Microplastics (MPs) are emerging contaminants, with wastewater treatment plants (WWTPs) as principal hotspots for their release into downstream systems, including constructed wetlands (CWs), a nature-based solution for water treatment. While non-buoyant MPs readily settle, buoyant MPs risk bypassing CWs and entering aquatic environments. Biofilm formation could influence MP transport by altering buoyancy, promoting sinking, and enhancing MP retention, yet its role in CWs remains unknown. This study, for the first time, quantifies the effects of MP polymer type, particle characteristics, exposure time, and seasonality on biofilm colonisation and its impact on terminal rising velocities of initially buoyant MPs in a UK-based CW receiving partially treated wastewater. Polypropylene (PP), expanded polystyrene (PS), and low-density polyethylene (LDPE) particles (3-5 mm) in spherical, beaded, and film shapes were incubated in situ over 12 months. Sampling followed two approaches: (1) a rolling bi-monthly schedule to capture seasonal variation, and (2) a long-term deployment with subsets retrieved every two months. Biofilm biomass was quantified by crystal violet staining, surface characteristics were captured by scanning electron microscopy (SEM), and terminal rising velocity experiments measured buoyancy changes. Biofilm growth showed strong seasonality, with peak biomass in late spring showing up to a 1972 % increase compared to winter. Despite widespread colonisation, changes in terminal rising velocity were minimal and largely non-significant (p < 0.05), indicating that biofilm formation alone is insufficient to retain initially buoyant MPs in CWs. These findings are crucial for deriving MP transport models and challenge assumptions that biofilm-induced density changes drive MP retention in CWs.

Brazilian soils have distinctive characteristics to European and North American soils which are typically used to investigate pesticide fate. This study aimed to compare soil-water partition coefficient (Kd), reversibility of adsorption and degradation half-life (DT50) of 5 pesticides covering a wide range of physico-chemical properties in contrasting Brazilian soils (Argissolo, Gleissolo, Latossolo and Neossolo) and a temperate (UK) alfisol soil, and to study their relationship with soil OM, clay and expandable clay content, CEC and pH. In addition, we used a novel laboratory test to evaluate sorption reversibility, the 3-Phase Assay (3PA). This stationary extraction system uses an organic solvent that sequesters compounds present in the soil aqueous solution and forces the diffusion of reversibly sorbed compounds. Kd (mL/g) values ranged from 4.85 to 79.26 for ametryn, 0.80 to 37.58 for clodinafop, 1.49 to 6.57 for fomesafen, 9.93 to 488.90 for thiabendazole and 2.52 to 6.77 for trifloxysulfuron. Desorption of the test compound (% applied radioactivity) ranged from 37.29 to 101.91 for ametryn, 32.05 to 100.67 for clodinafop, 34.73 to 73.33 for fomesafen and 4.15 to 66.87 for thiabendazole. Sorption reversibility was not assessed for trifloxysulfuron due to hydrolytic instability. DT50 (days) ranged from 29 to 90 for ametryn, 1 to 466 for clodinafop, 49 to 601 for fomesafen, 4 to 342 for thiabendazole and 22 to 38 for trifloxysulfuron. The data generated gives an overview of pesticide fate in Brazilian soils used for regulatory testing and is helpful for exposure risk assessment. The results showed that pesticide behaviour in Brazilian soils was not systematically different from those in European and North American soils. The 3PA was shown to be a reliable and simple method for assessing pesticide desorption in soil and could be adapted to assess pesticide bioavailability. The use of the 3PA allowed a more thorough explanation of the observed differences in degradation behaviour between the compounds.

Laboratory soil biodegradation studies required for approval of crop protection products (CPPs) are performed under continuous darkness, nullifying any potential contributions of algal and moss dominated biological soil crusts (BSC). There is growing evidence for metabolism of CPPs by phototrophic microorganisms under laboratory conditions, but limited data is available under field conditions. In this study we investigated the impact of the BSC on the fate of two 14C-fungicides under semi-field conditions using different light filters to alter formation of the BSC by exclusion/transmission of UV and photosynthetically active (PAR) wavelengths. Attenuation of PAR light significantly reduced formation of a BSC, which resulted in a significant slowing of the dissipation of benzovindiflupyr, which is known to be susceptible to phototrophic metabolism in aquatic systems, with 12-14 % more parent compound remaining at the end of the study when BSC development was impeded. For paclobutrazol, however, no significant difference in dissipation rate was observed. For both compounds there was significantly less non-extractable residue (NER) formation when BSC development was impeded (4-9 % reduction). Additionally, for both fungicides, the presence of a viable BSC resulted in 10-20 % more movement through the surface 5 mm of the soil, although this effect was limited to the period immediately after application and was likely due to increased porosity of the surface layer. This study confirms that the presence of phototrophs can significantly impact the environmental fate of CPPs on the surface of agricultural fields, either directly through metabolism or indirectly by altering the properties of the surface layer.

BackgroundArbuscular mycorrhizas (AM) are the most widespread terrestrial symbiosis and are both a key determinant of plant health and a major contributor to ecosystem processes through their role in biogeochemical cycling. Until recently, it was assumed that the fungi which form AM comprise the subphylum Glomeromycotina (G-AMF), and our understanding of the diversity and ecosystem roles of AM is based almost exclusively on this group. However recent evidence shows that fungi which form the distinctive 'fine root endophyte' (FRE) AM morphotype are members of the subphylum Mucoromycotina (M-AMF), so that AM symbioses are actually formed by two distinct groups of fungi.ResultsWe investigated the influence of nitrogen (N) addition and wheat variety on the assembly of AM communities under field conditions. Visual assessment of roots showed co-occurrence of G-AMF and M-AMF, providing an opportunity to compare the responses of these two groups. Existing 'AM' 18S rRNA primers which co-amplify G-AMF and M-AMF were modified to reduce bias against Mucoromycotina, and compared against a new 'FRE' primer set which selectively amplifies Mucoromycotina. Using the AM-primers, no significant effect of either N-addition or wheat variety on G-AMF or M-AMF diversity or community composition was detected. In contrast, using the FRE-primers, N-addition was shown to reduce M-AMF diversity and altered community composition. The ASV which responded to N-addition were closely related, demonstrating a clear phylogenetic signal which was identified only by the new FRE-primers. The most abundant Mucoromycotina sequences we detected belonged to the same Endogonales clades as dominant sequences associated with FRE morphology in Australia, indicating that closely related M-AMF may be globally distributed.ConclusionsThe results demonstrate the need to consider both G-AMF and M-AMF when investigating AM communities, and highlight the importance of primer choice when investigating AMF community dynamics.

An experimental study was conducted on how polymer density affects the transport and fate of microplastics in aquatic flows. For the first time, polypropylene (PP), polyethylene (PE), polymethyl methacrylate (PMMA), polyetheretherketone (PEEK), and polyvinyl chloride (PVC) were chemically stained and tested using solute transport techniques and velocities found among rivers in the natural environment (0.016 - 0.361 m/s). The movement of 3D-polymers with densities ranging from 0.9 - 1.4 g/cm(3) was quantified in a laboratory flume scaled to simulate open-channel flows in fluvial systems. Except for PP, in most conditions microplastics exhibited similar transport characteristics to solutes regardless of density and established solute transport models were successfully implemented to predict their transport and fate. Mass recoveries and ADE routing model demonstrated microplastic deposition and resuspension was associated with polymer density below critical velocity thresholds < 0.1 m/s. When density becomes the dominant force at these slower velocities, concentrations of denser than water microplastics will be momentarily or permanently deposited in channel beds and microplastics follow the classical Shields sediment transport methodology. This data is the first to provide microplastic suspension and deposition thresholds based on river velocity and polymer density, making a key contribution to research predicting microplastic fate and organismal exposure.

Heterotrophic bacteria and fungi are responsible for the decomposition of organic matter in soil. During this process, reactive oxygen species (ROS) can be produced directly and indirectly as extracellular byproducts of respiration. It is well known that nitrogen (N) cycle processes lead to the formation of volatile reactive nitrogen oxides (NOy), a group of climate-active gases that contribute to atmospheric chemistry and negatively impact human health. Primary microbial sources of NOy include ammonia-oxidising bacteria and denitrifying bacteria and fungi. Despite soil being a significant source of global NOy emissions, studies to date have primarily focused on N emissions from agricultural soils and there remains a large scope for investigating mechanisms of soil NOy production in a wider context in order to better constrain terrestrial and climate process models. Here, we propose a potential microbial mechanism involving ROS that could influence the production of NOy from soil. We utilised metagenomics and metatranscriptomics alongside continuous gas flux measurements and analysis of soil properties to evaluate the connection between microbial ROS and soil-sourced NOy. Our findings suggest that more NOy, particularly nitric oxide (NO), is produced in the presence of increased abundance of ammonia-oxidising and denitrifying taxa. NO can be lost to the environment or reacts with superoxide, an ROS produced via the enzymatic activity of soil organic matter (SOM) decomposers. This reaction produces peroxynitrite (ONOO-), which we have demonstrated to enhance nitrogen dioxide (NO2) emissions from soil. We have shown that the extent of NOy production through this pathway may be dependent on SOM composition, and the associated variability in carbon and nitrogen content. ### Competing Interest Statement The authors have declared no competing interest.

The genus Tetracladium has historically been regarded as an aquatic hyphomycete. However, sequencing of terrestrial ecosystems has shown that Tetracladium species might also be terrestrial soil and plant-inhabiting fungi. The diversity of Tetracladium species, their distribution across ecosystems, and the factors that shape community composition remain largely unknown. Using internal transcribed spacer (ITS) amplicon sequencing, we investigated the spatial distribution of Tetracladium in 970 soil samples representing the major ecosystems found across the British landscape. Species of the genus were found in 57% of the samples and across all vegetation types. The Tetracladium sequences we recovered included species common in aquatic ecosystems. However, we found five additional clades that clustered with environmental sequences previously found in terrestrial environments. The community composition of the Tetracladium OTUs was mainly related to vegetation type and soil pH. Strikingly, both taxon richness and overall abundance were highest in arable soils and showed positive relationships with soil pH. T. maxilliforme and a taxon of environmental sequences, Tetracladium group 1, was the biggest group, had the most relative abundance across ecosystems and was found in all vegetation types. Overall, this study provides insights into the community composition patterns of Tetracladium in terrestrial ecosystems and highlights the importance of vegetation characteristics in shaping Tetracladium communities.

The rhizosphere is a key interface between plants, microbes and the soil which influences plant health and nutrition and modulates terrestrial biogeochemical cycling. Recent research has shown that the rhizosphere environment is far more dynamic than previously recognised, with evidence emerging for diurnal rhythmicity in rhizosphere chemistry and microbial community composition. This rhythmicity is in part linked to the host plant's circadian rhythm, although some heterotrophic rhizosphere bacteria and fungi may also possess intrinsic rhythmicity. We review the evidence for diurnal rhythmicity in rhizosphere microbial communities and its link to the plant circadian clock. Factors which may drive microbial rhythmicity are discussed, including diurnal change in root exudate flux and composition, rhizosphere physico-chemical properties and plant immunity. Microbial processes which could contribute to community rhythmicity are considered, including self-sustained microbial rhythms, bacterial movement into and out of the rhizosphere, and microbe-microbe interactions. We also consider evidence that changes in microbial composition mediated by the plant circadian clock may affect microbial function and its significance for plant health and broader soil biogeochemical cycling processes. We identify key knowledge gaps and approaches which could help to resolve the spatial and temporal variation and functional significance of rhizosphere microbial rhythmicity. This includes unravelling the factors which determine the oscillation of microbial activity, growth and death, and cross-talk with the host over diurnal time frames. We conclude that diurnal rhythmicity is an inherent characteristic of the rhizosphere and that temporal factors should be considered and reported in rhizosphere studies.

Volatile reactive nitrogen oxides (NOy) are significant atmospheric pollutants, including NOx (nitric oxide [NO] + nitrogen dioxide [NO2]) and NOz (nitrous acid [HONO] + nitric acid [HNO3] + nitrogen trioxide [NO3] + ...). NOy species are products of nitrogen (N) cycle processes, particularly nitrification and denitrification. Biogenic sources, including soil, account for over 50% of natural NOy emissions to the atmosphere, yet emissions from soils are generally not included in atmospheric models as a result of a lack of mechanistic data. This work is a unique investigation of NOy fluxes on a landscape scale, taking a comprehensive set of land-use types, human influence, and seasonality into account to determine large-scale heterogeneity to provide a basis for future modeling and hypothesis generation. By coupling 16S rRNA amplicon sequencing and quantitative polymerase chain reaction, we have linked significant differences in functional potential and activity of nitrifying and denitrifying soil microbes to NOy emissions from soils. Further, we have identified soils subject to increased N deposition that are less microbially active despite increased available N, potentially as a result of poor soil health from anthropogenic pollution. Structural equation modeling suggests human influence on soils to be a more significant effector of soil NOy emissions than land-use type.

ABSTRACTSoil microbes play pivotal roles in global carbon cycling; however, the fundamental interactions between microbes and their infecting viruses remain unclear. This is exacerbated with soil depth, where the patterns of viral dispersal, ecology, and evolution are markedly underexplored. To investigate viral communities throughout the soil depth profile, we leveraged a publicly available metagenomic data set sampled from grassland soil in Northern California. In total, 10,196 non-redundant viral operational taxonomic units were recovered from soil between 20 cm and 115 cm below the surface. Viral prevalence was high throughout the soil depth profile, with viruses infecting dominant soil hosts, including Actinomycetia. Contrary to leading hypotheses, lysogeny did not dominate in the soil viral communities. Viral diversity was assessed at both the population level (i.e., macrodiversity) and strain level (i.e., microdiversity) to reveal diverse ecological and evolutionary patterns of virus-host interactions in surface and subsurface soils. Investigating viral microdiversity uncovered potential patterns of antagonistic co-evolution across both surface and subsurface soils. Furthermore, we have provided evidence for the potential of soil viruses to augment the remineralization of soil carbon. While we continue to yield a more comprehensive understanding of soil viral ecology, our work appeals to future researchers to further investigate subsurface viral communities.IMPORTANCESoil viruses can moderate the roles that their host microbes play in global carbon cycling. However, given that most studies investigate the surface layer (i.e., top 20 cm) of soil, the extent to which this occurs in subsurface soil (i.e., below 20 cm) is unknown. Here, we leveraged public sequencing data to investigate the interactions between viruses and their hosts at soil depth intervals, down to 115 cm. While most viruses were detected throughout the soil depth profile, their adaptation to host microbes varied. Nonetheless, we uncovered evidence for the potential of soil viruses to encourage their hosts to recycle plant-derived carbon in both surface and subsurface soils. This work reasons that our understanding of soil viral functions requires us to continue to dig deeper and compare viruses existing throughout soil ecosystems.

Physical interactions of microplastics within vegetation and turbulent flows of freshwater systems are poorly understood. An experimental study was conducted to investigate the underlying physical transport mechanisms of microplastics over submerged canopies across a range of flow conditions common in the natural environment. The effects of changing canopy heights were investigated by testing two model canopies of varying stem heights, simulating seasonal variation. This study determined and compared the mixing and dispersion processes for microplastics and solutes utilizing fluorometric tracing techniques. A hydrodynamic model was developed based on the advection‐dispersion equation for quantifying microplastic mixing in submerged canopies. Longitudinal dispersion coefficients for neutrally buoyant microplastics (polyethylene) and solutes were significantly correlated within submerged model vegetation irrespective of the complexity of the flow regime. Hydrodynamic and solute transport models were shown to be capable of robust predictions of mixing for neutrally buoyant microplastics in environmental flows over a canopy, facilitating a new approach to quantify microplastic transport and fate. We compare the mixing processes for microplastics and solutes then propose a hydrodynamic model for quantifying the mixing in submerged canopies.

The transport of microplastics within urban water systems remains poorly understood, with little prior research on their behaviour within manhole configurations. This study represents the first to measure and model the transport dynamics of microplastics within circular and square manholes under different hydraulic scenarios. The transport and fate of polyethylene (PE) was quantified and compared to solutes (Rhodamine WT dye) using energy losses, residence time distributions (RTDs), and mixing models within surcharging and overflowing manholes. The bulk mass of solute and PE concentrations followed similar flow paths across all conditions except for 17.3 ± 7.9 % of PE mass that was immobilized in a dead zone above the inlet pipe for manholes with a surcharge to pipe diameter ratio ≥2. Consequently, these microplastics only exit after a significant change in hydraulic regime occurs, causing microplastics to be at risk of being contaminated over a prolonged duration. No significant mixing differences for PE and solutes were found between manhole geometries. The deconvolution method outperformed the ADZ model with goodness of fit (Rt2) values of 0.99 (0.60) and 1.00 (0.89) for PE and solute mixing, respectively. This establishes the deconvolution method as the most accurate and appropriate model to accurately predict microplastic mixing in manholes and urban drainage systems.

Current literature suggests ecological niche differentiation between co-occurring Mucoromycotinian arbuscular mycorrhizal fungi (M-AMF) and Glomeromycotinian AMF (G-AMF), but experimental evidence is limited. We investigated the influence of soil age, water availability (wet and dry), and plant species (native Microlaena stipoides and exotic Trifolium subterraneum) on anatomical root colonisation and DNA profiles of M-AMF and G-AMF under glasshouse conditions. We grew seedlings of each species in soils collected from the four stages of a soil chronosequence, where pH decreases from the youngest to oldest stages, and phosphorus (P) is low in the youngest and oldest, but high in the intermediate stages. We scored the percentage of root length colonised and used DNA metabarcoding to profile fungal richness and community composition associated with treatment combinations. Soil age, water availability, and plant species were important influencers of root colonisation, although no M-AMF were visible following staining of M. stipoides roots. Soil age and host plant influenced fungal richness and community composition. However, response to soil age, potential host species, and water availability differed between M-AMF and G-AMF. Root colonisation of T. subterraneum by M-AMF and G-AMF was inversely correlated with soil P level. Community composition of M-AMF and G-AMF was structured by soil age and, to a lesser extent, plant species. Richness of M-AMF and G-AMF was negatively, and positively, correlated with available P, respectively. These findings are experimental evidence of ecological niche differentiation of M-AMF and G-AMF and invite further exploration into interactive effects of abiotic and biotic factors on their communities along successional trajectories.

Before agrochemicals can be registered and sold, the chemical industry is required to perform regulatory tests to assess their environmental persistence, using defined guidelines. Aquatic fate tests (e.g. OECD 308) lack environmental realism as they are conducted under dark conditions and in small-scale static systems, which can affect microbial diversity and functionality. In this study, water-sediment microflumes were used to investigate the impact of these deficiencies in environmental realism on the fate of the fungicide, isopyrazam. Although on a large-scale, these systems aimed to retain the key aspects of OECD 308 tests. Tests were carried out under both a non-UV light-dark cycle and continuous darkness and under both static and flowing water conditions, to investigate how light and water flow affect isopyrazam biodegradation pathways. In static systems, light treatment played a significant role, with faster dissipation in illuminated compared to dark microflumes (DT50s = 20.6 vs. 47.7 days). In flowing systems (DT50s = 16.8 and 15.3 days), light did not play a significant role in dissipation, which was comparable between the two light treatments, and faster than in dark static microflumes. Microbial phototroph biomass was significantly reduced by water flow in the illuminated systems, thereby reducing their contribution to dissipation. Comprehensive analysis of bacterial and eukaryotic community composition identified treatment specific changes following incubation, with light promoting relative abundance of Cyanobacteria and eukaryotic algae, and flow increasing relative abundance of fungi. We conclude that both water velocity and non-UV light increased isopyrazam dissipation, but the contribution of light depended on the flow conditions. These differences may have resulted from impacts on microbial communities and via mixing processes, particularly hyporheic exchange. Inclusion of both light and flow in studies could improve the extent they mimic natural environments and predict chemical environmental persistence, thus bridging the gap between laboratory and field studies.

Background Microbes are associated with nearly every plant surface and affect the host’s functioning. While many studies to date have investigated the composition of the plant microbiome and compared it with the plant’s metabolome (a measure of the host functioning), a holistic understanding which considers interactions both above and belowground is lacking. In this work, we hypothesise that soil is a reservoir for both root and leaf microbes, and that herbivory disrupts the regulation of the leaf microbial community. We further hypothesise that plant-associated microbial communities will correlate with the leaf metabolome, but the root microbiota will better correlate with the leaf metabolome than the leaf microbiota. To test these hypotheses, fungal and bacterial communities of herbivore-damaged and undamaged leaves, roots and the surrounding soil was characterised across 15 Plantago major populations sampled from across geographical and environmental gradients from Denmark. Microbial communities were then compared against untargeted and targeted (anti-herbivory/microbials) leaf metabolomic data. Results We found that the fungal communities showed significant inter-connectivity above and belowground, while bacteria were mainly specialised to each sample type. Herbivory had no effect on leaf microbiome or the metabolome. Fungal root endophytes (Glomeromycotina arbuscular mycorrhizal fungi) and leaf bacteria correlated most strongly with the overall leaf metabolome. However, there were specific pathogenic fungi found in the leaf associated with the abundance of specific metabolites. Conclusions In this study, demonstrate the complexity of plant metabolome-microbiome interactions, and the need to fully integrate the microbiome of different tissue types to fully understand plant’s secondary metabolome. The decreasing costs of -omics methods and high-performance computing costs has made it possible to perform multi-omic, multi-tissue approaches at large-scale, as in this work. It is therefore possible to identify novel plant-microbe interactions, even from plants sampled outside of controlled environments.

This article outlines the design and implementation of an internet-of-things (IoT) platform for the monitoring of soil carbon dioxide (CO2) concentrations. As atmospheric CO2 continues to rise, accurate accounting of major carbon sources, such as soil, is essential to inform land management and government policy. Thus, a batch of IoT-connected CO2 sensor probes were developed for soil measurement. These sensors were designed to capture spatial distribution of CO2 concentrations across a site and communicate to a central gateway using LoRa. CO2 concentration and other environmental parameters, including temperature, humidity and volatile organic compound concentration, were logged locally and communicated to the user through a mobile (GSM) connection to a hosted website. Following three field deployments in summer and autumn, we observed clear depth and diurnal variation of soil CO2 concentration within woodland systems. We determined that the unit had the capacity to log data continuously for a maximum of 14 days. These low-cost systems have great potential for better accounting of soil CO2 sources over temporal and spatial gradients and possibly flux estimations. Future testing will focus on divergent landscapes and soil conditions.

Fine root endophytes, recently reclassified as Mucoromycotinian arbuscular mycorrhizal fungi (M-AMF), are now recognized as functionally important as Glomeromycotinian AMF (G-AMF). However, little is known about the biogeography and ecology of M-AMF and G-AMF communities, particularly on a large scale, preventing a systematic assessment of ecosystem diversity and functioning. Here, we investigated the biogeographic assemblies and ecological diversity patterns of both G-AMF and M-AMF, using published 18S rDNA amplicon datasets and associated metadata from 575 soil samples in six ecosystems across China. Contrasting with G-AMF, putative M-AMF were rare in natural/semi-natural sites, where their communities were a subset of those in agricultural sites characterized by intensive disturbances, suggesting different ecological niches that they could occupy. Spatial and environmental factors (e.g., vegetation type) significantly influenced both fungal communities, with soil total‑nitrogen and mean-annual-precipitation being the strongest predictors for G-AMF and M-AMF richness, respectively. Both groups exhibited a strong spatial distance-decay relationship, shaped more by environmental filtering than spatial effects for M-AMF, and the opposite for G-AMF, presumably because stochasticity (e.g., drift) dominantly structured G-AMF communities; while the narrower niche breadth (at community-level) of M-AMF compared to G-AMF suggested its more susceptibility to environmental differences. Furthermore, co-occurrence network links between G-AMF and M-AMF were prevalent across ecosystems, and were predicted to play a key role in stabilizing overall communities harboring both fungi. Based on the macroecological spatial scale datasets, this study provides solid evidence that the two AMF groups have distinct ecological preferences at the continental scale in China, and also highlights the potential impacts of anthropogenic activities on distributions of AMF. These results advance our knowledge of the ecological differences between the two fungal groups in terrestrial ecosystems, suggesting the need for further field-based investigation that may lead to a more sophisticated understanding of ecosystem function and sustainable management.

AbstractBackgroundRecent studies demonstrated that microbiota inhabiting the plant rhizosphere exhibit diel changes in abundance. To investigate the impact of plant circadian rhythms on bacterial and fungal rhythms in the rhizosphere, we analysed temporal changes in fungal and bacterial communities in the rhizosphere ofArabidopsisplants overexpressing or lacking function of the circadian clock geneLATE ELONGATED HYPOCOTYL(LHY).ResultsUnder diel light–dark cycles, the knock-out mutantlhy-11and the gain-of-function mutantlhy-oxboth exhibited gene expression rhythms with altered timing and amplitude compared to wild-type plants. Distinct sets of bacteria and fungi were found to display rhythmic changes in abundance in the rhizosphere of both of these mutants, suggesting that abnormal patterns of rhythmicity in the plant host caused temporal reprogramming of the rhizosphere microbiome. This was associated with changes in microbial community structure, including changes in the abundance of fungal guilds known to impact on plant health. Under constant environmental conditions, microbial rhythmicity persisted in the rhizosphere of wild-type plants, indicating control by a circadian oscillator. In contrast, loss of rhythmicity inlhy-oxplants was associated with disrupted rhythms for the majority of rhizosphere microbiota.ConclusionsThese results show that aberrant function of the plant circadian clock is associated with altered rhythmicity of rhizosphere bacteria and fungi. In the long term, this leads to changes in composition of the rhizosphere microbiome, with potential consequences for plant health. Further research will be required to understand the functional implications of these changes and how they impact on plant health and productivity.

Background Soil microbes play pivotal roles in global carbon cycling, however the fundamental interactions between microbes and their infecting viruses remain unclear. This is exacerbated with soil depth, where the patterns of viral dispersal, ecology, and evolution are markedly underexplored. To investigate viral communities across soil depth, we leveraged a publicly available metagenomic data set sampled from grassland soil in northern California. Results 10,196 non-redundant vOTUs were recovered from soil sampled from 20 cm to 120 cm below the surface. Viral prevalence was high throughout the soil depth profile, with viruses infecting dominant soil phyla, including Actinomycetota . Contrary to leading hypotheses, lysogeny did not dominate in the soil viral communities. Viral diversity was investigated at both the population-level (i.e., macro diversity) and strain-level (i.e., micro diversity) to reveal diverse ecological and evolutionary patterns of virus-host interactions in surface and subsurface soil. Conclusions By investigating viral micro diversity in soil for the first time, we have uncovered patterns of antagonistic co-evolution across both surface and subsurface soils. Furthermore, we have provided evidence of soil viruses augmenting the remineralisation of soil carbon. While we continue to yield a more comprehensive understanding of soil viral ecology, our work appeals to future researchers to continue to investigate subsurface viral communities.