Abstract: To identify the important genes that may be involved in the metabolic process of Vicia faba to bentazone, and to clarify the metabolic detoxification mechanism, this study analyzed the leaf gene expression pattern of both a drug-resistant V. faba cv. VF4 and sensitive V. faba cv. Xiaohong after treatment with bentazone for 0 and 3 days using RNA-Seq technique. The sequencing results showed that a total of 46 892 242-52 525 260 Raw Reads were obtained, with 46 025 006-51 412 088 Clean Reads remaining after quality control. Wenn analysis showed that there were 512 common differentially expressed genes in all groups. KEGG enrichment showed that these 512 differentially expressed genes were mainly involved in isoflavone biosynthesis, peroxisome, diterpene biosynthesis, and other metabolic pathways. Further analysis showed that the expressions of CYP71D9, CYP81E8, MC5MAT1, MC5MAT2, IF3H, CXE12, CXE6, PODDCR, KAO2, CYP82G1, GA201-D, GST23, GSTs and GSTsF9 genes in the above metabolic pathways were up-regulated in drug-resistant V. faba after bentazone treatment, indicating that these genes may be involved in the resistance pathway of V. faba to bentazone. The expression profiles of these genes, as determined by qRT-PCR, were consistent with those obtained from transcriptome sequencing, confirming the reliability of transcriptome sequencing results. This study identifies the key genes potentially involved in the metabolic process of V. faba in response to bentazone, providing valuable materials for investigating the mechanism of bentazone tolerance. It also provides gene resources for the genetic breeding of herbicide-tolerant V. faba varieties.