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Rivers serve important functions for human society and are significantly impacted by anthropogenic nutrient inputs (e.g. organic and sulfur compounds). Reduced organic and sulfur compounds influence the nitrogen cycle as they are electron donors of microbial nitrate reduction. Water pollution caused by individual nutrients and the mechanisms have been studied, but how the variation in multiple nutrient loadings influences nitrate-reducing microorganisms is less understood. Two sets of microcosms were established and exposed to nitrate, along with either acetate or thiosulfate, at different times. Nutrient concentrations responded to the loading pollutant. The nutrient loading order was more important in shaping microbial community structure and microbial interactions through the exchange of growth-required substances. This indicated that upstream or historical nutrient inflows impacted current nitrate reduction by changing the seeding microbial community, highlighting the importance of river connectivity. Based on metatranscriptome analysis, although the order and type of nutrient loadings were equally important in regulating global transcriptomes, transcripts of enzymes for key metabolisms (nitrate reduction, sulfur oxidation, etc.) more actively responded to the nutrient type. The regulation of a small set of genes was sufficient to make the transition, while most transcripts were not degraded and regenerated. These insights are important for understanding the varying pollution status of rivers and for developing effective solutions, such as remediation.

The cyanobacterium Microcystis causes harmful algal blooms that pose a major threat to human health and ecosystem services, particularly due to the prevalence of the potent hepatotoxin microcystin (MC). With their pronounced EPS layer, Microcystis colonies also serve as a hub for heterotrophic phycosphere bacteria. Here, we tested the hypothesis that the genotypic plasticity in its ability to produce MC influences the composition and assembly of the Microcystis phycosphere microbiome. In an analysis of individual colonies of a natural Microcystis bloom, we observed a significantly reduced richness of the community in the presence of MC biosynthesis genes. A subsequent synthetic community experiment with 21 heterotrophic bacterial strains in co-cultivation with either the wild-type strain Microcystis aeruginosa PCC 7806 or the MC-free mutant ΔmcyB revealed not only a tug-of-war between phototrophic and heterotrophic bacteria, but also a reciprocal dominance of two isolates of the genus Sphingomonas and Flavobacterium. In contrast, an Agrobacterium isolate thrived equally well in both consortia. In substrate utilization tests, Sphingomonas showed the strongest dependence on Microcystis exudates with a clear preference for the wild-type strain. Genome sequencing revealed a high potential for complementary cross-feeding, particularly for the Agrobacterium and Sphingomonas isolates but no potential for MC degradation. We postulate that strain-specific functional traits, such as the ability to perform glycolate oxidation, play a crucial role in the cross-feeding interactions, and that MC is one of the determining factors in the Microcystis phycosphere due to its interference with inorganic carbon metabolism.

Fecal microbiota filtrate transfer is discussed as a safe alternative to fecal microbiota transfer (FMT) to treat ulcerative colitis. We investigated modulation of viral and bacterial composition during fecal microbiota filtrate transfer followed by FMT in six patients with active ulcerative colitis (where clinical activity improved in three patients after filtrate transfer) and combined 16S ribosomal RNA gene amplicon sequencing with a virome analysis pipeline including fast viral particle enrichment and metagenome mapping to detect frequencies of 45,033 reference bacteriophage genomes. We showed that after antibiotic treatment and during filtrate transfer, the bacterial community typically adopted a stable composition distinct to that before antibiotic treatment, with no change toward a donor community. FMT in contrast typically changed the bacterial community to a community with similarity to donor(s). There were no indications of an establishment of predominant donor viruses during filtrate transfer but a remodeling of the virome. In contrast, the establishment of donor viruses during FMT correlated with the predicted hosts established during such transfer. Our approach warrants further investigation in a randomized trial to evaluate larger therapeutic interventions in a comparable and efficient manner.

Bacterial species adapt to cold environments with diverse molecular mechanisms enabling their growth under low ambient temperature. The emergence of cold-adapted species at macro-evolutionary scale, however, has not been systematically explored. In this study, we performed phylogenetic analysis on the growth temperature traits in the genera that occupy broad environmental and host niches and contain known cold-adapted species. Our results demonstrate that in the genus Pseudomonas, cold-adapted species formed a distinct and conserved clade, whereas in Paenibacillus, cold-adapted species were sporadically distributed throughout the phylogenetic tree. The cold-adapted clade of Pseudomonas exhibited genome-wide signatures of adaptation and possessed clade-specific genes. This indicates that there are diverse evolutionary patterns in the divergence of cold-adapted species among different bacterial genera.

The intensification of agriculture has led to environmental degradation, including the loss of biodiversity. This has prompted interest in perennial grain cropping systems to address and mitigate some of these negative impacts. In order to determine if perennial grain cultivation promotes a higher microbial diversity, we assessed the endophytic microbiota of a perennial grain crop (intermediate wheatgrass, Thinopyrum intermedium L.) in comparison to its annual counterpart, wheat (Triticum aestivum L.). The study covered three sampling sites in a pan-European gradient (Sweden, Belgium, and France), two plant genotypes, three plant compartments (roots, stems, and leaves), and two sampling time points. We observed that the host genotype effect was mainly evident in the belowground compartment, and only to a lesser extent in the aboveground tissues, with a similar pattern at all three sampling sites. Moreover, intermediate wheatgrass roots harbored a different bacterial community composition and higher diversity and richness compared to their annual counterparts. The root bacterial diversity was influenced by not only several soil chemical parameters, such as the carbon:nitrogen ratio, but also soil microbial parameters, such as soil respiration and dehydrogenase activity. Consistent findings across time and space suggest stable mechanisms in microbiota assembly associated with perennial grain cropping, underscoring their potential role in supporting biodiversity within sustainable agricultural systems.

Harmful Algal Blooms (HABs) of the toxigenic dinoflagellate Karenia brevis (KB) are pivotal in structuring the ecosystem of the Gulf of Mexico (GoM), decimating coastal ecology, local economies, and human health. Bacterial communities associated with toxigenic phytoplankton species play an important role in influencing toxin production in the laboratory, supplying essential factors to phytoplankton and even killing blooming species. However, our knowledge of the prevalence of these mechanisms during HAB events is limited, especially for KB blooms. Here, we introduced native microbial communities from the GoM, collected during two phases of a Karenia bloom, into KB laboratory cultures. Using bacterial isolation, physiological experiments, and shotgun metagenomic sequencing, we identified both putative enhancers and mitigators of KB blooms. Metagenome-assembled genomes from the Roseobacter clade showed strong correlations with KB populations during HABs, akin to symbionts. A bacterial isolate from this group of metagenome-assembled genomes, Mameliella alba, alleviated vitamin limitations of KB by providing it with vitamins B₁, B₇ and B₁₂. Conversely, bacterial isolates belonging to Bacteroidetes and Gammaproteobacteria, Croceibacter atlanticus, and Pseudoalteromonas spongiae, respectively, exhibited strong algicidal properties against KB. We identified a serine protease homolog in P. spongiae that putatively drives the algicidal activity in this isolate. While the algicidal mechanism in C. atlanticus is unknown, we demonstrated the efficiency of C. atlanticus to mitigate KB growth in blooms from the GoM. Our results highlight the importance of specific bacteria in influencing the dynamics of HABs and suggest strategies for future HAB management.

Endophytes are microbes living within plant tissue, with some having the capacity to fix atmospheric nitrogen in both a free-living state and within their plant host. They are part of a diverse microbial community whose interactions sometimes result in a more productive symbiosis with the host plant. Here, we report the co-isolation of diazotrophic endophytes with synergistic partners sourced from two separate nutrient-limited sites. In the presence of these synergistic strains, the nitrogen-fixing activity of the diazotroph is amplified. One such partnership was co-isolated from extracts of plants from a nutrient-limited Hawaiian lava field and another from the roots of Populus trees on a nutrient-limited gravel bar in the Pacific Northwest. The synergistic strains were capable of increasing the nitrogenase activity of different diazotrophic species from other environments, perhaps indicating that these endophytic microbial interactions are common to environments where nutrients are particularly limited. Multiple overlapping mechanisms seem to be involved in this interaction. Though synergistic strains are likely capable of protecting nitrogenase from oxygen, another mechanism seems evident in both environments. The synergies do not depend exclusively on physical contact, indicating a secreted compound may be involved. This work offers insights into beneficial microbial interactions, providing potential avenues for optimizing inocula for use in agriculture.

Microbially induced carbonate precipitation (MICP) is a natural process with potential biotechnological applications to address both carbon sequestration and sustainable construction needs. However, our understanding of the microbial processes involved in MICP is limited to a few well-researched pathways such as ureolytic hydrolysis. To expand our knowledge of MICP, we conducted an omics-based study on sedimentary communities from travertine around the CO₂-driven Crystal Geyser near Green River, Utah. Using metagenomics and metaproteomics, we identified the community members and potential metabolic pathways involved in MICP. We found variations in microbial community composition between the two sites we sampled, but Rhodobacterales were consistently the most abundant order, including both chemoheterotrophs and anoxygenic phototrophs. We also identified several highly abundant genera of Cyanobacteriales. The dominance of these community members across both sites and the abundant presence of photosynthesis-related proteins suggest that photosynthesis could play a role in MICP at Crystal Geyser. We also found abundant bacterial proteins involved in phosphorous starvation response at both sites suggesting that P-limitation shapes both composition and function of the microbial community driving MICP.

Transport of organic matter (OM) occurs widely in the form of animal and plant detritus in global oceans, playing a crucial role in global carbon cycling. While wood- and whale-falls have been extensively studied, the in situ process of OM remineralization by microorganisms remains poorly understood particularly in pelagic regions on a global scale. Here, enrichment experiments with animal tissue or plant detritus were carried out in three deep seas for 4-12 months using the deep-sea in situ incubators. We then performed community composition analyses as well as metagenomic and metatranscriptomic analyses. The results revealed strikingly similar microbial assemblages responsible for decomposing animal and plant detritus. Genes encoding peptidases and glucoside hydrolases were highly abundant and actively transcribed in OM enrichments, which confirmed the roles of these enriched microbial assemblages in organic decomposition. Marinifilaceae, Desulfocapsaceae, Spirochaetaceae, and o-Peptostreptococcales were found to potentially contribute to nitrogen fixation. These core bacteria, acting as cosmopolitan anaerobes in decomposing fast-sinking particulate OM, may have been underestimated in terms of their role in deep-sea microbial-mediated biogeochemical cycles during conventional sampling and diversity survey.

The archaeal class Bathyarchaeia is widely and abundantly distributed in anoxic habitats. Metagenomic studies have suggested that they are mixotrophic, capable of CO₂ fixation and heterotrophic growth, and involved in acetogenesis and lignin degradation. We analyzed 35 Bathyarchaeia metagenome-assembled genomes (MAGs), including the first complete circularized MAG (cMAG) of the Bathy-6 subgroup, from the metagenomes of three full-scale pulp and paper mill anaerobic digesters and three laboratory methanogenic enrichment cultures maintained on pre-treated poplar. Thirty-three MAGs belong to the Bathy-6, lineage while two are from the Bathy-8 lineage. In our previous analysis of the microbial community in the pulp mill digesters, Bathyarchaeia were abundant and positively correlated to hydrogenotrophic and methylotrophic methanogenesis. Several factors likely contribute to the success of the Bathy-6 lineage compared to Bathy-8 in the reactors. The Bathy-6 genomes are larger than those of Bathy-8 and have more genes involved in lignocellulose degradation, including carbohydrate-active enzymes not present in the Bathy-8. Bathy-6 also shares the Bathyarchaeal O-demethylase system recently identified in Bathy-8. All the Bathy-6 MAGs had numerous membrane-associated pyrroloquinoline quinone-domain proteins that we suggest are involved in lignin modification or degradation, together with Radical-S-adenosylmethionine (SAM) and Rieske domain proteins, and AA2, AA3, and AA6-family oxidoreductases. We also identified a complete B12 synthesis pathway and a complete nitrogenase gene locus. Finally, comparative genomic analyses revealed that Bathyarchaeia genomes are dynamic and have interacted with other organisms in their environments through gene transfer to expand their gene repertoire.