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Heat shock factors (<i>HSF</i>s) are the key regulators of heat stress responses and play pivotal roles in tissue development and the temperature-induced regulation of secondary metabolites. In order to elucidate the roles of <i>HSF</i>s in <i>Cymbidium ensifolium</i>, we conducted a genome-wide identification of <i>CeHSF</i> genes and predicted their functions based on their structural features and splicing patterns. Our results revealed 22 <i>HSF</i> family members, with each gene containing more than one intron. According to phylogenetic analysis, 59.1% of HSFs were grouped into the A subfamily, while subfamily HSFC contained only two HSFs. And the <i>HSF</i> gene families were differentiated evolutionarily between plant species. Two tandem repeats were found on Chr02, and two segmental duplication pairs were observed on Chr12, Chr17, and Chr19; this provided evidence for whole-genome duplication (WGD) events in <i>C. ensifolium</i>. The core region of the promoter in most <i>CeHSF</i> genes contained cis-acting elements such as AP2/ERF and bHLH, which were associated with plant growth, development, and stress responses. Except for <i>CeHSF11</i>, <i>14</i>, and <i>19</i>, each of the remaining <i>CeHSF</i>s contained at least one miRNA binding site. This included binding sites for miR156, miR393, and miR319, which were responsive to temperature and other stresses. The <i>HSF</i> gene family exhibited significant tissue specificity in both vegetative and floral organs of <i>C. ensifolium</i>. <i>CeHSF13</i> and <i>CeHSF15</i> showed relatively significant expression in flowers compared to other genes. During flower development, CeHSF15 exhibited markedly elevated expression in the early stages of flower opening, implicating critical regulatory functions in organ development and floral scent-related regulations. During the poikilothermic treatment, <i>CeHSF14</i> was upregulated over 200-fold after 6 h of heat treatment. <i>CeHSF13</i> and <i>CeHSF14</i> showed the highest expression at 6 h of low temperature, while the expression of <i>CeHSF15</i> and <i>CeHSF21</i> continuously decreased at a low temperature. The expression patterns of <i>CeHSFs</i> further confirmed their role in responding to temperature stress. Our study may help reveal the important roles of <i>HSFs</i> in plant development and metabolic regulation and show insight for the further molecular design breeding of <i>C. ensifolium</i>.