Large genome databases have markedly improved our understanding of marine microorganisms1-5. Although these resources have focused on prokaryotes, genomes from many dominant marine lineages, such as Pelagibacter and Prochlorococcus, are conspicuously underrepresented. Here we present the Great Barrier Reef Microbial Genomes Database (GBR-MGD), comprising 5,283 prokaryotic genomes obtained from Great Barrier Reef seawater samples using Nanopore and Illumina sequencing, including a collection of high-quality genomes of underrepresented groups. We show that standard short-read assemblies miss these populations owing to a combination of strain heterogeneity and low-GC-percentage sequencing bias. The GBR-MGD also comprises 20 chromosome-level picoeukaryote and 808,585 viral genomes, including a newly described clade of marine Crassvirales. We demonstrate the utility of the GBR-MGD to identify indicator taxa that can reliably predict the effects of reef management practices, such as the establishment of marine protected zones.

Abstract Australia’s Great Barrier Reef is a biodiversity hotspot critical to ocean health, yet it faces increasing threats from climate change and localised impacts requiring effective conservation and management action. Rezoning of the Great Barrier Reef Marine Park in 2004 expanded No-Take Marine Reserves (NTMRs) to restrict extractive activities like fishing and collecting, creating one of the largest networks of marine reserves globally. Benefits like increased biomass of fisheries-targeted species and improved coral community health metrics have been reported, though the effects of zoning on water chemistry and seawater microbiology remain unexplored. Using data from the Great Barrier Reef Microbial Genomics Database, we investigated the structure of seawater microbiomes on 48 offshore reefs within NTMRs and fished reefs. A supervised classification method (MINT sPLS-DA) identified 350 indicator species that predict zoning with ∼71% accuracy (range 58–85%). Microbial communities broadly reflected reef states, with NTMR zones enriched in streamlined microbial oligotrophs ( Pelagibacter and SAR86) correlating with higher cover of hard coral, crustose coralline algae, and herbivore fish abundance under lower nutrient conditions. By contrast, fished reefs harbored opportunists (Flavobacteriales, especially UA16, and Pseudomonadales) associating with elevated nutrients and turf algae cover. Co-occurrence networks revealed stronger competitive interactions in fished reefs, where nutrient-responsive taxa may outcompete other microbes, underscoring the need to investigate how these shifts influence reef nutrient cycling and function. Our findings reveal ecosystem-wide effects of marine zoning beyond fish protection, with distinct seawater microbiomes between fished reefs and NTMRs, which will help build decision tools for more targeted reef health monitoring assessments.

The stability of the symbiotic relationship between coral and their dinoflagellate algae (Symbiodiniaceae) is disrupted by ocean warming. Although the coral thermal response depends on the complex interactions between host, Symbiodiniaceae and prokaryotes, the mechanisms underlying the initial destabilization of these symbioses are poorly understood. In a 2-month manipulative experiment, we exposed the coral Porites lutea to gradually increasing temperatures corresponding to 0–8 degree heating weeks (DHW) and assessed the response of the coral holobiont using coral and Symbiodiniaceae transcriptomics, microbial 16S rRNA gene sequencing and physiological measurements. From early stages of heat stress (< 1 DHW), the increase in metabolic turnover shifted the holobiont to a net heterotrophic state in which algal-derived nutrients were insufficient to meet host energy demands, resulting in reduced holobiont performance at 1 DHW. We postulate the altered nutrient cycling also affected the coral-associated microbial community, with the relative abundance of Endozoicomonas bacteria declining under increasing heat stress. Integration of holobiont stress responses correlated this decline to an increase in expression of a host ADP-ribosylation factor, suggesting that Symbiodiniaceae and Endozoicomonas may underlie similar endosymbiotic regulatory processes. The thermotolerance of coral holobionts therefore is influenced by the nutritional status of its members and their interactions, and this identified metabolic interdependency highlights the importance of applying an integrative approach to guide coral reef conservation efforts.

Seawater microbes (bacteria and archaea) play essential roles in coral reefs by facilitating nutrient cycling, energy transfer, and overall reef ecosystem functioning. However, environmental disturbances such as degraded water quality and marine heatwaves, can impact these vital functions as seawater microbial communities experience notable shifts in composition and function when exposed to stressors. This sensitivity highlights the potential of seawater microbes to be used as indicators of reef health. Microbial indicator analysis has centered around measuring the taxonomic composition of seawater microbial communities, but this can obscure heterogeneity of gene content between taxonomically similar microbes, and thus, microbial functional genes have been hypothesized to have more scope for predictive potential, though empirical validation for this hypothesis is still pending. Using a metagenomics study framework, we establish a functional baseline of seawater microbiomes across offshore Great Barrier Reef (GBR) sites to compare the diagnostic value between taxonomic and functional information in inferring continuous physico-chemical metrics in the surrounding reef. Integrating gene-centric metagenomics analyses with 17 physico-chemical variables (temperature, salinity, and particulate and dissolved nutrients) across 48 reefs revealed that associations between microbial functions and environmental parameters were twice as stable compared to taxonomy-environment associations. Distinct seasonal variations in surface water chemistry were observed, with nutrient concentrations up to threefold higher during austral summer, explained by enhanced production of particulate organic matter (POM) by photoautotrophic picocyanobacteria, primarily Synechococcus. In contrast, nutrient levels were lower in winter, and POM production was also attributed to Prochlorococcus. Additionally, heterotrophic microbes (e.g., Rhodospirillaceae, Burkholderiaceae, Flavobacteriaceae, and Rhodobacteraceae) were enriched in reefs with elevated dissolved organic carbon (DOC) and phytoplankton-derived POM, encoding functional genes related to membrane transport, sugar utilization, and energy metabolism. These microbes likely contribute to the coral reef microbial loop by capturing and recycling nutrients derived from Synechococcus and Prochlorococcus, ultimately transferring nutrients from picocyanobacterial primary producers to higher trophic levels. This study reveals that functional information in reef-associated seawater microbes more robustly associates with physico-chemical variables than taxonomic data, highlighting the importance of incorporating microbial function in reef monitoring initiatives. Our integrative approach to mine for stable seawater microbial biomarkers can be expanded to include additional continuous metrics of reef health (e.g., benthic cover of corals and macroalgae, fish counts/biomass) and may be applicable to other large-scale reef metagenomics datasets beyond the GBR.

Marine bacterioplankton underpin the health and function of coral reefs and respond in a rapid and sensitive manner to environmental changes that affect reef ecosystem stability. Numerous meta-omics surveys over recent years have documented persistent associations of opportunistic seawater microbial taxa, and their associated functions, with metrics of environmental stress and poor reef health (e.g. elevated temperature, nutrient loads and macroalgae cover). Through positive feedback mechanisms, disturbance-triggered heterotrophic activity of seawater microbes is hypothesised to drive keystone benthic organisms towards the limit of their resilience and translate into shifts in biogeochemical cycles which influence marine food webs, ultimately affecting entire reef ecosystems. However, despite nearly two decades of work in this space, a major limitation to using seawater microbes in reef monitoring is a lack of a unified and focused approach that would move beyond the indicator discovery phase and towards the development of rapid microbial indicator assays for (near) real-time reef management and decision-making. By reviewing the current state of knowledge, we provide a comprehensive framework (defined as five phases of research and innovation) to catalyse a shift from fundamental to applied research, allowing us to move from descriptive to predictive reef monitoring, and from reactive to proactive reef management.

The zoanthid Parazoanthus axinellae (Schmidt, 1862) is a widespread coral species in the Mediterranean coralligenous assemblages where two morphotypes are found: Slender and Stocky, differing in size, color, and preferred substrate. Due to these marked differences, Slender and Stocky morphotypes were hypothesized to be two species. Here, we used 2bRAD to obtain genome‐wide genotyped single nucleotide polymorphisms (SNPs) to investigate the genetic differentiation between Slender and Stocky morphs, as well as their population structure. A total of 101 specimens of P. axinellae were sampled and genotyped from eight locations along the Italian coastline. In four locations, samples of the two morphotypes were collected in sympatry. 2bRAD genome-wide SNPs were used to assess the genetic divergence between the two morphotypes (1319 SNPs), and population connectivity patterns within Slender (1926 SNPs) and Stocky (1871 SNPs) morphotypes. Marked and consistent differentiation was detected between Slender and Stocky morphotypes. The widely distributed Slender morphotype showed higher population mixing patterns, while populations of the Stocky morphotype exhibited a stronger genetic structure at a regional scale. The strong genetic differentiation observed between P. axinellae Slender and Stocky morphotypes provides additional evidence that these morphs could be attributed to different species, although further morphological and ecological studies are required to validate this hypothesis. Our study highlights the importance of resolving phylogenetic and taxonomic disparities within taxonomically problematic groups, such as the P. axinellae species complex, when performing genetic connectivity studies for management and conservation purposes. Graphical Abstract Schematic overview of the main genetic structuring patterns observed in this study. Coral polyps were colored to intuitively associate the reader to Parazoanthus axinellae morphotypes, with orange tones being attributed to the Stocky morphotype, and yellow tones to the Slender morphotype. Bidirectional arrows represent gene flow between coral individuals, with the number and thickness of arrows corresponding to the intensity of gene flow rates. The red dashed line represents the potential reproductive isolation between Slender and Stocky morphs

Savalia savaglia is an Atlantic-Mediterranean zoantharian species with a patchy geographic and bathymetric distribution. Due to its longevity, S. savaglia may form large-sized colonies which play a crucial role in the ecosystem as habitat formers. Despite its ecological importance, little is known about the population structure and intraspecific genetic diversity of this species. Using ddRAD-Seq genotyping, we obtained genome-wide single nucleotide polymorphisms (SNPs) from 50 S. savaglia individuals collected at different depths (8–60 m) and localities across the Mediterranean Sea (Marseille, Sardinia, Puglia and Montenegro) and eastern Atlantic (Portugal). Our molecular observations were discussed with the reproductive behaviour of the species to understand the observed patterns of connectivity and gene flow. These results highlight the presence of three main genetic clusters (Marseille; Sardinia; and Montenegro + Portugal + Puglia), with some of the Mediterranean individuals being genetically closer to the Atlantic population rather than to other Mediterranean populations. The strong linkage disequilibrium recorded across loci and the detection of clonal individuals in the shallow populations suggest that asexual reproduction seems to be the dominant reproductive strategy among the S. savaglia populations sampled at lower depths. Our work highlights the potential of genome-wide SNP data to study the reproductive behaviour in species such as S. savaglia that are difficult to investigate in the field. The genetic connectivity data obtained in this study can be used in the future to better guide the development of effective management and conservation plans.

Coralligenous assemblages are among the most species-rich and vulnerable habitats of the Mediterranean Sea. Nevertheless, data on connectivity patterns on species inhabiting these habitats, crucial to define management and protection priorities, are largely lacking. Moreover, unreliable species-level taxonomy can confound ecological studies and mislead management strategies. In the northwestern Mediterranean two Parazoanthus axinellae morphotypes differing in size, color and preferred substrate are found in sympatry. In this study, we used COI and ITS sequence polymorphism to assess (1) the genetic divergence between the two morphotypes, (2) their connectivity patterns and (3) their phylogenetic position within the Parazoanthidae. Specimens of P. axinellae were sampled in 11 locations along the northwestern Mediterranean; in 6 locations, samples of the two morphotypes were collected in sympatry. Small genetic diversity and structure were found within morphotypes, while marked and consistent differentiation was detected between them. Moreover, the less widespread morphotype appeared to be closer to Pacific species as P. juanfernandezii and P. elongatus. Our findings confirmed the limited knowledge on Parazoanthus species complex, and how this gap can have important implication for the conservation strategies of this widespread and valuable genus in the Mediterranean Sea.

Nutrient levels in coastal environments have been increasing globally due to elevated inputs of sewage and terrigenous sediments carrying fertilizers. Yet, despite their immense filtering capacities, marine sponges appear to be less affected by elevated nutrients than sympatric benthic organisms, such as corals. While the molecular-level stress response of sponges to elevated seawater temperatures and other toxicants has been defined, this study represents the first global gene expression analysis of how sponges respond to elevated nitrogen. Gene correlation network analysis revealed that sponge gene modules, coded by colours, became either highly upregulated (Blue) or downregulated (Turquoise, Black, Brown) as nitrogen treatment levels increased. Gene Ontology enrichment analysis of the different modules revealed genes involved in cell signalling, immune response and flagella motility were affected by increasing nitrogen levels. Notably, a decrease in the regulation of NF-kappaB signalling and an increase in protein degradation was identified, which is comparable to metabolic pathways associated with the sponge thermal stress response. These results highlight that Cymbastela stipitata can rapidly respond to changes in the external environment and identifies pathways that probably contribute to the ability of C. stipitata to tolerate short-term nutrient pulses.