［Purposes］ Bioinformatics was used to identify glutathione specific γ-glutamyl cyclotransferase 2(AmCHAC2) in Apis mellifera ligustica, and quantitative PCR was used to analyze the temporal and spatial expression characteristics of AmCHAC2 in A. mellifera ligustica. To explore the role of this gene in bee resistance to Nosema infection. ［Methods］ The amino acid sequence characteristics and domain composition of AmCHAC2 were analyzed by multiple sequence alignment. Online prediction of the protein structure, interacting protein network, cell localization characteristics and function. The phylogenetic tree was constructed to analyze the phylogenetic classification and evolutionary relationship of the protein. Temporal and spatial expression characteristics of AmCHAC2 in honeybee tissues were analyzed by real-time quantitative PCR. The expression characteristics of glutathione content in honeybee tissues were analyzed by kit. The content of reactive oxygen species in honeybee midgut was determined by ELISA. ［Findings］ AmCHAC2 was composed of 245 amino acid residues with a relative molecular weight of 28.3 kDa and a pI of 6.71. AmCHAC2 is a hydrophobic protein with no signal peptide and is localized in the cytoplasm. A large number of honeybees died after 14 days of infection with Noseama ceranae, and the survival rate was only 23%. Compared with the control group, the expression of AmCHAC2 was significantly up-regulated in the breast and midgut tissues of the bees infected with nosema, the content of GSH was significantly decreased, and the content of reactive oxygen species was significantly increased in the midgut, which triggered oxidative stress. ［Conclusions］ The upregulation of AmCHAC2 after infection with Nosema may play an important role in the resistance of A. mellifera ligustica to Nosema infection.