acid (AsA) and UV-B–absorbing compounds, increasing the activities of catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD), and increasing the energy accumulation of photosynthesis. All these results suggest that microwave radiation enhances plant metabolism and results in increased UV-B stress resistance. This is the first investigation reporting the use of microwave pretreatment to protect the cells of Isatis indigotica from UV-B-induced lesions.