【Objective】The Red pitaya (Hylocereus polyrhizus) HpSWEET7 gene, which that belongs to the sugar transporter SWEET (sugars will eventually be exported transporter) family, is was cloned, and its role in fruit sugar accumulation was investigatedis discussed, which provides a target gene for improving the fruit flavor and quality. 【Methods】 HpSWEET7 gene was isolated from ‘Zihonglong’, the sequence alignment and phylogenetic relationship were analyzed, the gene expression was measured analyzed by qRT-PCR, the subcellular localization was detected by fusing fluorescent protein, and the sugar transport activity was investigated by the yeast growth complementation and acidification assay. 【Results】The ORF (Open Reading Frame) of HpSWEET7 was 771 bp in length and encoded 256 amino acid residues. HpSWEET7 contained two MtN3_/saliva (Pfam No. PF03083) and seven transmembrane domains. HpSWEET7 was closely related to Arabidopsis thaliana AtSWEET6 and AtSWEET7, and belonged to the SWEET II class. The expression of HpSWEET7 in stem was significantly lower than that in fruit at 6 developmental stages; the expression level was the highest in fruit at 20 d after pollination, gradually decreased with fruit development and was the lowest in ripe fruit (30 d after pollination). The expression of HpSWEET7 was very weak in stems, and showed an increasing patterned in fruits from 10 d after pollination to 20 d, and reached the maximum value at 20 d after pollination; subsequently, it the expression decreased and was weak at fruit ripening stage (30 d after pollination). Subcellular localization assay showed that HpSWEET7 was located into the cell membrane. HpSWEET7 had showed the transport activities for glucose, fructose, mannose, sorbitol and galactose. The acidification test further proved that HpSWEET7 transported glucose and fructose. 【Conclusion】HpSWEET7 transports hexoses across the cell membrane, and participates in sugar accumulation during red pitaya fruit development.