Host vacuolar-type ATPase (VHA)-mediated carbon-concentrating mechanisms (CCMs)that promote photosynthesis have been observed in coral-, sea anemone-, and giant clam-
Symbiodiniaceae symbiosis. Similarly, analogous CCMs operate in some free-living secondary
endosymbiotic algae, where VHA is present in phagocytic-origin membranes surrounding the
chloroplasts. This thesis explored the potential role of VHA in the CCM of cultured
Symbiodiniaceae algae Breviolum psygmophilum and Miliolidium leei, which establish symbiosis
with corals and foraminifera, respectively. VHA subunit B (VHAB) transcripts were detected in B.
psygmophilum and symbiotic algae in the coral Stylophora pistillata. B. psygmophilum acclimated
to 60 and 400 µmol photons m-2 s-1 contained similar amounts of VHAB proteins per cell; however,
VHAB constituted a higher proportion of total protein under the high light condition. Respirometry
experiments indicated that VHA inhibition decreased O2 production in B. psygmophilum cultures
when illuminated by the light level they were acclimated to, suggesting a VHA-mediated CCM
under these conditions. However, the effects of pharmacological inhibitors varied across replicates
and times, suggesting that the CCM is highly responsive to light level, pH, inorganic carbon
concentration, algal circadian rhythm and growth phases. Effects of drug concentration and drug
incubation time should also be carefully considered. Unfortunately, B. psygmophilum ruptured in
the chemical fixative, which prevented establishing the subcellular localization of VHAB.
Successful immunolocalization of VHAB in M. leei that fixed properly revealed that VHAB
surrounded intracellular spherical structures resembling vacuoles but not the chloroplast. These
results highlight morphological and physiological differences between these two species within
Symbiodiniaceae.