Five genotypes of Breviolum Antillogorgia were grown at three temperatures where growth rate, carrying capacity, respiration, gross photosynthesis, and new photosynthesis were quantified. This work was conducted in laboratory growth chambers in Los Angeles, California, USA in 2021.
We collected symbionts from octocoral colonies in the genus Antillogorgia from Elbow Reef and Pickles Reef in the Florida Keys in September 2016 (11-18 m depth). Details of collection and genotyping are available in Pelosi et al. (2021). Briefly, we recovered five unique genotypes of Breviolum antillogorgium from the heterogenous mixture of strains within the colonies. Three of these genotypes (G1, G2, G3) were isolated and reared at 26 C for five years (~650-700 generations) prior to the start of the experiment; the other two genotypes (G4, G5) were isolated and subsequently reared at 30 C for the same period of time. In March 2021, we initiated a laboratory experiment to measure population dynamics and physiology of each genotype at three different temperatures. Every three days, we removed 50 micrometers from each culture and performed four replicate hemacytometer counts and used the mean as an estimate of cell density. On Day 27 of the experiment, we used microrespirometry plates to quantify changes in oxygen in the dark (respiration) and in the light (new photosynthesis).
We used the stock cultures of each genotype to initiate 15 new replicate 50 mL cultures at an initial density of 10,000 cells/mL. Five of these cultures of each genotype were maintained in a growth chamber set at 26 C (actual mean temperature +/- s.d. determined by HOBO Data Logger: 25.5 C +/- 0.51). Another five cultures of each genotype were grown in identical growth chambers set at 30 C (30.1 +/- 0.28) and 32 C (31.6 +/- 0.23). Lights were set on a 12:12 day:night cycle, with average day illumination of 4244 Lux (approximately 59 µmole m-2 s-1 based on a conversion of 1 lux = 0.014 µmole m-2 s-1). On Day 27 of the experiment, we removed two 2 mL from each culture grown at 26 C and used these samples to estimate rates of photosynthesis and respiration at each temperature. Replicate samples were placed in randomly assigned wells in a microrespirometry plate that quantified changes in oxygen concentrations over time (Loligo Systems, Viborg, Denmark) in the 26 C growth chamber. We also placed sterile f/2 samples in two wells in each plate to account for any background changes in oxygen concentration. Samples were dark-adapted for 10 min before measuring oxygen levels in each well every 15 sec for 10 min. Following this, we turned the lights on in the growth chambers, allowed samples two minutes to acclimate, and then again measured oxygen levels every 15 sec for 10 min. The microrespirometry plates were then moved to the 30 C growth chamber, and later the 32 C growth chamber, and allowed to acclimate for 15 minutes in each growth chamber before measurements were taken. We estimated respiration as the slope of a linear fit to declining oxygen levels over time in the dark, subtracting any background changes in oxygen. Similarly, we estimated net photosynthesis as the slope of a linear fit to increasing oxygen levels, accounting for background changes in oxygen in the light. We estimated gross productivity by adding the absolute value of respiration in each culture to net photosynthesis. We standardized respiration, net photosynthesis, and gross photosynthesis by the number of cells in each well, determined by replicate hemacytometer counts, as above.
Known Issues:
Genetic analyses revealed that genotype G3 was contaminated with cells of Genotype G1. The results from each genotype are presented here, but the results from G3 should be interpreted with caution.
terHorst, C., Coffroth, M. A. (2022) Physiological data from Breviolum antillogorgia genotypes grown in laboratory growth chambers in Los Angeles, CA in 2021.. Biological and Chemical Oceanography Data Management Office (BCO-DMO). (Version 1) Version Date 2022-05-24 [if applicable, indicate subset used]. doi:10.26008/1912/bco-dmo.874587.1 [access date]
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