Nutrient deficiency on cultivated plants may lead to human malnutrition when consumes these plants. So, the biofortification is a promising approach in increasing several crops in their contents of essentials nutrients for human health like selenium (Se). The current study focused on the using two different sources of Se-fertilizers for biofortifying broccoli under nutrient deficiency stress in sandy soil. The applied doses of selenium in each form were 10, 20, 30, and 40 ppm in the form of mineral and biological nano Se-fertilizers. The vegetative parameters and yield of broccoli were determined. In general, almost studied parameters recorded the highest values after applying the dose of 30 ppm in both mineral and nano Se-fertilizers. The studied parameters included fresh and dry weight of leaves and broccoli head, head diameter and its length gave the highest values when the dose of 30 ppm from both Se-fertilizers applied. The content of N, P, K, and Se nutrients significantly increased by increasing applied Se-doses till 40 ppm in both Se-fertilizers except P. The harvested yield of head broccoli was obtained after applied dose of 30 ppm in case of both Se-fertilizers, with an increase rate for mineral Se-fertilizer 60.1 and 57.2%, whereas were 51.8 and 47.4% for nano-Se fertilizer for both seasons, respectively. The biological Se-nanofertilizer may be is preferable for a safe biofortification. The most distinguished findings in the current study that producing biofortified broccoli rich in Se for human health can be achieved under grown in sandy soils or nutrient deficiency stress. Further studies are needed to answer more open questions regarding producing a safe and healthy food for human nutrition.