Supplementary material from "Coral calcifying fluid pH is modulated by seawater carbonate chemistry not solely seawater pH"

Reef coral calcification depends on regulation of pH in the internal calcifying fluid (CF) in which the coral skeleton forms. However, little is known about calcifying fluid pH (pH_CF) regulation, despite its importance in determining the response of corals to ocean acidification. Here, we investigate pH_CF in the coral Stylophora pistillata in seawater maintained at constant pH with manipulated carbonate chemistry to alter dissolved inorganic carbon (DIC) concentration, and therefore total alkalinity (A_T). We also investigate the intracellular pH of calcifying cells, photosynthesis, respiration and calcification rates under the same conditions. Our results show that despite constant pH in the surrounding seawater, pH_CF is sensitive to shifts in carbonate chemistry associated with changes in [DIC] and [A_T], revealing that seawater pH is not the sole driver of pH_CF. Notably, when we synthesize our results with published data, we identify linear relationships of pH_CF with the seawater [DIC]/[H⁺] ratio, [A_T]/ [H⁺] ratio and [CO₃^2-]. Our findings contribute new insights into the mechanisms determining the sensitivity of coral calcification to changes in seawater carbonate chemistry, which is needed for predicting effects of environmental change on coral reefs and for robust interpretations of isotopic palaeoenvironmental records in coral skeletons.