Heterotrimeric G-proteins are fundamental signal transducers highly conserved in plant species, which play crucial roles in regulating plant growth, development, and responses to abiotic stresses. Identification of G-protein members and their expression patterns in plants are essential for improving crop resilience against environmental stresses. Here, we identified eight heterotrimeric G-protein genes localized on four chromosomes within the barley genome by using comprehensive genome-wide analysis. Phylogenetic analysis classified these genes into four distinct subgroups with obvious evolutionary relationships. Further analysis on gene structure, protein motif, and structure indicated that G-proteins within each evolutionary branch exhibited similar exon-intron organization, conserved motif patterns, and structural features. Collinearity analysis showed no significant collinear relationships among those G-protein genes, indicating a unique evolutionary trajectory within barley. Moreover, cis-regulatory elements detected in the upstream sequences of these genes were involved in response to plant hormones and signaling molecules. Expression analyses revealed tissue-specific expression patterns and differential regulation in response to abiotic stresses. The expression patterns of G-protein genes were further validated using a quantitative real-time PCR (qRT-PCR) technique, indicating the reliability of transcriptomic data, as well as special responses to salt, drought, and waterlogging stresses. These findings may provide underlying mechanisms by which G-protein genes participate in salt tolerance of barley, and also highlight candidate genes for potential genetic engineering applications in improving crop resilience to salinity stress.