Light Quality Plays a Crucial Role in Regulating Germination, Photosynthetic Efficiency, Plant Development, Reactive Oxygen Species Production, Antioxidant Enzyme Activity, and Nutrient Acquisition in Alfalfa.

Light is a vital regulator of photosynthesis, energy production, plant growth, and morphogenesis. Although these key physiological processes are well understood, the effects of light quality on the pigment content, oxidative stress, reactive oxygen species (ROS) production, antioxidant defense systems, and biomass yield of plants remain largely unexplored. In this study, we applied different light-emitting diode (LED) treatments, including white light, red light, blue light, and a red+blue (1:1) light combination, to evaluate the traits mentioned above in alfalfa ( L.). Fluorescence staining showed that red light significantly triggered the oxidative stress indicators compared to blue and white light, while the combined red and blue light treatment significantly reduced the ROS (O, HO) intensity in alfalfa seedlings. Interestingly, the combined light treatment significantly boosted the seed germination rate (%), maximum photochemical quantum yield of PSII (Fv/Fm), leaf greenness (SPAD score), photosynthetic pigment levels (chlorophyll a, chlorophyll b, and carotenoids), and plant biomass yield in alfalfa seedlings. The red and/or combined (red+blue) light treatments significantly regulated antioxidant enzymes (SOD, CAT, APX, and GR) and the expression of genes related to the ascorbate-glutathione (AsA-GSH) pathway, including monodehydroascorbate reductase (), dehydroascorbate reductase (), ascorbate peroxidase (), and glutathione reductase (). These results indicate that light quality is crucial for regulating the morphological, physiological, and molecular traits linked to alfalfa improvement. These findings suggest a new approach to enhancing the adaptation, as well as the morphological and agronomic yield, of alfalfa and forage legumes through light-quality-mediated improvement.