Fig.1 Adaptive laboratory evolution improves freeze–thaw survival while maintaining baseline growth capacity in Saccharomyces cerevisiae. (IMAGE)

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Fig.1 Adaptive laboratory evolution improves freeze–thaw survival while maintaining baseline growth capacity in Saccharomyces cerevisiae.

Caption

 (A) Schematic overview of the ALE strategy. Three independent ancestral S. cerevisiae strains, DBY12007, PGY7, and PGY34, were subjected to repeated freeze–thaw selection. The resulting evolved lineages were designated ALE1, ALE2, and ALE3. Unless otherwise stated, ALE1–ALE3 refer to mixed evolved lineages collected at the indicated generation checkpoints rather than clonal isolates. (B) Viable cell density of ancestral and evolved lineages during ALE, expressed as CFU mL−1 normalized to OD600. (C) Doubling time of each lineage at different evolutionary stages under non-stress growth conditions. (D) Freeze–thaw survival rate of each lineage across the evolutionary trajectory. (E) Spot assay showing colony-forming capacity of ancestral and evolved lineages before and after freeze–thaw treatment across serial dilutions. Data in panels B–D are presented as mean ± SD from three independent biological replicates. Statistical comparisons were performed by two-way ANOVA followed by Tukey’s post hoc test; ***P < 0.001. Panel e shows a representative spot assay from independent biological replicates.

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