Early-branching cyanobacteria grow faster and upregulate superoxide dismutase activity under a simulated early Earth anoxic atmosphere

The evolution of oxygenic photosynthesis during the Archean (4-2.5 Ga) required the presence of complementary reducing pathways to maintain the cellular redox balance. While the timing of the evolution of superoxide dismutases (SODs), enzymes that convert superoxide to hydrogen peroxide and O₂, within bacteria and archaea is not resolved, the first SODs appearing in cyanobacteria contained copper and zinc in the reaction center (CuZnSOD). Here, we analyse growth characteristics, SOD gene expression (qRT-PCR) and cellular enzyme activity in the deep branching strain, Pseudanabaena sp. PCC7367, previously demonstrated to release significantly more O₂ under anoxic conditions. The observed significantly higher growth rates (p < 0.001) and protein and glycogen contents (p < 0.05) in anoxically cultured Pseudanabaena PCC7367 compared to control cultures grown under present-day oxygen-rich conditions prompted the following question: Is the growth of Pseudanabaena sp. PCC7367 correlated to atmospheric pO₂ and cellular SOD activity? Expression of sodB (encoding FeSOD) and sodC (encoding CuZnSOD) strongly correlated with medium O levels (p < 0.001). Expression of sodA (encoding MnSOD) correlated significantly to SOD activity during the day (p = 0.019) when medium O₂ concentrations were the highest. The cellular SOD enzyme activity of anoxically grown cultures was significantly higher (p < 0.001) 2 h before the onset of the dark phase compared to O₂-rich growth conditions. The expression of SOD encoding genes was significantly reduced (p < 0.05) under anoxic conditions in stirred cultures, as were medium O₂ levels (p ≤ 0.001), compared to oxic-grown cultures, whereas total cellular SOD activity remained comparable. Our data suggest that increasing pO₂ negatively impacts the viability of early cyanobacteria, possibly by increasing photorespiration. Additionally, the increased expression of superoxide-inactivating genes during the dark phase suggests the increased replacement rates of SODs under modern-day conditions compared to those on early Earth.