Synergistic effect of nitrate exposure and heatwaves on the growth, and metabolic activity of microalgae, Chlamydomonas reinhardtii, and Pseudokirchneriella subcapitata.

Aquatic biota are threatened by climate warming as well as other anthropogenic stressors such as eutrophication by phosphates and nitrate. However, it remains unclear how nitrate exposure can alter the resilience of microalgae to climate warming, particularly heatwaves. To get a better understanding of these processes, we investigated the effect of elevated temperature and nitrate pollution on growth, metabolites (sugar and protein), oxidative damage (lipid peroxidation), and antioxidant accumulation (polyphenols, proline) in Chlamydomonas reinhardtii and Pseudokirchneriella subcapitata. The experiment involved a 3 × 3 factorial design, where microalgae were exposed to one of three nitrate levels (5, 50, or 200 mg L NO) at 20 °C for 2 weeks. Subsequently, two heatwave scenarios were imposed: a short and moderate heatwave at 24 °C for 2 weeks, and a long and intense heatwave with an additional 2 weeks at 26 °C. A positive synergistic effect of heatwaves and nitrate on growth and metabolites was observed, but this also led to increased oxidative stress. In the short and moderate heatwave, oxidative damage was controlled by increased antioxidant levels. The high growth, metabolites, and antioxidants combined with low oxidative stress during the short and moderate heatwaves in moderate nitrate (50 mg L) led to a sustainable increased food availability to grazers. On the other hand, long and intense heatwaves in high nitrate conditions caused unsustainable growth due to increased oxidative stress and relatively low antioxidant (proline) levels, increasing the risk for massive algal die-offs.