Adaptive laboratory evolution is an effective method to industrial microbial genetic breeding. In this study, Lactobacillus casei Zhang which was evolved in LSM medium with gentamicin to 1 200 generations, was used as the research object, continue to evolve in LSM medium without gentamicin to 1 000 generations. The change of cell morphology, viable count, turbidity, resistance phenotypic and genotype were tracked and monitored. The results showed that:①the cell morphology, viable count and turbidity and resistance phenotypic were stable; ② Only 11 mutations were detected in the genome, including 4 synonymous mutations and 7 non-synonymous mutations. Therefore, it can be considered that the strain has a good genetic stability.