Much of our understanding of the cellular mechanisms underlying cnidarian‐algal symbiosis comes from studying the biological differences between the partners when they are engaged in symbiosis and when they are isolated from one another. When comparing the in hospite and ex hospite states in Symbiodiniaceae, the in hospite state is represented by algae sampled from hosts, and the ex hospite state is commonly represented by cultured algae. The use of cultured algae in this comparison may introduce nutrition as a confounding variable because, while hosts are kept in nutrient‐depleted conditions, culture media is nutrient rich and designed to facilitate algal growth. In this perspective, we reexamine how nutrition may be a confounding variable in studies that compare the biology of Symbiodiniaceae in hospite and in culture. We also suggest several innovations in experimental design to strengthen the comparison of the two lifestyles, including the adoption of nutritional controls, alternatives to culture for the representation of Symbiodiniaceae ex hospite, and the adoption of several proteomic approaches to find novel Symbiodiniaceae genes important for symbiosis.
Coral reefs are under threat from global climate change. Their decline is mainly caused by the fragility of their symbiotic relationship with dinoflagellate algae which they rely upon for their ecological success.
Many cnidarians rely on their dinoflagellate partners from the family Symbiodiniaceae for their ecological success. Symbiotic species of Symbiodiniaceae have two distinct life stages: inside the host, in hospite, and outside the host, ex hospite. Several aspects of cnidarian-algal symbiosis can be understood by comparing these two life stages. Most commonly, algae in culture are used in comparative studies to represent the ex hospite life stage, however, nutrition becomes a confounding variable for this comparison because algal culture media is nutrient rich, while algae in hospite are sampled from hosts maintained in oligotrophic seawater. In contrast to cultured algae, expelled algae may be a more robust representation of the ex hospite state, as the host and expelled algae are in the same seawater environment, removing differences in culture media as a confounding variable. Here, we studied the physiology of algae released from the sea anemone Exaiptasia diaphana (commonly called Aiptasia), a model system for the study of coral-algal symbiosis. In Aiptasia, algae are released in distinct pellets, referred to as egesta, and we explored its potential as an experimental system to represent Symbiodiniaceae in the ex hospite state. Observation under confocal and differential interference contrast microscopy revealed that egesta contained discharged nematocysts, host tissue, and were populated by a diversity of microbes, including protists and cyanobacteria. Further experiments revealed that egesta were released at night. In addition, algae in egesta had a higher mitotic index than algae in hospite, were photosynthetically viable for at least 48 hrs after expulsion, and could competently establish symbiosis with aposymbiotic Aiptasia. We then studied the gene expression of nutrient-related genes and studied their expression using qPCR. From the genes tested, we found that algae from egesta closely mirrored gene expression profiles of algae in hospite and were dissimilar to those of cultured algae, suggesting that algae from egesta are in a nutritional environment that is similar to their in hospite counterparts. Altogether, evidence is provided that algae from Aiptasia egesta are a robust representation of Symbiodiniaceae in the ex hospite state and their use in experiments can improve our understanding of cnidarian-algal symbiosis.
This protocol is designed for quantifying symbiont density during the onset ofsymbiosis and early proliferation. Other techniques should be used for quantifying high symbiont density.
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