Adaptive laboratory evolution of a minimal cell to low temperature

Temperature is a fundamental constraint on life. Most organisms adapt to temperature change through complex regulatory and stress-response systems. Whether such adaptation is possible in a genomically minimal cell remains unclear. Here, adaptive laboratory evolution was performed on the minimal genome bacterium JCVI-syn3B, shifting growth from 37{degrees}C to 25{degrees}C, a temperature at which the original strain cannot grow. After 40 serial passages, the evolved strains exhibited robust growth at 25{degrees}C. Multi-omics analyses revealed the adaptation mechanisms including increased mRNA turnover, enhanced DNA unwinding and stabilization of replication intermediates, elevated glycerolipid synthesis, and upregulation of division proteins associated with Z-ring assembly. Whole-genome transplantation was performed to distinguish genetic or non-genetic contributions, demonstrating that the cold-adapted phenotype was largely genetically encoded. These results indicated that even a minimal cell retains substantial evolvability and highlight its potential as a tractable research platform for cellular and evolutionary biology.