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ACO is one of the rate-limiting enzymes in the biosynthesis of ethylene, and it plays a critical role in the regulation of plant growth and development. However, the function of <i>ACO</i> genes in cotton is not well studied. In this study, a total of 332 <i>GhACOs</i>, 187 <i>GaACOs</i>, and <i>181 GrACOs</i> were identified in <i>G. hirsutum</i>, <i>G. arboretum</i>, and <i>G. raimondii</i>, respectively. Gene duplication analysis showed that whole-genome duplication (WGD) and tandem duplication were the major forces driving the generation of cotton <i>ACO</i> genes. In the promoters of <i>GhACOs</i>, there were cis-acting elements responding to stress, phytohormones, light, and circadian factors, indicating the possible involvement of <i>GhACOs</i> in these processes. Expression and co-expression analyses illustrated that most <i>GhACOs</i> were not only widely expressed in various tissues but also coexpressed with other genes in response to salt and drought stress. <i>GhACO106_At</i> overexpression in <i>Arabidopsis</i> promoted flowering and increased salt tolerance. These results provide a comprehensive overview of the <i>ACO</i> genes of cotton and lay the foundation for subsequent functional studies of these genes.