[Objectives]  To investigate the effect of imidacloprid on the growth, development and gene expression of Apis mellifera Ligustica drones. [Methods]  A. mellifera drone larvae were fed solutions containing 0.000 01, 0.000 1 or 0.001 μg/μL imidacloprid and their development and mortality were recorded daily. Larval weight was measured in the late larval stage (6 d) and differential expression of genes was analyzed in depth. [Results]  Imidacloprid treated drone larvae were lighter in weight than control larvae, significantly so at concentrations higher than 0.000 1 μg/μL. Mortality was also higher in the imidacloprid treatment group and increased with dosage. Analysis of differential gene expression indicated that 390 genes were up-regulated and 130 down-regulated. GO enrichment analysis revealed that the up-regulated genes were distributed in 55 GO teams, most of which are involved in cellular process, cells, cell parts, membranes, membrane parts and binding. Down-regulated genes were distributed in 48 GO teams, most of which are involved in cellular processes, cells and cell parts. There were also differentially expressed genes involved in reproduction, 21 of which were upregulated and 5 of which were down-regulated. KEGG pathway enrichment analysis showed that up-regulated genes were enriched in 159 pathways, among which the most enriched were involved in protein digestion and absorption and neuroactive ligand-receptor interaction. Down-regulated genes were enriched in 71 pathways, among which the most enriched were involved in lysosome, pancreatic secretion and neuroactive ligand-receptor interaction. [Conclusion]  Imidacloprid can inhibit the growth and development of A. mellifera drones, and can cause larval mortality. Analysis of gene expression indicates that imidacloprid affects the nervous, metabolic and reproductive, systems of drone larvae. These results highlight the importance of protecting bees from the toxic effects of pesticides.