Barnyardgrass () is one of the most troublesome herbicide-resistant weeds worldwide that interferes with rice growth and rice yield. Here we provide the data from a comparative proteomic analysis of leaves in resistant (R) and susceptible (S) biotypes of both with and without multi-herbicide treatment in two independent biological experiments using iTRAQ. The distribution of length and number of peptides, mass and sequence coverage of proteins were presented, and the repeatability of the replicates was analyzed. 1342 differential accumulated proteins were identified from 2248 unique peptides by searching uniprot database and data analysis. These results are associated with the research article "Quantitative proteomics reveals ecological fitness cost of multi-herbicide resistant barnyardgrass ( L.)" (X. Yang, Z. Zhang, T. Gu, M. Dong, Q. Peng, L. Bai, Y Li, 2017) [1].