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The nonexpressor of pathogenesis-related 1 (NPR1) family plays diverse roles in gene regulation in the defense and development signaling pathways in plants. Less evidence is available regarding the significance of the <i>NPR1</i>-like gene family in cotton (<i>Gossypium</i> species). Therefore, to address the importance of the cotton <i>NPR1</i>-like gene family in the defense pathway, four <i>Gossypium</i> species were studied: two tetraploid species, <i>G.</i><i>hirsutum</i> and <i>G. barbadense</i>, and their two potential ancestral diploids, <i>G. raimondii</i> and <i>G. arboreum.</i> In this study, 12 <i>NPR1</i>-like family genes in <i>G. hirsutum</i> were recognized, including six genes in the A-subgenome and six genes in the D-subgenome. Based on the phylogenetic analysis, gene and protein structural features, cotton NPR-like proteins were grouped into three different clades. Our analysis suggests the significance of cis-regulatory elements in the upstream region of cotton <i>NPR1</i>-like genes in hormonal signaling, biotic stress conditions, and developmental processes. The quantitative expression analysis for different developmental tissues and fiber stages (0 to 25 days post-anthesis), as well as salicylic acid induction, confirmed the distinct function of different cotton <i>NPR</i> genes in defense and fiber development. Altogether, this study presents specifications of conservation in the cotton <i>NPR1</i>-like gene family and their functional divergence for development of fiber and defense properties.