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Biochar and organic fertilizer are widely supported to maintain crop production and sustainable development of agroecosystems. However, it is unclear how biochar and organic fertilizer alone or in combination regulate soil functional microbiomes and their relationships to ecosystem multifunctionality (EMF). Herein, a long-term (started in 2013) field experiment, containing five fertilization treatments, was employed to explore the effects of biochar and organic fertilizer applications on the EMF (based on 18 functional indicators of crop productivity, soil nutrient supply, element cycling, and microbial biomass) and the functional microbiomes of bulk soil and rhizosphere soil [normalizing the abundances of 64 genes related to carbon (C), nitrogen (N), phosphorus (P), and sulphur (S) cycles]. Compared with single-chemical fertilization, biochar and organic fertilizer inputs significantly enhanced most ecosystem-single functions and, in particular, the EMF significantly increased by 18.7–30.1%; biochar and organic fertilizer applications significantly increased the abundances of soil microbial functional taxa related to C-N-P-S cycles to varying degree. The combined application of biochar and organic fertilizer showed a better improvement in these indicators compared to using them individually. Most functional microbial populations in the soil, especially the taxa involved in C degradation, nitrification, nitrate-reduction, organic P mineralization, and S cycling showed significantly positive associations with the EMF at different threshold levels, which ultimately was regulated by soil pH and nutrient availability. These results highlight the strong links between soil microbiomes and agroecosystem functions, as well as providing scientific support for inclusion of biochar in agricultural production and services with organic amendments.Graphical Abstract8-year field evidence revealed impacts of biochar and pig manure on soil functional microbiome and ecosystem functions.Biochar and pig manure inputs notably enhanced most ecosystem-single functions and the EMF increased by 18.7–30.1%.Biochar and pig manure inputs notably enriched soil functional microbes related to C-N-P-S cycles to varying degree.Increase in EMF was related to microbe-driven soil processes such as C degradation, nitrification, and Po mineralization.Inclusion of biochar in crop production with organic amendments could enhance agro-ecosystem functions and services.

The incredibly complex soil microbial communities at small scales make their analysis and identification of reasons for the observed structures challenging. Microbial community structure is mainly a result of the inoculum (dispersal), the selective advantages of those organisms under the habitat‐based environmental attributes, and the ability of those colonizers to sustain themselves over time. Since soil is protective, and its microbial inhabitants have long adapted to varied soil conditions, significant portions of the soil microbial community structure are likely stable. Hence, a substantial portion of the community will not correlate to often measured soil attributes. We suggest that the drivers be ranked on the basis of their importance to the fundamental needs of the microbes: (i) those that supply energy, i.e., organic carbon and electron acceptors; (ii) environmental effectors or stressors, i.e., pH, salt, drought, and toxic chemicals; (iii) macro‐organism associations, i.e., plants and their seasonality, animals and their fecal matter, and soil fauna; and (iv) nutrients, in order, N, P, and probably of lesser importance, other micronutrients, and metals. The relevance of drivers also varies with spatial and time scales, for example, aggregate to field to regional, and persistent to dynamic populations to transcripts, and with the extent of phylogenetic difference, hence phenotypic differences in organismal groups. We present a summary matrix to provide guidance on which drivers are important for particular studies, with special emphasis on a wide range of spatial and temporal scales, and illustrate this with genomic and population (rRNA gene) data from selected studies.

Natural and artificial marine surfaces are rapidly colonized by microscopic communities, including propagules of some macrofoulers, in a process called biofouling. These microbiomes play an important role in modulating the evolving microbial community, as well as the attachment and settlement of other invertebrate larvae. Microbiomes act as biochemical and biophysical interfaces in marine communities. This review explores the gene-level processes that underlie microbial functions relevant to biofouling and larval settlement, such as quorum sensing, extracellular polymeric substance (EPS), and innate immune system components, as well as biosynthetic and degradative processes that generate signaling molecules. We critically evaluate current knowledge on how microbial metabolites promote or inhibit larval recruitment in corals, barnacles, polychaetes, and bivalves, and how omics-based approaches are uncovering the functional potential of biofilm communities. We evaluate how these interactions influence ecosystem services, such as habitat structuring, reef resilience, and coastal infrastructure maintenance.

Deciphering the biology of natural microbial communities is limited by the lack of functional data. While transcriptomics enables gene expression profiling, mRNA levels often fail to predict protein abundance, the primary indicator of microbial function. Prior studies addressed this by calculating RNA-to-protein (RTP) conversion factors using conserved protein-to-RNA (ptr) ratios across bacterial strains, but their cross-species and cross-domain utility remained unknown. We generated comprehensive transcriptomic and proteomic data sets from seven bacteria and one archaeon spanning diverse metabolisms and ecological niches. We identified orthologous genes with conserved ptr ratios, enabling the discovery of RTP conversion factors that significantly improved protein prediction from mRNA, even between distant species and domains. This reveals previously unrecognized conservation in ptr ratios across domains and eliminates the need for paired proteomic data in many cases. Our approach offers a broadly applicable framework to enhance functional prediction in microbiomes using only transcriptomic data.

The impact of microbial communities, also known as the microbiome, on human health and the environment is receiving increased attention. Studying translated gene products (proteins) and comparing metaproteomic profiles may elucidate how microbiomes respond to specific environmental stimuli, and interact with host organisms. Characterizing proteins expressed by a complex microbiome and interpreting their functional signature requires sophisticated informatics tools and workflows tailored to metaproteomics. Additionally, there is a need to disseminate these informatics resources to researchers undertaking metaproteomic studies, who could use them to make new and important discoveries in microbiome research. The Galaxy for proteomics platform (Galaxy-P) offers an open source, web-based bioinformatics platform for disseminating metaproteomics software and workflows. Within this platform, we have developed easily-accessible and documented metaproteomic software tools and workflows aimed at training researchers in their operation and disseminating the tools for more widespread use. The modular workflows encompass the core requirements of metaproteomic informatics: (a) database generation; (b) peptide spectral matching; (c) taxonomic analysis and (d) functional analysis. Much of the software available via the Galaxy-P platform was selected, packaged and deployed through an online metaproteomics “Contribution Fest“ undertaken by a unique consortium of expert software developers and users from the metaproteomics research community, who have co-authored this manuscript. These resources are documented on GitHub and freely available through the Galaxy Toolshed, as well as a publicly accessible metaproteomics gateway Galaxy instance. These documented workflows are well suited for the training of novice metaproteomics researchers, through online resources such as the Galaxy Training Network, as well as hands-on training workshops. Here, we describe the metaproteomics tools available within these Galaxy-based resources, as well as the process by which they were selected and implemented in our community-based work. We hope this description will increase access to and utilization of metaproteomics tools, as well as offer a framework for continued community-based development and dissemination of cutting edge metaproteomics software.

Mediterranean-type ecosystems (METs) occur on five continents and represent some of the most climatically constrained yet biologically rich regions on Earth. In these environments, legumes and their nitrogen-fixing rhizobial symbionts—including widely distributed genera such as Rhizobium, Bradyrhizobium, and Ensifer—play a pivotal role in sustaining plant productivity, nutrient cycling, and ecosystem resilience. This review synthesizes current knowledge on the environmental regulation of legume–Rhizobium symbiosis specifically within Mediterranean-type ecosystems, focusing on how nitrogen (N) and phosphorus (P) availability, light conditions, and carbon allocation trade-offs shape symbiotic performance across the five Mediterranean-type regions of the world (California, central Chile, the Cape Region of South Africa, southwestern Australia, and the Mediterranean Basin). By integrating physiological, ecological, and biogeochemical perspectives, we highlight how the shared features of these regions—strong seasonal drought, chronic nutrient limitation (particularly P in southwestern Australia and the Cape Region), recurrent fires, and exceptionally high plant diversity—constrain and, at the same time, favor the ecological success of symbiotic legumes. Throughout the review, we use case studies from key legume genera such as Lupinus in Chile and southwestern Australia, Virgilia and other Cape legumes in South Africa, Acacia in Australian kwongan and woodlands, and Medicago and Cytisus in the Mediterranean Basin and California to illustrate how general principles of legume–Rhizobium ecology manifest under Mediterranean-type climatic and edaphic constraints. Beyond summarizing established mechanisms, we critically examine the limitations of current metagenomic approaches, which often provide descriptive inventories of soil microbial communities without linking microbial composition to functional outcomes. We argue that advancing the field requires integrated, hypothesis-driven research that combines multi-omic tools with plant eco-physiology, soil nutrient dynamics, and temporal replication. Finally, we outline key priorities for future research, including the integration of functional ‘omics’, the study of microbiome interactions beyond rhizobia, the development of predictive models for Mediterranean-type ecosystems under climate change, and the application of symbiotic principles to restoration and agroecological management. By bridging molecular, physiological, and ecosystem perspectives, this review provides a conceptual framework for understanding and enhancing legume–Rhizobium symbiosis across five continents in a rapidly changing world.

Sarracenia provide an optimal system for deciphering the host-microbiome interactions at various levels. We analyzed the pitcher microbiomes and metatranscriptomes of the parental species, and F1 and F2 generations from the mapping population ( Sarracenia purpurea X Sarracenia psittacina ) utilizing high-throughput sequencing methods. This study aimed to examine the host influences on the microbiome structure and function and to identify the key microbiome traits. Our quality datasets included 8,892,553 full-length bacterial 16s rRNA gene sequences and 65,578 assembled metatranscripts with microbial protein annotations. The correlation network of the bacterial microbiome revealed the presence of 3-7 distinct community clusters, with 8 hub and 19 connector genera. The entire microbiome consisted of viruses, bacterial, archaea, and fungi. The richness and diversity of the microbiome varied among the parental species and offspring genotypes despite being under the same greenhouse environmental conditions. We have discovered certain microbial taxa that are genotype-enriched, including the community hub and connector genera. Nevertheless, there were no significant differences observed in the functional enrichment analysis of the metatranscriptomes across the different genotypes, suggesting a functional convergence of the microbiome. We found that the pitcher microcosm harbors both rhizosphere and phyllosphere microbiomes within its boundaries, resulting in a structurally diverse and functionally complex microbiome community. A total of 50,424 microbial metatranscripts were linked to plant growth-promoting microbial proteins. We show that this complex pitcher microbiome possesses various functions that contribute to plant growth promotion, such as biofertilization, bioremediation, phytohormone signaling, stress regulation, and immune response stimulation. Additionally, the pitcher microbiome exhibits traits related to microbe-microbe interactions, such as colonization of plant systems, biofilm formation, and microbial competitive exclusion. In summary, the demonstrated taxonomical divergence and functionally convergence of the pitcher microbiome are impacted by the host genetics, making it an excellent system for discovering novel beneficial microbiome traits.

The endangered beluga ( Delphinapterus leucas ) population in the St. Lawrence Estuary (SLE) in eastern Canada, the largest estuary in the world, is declining. Elevated tissue concentrations of a wide range of environmental contaminants, for example, halogenated flame retardants (HFRs) including polybrominated diphenyl ethers (PBDEs), might play a role in the non-recovery of this population. In mammals, HFRs have been reported to impair the metabolic regulation, including amino acid and fatty acid pathways. In the present study, we collected both blubber and skin swab samples from tissue biopsies from 56 adult SLE belugas and analyzed their blubber for the concentrations of a comprehensive suite of PBDEs and other HFRs. Using 16S rRNA marker and shotgun metagenomic approaches using skin swabs, we investigated, for the first time, the SLE beluga skin microbiome and the SLE water microbiome, providing valuable comparative taxonomic and functional microbiome information. We found that belugas have a unique skin microbiome that is distinct from surrounding SLE water, regardless of the beluga sex or location in the SLE. We further characterized the core microbiome of SLE beluga skin and surrounding SLE water, and identified bacterial taxa and gene functional pathways associated with the skin microbiome that correlated with beluga blubber HFR concentrations. Namely, we identified the phylum Nitrospinae and candidate phylum PAUC34f as potential taxa of interest that are associated with blubber HFR concentrations. We hypothesize that the biodegradation of HFRs within the beluga blubber and skin results in an increase in local metabolite concentrations that leads to the proliferation of Nitrospinae and PAUC32f. This work demonstrates the utility of studying the core microbiome of the SLE beluga skin using a swab method that could be adapted to field sampling. Further studies of the temporal effects of contaminant exposure on SLE beluga skin and SLE water microbiomes is warranted for potentially better monitoring and protecting this marine mammal which is at risk.

Gut microbiota enable wood-feeding insects to digest recalcitrant diets. Two DNA-based analyses were performed. Amplicon sequencing of gut microbiota samples from Cryptocercus punctulatus showed inter-individual heterogeneity with visually distinct ordination patterns; however, no statistically significant differences were detected. Shotgun metagenomics was used to compare the taxonomic and functional profiles of C. punctulatus gut microbiota with those of other xylophagous Dictyoptera. Despite taxonomic differences, C. punctulatus microbiota revealed functional convergence with termites (Mastotermes darwiniensis and Nasutitermes sp.). Carbohydrate metabolism was performed by different bacterial phyla across all insects. All insect species possessed metabolic potential for cellulose, hemicellulose, pectin, and starch digestion, but lignin degradation capabilities were not detected. Termites showed higher abundance of chitin and xylan degradation pathways and nitrogen fixation genes, though nitrogen fixation was also present in Cryptocercus cockroaches. Genes for oxidative stress tolerance were present across all species but were most abundant in cockroaches, particularly, Cryptocercus. All insects harbored antibiotic resistance genes, with highest levels found in cockroaches. These findings indicate that metabolic requirements for wood digestion shape gut microbial community assembly across xylophagous insects, with distinct microbial taxa contributing to cellulose and hemicellulose breakdown. Moreover, the widespread presence of antibiotic resistance genes raises concerns about the potential transmission of antibiotic resistance within insect-associated microbiomes.