Abstract

Summary Coral microbiome transplantation has been proposed by Doering et al. (2021) as a tool to support microbiome-assisted coral restoration by transferring microbial communities from donor colonies to recipient corals. This protocol describes a low-technology workflow for harvesting and transplanting coral-associated microbiomes without requiring microbial isolation, culturing, or prior taxonomic characterization. The protocol is adapted from previous coral microbiome transplantation approaches and is intended for use in restoration or experimental settings where donor colonies can be selected based on measured performance traits, such as thermal tolerance. Donor colonies may be prescreened using acute thermal assays, such as the Coral Bleaching Automated Stress System (CBASS), to identify colonies with comparatively high thermal tolerance. Recipient corals are subjected to a microbiome-depletion step before inoculation to increase niche availability and facilitate establishment of donor-associated microorganisms. Donor microbiomes are then harvested as a coral tissue slurry and used to inoculate recipient corals under controlled incubation conditions prior to outplanting or downstream experimental use. This protocol provides a standardized field-compatible procedure for coral microbiome harvesting and transfer. It is designed to reduce technical barriers associated with microbiome-based interventions while maintaining a structured workflow for donor selection, recipient preparation, inoculation, and post-treatment handling. Overall objective To provide a standardized, low-technology, field-applicable workflow for harvesting donor-derived microbiomes from thermally tolerant coral colonies and transferring them to recipient colonies for experimental or restoration-oriented assessment of microbiome-assisted heat-stress resilience. Employed methods Thermal pre-screening of a coral population of interest using short-term heat-stress assays, such as CBASS, to identify comparatively tolerant donor colonies and susceptible recipient colonies. Collection of donor coral tissue using an airbrush to generate a donor-derived tissue slurry for microbiome harvesting. Induction of mucus release in recipient corals by inverted air exposure to reduce existing surface-associated microbial niche occupancy before inoculation. Incubation of recipient coral fragments in donor-derived, microbiome-enriched seawater to facilitate coral microbiome transplantation.

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