Polysaccharide is a quality marker of Polygonatum odoratum, which has significant pharmacological effects in immune regulation and anti-tumor. As one of the key enzymes in polysaccharide synthesis, sucrosesynthase (SUS) has always been an important research field to reveal the synthesis of plant polysaccharides. Based on transcriptom data of Polygonatum odoratum, the SUS gene family members were identified by bioinformatics, and their expression profiling were analyzed using qPCR. The results showed that eight PoSUS gene were identified, their protein ranged from 111 to 310 amino acid residues (aa) in length, and relative molecular weight varied from 12.81 kDa to 35.43 kDa, isoelectric point (pI) in the range of 5.83 to 9.18; phylogenetic analysis indicated that eight PoSUS genes were divided into 3 subfamily, and the subfamily Ⅲ included the largest PoSUS genes family member; subcellural localization analysis showed that most of PoSUS proteins were located in chloroplast. Additionally, expression patterns analysis revealed that PoSUS1 and PoSUS6 genes were preferably expressed in rhizome, and the transcript levels of PoSUS1 and PoSUS2 were higher in high-polysaccharide cultivar HN1 than those in a low-polysaccharide cultivar AH2. In addition, the CDS of PoSUS1 gene were cloned from HN1 and AH2, there are three amino acid difference between HN1 and AH2, which located in sucrose synthase domain. Our findings can laid a basis for the further study of functional analysis of SUS genes, and provided a theoretical basis for analyzing molecular mechanism of the formation mechanism of medicinal quality of Polygonatum odoratum.