The application of selenium (Se) can promote the growth of potatoes and increase the Se content of potato tubers. Nitrogen (N) can promote nutrient absorption and potato yield. However, the effects of Se and N on the yield and Se content of potato tubers have not been reported. Field experiments were conducted in 2019–2020 and 2020–2021. Three N levels, i.e., 0 kg N ha−1 (N0), 150 kg N ha−1 (N1) and 200 kg N ha−1 (N2), and three Se levels, i.e., 0 g Se ha−1 (Se0), 500 g Se ha−1 (Se1) and 1000 g Se ha−1 (Se2), were set up. Potato yield, root dry weight, root activity, leaf area index (LAI), net photosynthetic rate (Pn), malondialdehyde (MDA) content, antioxidant enzyme activity and Se content were investigated. N application increased root dry weight and activity, LAI, antioxidant enzymes activities, Se content and decreased MDA content. Pn, total potato yield and commercial potato yield first increased and then decreased with the increase in N application level. Se application increased root activity, Pn, antioxidant enzyme activities, total yield, commercial potato yield, commercial potato rate, Se content and decreased MDA content. In general, compared with the single application, the combination of N and Se fertilizers further promoted the growth of potatoes, increased the yield of potatoes and the Se content of potato pieces.