Aims Taking into account the relevance of biofortification of crop plants with selenium (Se) - an essential element for humans and animals -, this study assessed the effect of adding Se in the soil (as selenate) on: i) rice grains yield; ii) Se contents in polished rice grains; iii) physiological traits in rice leaves; and, iv) spatial distribution of Se and other elements in biofortified grains. Methods A field trial was conducted using five Se rates (12, 21, 38, 68, and 120 g ha −1 ). Physiologic and enzymatic evaluations were carried out in rice leaves. Rice grain yield and Se contents were assessed. Spatial distributions of Se and other elements were visualized mapping the biofortified rice grains with μXRF at Brazilian Synchrotron Light Source. Results Results showed that soil Se application was effective in producing rice grains with higher Se contents, yet no effects were verified on rice yield. Antioxidant enzyme activities and gas exchanges in rice leaves changed following the application of Se. The spatial distribution of different elements in biofortified grains varied, with Se being accumulated mainly in the rice endosperm. Conclusion Our results suggest that soil application of 47 and 36 g ha −1 of Se (as sodium selenate) may guarantee the production of rice grains with adequate Se levels for human consumption in Brazil and worldwide, respectively.