Biofortification can be exploited to enrich plants in selenium (Se), an essential micronutrient for humans. Selenium as selenate was supplied to two rocket species, Eruca sativa Mill. (salad rocket) and Diplotaxis tenuifolia (wild rocket), at 0–40 μM in hydroponics and its effects on the content and profile of sulphur (S)-compounds and other phytochemicals was evaluated. D. tenuifolia accumulated more total Se and selenocysteine than E. sativa, concentrating up to ~300 mg Se kg−1 dry weight from 10–40 μM Se. To ensure a safe and adequate Se intake, 30 and 4 g fresh leaf material from E. sativa grown with 5 and 10–20 μM Se, respectively or 4 g from D. tenuifolia supplied with 5 μM Se was estimated to be optimal for consumption. Selenium supplementation at or above 10 μM differentially affected S metabolism in the two species in terms of the transcription of genes involved in S assimilation and S-compound accumulation. Also, amino acid content decreased with Se in E. sativa but increased in D. tenuifolia and the amount of phenolics was more reduced in D. tenuifolia. In conclusion, selenate application in hydroponics allowed Se enrichment of rocket. Furthermore, Se at low concentration (5 μM) did not significantly affect accumulation of phytochemicals and plant defence S-metabolites.