［Purposes］ To identify transferrin of Apis mellifera ligustica (AmTsf) by bioinformatics, and to analyze the role of AmTsf in tolerance to toxicity of insecticide imidacloprid. ［Methods］ The amino acid sequence characteristics and domain composition of AmTsf were analyzed by multiple sequences alignment.Online tools predict the structure,cellular localization,interacting protein networks and biological functions of the protein. Phylogenetic tree was constructed to analyze the phyloclassification and evolutionary relationship of the protein.The response characteristics and functions of AmTsf to insecticide imidacloprid toxicity in bees were analyzed by real-time quantitative PCR combined with iron homeostasis. ［Findings］ Bioinformatics analysis predicted that AmTsf protein was hydrophobic, consisting of 712 amino acid residues,with a molecular weight of 78.7 kDa and pI of 6.77. A conserved amino acid residue of the signal peptide and transferrin family, primarily located in the nucleus and mitochondria. Experimental analysis found that exposure to environmental concentrations of imidacloprid caused mass bee deaths. Compared with the control group, 24 h exposure to imidacloprid resulted in excess iron ions in the head, thorax and midgut of bees. Importantly, the transferrin encoding gene AmTsf, which is closely related to the regulation of iron homeostasis, was synchronously up-regulated in all tissues with the increase of iron(p<0.05). ［Conclusions］ The results suggest that AmTsf may actively respond to imidacloprid toxicity tolerance in honeybees by regulating iron homeostasis.