Abstract: Trehalose-6-phosphate phosphatase (TPP) is a key enzyme in trehalose metabolism and plays an important role in plant growth and abiotic stress response. In order to excavate members of the TPP gene family involved in tobacco development and stress response, we identified 15 NtTPP genes in the common tobacco genome through bioinformatic methods, with 7 of the NtTPP genes located on chromosomes. Phylogenetic analysis revealed that the newly identified tobacco TPP family members could be divided into three subfamilies:CLASS I, CLASS II and CLASS III. Syntenic analysis revealed that there were eight homologous gene pairs between Arabidopsis and tobacco. Sequence analysis showed that the promoters of NtTPP genes contained various hormone- and stress-related elements. Furthermore, the expression of NtTPP genes was studied in different tissues and most of the NtTPP genes were expressed at the highest level in roots. The expression pattern analysis showed that 12 NtTPP genes were up-regulated under drought stress, and 13 NtTPP genes could be induced by salt treatments. This indicates that the NtTPP gene family members play an important role in drought and salt stress responses of tobacco.