Role of C₄ carbon fixation in Ulva prolifera, the macroalga responsible for the world’s largest green tides

Most marine algae preferentially assimilate CO₂ via the Calvin-Benson Cycle (C₃) and catalyze HCO₃⁻ dehydration via carbonic anhydrase (CA) as a CO₂-compensatory mechanism, but certain species utilize the Hatch-Slack Cycle (C₄) to enhance photosynthesis. The occurrence and importance of the C₄ pathway remains uncertain, however. Here, we demonstrate that carbon fixation in Ulva prolifera, a species responsible for massive green tides, involves a combination of C₃ and C₄ pathways_, and a CA-supported HCO₃⁻ mechanism. Analysis of CA and key C₃ and C₄ enzymes, and subsequent analysis of δ¹³C photosynthetic products showed that the species assimilates CO₂ predominately via the C₃ pathway, uses HCO₃⁻ via the CA mechanism at low CO₂ levels, and takes advantage of high irradiance using the C₄ pathway. This active and multi-faceted carbon acquisition strategy is advantageous for the formation of massive blooms, as thick floating mats are subject to intense surface irradiance and CO₂ limitation.