Environmental Microbiology

Pelagibacterales gMED is the dominant epipelagic genomospecies in the western Mediterranean Sea. We used the O-chain biosynthesis gene clusters, OBCs, as clonal barcodes to analyse strain-level population structure. In total, 385 OBC-defined clonal lineages were tracked across Mediterranean metagenomes spanning 14 years and depths from 5 to 90 m within the photic zone, with between 128 and 336 detected per metagenome. The relative conservation of dominant OBC types across years, seasons, and geographic locations indicated a persistently high and stable clonal diversity.

Competition for light has driven extensive pigment diversification among phytoplankton species, yet how this diversity shapes their spatiotemporal distribution in the field has been little studied so far. The cyanobacterium Synechococcus is an ideal model for addressing this issue, since this group has colonized most light spectral niches in marine environments. Here, we used an approach based on marker read recruitment from metagenomes to analyze the seasonal succession of Synechococcus pigment types (PTs) at two time-series stations off French coasts exhibiting contrasting oceanic regimes. Marked seasonality was observed at both sites. The shallow, permanently mixed English Channel site SOMLIT-Astan was characterized by an alternation between green-light specialists (PT 3a) peaking in spring, and chromatic acclimaters type A (PT 3dA) accounting for most of the Synechococcus community in winter. In contrast, the pigment diversity was much higher at the deep Mediterranean station BOUSSOLE. In the upper layer, the two main PTs were the blue light specialists (PT 3c), which dominated the community in summer and fall, and PT 3dA cells, which were more abundant in spring. The third most abundant PT was chromatic acclimaters type B (PT 3dB), which accounted for up to 15% of the surface community in late fall. Strikingly, PT 3dA was dominant at depth during most of the year. Multivariate analyses between PT abundances, clade abundances and environmental factors, notably water color indexes, suggested new associations between PTs to specific clades and ecological niches. This study provides novel insights for refining distribution models of Synechococcus PTs and phytoplankton groups in general.

Bacterial bloomers,populations that experience rapid and significant increases in abundance in response to environmental triggers, briefly dominate marine microbial communities, potentially impacting the ecosystem by channeling large amounts of nutrients and affecting carbon fluxes. Due to their ephemeral nature, bacterial bloomers are challenging to capture, and it remains unknown whether they are restricted to specific taxonomic groups or whether they exhibit recurrent patterns. We analyzed a decade-long time series from the Blanes Bay Microbial Observatory (BBMO, NW Mediterranean Sea) to investigate bacterial bloomers in two size fractions (free-living (0.2-3 um) and particle-attached (3-20 um) communities. We identified 57 Amplicon Sequence Variants (ASVs), less than 1% of the total bacterial richness, exhibiting recurrent or chaotic blooming-like behavior. Bloomers spanned diverse phyla, though some taxonomic coherence appeared within families containing multiple blooming taxa. Monthly sampling detected bloom events on average 4.6 +- 1.9 times per year across both size fractions. Once seasonality was accounted for, blooms showed weak associations with biological and physicochemical variables, likely a consequence of monthly sampling resolution. Nonetheless, a marked shift in the blooming community within the particle-attached size fraction coincided with ecosystem disturbances from the nearby harbour restoration, suggesting that bloomers may act as disturbance sentinels. Metagenomic data showed that blooms led to marked shifts in the community functional potential. Overall, our findings underscore the importance of investigating bloom dynamics to understand microbial contributions to biogeochemical cycles and stress the need for higher-frequency sampling to accurately capture these transient but ecologically relevant events.

In marine ecosystems, phytoplankton growth is frequently limited by iron, a micronutrient, due to its poor solubility from major sources such as atmospheric dust. Many phytoplankton cannot access dust-bound iron independently, and processes that solubilize this iron remain poorly understood. Here, we investigated whether bacterial partners can enhance phytoplankton growth under iron-limited conditions by facilitating utilization of dust-bound iron. Our study focused on Phaeodactylum tricornutum, a model diatom that is adapted to low iron growth conditions, grown in co-culture with bacteria isolated from its phycosphere. In iron-limited experiments using mineral dust as the sole iron source, the addition of Marinobacter significantly enhanced diatom growth compared to axenic controls, whereas Stappia significantly suppressed it. However, under iron-replete conditions, neither bacterium affected growth. These results indicated that under low-iron conditions, Marinobacter actively alleviates iron deficiency. Co-cultured bacterial cell abundances remained at least an order of magnitude lower than diatom cells. Marinobacter also enhanced algal growth within days of dust addition to established Fe-limited co-cultures, indicating its beneficial effect on P. tricornutum was not unique to a system in which it was newly introduced. Exometabolomic profiling comparing the axenic diatom and co-cultures revealed a suite of condensed aromatic organosulfur and peptide-like compounds associated with bacterial presence, as well as compounds that appeared to be unique to each co-culture, hinting at a molecular underpinning of each strains impact. Our findings demonstrate that low-abundance members of the phycosphere community can have a significant impact on host growth by modulating the accessibility of dust-bound Fe.

Cyanophages can influence the dynamics of toxic cyanobacterial blooms. However, cyanobacteria can become resistant to viruses through natural selection processes. Here, we investigate the acquisition of virus resistance in a toxic, freshwater, gas-vacuolate, bloom-forming cyanobacterium, Microcystis aeruginosa, under different nutrient concentrations. We find that gas-vacuolate M. aeruginosa subpopulations acquire virus resistance in low nitrogen cultures regardless of their phosphorus concentration, whereas non-vacuolate subpopulations do not. After resequencing susceptible and resistant M. aeruginosa variants, we identify a mutation in the transmembrane domain of a nitrogen-related transporter as the most likely genetic cause of the resistance. Infection experiments further reveal a larger viral burst size and higher phycocyanin content in gas-vacuolate cells compared to non-vacuolate ones. Based on these experimental results, we propose an ecological model in which lower nitrogen concentrations, higher light intensities and increased virus-host contact rates facilitate the evolution of virus resistance in upper lake layers during Microcystis-dominated blooms.

Chytrids are unicellular fungi that infect and degrade phytoplankton as parasites or saprotrophs. They impact not only food availability and quality in surface waters but also carbon cycling and sequestration. So far, their ecological significance has mostly been investigated for freshwater environments, whereas observations for marine environments are scarce -- even though chytrids can be highly abundant there, too (as shown for the Arctic Ocean). To test the chytrids' potential to control phytoplankton dynamics in the Arctic Ocean, we analysed metabarcoding and photosynthetic pigment data from two expeditions, Tara Polar Circle and MOSAiC; the latter providing a dense sampling transect across one year from the under-ice water column and sea ice samples. The phytoplankton communities of both environments were dominated by diatoms, with strong seasonal effects indicating blooms in the water column. Chytrids dominated fungal communities in both environments and revealed a strong cryo-pelagic coupling. They were especially abundant during the sea ice melt in water samples and in ice-associated (sympagic) samples, where they represented >2% and up to 61%, respectively, of all combined reads assigned to chytrids or phytoplankton. Co-occurrences of the two most abundant chytrid taxa with some of the most abundant diatom taxa and niche differentiation from other potential diatom parasites are consistent with the chytrids' critical role in controlling diatom blooms, especially in sympagic habitats.

Primary production in the Southern Ocean (SO) plays a critical role in regulating the global carbon cycle, yet the physiological mechanisms governing phytoplankton responses to iron (Fe) limitation and variable light remain poorly constrained. Using a custom made incubation system that simulated natural diel solar variability, we examined the interactive effects of Fe availability, light intensity, and photoperiod (continuous vs. variable) on three ecologically important SO phytoplankton: Fragilariopsis cylindrus, Phaeocystis antarctica, and Thalassiosira antarctica. Physiological, photophysiological, and proteomic measurements revealed that Fe availability was the dominant factor regulating growth, carbon production, photosynthetic performance and protein expression across all species. Distinct acclimation strategies emerged: F. cylindrus exhibited marked trade-offs between productivity and photoprotection under Fe stress, consistent with adaptation to stable, low-light, Fe-poor environments; P. antarctica maintained growth by flexibly modulating photoprotective and photosynthetic capacity, reflecting high plasticity suited to dynamic, open-ocean conditions; and T. antarctica expressed a balanced strategy, sustaining productivity and photoprotection simultaneously, characteristic of coastal bloom formers with higher Fe demand. Dynamic light regimes produced smaller, species-specific effects, influencing chlorophyll content and carbon storage primarily in T. antarctica. Correlation and z-score analyses demonstrated that Fe-rich photosynthetic proteins co-varied with biomass production, whereas photoprotective traits clustered independently, underscoring divergent energy-allocation strategies. Together, these results reveal how SO phytoplankton partition resources between productivity and photoprotection under shifting Fe-light regimes, providing mechanistic insight into their ecological niches.

Marine dissolved organic carbon is a chemically complex substrate pool that fuels heterotrophic bacteria, yet it remains difficult to determine which compounds are used by specific microbes. Bacterial transcriptomes offer a potential biosensor of substrate availability, but the reliability of this approach in chemically mixed substrates remains uncertain. Here, we evaluated the reliability of this transcriptional sensor approach using the model marine bacterium Ruegeria pomeroyi DSS-3 grown on either glucose or a defined mixture containing glycerol, benzoate, succinate, leucine, dimethylsulfoniopropionate, and trimethylamine N-oxide. Genome-wide transcription differed strongly between treatments, with the mixed-substrate treatment enriched in genes associated with C1 metabolism, sulfur oxidation, benzoate degradation, and motility. Across substrates, the most diagnostic transcriptional responses occurred at pathway entry points and first committed reactions, including glucose transport and Entner-Doudoroff metabolism, trimethylamine N-oxide transport and catabolism, and early steps of aerobic benzoate oxidation. In contrast, downstream metabolic genes were less substrate-specific, likely because multiple pathways converged on shared central metabolic intermediates. Transporter transcription was also less consistently diagnostic than expected, although substrate-binding subunits often showed the strongest responses within transporter complexes. Comparisons with previous single-substrate studies indicated that some transcriptional markers, particularly benzoate oxidation genes, remained detectable in the substrate mixture, whereas glycerol and succinate responses were weakened or lost. These findings show that transcriptomics can provide useful insight into bacterial substrate use, but interpretation is most robust when focused on experimentally validated transporters and early pathway genes, and when evaluated in the context of pathway connectivity, cellular physiology, and substrate mixture complexity.

Mangrove forests are major blue carbon ecosystems but are often characterized by low surface methane (CH4) emissions. Such low emissions, however, do not necessarily indicate weak methanogenesis, because CH4 production may be offset by internal CH4 consumption before reaching the atmosphere. Although previous community, genomic, and transcriptomic studies have implicated methylotrophic methanogenesis in mangrove sediments, direct taxon-resolved evidence linking methylated carbon assimilation to potentially active methanogens remains limited. Here, we combined methanogenic activity assays, DNA stable isotope probing (DNA-SIP), mcrA and 16S rRNA gene analyses, and phylogenetic comparisons to identify potentially active methanogens across saline-influenced mangrove soils. The results showed that CH4 production potentials were consistently dominated by methylotrophic pathways (1.86-2.78 g CH4 g-1 soil hr-1) across all sites. DNA-SIP, together with consistent community patterns in fresh soils, indicated the potential activity of methylotrophic and mixotrophic methanogens under saline conditions. Methanolobus-affiliated methanogens were associated with salinity, Na+, Cl-, and NH4+, whereas Methanosarcina and unclassified Methanosarcinaceae were linked to soil soluble organic carbon availability and water content, indicating niche differentiation among active methanogenic groups. Phylogenetic analyses incorporating reference sequences from diverse environments further showed that potentially active mangrove methanogens were dominated by saline-associated lineages. Together with our previous methanotrophic evidence from the same sites, these findings suggest that low CH4 emissions from mangrove blue carbon ecosystems can mask substantial internal CH4 cycling sustained by active methanogenesis and CH4 consumption.

The marine sponge holobiont, composed of the sponge host and its microbial symbionts, is a known source of abundant and diverse ether lipids (ELs). Apart from their structural role in the cytoplasmic membrane of archaea and some bacteria, ELs have often been linked to signaling functions and defense against pathogens. Despite the relevance of ELs, their biosynthesis, as well as the identity of their producers, remain elusive. Here, we report the analysis of potential ether lipid producing genes and gene clusters, detected in marine sponge metagenomes as well as public sponge genomes. We show that the sponge holobiont has the capacity to synthesize ELs via several pathways, and suggest the ability of the sponge holobiont to synthesize ELs under different O2 levels. Finally, targeted lipidome analysis confirmed that ELs are present in the lipid profiles of all of the studied sponge holobiont samples, and indicates that the biosynthesis of the plasmalogens detected is likely restricted to the sponge host itself, based on the detected hydrocarbon chain lengths. This work provides a basis for the challenging quest to decipher intricate EL biosynthesis in marine sponges and their associated microbes.

Filamentous phages infecting Gram-positive bacteria remain largely unexplored. Notably, only two filamentous phages, B5 and Philemon infecting Propionibacterium freudenreichii, have been described to date in the phage-rich dairy ecosystem. Although both were genomically characterized, only B5 was confirmed to be an infective filamentous single-stranded DNA phage. The aim of this study was to isolate and characterize new filamentous phages from Swiss-type cheese to investigate their diversity, structural features, host specificity, and potential adaptation to the dairy environment. Thirty raw and pasteurized milk cheeses from France were screened for phages infecting P. freudenreichii strains. Eleven phages were isolated, nine of which displayed a filamentous morphology. Named MINOG1 to MINOG9, these filamentous phages exhibited genomic features typical of this morphotype, including small single-stranded DNA genomes with collinear genes organized into functional modules. Comparison with B5 and Philemon revealed sequence divergence ranging from 0.1% to 7%. These phages also exhibited a diverse host range. To further explore phage-P. freudenreichii interactions, we screened the genomes of the strains used in this study, as well as additional genomes retrieved from the NCBI database, for CRISPR spacers predicted to target these filamentous phages. Numerous strains contained CRISPR spacers showing 79 to 100% identity to genomic regions of these phages. Two P. freudenreichii strains displayed markedly different phage resistance levels despite exact spacer-protospacer matches with phages B5, MINOG1, MINOG2, and MINOG8. Conversely, several strains were resistant to nearly all tested phages despite lacking CRISPR spacers targeting them suggesting the presence of additional defense systems in P. freudenreichii. IMPORTANCEFilamentous phages can play important roles in bacterial ecology by modulating host physiology, population dynamics, and bacterial adaptation to specific environments. However, filamentous phages infecting Gram-positive bacteria remain among the least explored bacterial viruses, and their diversity, ecology, and interactions with their hosts are still poorly understood. This knowledge gap is particularly relevant in dairy ecosystems, where phages are abundant and can influence microbial communities and fermentation processes. In characterizing nine new filamentous phages infecting Propionibacterium freudenreichii from Swiss-type cheeses, this study expands the known diversity of filamentous phages associated with Gram-positive bacteria and provides new insights into phage-host interactions and bacterial defense strategies in dairy-associated bacteria.

DNA chemical modifications, including methylation, are widespread in prokaryotes and mobilomes, including viruses, plasmids, and other extrachromosomal DNAs, and play important roles in their ecology and interactions. However, current knowledge of these modification systems and their association with interactions between hosts and mobilomes across communities, including those in extreme environments, is severely limited. Here, using single-molecule real-time sequencing and single-cell genome sequencing technologies, we conducted a culture-independent metaepigenomic analysis of microbial communities in hot spring biofilms. A total of 248, 332, and 465 genomes were constructed from diverse prokaryotes, viruses, and extrachromosomal circular DNAs, respectively, from 10 biofilm samples collected from 3 hot spring sites. In total, 1106 candidate methylated motifs and 3280 genes associated with the restriction-modification (RM) system, including DNA methyltransferases (MTases), were identified. In contrast to the varied methylated motifs, the nucleotide-level modification ratios were consistent with those of a common Escherichia coli genome, and an environment-dependent epigenomic preference attributed to the lack of C5-methylcytosine was observed, as supported by direct measurements of modified bases by liquid chromatography-tandem mass spectrometry. A systematic survey revealed various defense systems in the genome, and almost half of the MTase genes were estimated to be genetically involved in defense mechanisms against extracellular DNA, such as RM systems. The mobilomes and their predicted hosts shared epigenomic patterns within each interactive subnetwork, suggesting that mobilome DNA was modified by host MTase during the current infection or transfection, rather than serving as historical records. Our findings highlight that DNA modification shapes multiple ecological and evolutionary strategies in interactions between prokaryotes and mobilomes, and that epigenomes serve as a potential signature for accurate prediction of current host-phage interactions.

Symbiotic interactions between plants and nitrogen-fixing microorganisms are essential for sustainable agriculture, yet the molecular mechanisms underlying plant-cyanobacterium symbiosis remain poorly understood. In particular, the nature of the signalling mechanisms mediating partner recognition in associations involving Nostoc species is largely unknown. Recent proteomic analyses have identified proteins homologous to rhizobial Nod factors biosynthetic enzymes in Nostoc punctiforme, suggesting the existence of a Nod-like signalling system. However, the functional role of these components has not been experimentally validated. Here, we investigate the contribution of nod-like biosynthetic and regulatory genes to symbiosis by analysing mutants of N. punctiforme affected in genes with homology to nodB and nodD. Phenotypic characterization revealed that disruption of nodB-like genes does not impair free-living growth but affects early stages of plant association and colonization. Specifically, the nodB1 mutant is impaired in plant association and shows a mild defect in colonization, whereas the nodB3 mutant exhibits a severe defect in colonization. In contrast, nodD-like mutants exhibited altered symbiotic phenotypes, with specific regulators differentially affecting interaction and colonization efficiency in rice (Oryza sativa). In particular, mutation of nodD2 and nodD3 reduced plant association and severely compromised colonization in Oryza sativa, with a more pronounced phenotype in nodD3 mutant. Altogether, our results provide genetic evidence supporting the involvement of Nod-like components in cyanobacterial symbiosis and suggest the existence of a regulatory and biosynthetic module contributing to plant colonization. These findings shed new light on the evolution and diversity of symbiotic signalling mechanisms across plant-microbe interactions.

Lake Constance is a pre-Alpine, monomictic, oligotrophic lake situated at the southern end of Germany composed of two main water bodies: deep, oligotrophic Upper Lake Constance (ULC) and the shallow, more mesotrophic Lower Lake Constance (LLC). To date, no sequencing-based study exists of the seasonal succession of the microbial plankton in Lake Constance. Over one-year, microbial plankton communities were sampled biweekly from the top 20 m of the water column in both sites and separated into nanoplankton (NP) and picoplankton (PP). Communities were analysed using rDNA amplicon sequencing: NP samples were analysed by 18S rDNA, and PP samples by 18S and 16S rDNA sequencing. Temporal community diversity was compared between sites and the effect of two major environmental perturbations, winter vertical mixing in ULC and oxygen depletion of the bottom-water layer in LLC, on the community was examined. Despite strong environmental contrasts, microbial plankton communities exhibited conserved seasonal temporal dynamics across basins. In contrast, pronounced compositional shifts occurred during mixing and oxygen depletion events. Approximately 20% of detected taxa were positively associated with these events, with log fold changes reaching 9.82, reflecting rare or undetectable taxa outside these periods. Taxa favoured by these perturbations commonly exhibited high metabolic flexibility, including mixotrophy, fermentation, or anaerobic respiration, or possessed functional traits conferring tolerance to altered redox and mixing regimes. Our results suggest that the temporal dynamics of freshwater microbial plankton communities are driven by deterministic processes and highlight the profound impact of large, and less known, environmental changes on these communities.

The Eurasian spruce bark beetle Ips typographus, a major forest pest on Norway spruce (Picea abies), forms intimate associations with several types of microbial symbionts. While previous research has focused primarily on filamentous fungi, yeasts have remained largely unexplored. Here, we show that yeasts associated with I. typographus may contribute to host tree colonization by providing defensive benefits. Dominant yeasts (Yamadazyma, Kuraishia, Nakazawaea, and Wickerhamomyces), which are phylogenetically related to other insect-associated Saccharomycotina, significantly attract adult beetles. Moreover, several yeasts inhibit the growth of the pathogenic fungus Trichoderma harzianum in vitro, and beetle eggs benefit from the presence of Kuraishia capsulata by reduced fungal infection under semi-natural conditions. Strikingly, these effects are mediated by the yeasts' transformation of the tree's defensive stilbene glycosides into antimicrobial aglycones and phenolic acids that accumulate in beetle galleries. These findings reveal a previously unrecognized role of symbiotic yeasts in converting spruce defensive stilbene glycosides into antimicrobial aglycones and oxidative cleavage products that accumulate in beetle galleries, and might contribute to the survival of their bark beetle host.

Endosymbiosis is widespread throughout the tree of life. Understanding how the transition of a bacterial endosymbiont from facultative to host-dependent obligate life occurs is an important question for defining the origin of endosymbiosis. A novel gram-positive bacillus, Brevibacillus sp. TJ4 was isolated from the nitrogen-fixing yeast Rhodotorula mucilaginosa JGTA-S1, which houses several endobacteria within its cells. TJ4 can survive independently of yeast but exhibits genomic and metabolic features characteristic of an evolving endosymbiont, slowly assuming a host-dependent, obligate lifestyle. The TJ4 genome contains several incomplete pathways for carbohydrate, amino acid, vitamin, and cofactor metabolism, which is reflected in its increased reliance on host-derived nutrients and auxotrophy compared with that of other Brevibacillus spp. Comparative genomics revealed widespread genome rearrangements, loss of synteny, and multiple cross-genus and inter-kingdom horizontal gene transfer (HGT) events in TJ4 compared to other Brevibacillus spp. These HGTs include the acquisition of genes from bacteriophages and co-resident endobacteria of JGTA-S1. One such horizontally acquired gene, Type II 3-dehydroquinate dehydratase (AroQ), appears to have originated from the Rhodotorula host itself. This acquisition functionally restores the shikimate pathway in strain TJ4, as evidenced by the phylogenetic placement of AroQ from TJ4 within the clade of fungal AroQ homologs. Potential exploitation of the host JGTA-S1 appears to be a probable mode of endosymbiosis of TJ4, an evolving endosymbiont that we named Brevibacillus rhodotorulae sp. nov.

Biological N2O production from organic nitrogen is generally assumed to require canonical nitrification, which generates oxidized nitrogen that subsequently fuel denitrification. Whether this paradigm universally applies to nitrogen-rich microbial communities remains unclear. Here, we investigated N2O production across an industrial poultry manure composting process and found that substantial N2O formation occurred despite the apparent absence of canonical ammonia oxidation. Neither allylthiourea inhibition nor metagenomic analyses provided evidence for ammonia-oxidizing microorganisms or their activity. Instead, metagenomic analyses identified abundant bacterial nitric oxide synthase (bNos) genes, many of which were phylogenetically affiliated with Bacilli, the dominant bacterial group throughout composting. Physiological experiments with Bacillus isolates demonstrated a nitrification-independent route in which L-arginine was oxidized to NO2-/NO3-, consistent with bNOS-mediated NO formation followed by abiotic oxidation. Recovery of 15N-labelled N2O following 15NO2- addition established NO2- as an immediate precursor of aerobically produced N2O, confirming that the oxidized nitrogen generated through this alternative route subsequently fueled denitrification. Metagenomic analyses further revealed extensive denitrification potential but comparatively low nosZ abundance. Together, these findings identify a previously overlooked route linking organic nitrogen turnover to denitrification independently of canonical nitrification, thereby expanding current models of microbial N2O production in composts and potentially other protein-rich thermophilic environments.

Biological control strategies are increasingly being explored as sustainable alternatives for managing rice blast disease caused by Magnaporthe oryzae. In this study, we characterized three Bacillus pumilus isolates (DC01, DC09, and DC13) and evaluated their antifungal and plant-beneficial properties against M. oryzae. Whole genome sequencing revealed multiple biosynthetic gene clusters associated with the production of antimicrobial metabolites. All three isolates inhibited fungal growth in dual-culture assays, whereas heat-stable diffusible antifungal activity was primarily associated with the cell-free supernatants of DC09 and DC13. Exposure to bacterial supernatants disrupted fungal development, inducing abnormal hyphal morphology characterized by bulbous swelling, altered polarity, and increased branching in M. oryzae. Volatile organic compound assays further revealed that the DC isolates suppress fungal growth in the absence of physical contact. The isolates additionally inhibited the growth of other phytopathogenic fungi and selected human bacterial pathogens. All strains exhibited plant growth-promoting traits, including indole-3-acetic acid production and osmotic stress tolerance, whereas DC09 also displayed phosphate-solubilizing activity. Importantly, root inoculation with the DC isolates significantly reduced rice blast disease severity and induced expression of defense-associated genes involved in jasmonic acid/ethylene signaling and immune priming. Collectively, these findings identify the DC isolates, particularly DC09 and DC13, as promising multi-mechanistic biological control agents for sustainable rice blast management.

Microorganisms are the key drivers of carbon cycling in coastal marine sediment ecosystems, significantly influencing carbon storage and release during Gracilariopsis lemaneiformis cultivation. This study employed 16S rRNA sequencing, a high-throughput qPCR chip, and carbon isotope labeling to assess the impact of G. lemaneiformis cultivation on carbon cycling processes in coastal sediments. A comparative analysis was conducted between cultivated zones (GZ) of G. lemaneiformis and adjacent control zones (CZ). The results indicated that macroalgae cultivation significantly modified sediment-seawater exchange dynamics and accelerated carbon cycling within coastal marine sediment ecosystems. Furthermore, G. lemaneiformis cultivation increased the abundance of genes linked to polysaccharide degradation and carbon fixation pathways, thereby enhancing carbon cycling efficiency. The ecosystem multifunctional index, calculated based on carbon fixation gene abundance, was significantly higher in GZ compared to CZ. Incubation experiments using 13C-NaHCO3 demonstrated that cultivation markedly elevated the carbon fixation rate of sediment, emphasizing a higher potential for carbon sequestration in sedimentary environments cultivated with macroalgae. Additionally, cultivation significantly altered sediment microbial communities, simplifying their structural complexity. Key microbial taxa identified via k-core species analysis--including Subgroup10 of Desulfobacterota and MBNT15, correlated strongly with carbon fixation rates, indicating their pivotal roles in sediment carbon cycling processes. This study provides critical insights into how large-scale macroalgae cultivation influences coastal carbon dynamics and informs strategies for optimizing carbon management in aquaculture ecosystems.

Root-associated microorganisms play a pivotal role in helping plants adapt to drought stress. However, the underlying mechanisms of the rhizospheric microbiome under limiting soil moisture remain largely unresolved. Integrating total and active microbiome analyses enables a more accurate interpretation of microbial responses to climate change-associated water stress. We assessed the effect of reduced rainfall on two wheat genotypes, drought-tolerant (DT) and drought-sensitive (DS), using rainout shelters that allowed 100%, 75%, 50%, and 25% of natural precipitation to reach the crop. At the peak of the growing season, rhizosphere samples were collected for metagenomic (MG) and metatranscriptome (MT) sequencing. In parallel, rhizosphere volatile organic compounds (VOCs) were collected and analysed. Differential expression analysis of metatranscriptomic data using metagenomic abundance as a cofactor was performed by comparing all treatments to the 100% precipitation control. Our results demonstrate that particularly oxidative stress-related transcripts intensify in DS as rainfall decreases. Transcriptomic shifts primarily involved upregulation of transcripts associated with antioxidant (catalase, superoxide dismutase), heat shock proteins (Hsp10, Hsp60, DnaK/DnaJ, GroEL, GroES), as well as microbial functions related to osmoregulation, proline and glycine betaine (PutA, PutP, OpuBB), and plant growth-promoting traits such as auxin production, phosphate solubilization. Moreover, volatile organic compound (VOC) emissions differed significantly between the control and drought treatments, with higher emissions, particularly acetates, in the DS genotype than in the DT genotype. Overall, pronounced drought-induced shifts in active microbial functions and VOC emissions indicate high sensitivity and functional plasticity of the active microbiome, whereas the total microbiome remains robust under medium drought.