Table of Contents

• Coverage
• Dataset Description
  • Methods & Sampling
  • Data Processing Description
  • BCO-DMO Processing Description
  • Problem Description
• Related Publications
• Related Datasets
• Parameters
• Instruments
• Project Information
• Funding

Location
Samples were collected on shallow reefs (<10m) in the Florida Keys, USA. The temporary artificial reef was constructed adjacent to patch reefs within the Looe Key special preservation area. At the conclusion of the experiment, the reef was completely dismantled. The environment was well mixed by wave action within minimal tidal influence.

Collection and Analysis

Seawater samples were collected via VacuSIP and filtered through 0.2 µm membranes with a peristaltic pump to collect samples of ambient microbes. Total genomic DNA was extracted from cross sections of sponge tissue (~0.25 grams) and ½ of seawater filter membranes using Qiagen PowerSoil Powerlyzer extraction kit and following the manufacturers protocol. After DNA extraction, polymerase chain reaction (PCR) was performed following the 16S Illumina Amplicon protocol of the Earth Microbiome project (Caporaso, 2018) with barcoded 16S rRNA primers (515F and 806R; (Apprill et al. 2015, Parada et al. 2016) and PCR products were cleaned using AMPure beads (Beckman Coulter). DNA concentration in cleaned products was measured using a Qubit fluorometer (Qubit). The concentration of each sample was diluted to 4 nM and then all samples were pooled in equal volumes. Pooled samples were then sequenced on an Illumina MiSeq platform following standard Illumina protocols for sample preparation and loading except that custom sequencing primers from the EMP protocol were used. A 500 cycle V2 chemistry MiSeq kit was used during sequencing, which yielded paired-end 250 base pair (bp) amplicons.

Sample barcodes were input into the MiSeq to demultiplex samples. No additional data processing was done to these samples.

- Imported "PRJNA1179970_attributes_11_22.tsv" into the BCO-DMO System
- Removed columns without values
- Imported WoRMS report on all species included in the dataset as "easson_organisms_matched.txt" into the BCO-DMO System
- Join "easson_organisms_matched.txt" to "PRJNA1179970_attributes_11_22.tsv" on the species name, including the AphiaID and the LSID in the dataset
- Split the lat_lon field into Latitude and Longitude and removed the string components
- Convert West Long values to negative
- Exported file as "953999_v1_16s_reef_microbiome.csv"

NSF Award Abstract:
Coral reefs represent a paradox because, despite their immense productivity and biodiversity, they are found in nutrient-poor habitats that are equivalent to "marine deserts." High biodiversity is often associated with a division of resources that allows many types of organisms to coexist with minimal competition. Indeed, unlike many other organisms on coral reefs, sponges are adapted to efficiently remove bacteria, phytoplankton, and dissolved organic matter from seawater by filter-feeding. Sponges are a dominant component of coral reefs worldwide and in the Caribbean, where their biomass exceeds that of reef-building corals. For almost a quarter century, the success of sponges in the Caribbean has been linked to their filter-feeding ability. However, recent work demonstrated that coexisting sponges on Caribbean reefs host unique communities of bacteria that might allow sponges to access multiple pools of nutrients that are not available to other organisms. In this project, the investigators will test the hypothesis that ecologically dominant sponge species in the Caribbean have unique metabolic strategies that are mediated by their associations with microbes that live within the sponge body. This research will combine manipulative field experiments with a novel combination of modern analytical tools to investigate both filter-feeding by sponge hosts and the metabolic pathways of their microbes. This work will advance our understanding of the ecological and evolutionary forces that have helped shape the species present on Caribbean coral reefs. Additionally, this project will support three early-career investigators and provide training opportunities for graduate and undergraduate students at Nova Southeastern University, Appalachian State University, Stony Brook University, and Smithsonian Marine Station. The investigators will also develop innovative outreach programs that expand existing platforms at their institutions to increase public engagement and scientific literacy.

Marine sponges have been widely successful in their expansion across ecological niches in the Caribbean, with biomass often exceeding that of reef-building corals and high species diversity. However, whether this success is linked to efficient heterotrophic filter-feeding on organic carbon in the water column or to their evolutionary investment in microbial symbionts is yet to be fully elucidated. Microbial symbionts expand the metabolic capabilities of host sponges, supplementing heterotrophic feeding with inorganic carbon and nitrogen, mediating the assimilation of dissolved organic matter, and facilitating recycling of host-derived nitrogen. Despite these benefits, microbial symbiont communities are widely divergent across coexisting sponge species and there is substantial variation in host reliance on symbiont-derived carbon and nitrogen among host sponges; therefore, these associations likely mediate the ecological diversification of coexisting sponge species. The goal of this project is to test this transformative hypothesis by adopting an integrative approach to assess the individual components of holobiont metabolism (i.e., microbial symbionts and sponge host) in ten of the most common sponge species in the Caribbean. The investigators will isolate autotrophic and heterotrophic metabolic pathways and explore potential links between microbial symbiont community composition and the assimilation of particulate and dissolved organic matter (POM and DOM) from seawater. This project will elucidate whether Caribbean sponge species are on similar or divergent evolutionary trajectories, and will provide information that is critical for our understanding of how conditions in the Caribbean basin have shaped the evolution of benthic organisms.