31 March 2022

Heatwave

Danielle kept the heatwave ramp period a second day to experience a diel fluctuation in the heated treatement of 28.9 °C to 29.6 °C.

Daily measurements

Danielle recorded bleaching score and survivorship for the fragments and colonies and completed 2 daily measurements of light, temperature, salinity and pH. She also cleaned the tanks.

Respirometry - Photosynthesis and Respiration

To measure photosynthesis and respiration for our initial timepoint 1 fragments (n=24) we followed the Putnam Lab respirometer manual. Since our set up in Mo’orea has two six chamber respirometer stands, we did three separate runs with n=8 fragments with one blank on each stand for a total of n=10 chambers per run. We first took molecular samples of each fragment with one ~0.5-1 cm biopsy preserved in 1 mL of RNA/DNA shield and one ~0.5-1 cm biopsy snap frozen in liquid nitrogen and stored in the Gump Molecular -40 C freezer. We set our light levels to ~610 μmol photons m−2 s−1 (51% on the AI lights) following our PI cuvre calculations to measure photosynthesis for ~20 minutes or until a consistent reading was achieved and then we turn off the lights and conducted light-enhanced dark respiration for ~20 minutes or until a consistent reading was achieved. After each trial was completed, volumes were measured from the chambers and the physiological fragments were snap frozen and stored in the Gump Molecular -40 C freezer in whirl paks.

Data tracking

All daily measurements, bleaching score, and survivorship tracking for the colonies and fragments can be found on the Heatwave Google Drive and Gametogenesis GitHub. All respirometry data was uploaded to the timepoint 3 heatwave google drive folder and the Gametogenesis GitHub Repo.