Ocean Alkalinity Enhancement (OAE) does not cause cellular stress in a phytoplankton community of the sub-tropical Atlantic Ocean

Abstract. A natural plankton community from oligotrophic subtropical waters of the Atlantic near Gran Canaria, Spain, was subjected to varying degrees of ocean alkalinity enhancement (OAE) to assess the potential physiological effects, in the context of the application of ocean carbon dioxide removal (CDR) techniques. We employed 8.3 m³ mesocosms with a sediment trap attached to the bottom, creating a gradient in total alkalinity (TA). The lowest point on this gradient was 2400 μmol · L^-1, which corresponded to the natural alkalinity of the environment, and the highest point was 4800 μmol · L^-1. Over the course of the 33-day experiment, the plankton community exhibited two distinct phases. In phase-I (days 5–20), a notable decline in the photosynthetic efficiency (F_v/F_m) was observed. This change was accompanied by substantial reductions in the abundances of picoeukaryotes, small size nanoeukaryotes (nanoeukaryotes-1), and microplankton. The cell viability of picoeukaryotes, as indicated by fluorescein-di-acetate hydrolysis by cellular esterases (FDA- green fluorescence), slightly increased by the end of phase-I whilst the viability of nanoeukaryotes 1 and Synechococcus spp . did not change. Reactive oxygen species levels (ROS-green fluorescence) showed no significant changes for any of the functional groups. In contrast, in phase-II (days 21–33), a pronounced community response was observed. Increases in F_v/F_m in the intermediate OAE treatments of ∆900 to ∆1800 μmol · L^-1 and in chlorophyll-a (Chl-a), chlorophyll-c2 (Chl-c2) , fucoxanthin and divinyl-Chl-a were attributed to the emergence of blooms of large size nanoeukaryotes (nanoeukaryotes-2) from the genera Chrysochromulina, as well as picoeukaryotes. Synechococcus spp. also flourished towards the end of this phase. In parallel, we observed a total 20 % significant change in the metaproteome of the phytoplankton community. This is considered a significant alteration in protein expression, having substantial impacts on cellular functions and the physiology of the organisms. Medium levels of ∆TA showed more upregulated and less downregulated proteins than higher ∆TA treatments. Under these conditions, cell viability significantly increased in pico and nanoeukaryotes-1 in intermediate alkalinity levels, while in Synechococcus spp., nanoeukaryotes-2 and microplankton remained stable. ROS levels did not significantly change in any functional group. The pigment ratios DD+DT : FUCO, and DD+DT : Chl-a increased in medium ∆TA treatments, supporting the idea of nutrient deficiency alleviation and the absence of physiological stress. Taken all data together, this study shows that there is minimal evidence indicating a harmful impact of high alkalinity on the plankton community. The OAE treatments did not result in physiological fitness impairment, thus OAE did not cause cellular stress in the phytoplankton community studied.