ABSTRACT Exogenous selenium (SE) application during lettuce (Lactuca sativa L.) production may prevent Se deficiency. Low concentrations of Se have beneficial effects on plant cell metabolism and application of Se can increase growth, yield, and quality and reduce nitrate concentration in lettuce. In the present study, biosynthetic synthesis of selenium (Se) nanoparticles (NPs) was carried out via Se ions reduction during exposure to rosemary (Salvia rosmarinus Spenn.) extract. Using scanning electron microscopy (SEM), the shape and size of the Se NPs were determined. To investigate effects of concentrations of sodium selenite (1, 2, and 4 mg∙L−1) and Se NPs (1, 2, and 4 mg∙L−1) on morphological and physiochemical traits of lettuce. Most nutrient solutions, especially 2 mg∙L−1 Se NPs, increased plant height, leaf number, fresh weight, chlorophyll a, total chlorophyll, and nitrate reductase activity of plants. Application of 2 and 1 mg∙L−1 Se NPs, caused a decrease in the level of nitrate accumulation. Under nutrient treatments, concentrations of P, K, Fe, Zn, and Se, and concentration of N decreased. Foliar application of biosynthetic synthesized Se NPs, may have potential for improved quantity and reduced nitrate accumulation in lettuce.