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As an important hormone response gene, Gretchen Hagen 3 (GH3) maintains hormonal homeostasis by conjugating excess auxin with amino acids during plant stress-related signaling pathways. <i>GH3</i> genes have been characterized in many plant species, but they are rarely reported in potato. Here, 19 <i>StGH3</i> genes were isolated and characterized. Phylogenetic analysis indicated that StGH3s were divided into two categories (group I and group III). Analyses of gene structure and motif composition showed that the members of a specific StGH3 subfamily are relatively conserved. Collinearity analysis of <i>StGH3</i> genes in potato and other plants laid a foundation for further exploring the evolutionary characteristics of the <i>StGH3</i> genes. Promoter analysis showed that most <i>StGH3</i> promoters contained hormone and abiotic stress response elements. Multiple transcriptome studies indicated that some <i>StGH3</i> genes were responsive to ABA, water deficits, and salt treatments. Moreover, qRT-PCR analysis indicated that <i>StGH3</i> genes could be induced by phytohormones (ABA, SA, and MeJA) and abiotic stresses (water deficit, high salt, and low temperature), although with different patterns. Furthermore, transgenic tobacco with transient overexpression of the <i>StGH3.3</i> gene showed positive regulation in response to water deficits by increasing proline accumulation and reducing the leaf water loss rate. These results suggested that <i>StGH3</i> genes may be involved in the response to abiotic stress through hormonal signal pathways. Overall, this study provides useful insights into the evolution and function of <i>StGH3s</i> and lays a foundation for further study on the molecular mechanisms of <i>StGH3s</i> in the regulation of potato drought resistance.