Dataset: Exudate Uptake Incubations - Macronutrient Data

Experiments were conducted to monitor responses of reef seawater microbial communities to concentrated exudates from P. astreoides and G. ventalina. After the organism incubations were conducted, excess filtrate (2 l) from 3 of the 6 colony/fragment incubations (selected randomly) were pooled into an acid-washed, 10 l carboy and mixed. Excess filtrate (~2 l) from the three control incubations were also pooled into a second, acid-washed, 10 l carboy and mixed.

For the P. astreoides experiment, surface seawater was collected from the offshore site and coarsely filtered through a GF/A filter (1.6 µm nominal pore size) using peristalsis to remove larger cells and minimize heterotrophic grazing, but retain bacteria and archaea. Approximately 2.4 l of this inoculum was added separately to each of the 10 l carboys (pooled coral and control filtrate) to create a 5:2 ratio of filtrate: inoculum. After this addition, each carboy was mixed and a suite of samples were collected for different analyses including cell counts, inorganic and organic macronutrient quantification, and 16S rRNA gene sequencing. This collection marked the first timepoint (0 hours [hrs]) for the exudate uptake experiment. For the G. ventalina experiment, reef seawater inoculum was collected from Tektite reef (Table S1).

For each experiment, coral metabolite or control filtrate seawater were transferred into separate 1 l acid-washed polycarbonate bottles (6 bottles per treatment). Within each treatment (coral and control), 3 of the 6 bottles were blackened to block light. The bottles were placed into a flow-through seawater table. PAR readings in the seawater table for the P. astreoides and G. ventalina experiments ranged from 250 – 1000 and 164 - 530 µmol quanta m -2s -1, respectively, with variation caused by shading and cloud cover. Over the course of 48 hours, samples were collected for cell counts (1 ml) at all timepoints (0, 12, 24, 36, and 48 hrs), macronutrient analyses (30 – 40 ml) at 0 and 48 hrs, and microbial community analyses (60 – 300 ml) at 0, 24, and 48 hrs. 

Samples (40 ml) collected for total organic carbon (TOC) and total nitrogen (TN) analyses were aliquoted into combusted, borosilicate EPA vials and acidified using 75 µl of phosphoric acid. Samples were stored at room temperature for two weeks and then refrigerated at 4 °C until analysis.

For inorganic nutrient analyses, seawater (30 ml) was allocated into acid-washed, polypropylene bottles (Nalgene) and frozen at -20 °C.