ABSTRACT: BACKGROUND:Shade stress, a universal abiotic stress, suppresses plant growth and production seriously. However, little is known regarding the protein regulatory networks under shade stress. To better characterize the proteomic changes of maize leaves under shade stress, 60% shade (S) and supplementary lighting (L) on cloudy daylight from tasseling stage to physiological maturity stage were designed, the ambient sunlight treatment was used as control (CK). Isobaric tag for relative and absolute quantification (iTRAQ) technology was used to determine the proteome profiles in leaves. RESULTS:Shading significantly decreased the SPAD value, net photosynthetic rate, and grain yield. During two experimental years, grain yields of S were reduced by 48 and 47%, and L increased by 6 and 11%, compared to CK. In total, 3958 proteins were identified by iTRAQ, and 2745 proteins were quantified including 349 proteins showed at least 1.2-fold changes in expression levels between treatments and CK. The differentially expressed proteins were classified into photosynthesis, stress defense, energy production, signal transduction, and protein and amino acid metabolism using the Web Gene Ontology Annotation Plot online tool. In addition, these proteins showed significant enrichment of the chloroplasts (58%) and cytosol (21%) for subcellular localization. CONCLUSIONS:60% shade induced the expression of proteins involved in photosynthetic electron transport chain (especially light-harvesting complex) and stress/defense/detoxification. However, the proteins related to calvin cycle, starch and sucrose metabolisms, glycolysis, TCA cycle, and ribosome and protein synthesis were dramatically depressed. Together, our results might help to provide a valuable resource for protein function analysis and also clarify the proteomic and physiological mechanism of maize underlying shade stress.