| Summary: | <i>CONSTANS</i>-like (<i>COL</i>) genes play important regulatory roles in flowering, tuber formation and the development of the potato (<i>Solanum tuberosum</i> L.). However, the <i>COL</i> gene family in <i>S. tuberosum</i> has not been systematically identified, restricting our knowledge of the function of these genes in <i>S. tuberosum</i>. In our study, we identified 14 <i>COL</i> genes, which were unequally distributed among eight chromosomes. These genes were classified into three groups based on differences in gene structure characteristics. The COL proteins of <i>S. tuberosum</i> and <i>Solanum lycopersicum</i> were closely related and showed high levels of similarity in a phylogenetic tree. Gene and protein structure analysis revealed similarities in the exon–intron structure and length, as well as the motif structure of COL proteins in the same subgroup. We identified 17 orthologous <i>COL</i> gene pairs between <i>S</i>. <i>tuberosum</i> and <i>S. lycopersicum</i>. Selection pressure analysis showed that the evolution rate of <i>COL</i> homologs is controlled by purification selection in <i>Arabidopsis</i>, <i>S</i>. <i>tuberosum</i> and <i>S. lycopersicum</i>. <i>StCOL</i> genes showed different tissue-specific expression patterns. <i>StCOL5</i> and <i>StCOL8</i> were highly expressed specifically in the leaves of plantlets. <i>StCOL6</i>, <i>StCOL10</i> and <i>StCOL14</i> were highly expressed in flowers. Tissue-specific expression characteristics suggest a functional differentiation of <i>StCOL</i> genes during evolution. Cis-element analysis revealed that the <i>StCOL</i> promoters contain several regulatory elements for hormone, light and stress signals. Our results provide a theoretical basis for the understanding of the in-depth mechanism of <i>COL</i> genes in regulating the flowering time and tuber development in <i>S. tuberosum</i>.
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