BackgroundDendrobium officinaleKimura et Migo (D. officinale) is a well-known traditional Chinese medicine with high content polysaccharides in stems. The SWEET (Sugars Will Eventually be Exported Transporters) family is a novel class of sugar transporters mediating sugar translocation among adjacent cells of plants. The expression patterns of SWEETs and whether they are associated with stress response inD. officinaleremains uncovered.ResultsHere, 25SWEETgenes were screened out fromD. officinalegenome, most of which typically contained seven transmembrane domains (TMs) and harbored two conserved MtN3/saliva domains. Using multi-omics data and bioinformatic approaches, the evolutionary relationship, conserved motifs, chromosomal location, expression patterns, correlationship and interaction network were further analyzed.DoSWEETswere intensively located in nine chromosomes. Phylogenetic analysis revealed that DoSWEETs were divided into four clades, and conserved motif 3 specifically existed in DoSWEETs from clade II. Different tissue-specific expression patterns ofDoSWEETs suggested the division of their roles in sugar transport. In particular,DoSWEET5b,5c, and7ddisplayed relatively high expression levels in stems.DoSWEET2band16were significantly regulated under cold, drought, and MeJA treatment, which were further verified using RT-qPCR. Correlation analysis and interaction network prediction discovered the internal relationship ofDoSWEETfamily.ConclusionsTaken together, the identification and analysis of the 25DoSWEETsin this study provide basic information for further functional verification inD. officinale.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12864-023-09419-w.