Soil Biology and Biochemistry

Decomposition of organic matter is a key process in soils contributing to carbon and nutrient cycling. To identify management strategies for agroecosystems that reduce nutrient losses while maximizing plant growth, it is important to understand which parameters determine decomposition rates. This study therefore investigated how the presence of winter wheat (Triticum aestivum var. Asory) affects decomposition in a controlled Ecotron setup with two soil types with varying organic matter content across three simulated climates (2013, 2068, 2085). Using the tea bag index, interstitial soil pore water analyses, microbial biomass quantification, bacterial and fungal gene abundance, and soil respiration measurements, we tested the hypotheses that plant exudates would enhance decomposition rate and microbial biomass, while plant nitrogen uptake would deplete soil nitrate, potentially mitigated by fertilization. Contrary to expectations, decomposition rates were lower in planted than in unplanted soils, suggesting resource competition between plants and microbes. No significant differences were observed in microbial biomass or respiration due to plant presence, and fertilization effects on nitrate or microbial mineralization were undetectable, likely due to rapid turnover of organic molecules including uptake by plants and microbes. Mechanistically, fungi and soil humidity were more important for decomposition than bacteria or temperature. The findings corroborate climate impacts on decomposition but also indicate microbial resilience and highlight the potential of management strategies like cover crops, adjusted planting dates and crop residual management which can contribute to healthy soils by sustaining carbon and nutrient cycling.

Arctic winters are long and cold and have traditionally been considered a period of limited biological activity. However, the seasonal dynamics of microbial community composition and functional potential during winter remain poorly understood. Here, we investigated taxonomic (bacteria, fungi, archaea) and functional (fungal guilds and nitrogen cycling genes) dynamics throughout a full year at two Arctic tundra heath sites with contrasting snow regimes. A steep drop in microbial abundances in early to mid-winter, likely linked to freeze-thaw events, coincided with shifts in soil pH and elevated community turnover. Saprotrophic and root-associated fungi were more abundant in the cold-season, while inorganic nitrogen cycling groups were more abundant in summer and declined toward winter despite high bacterial abundance. This indicates sustained organic matter cycling during the winter and expanded inorganic nitrogen cycling in the summer. Functional gene ratios further suggested a higher early-winter nitrogen loss potential via nitrous oxide and greater late-winter nitrogen retention. Site-specific differences in snow regime altered the timing and magnitude of these dynamics. Together, our results demonstrate that winter represents a critical and dynamic period for microbial community restructuring with important implications for nitrogen turnover in Arctic tundra soils.

The soil microbiome is key to plant health and nutrient acquisition, and viruses likely play important but largely unknown roles in these processes. To interrogate bulk and rhizosphere soil viral biogeography, we collected samples over a tomato growing season in California from an experiment testing arbuscular mycorrhizal fungi (AMF) treatment. We generated 78 viromes, 16S rRNA gene, and ITS1 amplicon datasets, and 33 rhizosphere metatranscriptomes. Of 67,038 DNA viral species genomes (vOTUs), 25% were previously identified, predominantely in agricultural systems, suggesting habitat filtering and greater viral homogeneity across agricultural compared to natural soils globally. Rhizospheres had significantly higher DNA viral richness than bulk soils, whereas no significant richness differences were observed for other biota. 60% of vOTUs were shared between compartments, compared to only 21-23% of bacterial and fungal taxa. Although bulk soil viral biogeography resembled that of prokaryotes, with significant structuring by moisture content, greater virome similarity between high-moisture bulk soils and rhizospheres suggests that conditions with high host activity selected for similar viral communities. In rhizospheres, while bacterial and fungal communities differed most over time, DNA and RNA viral communities differed most by sampling location, matching prokaryotic transcriptional patterns and further implicating host activity in viral biogeography. Similarly, AMF treatment induced changes in the prokaryotic transcriptome but, across biota, only significantly affected DNA viral communities. Overall, results indicate strong viral responses to spatiotemporally localized conditions, with viral biogeography reflecting both dispersal opportunities (high between neighboring bulk and rhizosphere soils, low across fields) and selection via local host activity.

Soil microbes support life on Earth by regulating the availability of nutrients in soils, yet we lack a fundamental, baseline knowledge of which fungi and bacteria are associated with specific soil nitrogen (N) cycling processes across ecosystems. We identified functional and taxonomic groups of fungi and bacteria that are associated with net ammonification and nitrification rates in soils from diverse ecosystems across the United States, including the environmental contexts where these relationships exist. To accomplish this, we co-analyzed soil, microbial, plant, and climatic data from 19 sites across the U.S. National Ecological Observatory Network (NEON). Distinct microbial groups were associated with net ammonification versus nitrification rates, highlighting the need to measure and model these two processes separately. The relative abundance of several microbial groups known for their N-decomposition abilities (i.e., Acidobacteriae, Bacteroidia, Saccharomycetes yeasts, ectomycorrhizal fungi) were positively associated with net ammonification rates across diverse environmental conditions. Meanwhile, pathogenic fungi, copiotrophic bacteria, and bacterial classes containing denitrifying bacteria were positively associated with net nitrification rates in many wet, hot, and high-N environments. These results deepen our understanding of soil microbiome ecology and represent a practical starting point to develop microbial-explicit biogeochemical cycling models at large spatial scales.

Plant species loss and nitrogen fertilization affect grassland biodiversity. However, their interactive effects on plant communities, soil properties, and the soil microbiome remain insufficiently understood. We analyzed how the removal of plant species, with and without urea addition, influenced plant diversity, soil properties, and soil bacterial communities in a Tibetan Plateau grassland. Continuous plant species removal and urea addition over seven years modified plant beta-diversity equally strong, while urea exerted a stronger negative effect on plant alpha-diversity. Both, plant species removal and urea addition caused soil acidification and an increase in NO2-/NO-, while dynamics in TOC, TON and TOC: TON were mainly driven by the growing season. Structural equation modeling identified soil acidification via urea addition as the most important indirect driver that negatively affected bacterial alpha-diversity and shifted bacterial beta-diversity. Urea addition also exerted direct negative effects on bacterial alpha- and beta-diversity, causing repression of oligotrophic (Acidobacteriota, Chloroflexota, Planctomycetota, Gemmatimonadota) and stimulation of copiotrophic (Bacillota, Bacteroidota, Pseudomonadota) bacterial taxa. Plant species removal caused slight increases in bacterial alpha-diversity, paralleled by less diverse but more even plant communities. We show that soil acidification by urea fertilization outweighs plant species loss in its negative effect on bacterial soil biodiversity in Tibetan grasslands.

Microbial communities can shift under drought in ways that enhance plant performance during drought ("microbe-mediated acclimation"). However, it is also possible for microbial communities to shift in ways that worsen the effects of drought ("mal-acclimation"). It is unclear how and where microbe-mediated acclimation vs. mal-acclimation occurs, or if there are types of soils or microbial communities that are more likely to harbor microbes that enhance plant acclimation and limit mal-acclimation. We tested for microbe-mediated plant acclimation/mal-acclimation to drought in soils from 21 maize farms in the midwestern United States, spanning a range of climate, soil types, and management practices. We first conditioned soil microbial communities to drought vs. well-watered conditions in a greenhouse and then tested for microbe-mediated acclimation by growing maize in soils inoculated with the conditioned microbial communities under drought and well-watered conditions. Drought-conditioned soils did not enhance plant performance under drought. In fact, one third of the farms exhibited mal-acclimation, especially under well-watered conditions where wet-conditioned soils reduced plant performance in well-watered contemporary conditions. Farm management practices, climate, soil texture, and microbial diversity generally did not predict when this microbe-mediated mal-acclimation occurred. Overall, these results suggest that in agricultural soils, microbes may frequently impede-rather than facilitate-plant acclimation to soil moisture levels. Open research statementThe plant and soil data used in this study are available via the Environmental Data Initiative repository at https://doi.org/10.6073/pasta/f4a0db3a076cf6d8cef908947f82736e. The bacterial and fungal amplicon sequence data are available via the European Nucleotide Archive under accessions PRJEB110071 and PRJEB109827, respectively.

Plant invasion can modify soil microbial communities and ecosystem processes through plant-soil feedbacks, yet it remains unclear whether these effects are expressed mainly through taxonomic turnover or through shifts in microbial function and interaction structure. We tested how soil legacy generated by the invasive Conyza bonariensis, the native Helminthotheca echioides, or unplanted control soil influenced short-term microbial responses to standardized amendments and plant-derived inputs. In Experiment 1, conditioned soils were amended with water, cellulose, or ammonium and analyzed for extracellular enzyme activity, qPCR-based gene abundance, bacterial community composition, and family-level co-occurrence networks. In Experiment 2, the same soil legacies were exposed to water, glucose, or sterile root exudates from native or invasive plants. Native- and invasive-conditioned soils differed significantly in composition, but they were not consistently distinguished by strong indicator taxa, indicating that legacy effects were expressed mainly through redistribution of shared taxa rather than community turnover. In contrast, functional responses were clearer: enzyme activity and nirS abundance showed strong soil-legacy dependence, and network analysis revealed that invasive-conditioned soil supported a denser, more positive, and more compact family-level association structure than native-conditioned soil. In Experiment 2, invasive root exudates produced stronger short-term functional-based differentiation among soil legacies than native exudates, especially for extracellular enzymes. Together, the two experiments show that plant invasion can leave a persistent belowground legacy that is expressed primarily through functional filtering and network rewiring of a broadly shared microbiome, rather than through major taxonomic turnover alone.

The contribution of soil chemistry to plant growth and resilience, including presence of phytohormones, is increasingly recognized. However, comprehensive characterization of soil phytohormones remains limited by chemical complexity of soil matrices, diversity and low- abundance of metabolites. To enable further discoveries we developed and validated performance of a liquid chromatography-mass spectrometry method with solid phase extraction, integrating targeted and untargeted hormonomic approaches for comprehensive soil phytohormone profiling. Method performance was evaluated for sixteen plant growth-regulating compounds and precursors, including abscisic acid, auxins, cytokinins, gibberellic acid, jasmonic acid, salicylic acid, karrikins, melatonin, serotonin, and tryptophan. The method demonstrated strong linearity (R{superscript 2} = 0.989-0.999), high sensitivity (limits of detection and quantification 0.1-50.2 and 1.4-167.3 pg on-column, respectively), and acceptable precision (1.3-9.6% intraday; 3.4-34.8% interday). Soil composition had a significant effect on recovery, with recovery being poor in some soils such as clay-rich soils; however, recovery for most phytohormones were within 20% of the matrix- adjusted spiked value. Validation results confirm that the method is suitable for use and was then used to quantify analytes in representative soil types. Integration of untargeted analysis expanded coverage to 250 additional putative phytohormones and hormone-related metabolites, revealing chemical signatures potentially associated with plant community composition. The method is robust across these soils spanning sandy, peat-rich, and clay-rich textures. This approach provides a versatile framework for investigating belowground phytohormone dynamics and their roles in plant physiology, resilience, and soil-plant feedbacks.

O_LIPhyllosphere microbiomes are subject to microbial import from various sources and undergo substantial changes during phenological changes of plants. However, these processes are still poorly understood for forest canopies. We propose that phenology-driven changes in host properties, and rainwater-mediated, within-canopy transport shape the phyllosphere microbiome in temperate forests. Leaves and throughfall samples were collected from oak, ash and linden trees at top, mid, and bottom canopy positions at the Leipzig canopy crane facility (Germany) at time points representing early, mid and late phenological stages. Bacterial community composition was assessed by 16S rRNA gene amplicon sequencing. C_LIO_LIPhenological stages explained 19% of phyllosphere bacterial community variation, followed by tree species identity (12%) and canopy position (2%). Later phenological stages exhibited more homogeneous and functionally redundant phyllosphere communities along with a strong decline of plant pathogens and increasing potential for microbially mediated biocontrol mechanisms. Throughfall transported up to 1011 bacterial cells per litre with maximum bacterial fluxes at the canopy top. C_LIO_LIOur findings demonstrate that in temperate forests, phenology-driven effects on the phyllosphere microbiome are far more important than tree species specific effects. Extent and selectivity of throughfall-mediated mobilization may play a crucial role for the spatial heterogeneity of microbial communities in tree crowns. C_LI

Invasive plants can reshape ecosystems by altering soil biogeochemistry and microbial functioning under global change. Competitive interactions between the invasive Conyza bonariensis and the native Helminthotheca echioides were evaluated under warming, nitrogen enrichment, and elevated CO2, together with rhizosphere microbial function in solitary versus competitive growth. Plants were grown alone or in interspecific competition under elevated temperature (27 vs 29 {degrees}C), ammonium-nitrate fertilization versus no fertilization, and ambient versus elevated CO2 (400 vs 720 ppm). Plant traits and relative growth rate (RGR) were measured alongside potential extracellular enzyme activities (EEA) of -D-glucosidase (C acquisition) and N-acetyl-{beta}-D-glucosaminidase (NAGase; N acquisition) and functional gene abundances (nirS and bacterial amoA). To relate enzyme signals to plant demand and microbial biomass, we calculated a growth-normalized rhizosphere investment metric (Specific Rhizosphere Index; SRI) and a biomass-normalized investment metric (Specific Enzyme Activity; SEA). Competition effects were summarized as {Delta}SRI and {Delta}Tax (change from alone to competition) to quantify how competition altered growth- and biomass-normalized investment. Plant responses were driver- and context-dependent. Elevated CO2 produced the largest changes in growth traits, especially for the invasive species. Warming effects were modest in solitary plants but became apparent under competition, where elevated temperature reduced competitive suppression via increased invasive leaf production and reduced constraints on native leaf expansion. Fertilization caused comparatively small shifts in plant endpoints. Microbial responses depended strongly on soil conditioning history. Potential EEA showed limited shifts with warming and fertilization, whereas elevated CO2 enhanced NAGase mainly in invasive-conditioned soils and increased nirS across soils. Despite overlap in ecoenzymatic stoichiometry, SRI and {Delta}Tax revealed treatment- and legacy-dependent patterns in how competition re-scaled microbial C and N acquisition relative to plant growth and microbial biomass. Together, these results indicate that global change can decouple plant growth from enzymatic investment and reconfigure invasive-native interactions through shifts in above-belowground coupling.

Water scarcity is an increasing constraint on agricultural productivity and demands scalable strategies that improve crop performance under reduced irrigation. As soil microorganisms regulate key processes at the soil-plant interface, microbial inoculants may help sustain plant growth and physiological function during water limitation. Here, we assembled five functionally diverse microbial consortia containing taxa selected to support rhizosphere colonization, soil structural stabilization, and fungal-mediated nutrient and water foraging. These consortia were evaluated in greenhouse trials with lettuce and spinach grown under full irrigation or a 30% deficit irrigation regime (70% of crop water requirement). Crop responses were assessed using yield, harvest delay, root length, wilting incidence, chlorophyll content, and Water Band Index (WBI). Across both crops, microbial consortium treatments improved performance under deficit irrigation relative to untreated water-stressed controls. In lettuce, yield increased by 3-9%, while in spinach yield increased by 4-13%, with several treatments restoring performance to levels not significantly different from the fully irrigated control. Microbial treatments also reduced harvest delay by an average of three to four days, improved root length, lowered wilting incidence, and reduced WBI, indicating reduced plant water stress. In several cases, these physiological responses approached those observed under full irrigation despite 30% lower water input. Higher application rates (500 vs 250 g h-1) generally produced stronger responses, although this trend was not always statistically significant. Together, these results show that complex microbial consortia can buffer the negative effects of deficit irrigation and improve crop performance in leafy greens. These findings support the development of microbial inoculants as biologically based tools to enhance agricultural resilience under increasing water scarcity. TeaserMicrobial soil inoculants help crops maintain yield and harvest synchrony under reduced irrigation.

Bacterial interactions-whether positive or negative - are crucial for the functioning of microbial communities. Though bacterial interactions are mainly expected to be negative, the sign and strength of interactions are predicted to be context dependent, with interactions typically being more positive in more stressful and nutrient-poor conditions. However, systematic studies investigating how the environment affects interactions between multiple taxa are lacking. Here, we determine if interactions between a panel of natural soil isolates change in response to the environment in which they are grown, with two different artificial media used (one simple and one complex) and a more ecologically relevant soil wash. To maximise natural variation in interactions, we collected multiple isolates from multiple sites: co-occurring (sympatric) isolates were predicted to show more negative interactions than allopatric isolates because of greater overlap in resource use. Pairwise interactions were in general negative, but more negative when grown in a complex lab-derived medium (Tryptic Soy Broth). Mutually beneficial interactions were most common in a simple resource medium (M9 minimal media) and exploitative interactions were most frequent in a soil broth. These patterns were independent of whether species originated from the same or a different site. The study supports the prediction that nutrient rich environments promote more negative interactions, and that measuring interactions of soil isolates in standard lab media is likely to misrepresent interactions occurring in natural environments.

Emergence of multi-drug resistant (MDR) pathogens is facilitated by the mobilization of resistance genes from bacteria in animal and environmental habitats, a process often mediated by plasmids. While fertilization of agricultural soils with manure is hypothesized to serve as a pathway for transferring antimicrobial resistance plasmids to soil and crop bacteria, evidence is limited. In this study, we aimed to determine whether MDR-plasmids from manure transfer in soil, leading to the formation of long-term agricultural resistance reservoirs. To this end, we introduced a known MDR plasmid to agricultural soil where barley was subsequently grown and monitored spread of the plasmid over the course of a growing season (up to 190 days). Our experimental design mimicked conventional agricultural practices at a microcosm scale. A digital droplet PCR approach indicated plasmid transfer in the rhizosphere, which was confirmed by a targeted Hi-C method (termed Hi-C+). This demonstrated transfer of the plasmid to soil bacteria 10 days after barley planting but was not observed afterwards. The new plasmid hosts could not be identified, as plasmid-associated host Hi-C reads were absent from existing databases. This implies these hosts were rare and likely unculturable members of the soil microbiome. Our findings demonstrate that plasmid transfer from manure to soil can occur under conditions reflecting those found in agricultural settings. Furthermore, rare and uncharacterized members of the soil microbiomes may participate in acquiring MDR plasmids from manure bacteria, raising important questions about their role in spreading resistance plasmids.

In tundra ecosystems, moss-associated microbes are a major source of new nitrogen, yet the relative contributions of environment, host identity, and microbiome composition to variation in nitrogen-fixation rates are difficult to disentangle. To test how environmental change alters moss microbiomes and nitrogen-fixation rates, we used a one-year reciprocal transplant experiment between two Alaskan tundra sites that differ by 5{degrees}C in mean annual temperature. Intact moss cores containing one of three moss species, Hylocomium splendens, Aulacomnium turgidum, and Pleurozium schreberi, were transplanted between sites or returned to their home site. After one year, we quantified nitrogen-fixation rates using 15N incubation and characterized bacterial communities using 16S rRNA gene amplicon sequencing. H. splendens showed consistently low nitrogen-fixation rates with little transplant response, whereas P. schreberi and A. turgidum home and transplant tundra cores generally exhibited higher rates at the cooler, more northern site regardless of origin. In contrast, bacterial community structure changed little following transplantation, with composition driven primarily by moss species. Only in cyanobacteria and some heterotrophic bacterial lineages did we find subtle ASV-level changes. The absence of an association between microbial composition and nitrogen fixation, together with the heterogeneity among moss species, suggests that over short timescales, host physiology and microenvironment play a larger role in the variation of nitrogen-fixation rates than community turnover. The fact that short-term shifts in moss-associated nitrogen-fixation rate are driven primarily by host species identity, rather than microbiome restructuring, has important implications for near-term predictions of nitrogen inputs under Arctic climate change.

Phytophthora species are globally significant soilborne oomycetes responsible for widespread ecosystem decline. Standard soil sampling protocols, originally developed for qualitative baiting assays, typically require collecting substantial soil volumes in order to capture viable propagules. While effective for culture-based detection, these protocols are labour-intensive and can damage the shallow root systems of sensitive host species such as New Zealand kauri (Agathis australis). Phytophthora agathidicida (PA), the pathogen associated with kauri dieback disease, is routinely surveyed using these methods. However, quantitative data describing the vertical distribution of PA in natural forest soils are lacking. Consequently, it remains unclear whether extensive depth sampling is necessary to ensure consistent molecular detection. In this study, we applied a quantitative oospore DNA (oDNA) qPCR assay to characterise the fine-scale vertical distribution of PA across four soil depth increments (0-5, 5-10, 10-15, 15-20 cm) from 12 kauri trees representing a range of disease stages. Results revealed distinct vertical stratification, with PA DNA concentrations peaking within the upper 0-10 cm of soil in non-symptomatic and possibly symptomatic trees. In symptomatic trees, the absolute peak occasionally reached 10-15 cm, while pathogen signals remained consistently detectable within the top 10 cm. Field validation from an additional eight trees confirmed that targeted 0-10 cm "shallow" sampling yielded higher PA concentrations than deeper sampling protocols. These findings provide a data-driven basis for refining soil sampling strategies, enabling more sensitive molecular detection while minimising disturbance and logistical effort in fragile ecosystems. IMPORTANCEPhytophthora species are among the most destructive soilborne pathogens globally, requiring robust diagnostic protocols for both agricultural and conservation settings. Traditional sampling frameworks were established to meet the biological requirements of baiting assays, which often necessitate collecting large soil volumes from broad depth profiles to ensure the capture of viable, infectious propagules. However, these extensive requirements are labour-intensive and can cause significant soil disturbance in sensitive forest ecosystems. Using P. agathidicida as a model, this study provides a high-resolution quantitative assessment of how pathogen DNA is distributed vertically across different disease stages. We demonstrate that while absolute peak abundance can shift within the 0-15 cm range as infection progresses, the pathogen signal remains consistently detectable within the top 10 cm. This evidence-based approach suggests that targeted, shallow sampling enhances sensitivity by reducing signal dilution, offering a lower-impact path for monitoring soilborne oomycetes worldwide.

The Imperial Palace in Tokyo serves as a significant reservoir of biodiversity within the urban landscape; however, its soil microbial communities remain uncharacterized despite decades of macro-biological surveys. This study presents the first dataset profiling the soil microbiome of the Imperial Palace Outer Gardens, utilizing both 16S rRNA amplicon and shotgun metagenomic sequencing to fill this knowledge gap. We collected bulk soil samples from four distinct sites, including pond sediments and soils beneath ginkgo and pine trees, to capture a range of environmental conditions within this conserved greenspace. Both 16S rRNA amplicon sequencing and shotgun metagenomic sequencing revealed that Pseudomonadota and Actinomycetota were the predominant phyla across all samples. Notably, sites with monoculture vegetation, such as those beneath pine trees, exhibited lower microbial diversity than other locations. Functional annotation identified core metabolic pathways and detected specific antimicrobial resistance and virulence factor genes in selected samples. These datasets provide a critical baseline for future research into urban ecosystem dynamics, soil health, and the intersection of environmental conservation and public health.

Modern agriculture faces the dual challenge of increasing food production while reducing reliance on synthetic inputs that degrade soil ecosystems and compromise long-term sustainability. Algal biomasses have emerged as promising biostimulants, yet their capacity to selectively modulate soil microbiomes and plant growth-promoting bacterial (PGPB) functions remains poorly understood. Here, we evaluated 17 phylogenetically and biochemically diverse macro- and microalgal extracts to determine their effects on soil microbial communities, bacterial functional traits, and tomato (Solanum lycopersicum) performance. Algal supplementation selectively restructured microbial communities without disrupting overall diversity, promoting taxa associated with plant-beneficial functions, including Bacillus, Pseudomonas, and Actinobacteria. In soil microcosms, specific treatments increased culturable bacterial abundance by up to [~]200-fold relative to the initial soil. Functional assays revealed strong extract- and strain-dependent responses. Siderophore production and ACC-associated activity were the most consistently stimulated traits, whereas auxin production, biofilm formation, and proline synthesis showed more variable or context-dependent responses. Notably, Ulva sp. (AP11.2) enhanced siderophore production across the majority of isolates, with over four-fold increases in individual strains, while Arthrospira-derived extracts (NG4.1, N14.1) consistently promoted bacterial growth across multiple taxa. In contrast, extracts such as Nannochloropsis sp. (NG6.1) and Tetraselmis sp. (NG5.1) induced more selective or inhibitory responses, highlighting extract-dependent functional trade-offs. Integration of biochemical and biological datasets identified fatty acid composition as a key axis associated with microbial functional responses, whereas volatile organic compound profiles showed weaker and less consistent associations. These microbiome and functional shifts translated into improved plant performance, with algal treatments increasing tomato growth and reducing mortality by approximately 20% under non-sterile soil conditions characterized by pathogen-associated pressure. Together, these findings demonstrate that algal extracts act as selective modulators of soil microbiomes, enhancing specific bacterial functions and improving plant performance in a context-dependent manner. This work provides a mechanistic framework for the development of targeted algal-based biostimulants aimed at reducing agrochemical inputs and advancing microbiome-informed agriculture.

BackgroundMost land plants depend on the ancestral arbuscular mycorrhizal (AM) symbiosis for phosphorus (P) acquisition. However, several plant lineages have independently lost this symbiosis, raising fundamental questions about how these non-mycorrhizal plants meet their nutritional requirements without this crucial partnership. ResultsComparative genomic analyses confirmed that Cyperaceae, Caryophyllaceae, and Brassicaceae lack genes essential for AM symbiosis, indicating that these lineages independently abandoned this association 90-122 million years ago. Field surveys of 42 wild populations across seven sites revealed that while non-mycorrhizal plants generally maintain shoot P levels comparable to those in AM neighbors, lower shoot P levels can be observed in low P soils. To identify fungal taxa potentially associated with P nutrition in non-mycorrhizal plants, we applied a machine-learning approach to predict plant P-accumulation from root microbiome composition. The model explained substantial variance in plant P-accumulation (57-69%), and identified 85 fungal taxa as key predictors of shoot P-accumulation, predominantly belonging to the Helotiales (28%) and Pleosporales (23%) orders. Experimental validation of two phylogenetically distant Helotiales lineages (Tetracladium maxilliforme OTU29 and Helotiales sp. OTU7), using isotopic tracing, demonstrated their capacity to enhance plant growth and transfer P (and N) to their native non-mycorrhizal hosts under P-limiting conditions. ConclusionsOur findings suggest that non-mycorrhizal plants engage in nutritional partnerships with diverse Helotiales lineages that could collectively contribute to their mineral nutrition. However, given the widespread distribution of these Helotiales fungi, including in roots of AM plants, they may play a broader role in plant nutrition, i.e. also in mycorrhizal hosts. This study provides proof of concept for a novel framework integrating machine-learning predictions with experimental validation to identify functionally important microbial partnerships in natural plant communities.

The contribution of tree foliage to atmospheric methane (CH4) and nitrous oxide (N2O) fluxes remains a major uncertainty in global GHG budgets. We made repeated in situ measurements of foliar CH4 and N2O fluxes across 25 temperate tree species interplanted at a forest restoration site using high-resolution laser spectroscopy. Tree foliage was consistently a net CH4 sink and a net N2O source in all species. Foliar CH4 oxidation increased by [~]33% in fall relative to spring and was [~]3-fold higher in shade-tolerant than shade-intolerant angiosperm species. Species differences accounted for most of the variability in fluxes, while correlations with soil emissions were comparatively weak. Microbial DNA sequencing revealed that the highest CH4-oxidizing angiosperm species (Tilia americana) harbored abundant Type I methanotrophs, whereas the lowest-oxidizing species (Prunus virginiana) had nearly 100-fold lower methanotroph abundance, with a foliar microbial community dominated by facultative methylotrophs. Global warming potential (GWP) scaling indicates that foliar CH4 uptake overwhelmingly dominates the net climate forcing effect. Our results suggest that the large and predictable differences in foliar CH4 uptake among tree species and associated differences in foliar microbial communities are of importance in understanding and potentially enhancing the global terrestrial CH4 sink.

Succession is an ecosystem building process in which a habitat and its community interact predictably by increasing diversity, habitat engineering, and ultimately reaching a climax community, where other ecological processes influence its dynamic. Key to succession is the establishment of primary producing habitat forming species, which drives niche differentiation leading to increasing diversity. Here, we use the primary colonizing and habitat forming seagrass, Halophila ovalis, to demonstrate that it drives bacterial succession in a meadow ecosystem, and its microbiome, both rhizoplane and phylloplane, are under host selection. Many of the characteristics attributed to plants for habitat modification are microbial processes such as nitrogen fixation and sulfide detoxification and succession is often extrapolated to such processes. To determine if succession (increasing diversity) or selection (reducing diversity) drives changes in diversity (16S rRNA gene) or habitat modifying processes (nifH, soxB, aprA, dsrA), molecular analysis was performed along chronosequences (as a proxy for succession) of seagrass patches. Bacterial communities were sampled within the meadow ecosystem and the microbiomes of H. ovalis (both rhizoplane and phylloplane). Genes involved in biogeochemical cycling are differentially impacted within the microbiome and meadow sediments, with only nifH under succession. All genes from all niches sampled for community analysis are under directional community trajectories, despite being subjected to distinct ecological processes, signifying that many ecological processes, including succession and host association, drive community assemblage.