Soil enzymes play critical roles in the decomposition of organic matter and nutrient cycling.In agricultural soils, enzyme activity is mainly affected by fertilization management such as chemical and organic fertilizers as well as temperature.In this study, the effects of fertilization regimes (i.e., organic vs. conventional) on soil enzyme activity for rice paddy under different temperature (ambient vs. elevated) were investigated through pot experiments using temperature gradient chambers.Rice plants were grown under different fertilization and temperatures for 121 days, and soil samples were collected in the middle-season drainage and the harvest seasons and analyzed for β-1,4-glucosidase (BG), β-1,4-N-acetylglucosaminidase (NAG), and acid phosphatase (AP), which involve carbon (C), nitrogen (N), and phosphorus (P) cycling, respectively.Unexpectedly, enzyme activity was rarely affected by fertilization treatments and decreased by elevated temperature, which is not in agreement with the theoretical relationship between temperature and enzyme activity.The decreased enzyme activity by elevated temperature was consistent with decreased soil water content of the soils caused by increased evapotranspiration, suggesting decreased substrate diffusion might restrict microbial and thus enzyme activity.The higher enzyme activity in the harvest than that in the middle growing season could be also attributed to the higher soil water content in the harvest season compared with the middle growing season.In addition, increased rice biomass in the later season should have provided more substrate to microbes through diffusion under high soil water conditions.Our results suggest that soil moisture may be a key factor affecting soil enzyme activity by affecting substrate diffusion.