Fragments (~5cm) of Acropora cervicornis genotype ML-50 (Coral Sample Registry (Moura et al., 2021) accession: fa13971c-ea34-459e-2f13-7bfddbafd327) were collected from the Mote Marine Laboratory in situ coral nursery in Looe Key (24.5627,-81.4005) between April and June of 2019.

Prior to experimental manipulation, fragments were allowed to acclimate to aquarium conditions for seven days. Nutrient enrichment was performed four times a day for 42 days (six weeks). Coral fragments were sacrificed for sampling: prior to nutrient exposure (T0), and after six weeks. Using sterile bone cutters, tissue was scraped from each fragment (avoiding the apical tip) and added directly to 2 mL tubes containing 0.5ml DNA/RNA shield (Zymo Research) and Lysing Matrix A (MP Biomedicals, 0.5 g garnet matrix and one 1/4" ceramic sphere). Tubes were immediately preserved at -80°C until further processing. Total RNA was extracted from 500 μl of tissue slurry using the E.Z.N.A.® DNA/RNA Isolation Kit (Omega Bio-Tek) and then stored at -80°C until further processing. Residual DNA contamination was removed from RNA isolates using the RQ1 RNase-Free DNase (Promega). Ribosomal RNA was removed using equal parts ‘plant leaf’, ‘human/mouse’ and ‘bacteria’ Ribo-Zero kits (Illumina). RNA quality and concentration were verified by BioAnalyzer (Agilent Technologies, Santa Clara, CA) and quantitative PCR, respectively. cDNA library prep and sequencing was performed at Oregon State University’s Center for Quantitative Life Sciences (CQLS) Core Laboratories with the HiSeq 3000 platform. Three biological replicates for each treatment were sequenced (n=9).

Instruments:

Mote Marine Lab flow-through seawater raceways and 4.7L tanks: held corals during experiment
boiler and chiller: controlled aquarium temperature
Sterile bone cutters: used to collect coral tissue from each fragment
-80°C Freezer: used to keep samples at a low temp prior to processing
BioAnalyzer (Agilent Technologies, Santa Clara, CA): used to verify RNA quality and concentration
Oregon State University’s Center for Quantitative Life Sciences (CQLS) Core Laboratories with the HiSeq 3000 platform: used for sample sequencing

Organism identifiers (ScientificName, NCBI Taxonomy ID, Life Science Identifier (LSID):
Acropora cervicornis, NCBI:txid6130, urn:lsid:marinespecies.org:taxname:206989
'Candidatus' Aquarickettsia rohweri, NCBI:txid2602574, N/A

928636_v1_ca-aquarickettsia-rohweri-transcriptomes.csv