Abstract: To investigate the putative mechanisms mediated by HMA genes in Capsicum annuum L., and divided them into two subfamilies based on sequence alignment and phylogenetic analysis and six Capsicum annuum L. HMA genes were identified. Chromosome mapping, gene structure analysis, and motif composition assessment of the HMA gene family in Capsicum annuum L. species revealed that these genes exhibit a relatively conserved nature during their evolutionary history. Promoter analysis showed that most of the Capsicum annuum L. HMA genes contained cis-acting elements associated with abiotic stress. Transcriptome data analysis showed that the Cu/Ag subgroup（CaHMA2—CaHMA6） genes were expressed in all tissues and developmental stages of plants, while the genes of the Zn/Co/Cd/Pb subgroup were not expressed in all growth stages of Capsicum annuum L.. The expression of the gene family members under cadmium（Cd） and copper stress was verified by qRT-PCR. It was found that the expression of CaHMA5 in the Cu/Ag subgroup was significantly up-regulated under two heavy metal stresses, indicating that CaHMA5 played an important role in cadmium and copper stress. Finally, through the yeast cadmium tolerance test, it was found that CaHMA5 could improve the cadmium tolerance of yeast and restore its growth, suggesting that CaHMA5 had Cd²⁺ transport function.This study provided a theoretical basis for further analysis of the function of CaHMA5 and the cultivation of new Capsicum annuum L. varieties.