Arbuscular mycorrhiza fungi (AMF) can colonize the roots of Amorpha fruticosa, a perennial leguminous woody shrub, and form arbuscular mycorrhiza (AM). AMF have significant promoting effects on A. fruticosa growth as the intensity of fungal colonization increases. Taking AMF-A. fruticosa symbionts as the experimental material, gel-free isobaric tags for relative and absolute quantification (iTRAQ) coupled with two-dimensional liquid chromatography-tandem mass spectrometry (LC-MS/MS) were used to investigate the expression of A. fruticosa mycorrhizal proteins at the maturation stage. A total of 3,473 proteins were identified, of which 77 showed dramatic changes in their root expression levels; 33 increased, and 44 decreased. We also found nine AMF proteins that were expressed with AMF treatment. The 77 proteins were classified according to function. Plant proteins were assigned into 11 categories: metabolism-related (32%), protein folding and degradation-related (22%), energy-related (10%), protein synthesis-related (8%), stress and defense-related (24%), transcription-related (6%), membrane and transport-related (4%), cellular structure-related (2.5%), signaling transduction-related (11%) and unknown proteins (5%). The results of the study provide a foundation for further investigation of the metabolic characteristics and molecular mechanisms of AM.