Abstract
Non-photochemical quenching (NPQ) is a key photoprotective mechanism in Symbiodiniaceae, enabling photosystem II (PSII) to dissipate excess excitation energy under stress. The balance between regulated (ΦNPQ) and unregulated (ΦNO) energy dissipation influences thermal tolerance, yet the temperature thresholds at which this balance shifts remain poorly defined. Here, we used the Phenoplate, a high-throughput fluorometric platform integrating rapid light curves with controlled temperature ramping, to examine short-term thermal responses in Breviolum minutum across 6–71 °C. We identified a sharp transition at 41 °C where ΦNPQ collapsed and was replaced by ΦNO, indicating loss of regulated photoprotection. This switch coincided with a pronounced drop in PSII effective quantum yield (ΦII) and substantial reductions in cell density, marking a thermal Achilles’ heel in the photoprotective capacity of this species. Despite this regulatory breakdown, a fraction of cells persisted for at least three days post-exposure. These results demonstrate that B. minutum maintains regulated photoprotection up to a discrete threshold, beyond which unregulated becomes the dominant pathway and survival is compromised. Identifying such thermal inflection points in coral symbionts provides mechanistic insight into their vulnerability under acute heat stress and may inform early-warning indicators for coral bleaching susceptibility.