All data generated for this project are associated with NCBI BioProject PRJNA950338.
Functional characterization with metatranscriptomics & community characterization with metabarcoding
Vent plume seawater sampling was performed using a Seabird 911+ CTD rosette including a Wet Labs C-Star transmissometer along an ~3 km transect following an axial ridge along the Mid-Atlantic Ridge so that sampling was conducted south of, over, and north of the Moytirra vent. Hydrothermal plumes were detected on the first CTD cast at ~2450m based upon a change in beam transmission (turbidity) at which point Niskin bottle sampling began, firing bottles approximately every 200m. Once all bottles were filled, the CTD was retrieved, emptied, and redeployed for a total of two casts. Samples were taken both within (n=14), below (n=1), and above (n=1) turbidity peaks to characterize both within and out of plume biology along the transect.
Samples were filtered across three 47mm 0.2 µm Whatman ME filters using Sentino® Microbiology Peristaltic Pumps equipped with 500 mL Sentino® Magnetic Filter Funnels and disposable sterile fluid paths. Total volume ranged between 1-2 L per filter per sample.After all volume was filtered, each filter was placed in a 2 mL cryovial and frozen at -80°C. Samples were stored at this temperature or colder until extraction and sequencing.
DNA and RNA were simultaneously extracted using the MAcherey-Nagel NucleoMag DNA/RNA Water kit following the manufacturer's instructions with the following modification - lysis buffer volume was increased to 1 mL and the resulting lysate was split between two tubes (for DNA and RNA).