Laccase (LAC, EC 1.10.3.2) is integral to the formation of lignin synthesis, flavonoid production, and responses to both biotic and abiotic stresses. While recent studies have characterized numerous LAC gene families and their functions across various plants, information regarding LAC genes in woodland strawberry (Fragaria vesca) remains limited. In this study, we identified a total of 57 FvLAC genes in the Fragaria vesca genome, which were phylogenetically categorized into five distinct groups. Analysis of the gene structures revealed a uniformity in the exon–intron structure among the subgroups, while conserved motifs identified unique motifs specific to certain subgroups, suggesting functional variations. Chromosomal localization studies indicated that FvLACs are distributed across seven chromosomes, and collinearity analysis demonstrated that FvLACs exhibit collinearity within the species. Additionally, cis-acting element analysis suggested that FvLAC genes are involved in stress responses, hormone responses, light responses, and the growth and development of plants. qRT-PCR demonstrated that FvLACs responded to salt, drought, and hormone stresses, with the expression levels of FvLAC24, FvLAC32, and FvLAC51 continuously increasing under these stress conditions. Furthermore, transgenic yeast experiments revealed that FvLAC51 enhanced yeast tolerance to both salt and drought stresses, while FvLAC24 and FvLAC32 negatively regulated yeast tolerance under these same conditions. These findings provide a theoretical foundation for further investigation into the functions of FvLAC genes in woodland strawberry.