Exploring stress tolerance mechanism of evolved freshwater strain Chlorella sp. S30 under 30 g/L salt

[Display omitted] •Salt tolerance strain, Chlorella sp. S30, was obtained after 138 days.•30 g/L salt caused loss to photosynthesis and oxidative phosphorylation.•Salt tolerance mechanism was related to antioxidant enzymes and ABC transporters.•Up-regulations of C3, C4 and CAM pathways were beneficial to CO2 fixation. Enhancement of stress tolerance to high concentration of salt and CO2 is beneficial for CO2 capture by microalgae. Adaptive evolution was performed for improving the tolerance of a freshwater strain, Chlorella sp. AE10, to 30 g/L salt. A resulting strain denoted as Chlorella sp. S30 was obtained after 46 cycles (138 days). The stress tolerance mechanism was analyzed by comparative transcriptomic analysis. Although the evolved strain could tolerate 30 g/L salt, high salinity caused loss to photosynthesis, oxidative phosphorylation, fatty acid biosynthesis and tyrosine metabolism. The related genes of antioxidant enzymes, CO2 fixation, amino acid biosynthesis, central carbon metabolism and ABC transporter proteins were up-regulated. Besides the up-regulation of several genes in Calvin-Benson cycle, they were also identified in C4 photosynthetic pathway and crassulacean acid metabolism pathway. They were essential for the survival and CO2 fixation of Chlorella sp. S30 under 30 g/L salt and 10% CO2.