Soil Biology and Biochemistry

AbstractSoil microbes have sophisticated mechanisms to detect and respond to short pulses of C inputs, often involving changes in gene-expression. We studied gene transcription in a soil microbial community before, and 8, 24, and 48h after glucose addition (0.7 mg C g-1 dry soil) to understand how microbes react to periods of substrate excess and subsequent starvation. The relative transcript abundance of genes associated with energy metabolism and biosynthesis of amino acids, lipids, nucleotides, and cell wall components increased 8h after glucose addition. By 24 and 48h, the abundances of these transcripts reversed. Transcript abundance for genes associated with degradation of lipids, nucleotides, and (hetero)cyclic hydrocarbons decreased at 8h, but increased 24 and 48h after glucose addition. Simultaneously with a rise in transcripts for energy production and biosynthesis at 8h, transcription of regulatory genes for the exponential growth phase and ribosome assembly and maturation increased. In contrast, at 24 and 48h, transcript abundance for genes associated with ribosomal hibernation, sporulation, and regulation of the stationary phase increased, while transcripts for regulators for the exponential phase, and ribosome activation decreased. Based on changes in transcript abundance of phosphoenolpyruvate carboxylase and pyruvate carboxylase, it appeared that 8h after glucose addition glycolytic activity was high, however, gluconeogenesis returned at 24 and 48h. High levels of transcripts for nrtC-ntrB indicated N limitation 8 and 24h after glucose addition. Transcripts associated with Type VI Secretion Systems increased 24 and 48h after start of the experiment, suggesting a short lag between primary consumers and predatory bacteria. These results illustrate how metatranscriptome analysis can be used to study the ecophysiology of soil microbes providing details on the timing of exponential and stationary phase processes, coordination between anabolism and catabolism, and emerging nutrient limitations in natural soil communities. Research HighlightsO_LIWe studied gene transcription of a soil microbial community after glucose addition C_LIO_LITranscript abundances for biosynthesis and energy production initially increased, while those for degradation decreased C_LIO_LITranscripts of regulators and sporulation genes indicated start of stationary phase at 24h C_LIO_LINitrogen limitation induced transcription of nitrogen stress genes C_LI

Soil microbial growth and respiration play a critical role for soil organic carbon dynamics. Yet, we lack understanding of the main controls of soil microbial carbon metabolism at large scales. Here, we investigated whether and how the chemical composition of microbially available organic matter affects soil microbial carbon metabolism across soil systems. We linked soil microbial growth and respiration rates as well as carbon use efficiency (quantified with 18O stable isotope probing) to the chemical composition of extractable organic matter (characterized with reversed-phase liquid chromatography coupled to Fourier-transform ion cyclotron resonance mass spectrometry) along a geoclimatic gradient of 33 Chilean temperate grassland soils. We found that biomass-normalized rates of growth and respiration were primarily positively linked to aliphatics such as carbohydrate-, proteinaceous- and amino sugar-like compounds, and secondarily to unsaturated lignin-like compounds. Respiration was positively linked to compounds with carbon in a reduced oxidation state, suggesting carbon-conserving catabolism, while growth appeared unrelated to the oxidation state of carbon. This suggests that other mechanisms than mere energetic constraints control microbial growth rates in aerated soils. Our findings demonstrate that information on the chemical composition of bioavailable organic matter can provide insights into the processes that govern the fate of carbon across different ecosystems. Key pointsO_LIWe investigated if bulk soil microbial growth (18O stable isotope probing) and respiration is linked to the chemical composition of extractable organic matter (LC-FT-ICR MS) along a geoclimatic gradient of temperate grasslands. C_LIO_LIHigher rates of microbial carbon turnover were positively linked to aliphatic and unsaturated compounds. C_LIO_LISpecific (i.e., biomass-normalized) respiration was positively linked to compounds with carbon in a reduced oxidation state, suggesting carbon-conserving catabolism. C_LIO_LISpecific growth was unrelated to the oxidation states of substrate carbon, suggesting that soil microbial substrate use for anabolism may not be determined by direct energetic constraints. C_LI

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.

Soil organic matter (SOM) consists of diverse biochemical constituents, spanning a spectrum of chemical complexity, and the relative abundance of these substrates influences microbial metabolism and soil carbon persistence. However, mechanistic controls governing these processes and how they may be affected by environmental change remains incomplete. This study aims to assess (1) the molecular-level changes that occur across stages of root decomposition, from undecayed plant root litter to 1-year decomposed root litter, to mineral SOM and (2) how these changes are altered by anthropogenic nitrogen (N) deposition by using SOM biochemical and microbiome datasets and a long-term field experiment. N deposition did not significantly alter undecomposed or 1-year decomposed root litter, but did alter decomposing microbial communities and mineral SOM biochemical composition, specifically in lignin- and lipid-derived compounds. Taken together, this restructuring of microbial communities and alteration of SOM biochemistry likely contributed to the previously observed reduction in SOM decomposition.

High tunnels and open-field systems differ markedly in soil physicochemical properties, yet their effects on belowground microbiomes remain poorly understood. We characterized bacterial and fungal communities in paired high-tunnel and adjacent field soils from 100 small-scale vegetable farms across Minnesota, integrating amplicon sequencing of 16S rRNA and ITS2 regions with soil nutrient data, arbuscular mycorrhizal fungi (AMF) spore counts, and microbial co-occurrence networks. High-tunnel soils had higher pH, organic matter, and multiple macronutrients (notably P, K, and N forms) and lower bulk density than fields, reflecting intensive organic amendments and reduced leaching. Despite these differences, bacterial and fungal alpha diversity did not differ between environments, whereas beta diversity analyses revealed strong shifts in community composition. High tunnels were enriched in salt- and stress-tolerant bacterial phyla (Firmicutes, Deinococcota, Patescibacteria, Halanaerobiaeota, Halobacterota) and saprotrophic fungal groups (Mortierellomycota, Ascomycota, Basidiomycota, Mucoromycota), while several oligotrophic or symbiotic taxa, including Acidobacteriota and Glomeromycota, declined. Glomeromycota relative abundance was negatively correlated with high soil phosphorus, whereas AMF spore densities did not decline, suggesting suppression of active mycorrhizal symbioses rather than propagule loss under high-nutrient conditions. Co-occurrence network analyses showed that bacterial and fungal networks in high tunnels were less dense, more modular, and exhibited higher ratios of positive to negative associations than field networks, consistent with stress-induced shifts toward more facilitative interactions. Collectively, our results indicate that high-tunnel production homogenizes soil microbiomes and selects for stress- and high-nutrient-adapted taxa, with potential consequences for nutrient cycling, AMF function, and long-term agroecosystem outcomes.

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 microflora is fundamental to ecosystem functioning, yet their contribution in Miyawaki afforestation, a globally implemented ecological engineering approach, remains poorly characterized. In the present study, we examined the bacterial taxonomic diversity and their functional potential in a peri-urban Miyawaki mini-forest and compared with a nearby grassland the pre-existing ecosystem across dry and wet seasons. The Miyawaki plantation comprised of highly diverse native trees, sub-trees and shrubs spanning evergreen and deciduous varieties, potentiating nitrogen-fixation, diverse litter generation and rooting strategies resulting in pronounced functional heterogeneity. Notably relative to grassland, the Miyawaki forest was intensively managed and supplemented with organic amendments, and supportive irrigation, buffering the seasonal moisture stress. Using 16S rRNA amplicon sequencing of soil eDNA, we characterized seasonal variation in soil bacterial communities in both the systems. The observed soil bacterial community organization in forest as compared to the unmanaged grassland indicates combined influence of vegetation structure, dense canopy cover, continuous litter generation and root exudates. Microbial assemblages in the forest specialised in heterotrophic complex carbon degradation, biofilm formation, exopolysaccharide production and sporulation pathways which suggests adaptive abilities to anoxic microsites and other stressful conditions. In contrast, grassland soils harboured less diversified bacterial communities dominated by phototrophic and oxidative stress adaptation pathways consistent with sun lit, non-irrigated and moisture-variable conditions. Nonetheless, functional divergence in dry season reflects temporal reorganization of microbial communities marking a gradual trend towards soil ecosystem development. Together, these findings establish microbial baseline for Miyawaki forests revealing how tree-dense mini-forests restructure soil bacterial communities relative to grasslands highlighting the value of identifying soil microbial indicators for critically evaluating urban afforestation outcomes over extended time scales to inform sustainable design and policy. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=113 SRC="FIGDIR/small/704982v1_ufig1.gif" ALT="Figure 1"> View larger version (56K): org.highwire.dtl.DTLVardef@15e1bb1org.highwire.dtl.DTLVardef@16c1dddorg.highwire.dtl.DTLVardef@11c99ecorg.highwire.dtl.DTLVardef@bd6d1d_HPS_FORMAT_FIGEXP M_FIG C_FIG Schematic representation of the study depicting the Miyawaki forest and nearby grassland.

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.

AimsHarnessing rhizosphere processes offers a valuable opportunity to optimize nutrient use efficiency in agroecosystems. In nutrient-limited soils, plants discharge part of photosynthate surplus via root exudation, including carboxylates, which may enhance mineral dissolution and nutrient mobilization. We aimed to assess how plant responses to nutrient limitation translated into changes in exudate profiles, and how these exudates, in turn, drive bioweathering across soils of contrasting mineralogy and weathering degree. MethodsWe conducted a hydroponic experiment with Lupinus albus grown in a phosphorus (P) gradient over seven weeks. We measured plant biomass and root traits, performed a metabolomics analysis and quantified seven carboxylates in root exudates using gas chromatography-mass spectrometry. To assess bioweathering across contrasted soil domains, we conducted batch dissolution tests with exudates using three soil horizons--each with distinct physicochemical properties: enriched in organic matter, iron oxides, or primary silicates. ResultsAt the intermediate level of P supply, shoot biomass was comparable to that under high P, but plants produced more root biomass and a higher total carboxylate exudation rate. Despite low carboxylate concentrations (<100 ppb), exudates promoted the dissolution of Ca, Mg, Si, Fe, P and K in all soils. Yet, the degree of element released varied among soil types. ConclusionThese findings highlight the importance of root exudates in enhancing mineral dissolution, with effects dependent on soil physicochemical properties. The results suggest that managing agroecosystems under moderate nutrient limitation could be a sustainable strategy to increase root-to-shoot ratios, enhance bioweathering and nutrient release in rhizosphere.

Widely accepted climate predictions indicate that drylands will expand to cover more than half of the Earths terrestrial surface by the end of the 21st century. In these environments, harsh conditions including nutrient and water limitations restrict plant and animal life, thereby increasing the importance of soil microbial communities in nutrient cycling and ecosystem functioning. The Namib Desert is a distinctive dryland ecosystem characterised by a steep natural aridity gradient, transitioning from a coastal hyperarid zone influenced by frequent fog deposition to an inland arid region receiving seasonal rainfall. This study investigates the impact of water availability and moisture regime on microbial trace gas oxidation and community composition across this aridity gradient. Quantitative analyses revealed that total microbial abundance and activity indicators, including ATP concentrations and respiration rates, were significantly (p < 0.005) reduced in hyperarid soils compared to their arid counterparts. In contrast, hyperarid fog-dominated soils exhibited significantly (p < 0.0005) elevated rates of atmospheric hydrogen oxidation, even in the absence of water inputs. We propose that sustained high-affinity hydrogen oxidation, coupled with rapid microbial resuscitation following wetting events, supports shallow sub-surface microbial communities in the Namib Desert, particularly in the coastal hyperarid zone. Together, these findings challenge current understanding of the lower limits of microbial activity and reveal alternate metabolic pathways that enable microbial persistence in hyperarid hot desert soils. ImportanceDrylands are expanding globally, yet the mechanisms that allow microbial life to persist under extreme and sustained water limitation remain poorly understood. This study demonstrates that atmospheric trace gas oxidation, particularly high-affinity hydrogen oxidation, supports active and resilient microbial communities in hyperarid soils of the Namib Desert, even in the absence of liquid water inputs. By revealing how microbes may couple trace gas metabolism to energy and water generation, our findings provide new insight into the lower limits of microbial activity in dry hot desert soils and highlight the need to investigate how microbes persist and sustain soil ecosystem functioning.

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.

Fire can be a major pulse disturbance to soil microbial communities. Yet regular burning is a natural and essential process that maintains biodiversity and the unique attributes of rare and imperiled fire-dependent ecosystems. Most studies of fire effects on soil microbial communities typically focus on short-term (<1 year) responses following a single fire event. Here we examined the longer-term effects of repeated prescribed fire at the Albany Pine Bush--a fire-dependent, inland pitch pine (Pinus rigida) barren ecosystem of the northeastern US. We observed that this long-term fire management (i.e., a fire interval of approximately every 5 to 13 years over the past 30 years) has led to substantial depletion of soil nitrogen, specifically nitrate. However, we found no lasting shifts in the higher-level taxonomic composition of soil prokaryotic communities. Instead, metagenomic analysis revealed significant changes in the nitrogen-cycling functional potential, specifically, decreased dissimilatory nitrate reduction and denitrification potential in repeatedly burned soils. Collectively, these data suggest fire-induced geochemical changes persist under repeated burning, potentially driving substantial shifts in soil microbial functional diversity. Our study reveals strain-level changes that would be missed when examining only higher-level taxonomic patterns. Where fire is repeatedly applied, fire-induced shifts in soil microbial communities can persist well beyond a few weeks after burning--challenging prevailing views of short-lived belowground effects of prescribed burns.

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.

Climate change is increasing the frequency of wildfires in ecosystems that historically rarely burn, such as wet heaths and peatlands, thereby threatening carbon storage, biodiversity, and ecosystem functioning. We conducted a three-year, multi-level study to assess early post-fire recovery trajectories of soil physicochemical properties, vegetation, and soil microbial communities in a wet peatland-heathland mosaic affected by a flaming wildfire. Using a paired-plot design of burned and adjacent intact plots, we observed immediate spikes in bioavailable nitrogen (NH, NO-) and phosphorus (POlsen) and a reduction in soil moisture in burned plots, yet two years later these parameters had normalized, indicating rapid abiotic recovery. Vegetation was also strongly altered in the year of the fire, quantifiable by a distinct destruction of herb, moss, tree and litter cover. Although initial regrowth was dominated by a relatively fast resprouting of the graminoid Molinia caerulea, its absolute cover in burned plots never exceeded its cover in intact plots, suggesting this species did not expand post-fire. More typical peatland and wet heath species, including ericoid shrubs and Sphagnum mosses, recovered more gradually but largely returned to pre-fire levels within the timespan of our study, highlighting high vegetation resilience. Soil microbial communities showed contrasting responses. Prokaryotic communities shifted immediately after burning but largely recovered within one year. Fungal communities, however, exhibited stronger and more persistent changes and followed a distinct recovery trajectory shaped by succession of immediate and delayed fungal responders. Overall, pyrophilous and fire-tolerant fungi, such as Coniochaeta spp., increased, as did many presumably generalist or opportunistic saprotrophs. Litter and wood-associated saprotrophs as well as many mycorrhizal taxa, however, declined. Ongoing fungal shifts occurred even after soil chemistry and vegetation had largely returned to baseline, reflecting a temporary decoupling between above- and belowground communities that may have cascading effects on ecosystem functioning. In conclusion, our results reveal differential recovery trajectories across the soil-microbiome-vegetation interface and highlight that seemingly rapid abiotic and aboveground biotic recovery can mask prolonged microbial disruptions. We emphasize the importance of multi-level assessments for understanding ecosystem resilience. HighlightsO_LISoil physicochemistry, vegetation and prokaryotes recovered rapidly after a peatland wildfire C_LIO_LIFungal communities lagged behind and followed a slower recovery trajectory C_LIO_LIThe timing and duration of fungal responses to fire varied across taxa and included immediate or delayed as well as short-lived or persistent responders C_LIO_LIThere was a mismatch between vegetation and fungal recovery trajectories, evidenced by a transient post-disturbance decoupling between above- and belowground biotic communities C_LIO_LIPresumed aboveground recovery can mask prolonged belowground disruptions, with potential implications for decomposition, nutrient cycling, and plant-microbe interactions C_LI

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.

Photorhabdus bacteria are potent insect-killing microbes associated with entomopathogenic nematodes and offer opportunities for environmentally benign pest control. They can be applied as foliar sprays or soil drenches without their nematode vector, resulting in massive amounts of Photorhabdus cells and their (toxic) metabolites introduced into the soil. However, their effects beyond the target organisms are unknown. To fill this knowledge gap, we investigated the soil legacy effects of Photorhabdus cells and their metabolites on soil microbial communities, plant performance and resistance to herbivores. To this end, we first conditioned soils with i) mechanically killed (MK) or Photorhabdus-infected insect larvae, ii) aqueous extracts of MK or Photorhabdus-infected insect larvae, iii) cell-free Photorhabdus supernatants, iv) autoclaved soil complemented with live soil previously conditioned with MK or Photorhabdus-infected insect larvae. We then grew maize plants in these soils and measured plant biomass, profiled soil microbial communities and plant metabolites, and evaluated plant resistance against two pest insects Diabrotica balteata and Spodoptera frugiperda. We found that conditioned soils increased plant biomass by 10-26% relative to controls and significantly altered soil bacterial and nematode communities, and to a lesser extent, fungal communities. Re-inoculating conditioned soil microbiota into autoclaved soils recreated the plant growth-promoting effects, indicating microbial-mediated mechanisms. Additionally, plants grown in soils conditioned with Photorhabdus-infected insect cadavers were often more resistant to herbivorous insect attack, in a strain-specific manner. On average, D. balteata and S. frugiperda larvae gained 10-20% and 10-59% less weight, respectively, when fed on plants grown in conditioned soils than on plants grown in control soils. The plant metabolic profiles of plant leaves and roots also varied with resistance levels. We conclude that Photorhabdus metabolites modulate soil microbial communities towards a structure that enhances plant growth and triggers systemic responses against herbivores.

Microplastics are increasingly recognized as emerging contaminants in terrestrial ecosystems, yet their mechanistic impacts on soil multifunctionality remain poorly understood. Here, we evaluated the influence of two microplastic polymers, polyethylene terephthalate and polypropylene, on soil functioning by subjecting soils to a gradient of concentrations of these microplastics, and measuring six variables representing soil physical, chemical, and biological functions. A statistical framework combining multi-model inference with threshold detection and machine learning was implemented in this study to identify the main pathways of soil multifunctional change. Most significant responses followed nonlinear trends and threshold shifts, primarily in physical properties, indicating that microplastic stress first impacts soil structure before cascading to chemical and biological processes. We identified two system-level thresholds at 0.3% PP and 0.55% PET w/w; while random forest highlighted water-stable aggregates as the dominant predictor of overall soil multifunctionality. Our findings provide new quantitative evidence of complex soil multifunctionality responses to microplastic pollution. Most importantly, physical deterioration emerged as an early-warning signal of microplastic disturbance, thereby advancing our understanding of microplastic pollution on soil systems.

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