The enlarged corm of Chinese water chestnut (Eleocharis dulcis) is the main edible part, and the related genes involved in the regulation of the corm enlargement process remain unclear. COL5 (CONSTANS-Like 5) gene is a member of COL gene family, which may play an important role in the development of plant storage organs. The transcript fragment with high homology to COL gene in other plants was screened from the Chinese water-chestnut transcriptome databases and cloned its cDNA sequence and full-length DNA, and then the bioinformatics analysis and spatiotemporal expression pattern analysis were analyzed to study the expression pattern and possible role of Chinese water-chestnut COL5 during corm enlargement. The results showed that the cloned COL5 gene related to Chinese water-chestnut enlargement had an ORF of 1017 bp, encoding 338 amino acids, and the DNA sequence was 1275 bp, containing an intron of 257 bp, which was named CwCOL5 (ON934922). Bioinformatics analysis showed that the CwCOL5 protein with the formula C₁₅₉₂H₂₅₁₅N₄₇₇O₅₀₈S₁₇, the relative molecular weight of the predicted protein was 37 010.39 Da, and a theoretical isoelectric point of 5.89. The total average hydrophilic value (gravy) was -0.462, the instability coefficient was 45.63, which was a hydrophilic unstable protein. There were 31 phosphosites, no transmembrane structure, and no signal peptide, and additionally subcellular prediction analysis showed that it was located in the nucleus. CwCOL5 contained two B-box Zinc fingers and a highly conserved CCT domain. Phylogenetic analysis showed that CwCOL5 was closely related to the COL protein of Nelumbo nucifera and Arabidopsis. The results of PCR showed that CwCOL5 had a high expression in the leaves and water-chestnut peel, and the expression of CwCOL5 increased rapidly in the early stage of corm enlargement. The CwCOL5 gene belongs to a typical transcription factor family of zinc finger protein, and the gene may be involved in the process of corm enlargement of Chinese water-chestnut. In this study, COL5 gene was cloned from the corm of Chinese water-chestnut and its expression pattern was analyzed, which would provide a theoretical basis for molecular mechanism of plant corm enlargement.