Purpose: The enrichment of health-promoting compounds in plants and foods has received significant research attention over the past years, leading researchers to use cutting-edge technologies like elicitation in agriculture and food production systems. Engineered nanoparticles (NPs) have been shown to function as effective elicitors, enhancing the synthesis of secondary metabolites. Although carbon dots (CDs) are at the forefront due to their favorable characteristics, such as being green, biocompatible, and low toxicity, their functions as elicitors have not been thoroughly investigated. The aim of this study was, therefore, to investigate the potential effect of sugar beet molasses carbon dots (SBM-CDs), characterized by their endogenous food-borne nature as elicitors, on the agronomic and bioactive compounds of wheatgrass juice obtained from hydroponically cultivated wheatgrasses. Methods: Wheatgrasses were grown with and without SBM-CDs extracted from molasses at 50–200 mg L− 1 concentrations through a nutrient solution in a hydroponic system. After 7 days, wheatgrass juice was obtained by squeezing wheatgrass. The effects of SBM-CDs were investigated by assessing the agronomic parameters and bioactive compounds of wheatgrass juice. Results: The amount of β-carotene, vitamin E, vitamin C, and chlorophyll a increased by 150%, 84%, 25%, and 89%, respectively, with the application of 200 mg L− 1 SBM-CDs (p < 0.01) in comparison with the control group (the application without SBM-CDs). Besides, this application resulted in a 34% increase in the total quantity of tested phenolic compounds. Conclusions: These data suggest that our biomass-derived renewable CDs may be a novel category of elicitors for enhancing the production of bioactive compounds in wheatgrass.