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Global warming is a serious threat to food security and severely affects plant growth, developmental processes, and, eventually, crop productivity. Respiratory metabolism plays a critical role in the adaptation of diverse stress in plants. Aconitase (ACO) is the main enzyme, which catalyzes the revocable isomerization of citrate to isocitrate in the Krebs cycle. The function of ACO gene family members has been extensively studied in model plants, for instance Arabidopsis. However, their role in plant developmental processes and various stress conditions largely remained unknown in other plant species. Thus, we identified 15 ACO genes in wheat to elucidate their function in plant developmental processes and different stress environments. The phylogenetic tree revealed that <i>TaACO</i> genes were classified into six groups. Further, gene structure analysis of <i>TaACOs</i> has shown a distinctive evolutionary path. Synteny analysis showed the 84 orthologous gene pairs in <i>Brachypodium distachyon, Aegilops tauschii, Triticum dicoccoides, Oryza sativa</i>, and <i>Arabidopsis thaliana</i>. Furthermore, Ka/Ks ratio revealed that most <i>TaACO</i> genes experienced strong purifying selection during evolution. Numerous cis-acting regulatory elements were detected in the <i>TaACO</i> promoters, which play a crucial role in plant development processes, phytohormone signaling, and are related to defense and stress. To understand the function of <i>TaACO</i> genes, the expression profiling of <i>TaACO</i> genes were investigated in different tissues, developmental stages, and stress conditions. The transcript per million values of <i>TaACO</i>s genes were retrieved from the Wheat Expression Browser Database. We noticed the differential expression of the <i>TaACO</i> genes in different tissues and various stress conditions. Moreover, gene ontology analysis has shown enrichment in the tricarboxylic acid metabolic process (GO:0072350), citrate metabolic process (GO:0006101), isocitrate metabolic process GO:0006102, carbohydrate metabolic (GO:0005975), and glyoxylate metabolic process (GO:0046487). Therefore, this study provided valuable insight into the ACO gene family in wheat and contributed to the further functional characterization of <i>TaACO</i> during different plant development processes and various stress conditions.