The heat shock transcription factor (HSF) family is closely associated with plant growth, development and stress responses. To explore the function of the barley HSF gene family, this study used bioinformatics methods to identify HvHSF genes at the whole genome level and analyze the physicochemical properties, subcellular localization, evolutionary relationships, and collinearity relationships of the HvHSF proteins, as well as the gene structure, promoter cis-acting elements, and protein interaction networks of HvHSF. The results showed that the molecular weight of 29 HvHSF proteins ranges from 10.24 kDa to 56.59 kDa, classified as relatively unstable hydrophilic proteins, with a predicted localization in the nucleus. Evolutionary analysis showed that barley HvHSF genes can be divided into A, B and C groups and are more closely related to rice. Collinearity analysis revealed that HvHSF5d and HvHSF7b are collinear gene pairs. Domain and gene structure analyses revealed that closely related HvHSF genes have similar domain structures and gene structures. Promoter cis-acting element analysis showed that the HvHSF promoter contains various cis-acting elements related to growth and development, as well as stress responses. Protein interaction network predictions indicated that HvHSF may be involved in plant stress response regulation and H₂O₂ scavenging. Through analysis of the barley stress transcriptome combined with qRT-PCR validation, it was found that HvHSF exhibited differential expression under salt and alkali stresses. The expression levels of HvHSF1a, HvHSF1b, HvHSF2a, HvHSF3c and HvHSF5b were increased under salt and alkali stresses. However, HvHSF6 performed increased expression level under salt stress, but decreased expression level under alkali stress. This study confirmed that the HSF gene family responds to salt and alkali stresses and may contribute to the regulation of salt-alkali tolerance.