The study was performed to analyze the impact of seed pretreatment by static magnetic field (SMF) of 200 mT for 1 h on photosynthetic performance of soybean (Glycine max) seedlings under ambient (aUV-B) and supplemental ultraviolet-B (a+sUV-B) stress. Ambient and supplemental UV-B were found to decrease the plant growth, chlorophyll concentration, PSII efficiency, selected JIP-test parameters such as Fv/Fm, φEo, ΔV(I–P), PIABS, PItotal, and rate of photosynthesis in the leaves of soybean seedlings emerged from untreated (UT) seeds. aUV-B and a+sUV-B were observed to increase the synthesis of UV-B-absorbing substances (UAS), reactive oxygen species (ROS) like superoxide radical (O2·−) and hydrogen peroxide (H2O2), antioxidants like ascorbic acid and α-tocopherol and decrease the nitrate reductase (NR) activity; subsequently, it results in a decreased rate of photosynthesis, biomass accumulation, and yield. However, our results provided evidence that SMF pretreatment increased the tolerance of soybean seedlings to UV-B radiation by increased NO content and NR activity; higher efficiency of PSII, higher values of φEo, ΔV(I–P), PIABS, and PItotal, decreased intercellular CO2 concentration, lower amount of UAS, ROS, and antioxidants that consequently improve the yield of soybean plants under aUV-B as well as a+sUV-B stress. Thus, our results suggested that SMF pretreatment mitigates the adverse effects of UV-B stress by the enhancement in photosynthetic performance along with higher NO content which may be able to protect the plants from the deleterious effects of oxidative stress caused by UV-B irradiation.