Applied Soil Ecology

Low-density polyethylene (LDPE) microplastics (MPs) are the most frequently sampled type of microplastic in agricultural soils, potentially threatening the soil environment. The majority of MPs that have been investigated are produced from standard polymer formulations, for which the nature of the added compounds is often unknown. Furthermore, standard ecotoxicity tests performed on model species are insufficient for assessing the ecological consequences of MPs contamination in soil. This study examined the responses of multiple keystone species to exposure to MPs in interaction with various additives. No significant effects on their growth were observed when organisms were exposed to MPs alone. However, significant reductions in growth occurred when organisms interacted within uncontaminated soil: the introduction of plants reduced potworm biomass by 49 {+/-} 4.1 % while the introduction of potworms reduced earthworm biomass by 41 {+/-} 5.2%. In MP-contaminated soil containing plants, the average individual biomass of potworm increased significantly from 1.16 {+/-} 0.09 mg in uncontaminated conditions to 2.01 {+/-} 0.27 mg. This suggests that MPs limited the negative effects of interactions. Similar patterns were observed for the potworm-earthworm interaction. MPs containing the highest concentrations of additives induced the strongest biological responses. Analysis of soil parameters revealed that these impacts are likely linked to the disruption of nitrogen cycling. Therefore, it is imperative to comprehensively address the interactions between soil organisms and the influence of additives on plastic ecotoxicity in order to better assess the ecological risk posed by MPs.

Biochar amendments are increasingly applied to improve soil physical functioning and support carbon dioxide removal, but their effects on intrinsic soil thermal properties remain poorly characterised. We conducted the first global systematic meta-analysis of 19 independent studies, 231 control-biochar comparisons, and 529 property-specific effect sizes to test how biochar changes soil heat transfer and storage. Biochar reduced thermal conductivity by 17.6% (95% CI, -22.7 to -12.2), thermal diffusivity by 11.0% (-14.5 to -7.3), and volumetric heat capacity by 8.3% (-12.3 to -4.1). Gravimetric heat capacity showed no significant overall response (+3.3%; -7.6 to 15.4) but was supported by fewer studies. Negative responses were directionally consistent for thermal conductivity, diffusivity, and volumetric heat capacity. Moderator analyses showed that responses were most consistently associated with post-application bulk density and changes in bulk density, while application rate modulated response magnitude and soil texture constrained context dependence. Co-variation among thermal conductivity, thermal diffusivity, and volumetric heat capacity matched expected physical dependencies, indicating coordinated structural reorganisation rather than independent shifts in isolated parameters. These estimates describe intrinsic conductive and storage properties; field-scale soil temperature responses may also be modified by albedo, evaporation, vegetation, and surface energy balance. Improved integration of soil thermal measurements with moisture dynamics, structural changes, and carbon cycling is essential to accurately represent biochar effects in soil and land-surface models.

Soil harbours a wide diversity of microbes responsible for essential functions, such as depolymerizing the C and N in organic matter through the production of exoenzymes. Some of these exoenzymes are universal, whereas others are specific to certain microbes. We hypothesized that higher microbial alpha diversity is associated with greater depolymerization capacity, specifically for protein and cellulose depolymerization, which will result in more N being mineralized. We therefore diluted two soil microbial communities, one from a forest soil and one from an agricultural soil, to create a diversity gradient. After nine weeks, we transferred these communities to a synthetic soil in which microbial necromass was the only nitrogen source. Before the transfer and two weeks after, we quantified protease, deaminase and {beta}-glucosidase potential activity, characterized the bacterial and fungal communities, and measured the quantity of nitrogen mineralized. The dilution had very little effect on the processes measured, with no clear trend. For identical alpha diversity values, some communities had high process rates, while other not. It appeared that these communities varied widely, a side effect of the dilution approach, and that this variation was significantly linked to process rates. This shows that community composition (beta diversity) is more strongly related to enzymatic potential and mineralization than species richness (alpha diversity) following necromass addition. In conclusion, the relationship between diversity and depolymerization of microbial necromass is not simply a matter of a linear decrease along with diversity but is rather linked to how reduced diversity results in more stochastic microbial communities. Highlights- Community composition (beta diversity) influence more microbial necromass depolymerization than species richness - Abundance of specific microbes explained ammonification and nitrification processes - Mineralization rates is different between crop and forest soil

Increasing global droughts exert large but poorly understood effects on the microbial communities and ecology of soil. Microbial communities generally show resilience and return to pre-drought conditions when short-term droughted soils are rewet; soils exposed to long-term drought, however, often show a lag upon rewetting, after which microbial communities may or may not return to their pre-stressed conditions. Though short-term droughts have been widely studied, long-term drought manipulation experiments remain rare, especially those that compare microbial response to short-term and long-term drought in tandem. We conducted a 1000-day drought simulation in controlled laboratory conditions with soil cores collected from a tidal freshwater ecosystem in Washington state, USA, and subsequently exposed them to rewetting for two weeks. We also included short-term (30-day and 90-day) drought and rewet treatments to directly compare microbial community and organic matter responses across drought durations. We found distinct microbial taxa belonging to Firmicutes and Actinobacteria enriched after the 1000-day drought, but not after the short-term droughts. While we hypothesized that the microbial community would recover from a short-term drought after rewetting to resemble pre-drought conditions, our results revealed community dissimilarities between rewet and pre-drought conditions across all drought durations. These findings suggest unique microbial life history strategies within certain microbial phyla that make them successful colonizers during an extended drought period, and the influence of environmental and physiological context on microbial responses to rewetting. ImportanceDroughts are increasing in frequency and intensity globally with severe implications for ecosystem services and soil functions. It is important to understand how long-term drought impacts soil microbial communities and organic matter chemistry to better predict future ecosystem responses to sustained moisture deficit conditions. We subjected soils to short-term (30 and 90 days) and long-term (1000 days) drought treatments and subsequently rewetted them to understand microbiome recovery to pre-drought conditions. Our results showed that prolonged drought drastically changes the microbial community and soil organic matter profile compared to short-term drought. While we expected the soil microbiome to recover upon rewetting after short-term drought, our results showed an altered microbiome composition, compared to pre-drought conditions, for both short-and long-term drought, suggesting microbial responses to soil rewetting was independent of drought duration imposed. These results provide important insights into soil biological and chemical functions that remain sensitive to change under fluctuating soil moisture conditions and future drought scenarios.

The reintroduction of extinct or endangered species to restore ecosystem function is an essential aspect of rewilding. The Wilder Blean Project at West Blean and Thornden Woods in Canterbury, UK, is committed to rewilding natural processes and enhancing biodiversity in one of England's oldest and largest areas of ancient woodland. The introduction of European bison (Bison bonasus) is an important part of the project. However, how the reintroduction of large herbivores influences local biodiversity and ecosystem functions during the early stages of rewilding remains poorly understood. Soil samples were collected from the same sampling sites before and two years after bison were reintroduced and profiled by metagenomic sequencing using Oxford Nanopore Technologies sequencing platforms. The results showed that the alpha diversity of soil organisms did not change significantly before and after the introduction of European bison, while beta diversity showed modest shifts in community composition. The relative abundance of some nitrogen-fixing and photosynthetic microbial genera showed declines in the 2024 Bison Area, while the mycorrhizal fungus genus Rhizophagus was significantly less abundant than in the 2024 Control Area. Despite relatively stable taxonomic diversity, functional composition differed significantly between the 2022 and 2024 Bison areas and among the 2024 rewilding treatments, revealing a decoupling between taxonomic diversity and functional composition. Amino acid synthesis pathways and carbon metabolism pathways were significantly enriched. These findings highlight the potential of long-read Oxford Nanopore metagenomics to reveal functional shifts that may not be apparent from taxonomic diversity alone. Although these early-stage responses cannot yet predict long-term rewilding trajectories, continued longitudinal monitoring integrating microbial, soil physicochemical, and ecosystem-level measurements will be essential to determine the persistence and ecological significance of these functional shifts.

Soil microbial communities have a variety of mechanisms to deal with emerging drought stress. One well-documented mechanism is increased microbial production of extracellular polymeric substances (EPS), which can potentially change the soil density and water holding capacity. Yet little is known about how microbial diversity influences the functional capacity of EPS formation and the resulting outcomes in water dynamics. To understand more about communal microbiome EPS production, we set up sterile mesocosms where we examined the effects of microbial diversity (high or low treatments) and nutrient input (supplement or deficient treatments) on these processes. To capture the microhydrology of the mesocosms, we measured water holding (WH), infiltration, evaporation, and soil properties we believe microbes are altering (EPS, soil aggregation). Our hypothesis stated that if diversity was artificially manipulated, then soil-water properties will be altered via production of EPS. We predicted that low diversity systems would have lower functional diversity, leading to less EPS production, moisture storage, and minimal changes from inert soil media. As predicted, we found that the high-diversity systems had a higher water retention and lower rates of water loss over time than low-diversity systems. This trend was magnified in the nutrient-supplemented treatment, suggesting that EPS production and subsequent water-holding traits are emergent features of the microbiome. Unexpectedly, we observed a correlation between the amount of water retained and the quantity of lipid EPS produced. This suggests that EPS composition, rather than quantity, is determinative of a biofilms function. In conclusion, it appears that microbial diversity influences soil properties that are important to moisture retention within these systems. To date, the role that microbes and their diversity play in soil hydrology has been severely understudied, so this work aims to build ecological understandings of these systems. These findings are valuable, for if we learn how microbes manipulate soil moisture, we can apply these functions to advance sustainable agricultural practices and enhance ecosystem resilience to water scarcity in arid regions. Open Research StatementUpon publication data, and code will be made available through Zenodo. Sequencing data will be uploaded to NCBI SRA.

Plants form holobionts by associating with diverse microbiota. Self-organization gives rise to emergent properties of the holobiont, such as increased resistance to pathogens. However, the local factors contributing to the self-organization are not well understood. We hypothesized that nematode communities and their associated microbiota govern the rhizobiome of the model plant holobiont tomato in terms of its suppression of root invasion by the parasite Meloidogyne hapla, and that the soil legacy influences the suppressive potential mediated by these biota. In pot experiments, a resistant tomato holobiont was favored by assembly in the presence of a nematode community conditioned by tomato plants, compared to oilseed rape or fallow soil. Nematode communities conditioned by tagetes could enhance resistance even better than tomato. Microbiota from crushed tomato-conditioned nematode communities increased resistance of the tomato holobiont, compared to microbiota of nematode communities conditioned by maize, or heat-inactivated microbiota. The 0.2 micrometre filtered microbiota from crushed nematodes had the same effect, suggesting a role of nematode-associated bacteriophages in holobiont assembly. The results indicate that soil nematodes and their associated microbiota play a role in the local organization and stabilization of plant holobionts. They can influence the resistance of plants that subsequently grow in the same soil. From an applied perspective, crop rotation schemes that alter nematode-microbiota communities could be harnessed to engineer crop holobionts.

Nutrient limitation is an important control on heterotrophic microbial activity that helps to stabilize the massive stocks of organic carbon held in ombrotrophic peatlands. Minerals contained in atmospheric dusts are critical nutrient sources for peatlands, yet the role of dust in supporting the below-ground microbial processes that underpin primary productivity is largely unknown. We investigated how mineral dust generated from mining waste rock (<20 {micro}m) influences element bioavailability and subsurface microbial functioning using flow-through soil mesocosms. The bioavailability of base cations (Ca, Mg, K), transition metals (Fe, Al, Ni, Cu), Al, and P was tracked over two months at three soil depths (0-6, 6-12, and 12-18 cm) using an extended sequential extraction method. We also analyzed microbial community composition (16S rRNA and ITS amplicon sequencing) and mineralization capacity (exoenzyme assays and carbon substrate incubations). After two months, the concentration of metals in the peat increased substantially after dust application, but the mobility and bioavailability varied by element. Responses of microbial communities to dust application were highly dependent on depth from the surface. Carbon substrate incubations revealed enhanced mineralization capabilities in soil from the surface zone (0-6 cm), but a relatively low stimulation of exoenzymes. Soil pH and phosphorus mobility were also impacted near the site of dust application, while acid phosphatase activity was lower throughout the column. In the middle zone (6-12 cm), the activities of {beta}-glucosidase, {beta}-xylosidase, and NAGase were higher with dust exposure. Measured microbial activity mostly remained unchanged in the lowest depth (12-18 cm). We observed increases in the relative abundances of putative saprotrophic fungi throughout the mesocosm profile. Results from this experiment show that the deposition and weathering of mineral dust can induce a complex set of changes to the capacity and nature of microbial carbon mineralization within a shallow layer of peat.

Background and aimsExcess nitrate (NO3-), from fertilizer overuse and intensive agriculture, can pollute water and contribute to greenhouse gas production (nitrous oxide - N2O). Plant metabolites from pastural herbs such as Plantago lanceolata (plantain) can inhibit microbial nitrification of ammonium to NO3- (biological nitrification inhibition - BNI) and change soil nitrogen cycle dynamics (lower potential nitrification rate - PNR). The main aim was to investigate differential plant metabolite expression associated with BNI and lowered PNR in different soil types. MethodsSix plantain cultivars were tested for BNI potential and screened for metabolites that correlated with inhibition of the ammonia oxidising bacterium (AOB) Nitrosospira multiformis. PNR and microbiome change was then investigated in four different New Zealand soils under the plantain cultivar Agritonic and ryegrass cultivar One50. ResultsPNR under plantain was 11 to 41% lower than fallow soil while PNR under ryegrass was 0 to 39% lower. In addition to verbascoside and aucubin, plantain metabolites associated with lower PNR included plantamajoside, riboflavin 3- and 5-sulfate, plantagoguanidinic acid. Chlorogenic acid was associated with lowered PNR under ryegrass. PNR reductions, microbiome structure and the ratio of ammonia oxidising archaea (AOA) relative to AOB was modulated by soil type. ConclusionPlantain and ryegrass lowered the PNR in four different soils and was correlated with metabolites beyond just aucubin and verbascoside. Based on candidate BNI-associated metabolites identified, it was hypothesised that lowered PNR is likely indirect through mechanisms such as chelation and appears to be dependent on both plant physiology and soil physicochemistry.

Soil microbes are considered central in litter decomposition and soil carbon formation. Microbial activity and abundance are controlled by microbivores, which are themselves preyed upon by top predators. However, the role of top predators in carbon cycling is rarely studied, especially in bacterivore-dominated food webs. Here we tested how trophic cascades, consisting of microbes, microbivores (bacterivore-dominated nematode communities) and top predators (nematode-feeding mites) impact carbon cycling and associated microbial pools and processes. We found that our model top predator decreased the abundance of fungivorous nematodes and had cascading effects on microbiome composition, notably increasing Gram-positive bacterial biomass, thus promoting the bacterial energy channel. These trophic cascades propagated to carbon cycling, decreasing heterotrophic respiration by 10 % while maintaining litter decomposition rates. Taken together, our results suggest that top predators increase carbon cycling efficiency and highlight the importance of complex trophic interactions, including trophic cascades, in determining soil carbon cycling.

The perennial grass Miscanthus x giganteus (miscanthus) offers a sustainable alternative to traditional biomass feedstocks while improving key soil health parameters, including aggregation. Aggregate stability results from dynamic soil-plant-microbe interactions, yet the relative importance of each factor remains an active research question. Building on previous observations that miscanthus alters soil structure to improve water-holding capacity and aggregate stability, we characterized the communities of soil bacteria and arbuscular mycorrhizal fungi (AMF) across three sites in Iowa, USA, comparing miscanthus to annual maize (Zea mays L.) and non-cropped perennial turfgrass (Poa spp.). We examined whether microbiomes co-varied with soil aggregation and, if so, whether plant cover identity or life history categorization better explained the observed patterns. Bacterial and AMF communities varied across sites and plant types, with signals that life history and plant cover identity both mattered. Aggregate stability aligned with a perennial-annual divergence in microbial beta diversity, while finer-scale differences in community composition and network structure were plant-specific. Soils under perennial plants were enriched in microbial groups positively correlated with aggregate stability; we identified 61 bacterial and 8 AMF "architect" taxa for future study. Within- and cross-kingdom co-occurrence network analysis revealed greater complexity under perennial plants: 1.9-fold more network links in miscanthus bacteria-bacteria networks than in maize, and 1.7-fold more in turfgrass AMF-AMF networks. Miscanthus fundamentally shapes microbial interactions, particularly among bacteria, relating to improved soil physical structure. Understanding these soil-plant-microbe feedbacks advances the development of biomass feedstocks with a portfolio of soil health benefits for next-generation biofuels and bioproducts. IMPORTANCEPerennial bioenergy crops can provide the raw material for biofuels and bioproducts while simultaneously improving soil health. Miscanthus x giganteus (miscanthus) efficiently stabilizes soil aggregates, potentially leading to higher water retention and erosion resistance. Understanding the microbial contributions to these outcomes is key to building resilient, sustainable bioenergy systems. This study highlights the connections between communities of soil microbes--bacteria and arbuscular mycorrhizal fungi--across three sites and three plant covers, including miscanthus, maize, and turfgrass. We identify a guild of potential "microbial architects" linked to soil aggregation and show more interconnected microbial networks under the perennial plant covers compared to annual maize. These insights shed light on the interactions between soil biological communities and soil physical and chemical properties. More broadly, the results may inform efforts to harness plant-associated microbiomes for sustainable biomass production.

Modern agriculture is characterized by substantial fertilizer nitrogen (N) losses from soils, resulting in low crop N-use efficiency. Biological nitrification inhibitors (BNIs) are studied as a strategy to improve N retention in soils by suppressing nitrification. However, the impacts of applying exogenous BNIs to crops with unknown intrinsic BNI capacity remain poorly understood. In this study, we evaluated the impacts of adding three BNIs (methyl 3-(4-hydroxyphenyl) acrylate [MHPA], 6-methoxy-2(3H)-benzoxazolone [MBOA], and limonene), their mixture, and the synthetic nitrification inhibitor 3,4-dimethylpyrazole phosphate (DMPP) on barley (Hordeum vulgare L.) growth, plant and soil N dynamics, and soil microbial communities. Using a rhizobox system with planted and bare-soil compartments, combined with 15N isotope tracing and molecular microbial community analyses, we assessed the spatio-temporal dynamics of N transformations, losses, plant N uptake, and microbial community responses in an alkaline agricultural soil. Independent of inhibitor application, the applied fertilizer N was lost primarily through NO- leaching (3-9% of the applied N). In contrast, N2O emissions represented only 0.001-0.028% of the applied N and varied with inhibitor type. MHPA increased dissolved inorganic N soil pools without affecting plant biomass or 15N uptake or strongly shifting microbial community composition. MBOA reduced NO3- concentrations in soil pore water without influencing plant growth or N uptake but shifted soil microbial community composition. In contrast, limonene reduced plant growth and 15N uptake and most significantly altered microbial community composition, without significantly changing N availability. Applying a BNI mixture, as well as limonene alone, was detrimental to plant growth and 15N uptake. DMPP showed only minor effects on N pools, plant growth, plant N uptake and microbial community composition. Overall, our results reveal both the potential and limitations of exogenous BNI application for improving N retention in crop systems.

The use of nitrogen (N) fertilizers to meet global food demands is expected to continue rising. However, up to 70% of N applied to agricultural soils is lost through microbially mediated processes such as nitrification. Inhibiting nitrification is thus a key strategy to reduce N losses and improve fertilizer N use efficiency. Various plant-derived compounds, termed biological nitrification inhibitors (BNIs), have been shown to reduce accumulation of nitrification products, intermediates, and byproducts (nitrite, nitrate, nitric and nitrous oxides). However, the mechanisms by which BNIs affect nitrifiers, along with their specificity and persistence in soil are not well understood. Here, we evaluated the effects of three BNIs: methyl 3-(4-hydroxyphenyl) acrylate (MHPA), 6-methoxy-2(3H)-benzoxazolone (MBOA), and limonene, on ammonia-oxidizing, total microbial, and fungal communities in two soils with contrasting pH. Their persistence in each soil was also evaluated. Although ammonia-oxidizing archaea initially dominated nitrifier communities in both soils, their bacterial counterparts significantly increased after mineral N addition but also were more sensitive to BNI application. Limonene and the synthetic inhibitor DMPP stimulated ammonium immobilization, as total soil mineral N was significantly reduced. Limonene and MHPA had the strongest off-target effects, increasing the relative abundance of hydrocarbon-degrading bacteria and potential fungal pathogens, respectively. In contrast, MBOA inhibited nitrification with minimal off-target effects. Among the tested BNIs, MBOA was also the most persistent in the high-pH, high-nitrification-rate soil. Our results show that MBOA is a promising biological inhibitor and highlight the importance of understanding BNIs ecological effects to develop targeted and sustainable N management strategies.

Nitrous oxide (N2O) is a potent greenhouse gas and is largely produced by incomplete denitrification. Although we know many of the microbial species that denitrify, we are still unable to reliably predict N2O production from soils. Recent work in microbial ecology has shown that when key microbes are considered as members of functional ensembles rather than isolated, the predictive power linking their activity to emergent properties increases dramatically. We hypothesized that the active microbial community during high N2O production would be taxonomically distinct from the inactive portion and increases in N2O production rates would correlate more strongly with increased abundance across multiple active taxa than with dominance by a single active species. We conducted a microcosm experiment where agricultural soil was incubated in anaerobic vials for up to 15 hours while tracking N2O production. Using bioorthogonal non-canonical amino acid tagging paired with fluorescence-activated cell sorting and 16S rRNA amplicon sequencing (BONCAT-FACS-Seq), we probed the active subset of the microbial community throughout the incubation period. Analysis of 16S rRNA gene amplicons revealed that the active and inactive fractions contained distinct taxa, and the taxonomic composition of the active fraction shifted over time. We found that less than 1% of the microbial community was responsible for N2O flux rates as high as 3.84 {micro}g N2O-N g dry soil-1 hr-1. The level of activity (median fluorescent intensity of active cells) correlated well with N2O production rates. The Ensemble Quotient Optimization for Microbiomes (mEQO) tool was used to identify an ensemble of eight organisms whose combined abundance best correlated with N2O fluxes. Overall, our results reveal that N2O fluxes are driven not by changes in a single taxon but by shifting ensembles of active microorganisms whose combined functional potential supports consistent emissions. This study applied a novel conceptual and methodological framework with a distinct focus on the active microbial community, rather than the entire community; if our observation that N2O flux rates are correlated with an ensemble of organisms is broadly confirmed, then framing denitrification as a community trait may increase predictability of this key process.

Native ash trees are destined for functional extinction in North America due to the spread of the non-native emerald ash borer. Yet, the consequences of ash loss for soil fungi are unclear. To address this, we employed a factorial study of forest soil fungi in two hydropedological soil types beneath four canopy tree species -- including white ash (Fraxinus americana). Sporocarp surveys and community DNA metabarcoding from soil samples revealed patterns in fungal communities related to canopy tree species but not soil type. Ash trees supported a particularly rich soil fungal community that was distinguishable from communities beneath beech, birch, and maple. We identified over 100 fungal taxa (OTUs) that are at risk of decline or loss from the studied forest, due to their association with ash. Our results indicate that canopy tree species influence soil fungi much more broadly than just the species with which they have mycorrhizal associations.

Smallholder farming systems in sub-Saharan Africa are constrained by declining soil fertility, erosion, and rising fertilizer costs, creating an urgent need for scalable inputs that sustain yields while maintaining soil health. While there is some evidence that microbial inoculants may offer a promising complement to conventional fertility management, field-scale evidence in tropical cereal and tuber systems remains limited. Here, we evaluated a multi-species inoculant composed of 20-22 Bacillus and Streptomyces species on potato and maize across four sites in Rwanda over two growing seasons (2025A and 2025B). Treatments included the inoculant applied at two rates (150 and 250 g ha-1), both alone and in combination with standard fertilization (inorganic fertilizer plus manure), alongside untreated and fertilized controls. Co-application of the inoculant with standard fertilization increased yield and plant biomass beyond fertilization alone, with gains of 6-51% for maize and 3-58% for potato. However, while the inoculant applied alone outperformed untreated controls, it generally did not match standard fertilization. Responses were strongest and most consistent for large-grade potato tubers, and application rate interacted with crop type, whereby the lower dose maximized marketable tuber yield, while maize showed a positive dose-response for grain and biomass. Yield increases were not accompanied by reductions in crop nutrient density, which was instead governed by site-level differences. Altogether, these results indicate that multi-species microbial inoculants are an effective complement to existing fertility practices that may offer, pending further research, a potential pathway to partial fertilizer replacement while sustaining productivity and nutritional quality in smallholder tropical agriculture.

Crop species may shape soil chemical properties and root-associated microbiota, but direct comparisons between contrasting crops remain limited. We compared soils and root-associated bacterial communities of Welsh onion (Allium fistulosum) and sweet potato (Ipomoea batatas) under the same field context. Sweet potato soil showed significantly lower electrical conductivity, inorganic nitrogen, and Mg saturation than control soil. Root-associated communities differed between crops, whereas alpha diversity did not. Proteobacteria-related taxa were more represented in Welsh onion roots, whereas Actinomycetia-related taxa were more represented in sweet potato roots, providing a basis for future studies on crop-specific soil microbial management.

Soil bacterial communities exhibit biogeographic patterns along environmental gradients, yet why some environmental factors contribute more strongly to community turnover than others remains poorly understood. Here, we tested whether this variation can be explained by the phylogenetic depth at which bacterial responses to each environmental factor are conserved. Across 40 forest sites in Japan spanning multiple soil and climatic gradients, environmental factors whose bacterial responses were conserved at deeper phylogenetic levels contributed more strongly to bacterial community turnover. We further asked whether phylogenetic clades that share similar environmental responses represent ecologically meaningful units for understanding bacterial community responses. Using soil pH as a focal test case, we found that response-defined clades improved prediction of taxon-level abundance shifts and community-level compositional shifts compared with models that treated taxa as independent units. Together, these findings show that the phylogenetic depth of bacterial environmental responses links trait conservation, community turnover and soil bacterial biogeographic patterns. Significance StatementSoil bacterial communities form biogeographic patterns along environmental gradients, but it remains unclear why some environmental factors drive stronger community turnover than others. This study shows that the strength of bacterial community turnover can be predicted from the phylogenetic depth at which bacterial responses to environmental factors are conserved. Across forest soils in Japan, deeply conserved bacterial responses were linked to stronger community turnover, and clades sharing conserved responses improved prediction of both taxon- and community-level shifts. These findings identify phylogenetically conserved response structure as an organizing principle for understanding and predicting soil bacterial biogeographic patterns.

Background and AimsAnthropogenic global change is altering the environmental stressors facing plants and their microbial symbionts. Changes in drought and temperature have received wide attention, but how other pervasive human impacts - land conversion for agricultural development and urbanization, and changes in nutrient conditions and pollutants - impact plant- microbe symbioses is relatively unknown. Here, we investigated how these anthropogenic global change drivers influence historic changes in the prevalence of widespread symbionts of grasses, Epichloe fungal endophytes. MethodsWe examined 8,739 seeds from 1,951 herbarium specimens collected between 1895 and 2019 for the presence of seed-transmitted Epichloe fungal endophytes in three grass host species (Agrostis hyemalis, Agrostis perennans, and Elymus virginicus). We hypothesized that the symbiosis provides fitness benefits under anthropogenic stresses (i.e. increased nitrogen deposition and land use change) that should translate to increased prevalence of the interaction among specimens exposed to those stresses. Key ResultsAnthropogenic stresses had contrasting effects on endophyte prevalence. Notably, among Agrostis host species, high nitrogen deposition was associated with high endophyte prevalence and with increasing trends in prevalence through time. We also found that highly urbanized landscapes were associated with reduced prevalence and negative temporal trends in endophyte prevalence across species. We also identified a weak positive relationship between agricultural land cover and average endophyte prevalence for Elymus virginicus, though temporal trends in prevalence did not differ between high and low levels of agricultural land cover. ConclusionsAnthropogenic stressors influenced endophyte prevalence in diverse ways. While we found increasing prevalence in the face of nitrogen deposition, a sign of the potential resilience of the symbiosis, urban land cover was associated with declining endophyte prevalence, a sign that anthropogenic activity may contribute to a breakdown of the symbiosis.

Soils are recognized as reservoirs of antibiotic resistance genes (ARGs) with the potential to transfer to clinical pathogens, creating antimicrobial resistance (AMR) that poses a threat to human health. While large-scale AMR surveys have profiled how diverse biomes shape soil resistomes, less is known about the influence of specific soil properties. Here, we combined metagenomics and 16S rRNA amplicon sequencing with isolate-based approaches to investigate drivers of soil AMR across a tropical watershed from beach to mountaintop in Waimea Valley, Oahu, Hawai{square}i. We leveraged functional- and taxonomic-classification of resistances to unravel how soil properties interact with bacterial taxa to structure resistomes. Metagenomic- and isolate-resistomes showed remarkable consistency, including a general gradient of increasing AMR from ridge to beach. Resistome functional composition was significantly correlated with total bacterial community structure. The relationship between resistances and soil properties was primarily dictated by taxonomic composition of each resistance. Rifampin- and Vancomycin-ARGs associated with Actinomycetes negatively correlated with soil physical properties, while resistant genes and isolates from Gammaproteobacteria positively correlated with enzymatic activity metrics. These findings indicate that soil properties structure the resistome indirectly through taxonomic filtering of microbial hosts and challenge the notion that AMR is decoupled from phylogenetic relatedness.