While honey bees play a vital role in global crop production, they face increasing exposure to xenobiotic chemicals during commercial pollination. Multidrug-resistance (MDR)-type ATP-binding cassette (ABC) transporters provide the first line of defense against xenobiotic chemicals and are upregulated in resistant pest organisms. While previous studies in bees have focused on the role of metabolic enzymes in insect detoxification, the presence and function of ABC transporters across the hive caste system remains largely unexplored. This study investigated the gene expression profiles of 12 ABC transporters known to be involved in chemical detoxification in arthropods across ten honey bee castes and life stages using quantitative real-time PCR. Protein homology to known MDR transporters in humans and Drosophila was inferred from BLAST and through phylogenetic analysis. Seven ABC genes that showed increased gene expression during worker bee development were identified as MDR-like transporters (AmeABCB1, AmeABCB6, AmeABCC1, AmeABCC4a-c, AmeABCG1); and their expression levels were further investigated in reproductive caste members (drone larvae, adult drones, queen ovaries, and queens). Significant variations were observed in defense gene expression among all castes suggesting reduced chemical defense capabilities in queens as evidenced by a dramatically reduced expression of five MDR-like transporter genes in queen bees relative to worker eggs: ABCB1 (4-fold), ABCC1 (2-fold), ABCC4a (2-fold), ABCC4b (3-fold), and ABCC4c (2-fold). Although our findings suggest that drones and queens are more vulnerable to direct xenobiotic exposure compared to workers, further research is required to better understand the different hive members’ responses to chemical threats.