Tonoplast intrinsic proteins (TIPs), localizing on the tonoplast or the vacuole formed body, belong to one subclass of aquaporins and are known to play important roles in plant abiotic stress responses. In the present study, 23 GmTIP genes were identified on the genomic level, using soybean genome database. Chromosomal distribution analyses suggested GmTIP genes were distributed on 17 chromosomes. Protein multiple sequence alignment analyses showed that all GmTIP proteins contained six representative trans-membrane domains (TM1 to TM6) and two conserved NPA (Asp-Pro-Ala box) domains. Protein feature analyses indicated that the number of amino acids of GmTIPs ranged from 237 to 255, the molecular masses ranged from 27.0 to 27.2 KDa and the isoelectronic points ranged from 5.08 to 10.01. Evolutionary relationship analyses showed that GmTIPs were divided into five clades (TIP1 to TIP5), which was consistent with Arabidopsis AtTIPs. And each clade was composed of the homologous TIPs from soybean and Arabidopsis, which may be functionally conserved. Furthermore, the responsiveness of 5 GmTIP candidate genes in response to abiotic stresses (drought and salt) and hormone signals (abscisic acid (ABA) and amino cyclopropane carboxylic acid (ACC)) were investigated using qRT-PCR. The results revealed that these environmental stresses and exogenous hormones could induce the up-regulation or down-regulation of these tested GmTIP genes in leaves or roots. Among them, GmTIP2;6 was most significantly up-regulated in roots after drought or salt treatment. However, for GmTIP2;1, GmTIP2;2 and GmTIP4;1, drought treatment provoked their dramatic down-regulation expression in roots and their dramatic up-regulation expression in leaves . For ACC hormone treatment, GmTIP2;2 was strong up-regulation in roots. These findings will facilitate further study of the functional roles and molecular mechanisms of GmTIP genes during abiotic stress responses and promote the application of GmTIP genes in molecular breeding of vegetable soybean.