AbstractAcid rain, which has negative impacts on the vegetation of ecological systems, is widespread in Northern and Southern China. However, relatively little is known about the effects of acid rain on the growth and yield of economically important tree species in China. To address this issue, we studied the responses of mulberry seedlings to simulated acid rain (SAR) at different pH values. At pH 4.5, SAR induced increased antioxidant activities, total antioxidant capacity, and the accumulation of reactive oxygen species (OFR) relative to controls. However, the growth of the seedlings under SAR treatments at pH 4.5 and pH 5.6 was greater than controls. No significant differences in photosynthesis and chlorophyll a fluorescence quenching parameters were observed between the SAR treatments at pH 4.5 and pH 5.6 and controls. However, the SAR treatment at pH 3.5 resulted in altered leaf surface characteristics and changes to chloroplast ultrastructure, together with an increase in membrane electrical conductivity and an accumulation of OFR and malondialdehyde. In contrast, leaf antioxidant enzyme activities were decreased, together with electron transport parameters and photosynthesis. Taken together, these results show that the effects of acid rain on the growth and leaf physiology of mulberry seedling are dependent on pH. Moreover, mulberry seedlings had a high tolerance to acid rain at pH 4.5.