This study examined the potential response mechanisms of Ligilactobacillus salivarius AR612 to glucose stress through whole-genome and comparative transcriptome analysis. We obtained the basic genome information of L. salivarius AR612. The full genome length of L. salivarius AR612 was 1970245 bp, with a GC content of 33.01% and 1894 coding genes. Moreover, we identified many genes associated with genetic adaptations to various stress factors, including temperature, pH, osmotic pressure, bile salts, and oxidative stress. Physiological analysis revealed that the growth and morphology of AR612 changed significantly under glucose stress, with a decrease in the maximum growth and irregular cell morphology. Furthermore, a comparison of transcriptome data indicated that glucose stress induced changes in the number of differential genes. Moreover, AR612 could respond to extracellular glucose stress by changing the expression of genes related to cell morphology, carbohydrate metabolism, amino acid metabolism, fatty acid synthesis, and nucleotide metabolism. This study provides valuable theoretical insights for future research on the adaptation of L. salivarius AR612 to nutritional stress and its application in industrial processes.