Iron (Fe) is a vital element for plants and other organisms, involving in several physiological processes including respiration, chlorophyll biosynthesis, and photosynthesis. Unfortunately, how Fe accumulation regulates in response to light quality has not been well established in plants. Therefore, the aim of the study was to explore the mechanism of Fe homeostasis by light quality. In this study, we found morpho-physiological attributes were significantly improved in response to blue (λmax: 450) compared to white (λ max: 500) and red (λmax: 660) light. The root-shoot length, plant biomass, photosynthesis efficiency (Fv/Fm) and leafgreen (SPAD) significantly declined in response to white and red light. However, these parameters were improved and iron deficiency was substantially alleviated by blue light exposure in alfalfa seedlings. This study might be useful to the forage breeders and farmers for improving alfalfa yield and nutritional benefits.