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Multi‐omics of the Soil Microbiome

TheArctic is characterized by vast amounts of carbon stored in permafrost and isan important focal point for the study of climate change as increasing temperature may accelerate microbially‐mediated release of carbon stored inpermafrost into the atmosphere as CO 2 and CH 4 . Yet surprisingly, very little is known about the vulnerability of permafrost andresponse of microorganisms in the permafrost to their changing environment. To address these knowledge gaps we used omics technologiesto explore permafrost microbial communities, to reveal hitherto unknown soilmicrobial populations and to link them to specific soil biogeochemical processes. For these projects we employed an omics pipeline that includes metagenomics, metatranscriptomics and metaproteomics to determine the phylogenetic and functional gene compositions and their expression in complex, non‐sterile soil samples collected from the field at several locations in Alaska. These studies have presented several challenges due to the unprecedented large size of the datasets and the insufficiency of available bioinformatic tools & databases to analyze, correlate and integrate thedata. To address these challenges we developed a novel functional gene databaseand used different assembly and analysis algorithms to determine key featuresin the datasets. The data reveal the microbial diversity, composition andfunctional potential in permafrost and how the microbial communities respond toclimate change impacts. Support or Funding Information Microbiomes in Transition, LDRD, Pacific Northwest National Laboratories, Richland, Washington

Jansson, Janet K.↗

Soil Water Percolation Chemistry, April 2017 to March 2019, BR-Ma2, Manaus

Soil water percolation chemistry including major cations, anions and isotope geochemistry. Samples were collected from six passive wick flux meters across three topographic positions (valley, slope and plateau). Data has been processed to compute the monthly mean for each topographic position where two flux meters were installed at each topographic position. All samples were processed at the Geology, Geochemistry and Geomaterials Research Laboratory (GGRL) in Los Alamos, NM USA. The metadata tab included in the .csv has additional information on locations where the sensors were installed, and other installation/maintenance details. Contact ksolander@lanl.gov if you need to use this dataset for additional information.

54 ENVIRONMENTAL SCIENCES↗

The microbiome structure of decomposing plant leaves in soil depends on plant species, soil pore sizes, and soil moisture content

Microbial communities are known as the primary decomposers of all the carbon accumulated in the soil. However, how important soil structure and its conventional or organic management, moisture content, and how different plant species impact this process are less understood. To answer these questions, we generated a soil microcosm with decomposing corn and soy leaves, as well as soil adjacent to the leaves, and compared it to control samples. We then used high-throughput amplicon sequencing of the ITS and 16S rDNA regions to characterize these microbiomes. Leaf microbiomes were the least diverse and the most even in terms of OTU richness and abundance compared to near soil and far soil, especially in their bacterial component. Microbial composition was significantly and primarily affected by niche (leaves vs. soil) but also by soil management type and plant species in the fungal microbiome, while moisture content and pore sizes were more important drivers for the bacterial communities. The pore size effect was significantly dependent on moisture content, but only in the organic management type. Overall, our results refine our understanding of the decomposition of carbon residues in the soil and the factors that influence it, which are key for environmental sustainability and for evaluating changes in ecosystem functions.

16S rDNA↗

Microbial regulation of the soil carbon cycle: evidence from gene–enzyme relationships

A deficit of empirical evidence for the microbial regulation of ecosystem processes, including carbon (C) degradation, hinders our ability to develop a framework to directly incorporate the genetic composition of microbial communities in the enzyme-driven Earth system models. Herein we evaluated the linkage between microbial functional genes and extracellular enzyme activity in soil samples collected across three geographical regions of Australia. We found a strong relationship between different functional genes and their corresponding enzyme activities. This relationship was maintained after considering microbial community structure, total C and soil pH using structural equation modelling. Findings showed that the variations in the activity of enzymes involved in C degradation were predicted by the functional gene abundance of the soil microbial community (R 2 >0.90 in all cases). Our results provide a strong framework for improved predictions on soil C dynamics that could be achieved by adopting a gene-centric approach incorporating the abundance of functional genes into process models.

59 BASIC BIOLOGICAL SCIENCES↗

Distribution and migration mechanism of fluoride in groundwater in the Manas River Basin, Northwest China

Elevated fluoride (F) concentration in groundwater is posing a public health risk in the Manas River Basin (MRB), Northwest China. Based on the characterization of regional groundwater flow, 90 groundwater samples from aquifers were analyzed, along with top-soil leachate and pore-water samples from aquitards. Stable oxygen (δ{sup 18}O) and hydrogen isotopes, radiocarbon and hydrochemical analyses of the groundwater and pore-water samples were conducted to trace groundwater hydrological and hydrochemical processes and thereby understand the distribution and migration mechanism of F. The groundwater is recharged by meteoric precipitation through vapor condensation processes in the Tianshan Mountains. The F concentration in groundwater samples from this basin ranged from 0.11 to 48.15 mg/L (mean 2.56 mg/L). In 37 of the 90 groundwater samples, the F concentrations were above the safe level for drinking water. The F concentrations progressively increased with the residence time and well depths in the northwest of the alluvial-fluvial plain, where groundwater is overexploited for agricultural and domestic use. Positive correlations between F and sodium (Na)/calcium (Ca) indicate that the enrichment and migration of F are influenced by cation exchange processes under high-Na and alkaline pH conditions. The relationships between δ{sup 18}O and F and chloride (Cl) concentrations were nonlinear due to leaching and mixing processes. This shows that vertical leaching by irrigation return flow and mixing with pore water are the dominant processes driving the migration of F in the groundwater flow system of MRB, in addition to geochemical processes.

54 ENVIRONMENTAL SCIENCES↗

Transformative Impacts of Laser-Induced Breakdown Spectroscopy on Environmental and Biological Research at Oak Ridge National Laboratory

This manuscript will present an advancement of transformative research that has been conducted at Oak Ridge National Laboratory (ORNL) over a 25-year period (2000–2025) on a variety of environmental and biological matrices. These investigations derived a fundamental understanding of how elemental detection and analysis of these matrices led to the knowledge and discovery of natural processes in plants and the environment. Each project led to the initiation of a new research area which unearthed awesome and novel breakthroughs. Highlights are listed below: 1. The preliminary research at ORNL centered on the detection of aerosols utilizing Laser-induced Breakdown Spectroscopy (LIBS) technology. The Clean Air Act Amendment (CAAA) of 1990 highlighted the importance of identifying hazardous air pollutants (HAPs) due to their impact on environmental and human health, thereby underscoring the need to detect various toxic elements. Research in aerosol chemistry aimed to identify these harmful elements released by factories during periods of increased emissions in their manufacturing processes. LIBS emerged as the most effective method for real-time, in situ measurements of metal species in both gaseous and aerosol phases. 2. An understanding of the presence of total carbon in soils gives perspective on how to develop carbon sequestration strategies. The recognition that carbon sinks can evolve back to carbon sources to emit back to the atmosphere was an important consideration. Also, the concentration of carbon in soil indicates the health of land areas for growing crops successfully. 3. The direct detection of most of the elements in a wood sample in a single emission spectrum, without sample preparation, encouraged the research to use the LIBS technique for preservative treated wood coupled with use of multivariate statistical methodology. Additionally, it encouraged the researchers to try to differentiate natural woods from different parts of the country, and it was successfully demonstrated that LIBS coupled with MVA analysis could differentiate wood of different species from each other and of similar species grown in different environments based on their elemental spectra. This was a breakthrough since it revealed a systematic approach to connect elemental scarcity and abundance to either drought or typical rainfall conditions for the hardwood trees grown in specific areas. 4. Furthermore, the research progressed to reveal physiological and developmental processes contributing to biomass production such that the variation in leaf elemental composition increases our understanding of terrestrial nutrient cycles, as well as tracking the transfer of toxic elements from soils to living organisms. 5. Recently another breakthrough viz., ionomics initiated the correlation of elements to specific genes, uncovering the function that the element performed in the plant. More recently, this has been extended from plants to fungi as well as fungi growing in symbiotic relations with plants.

09 BIOMASS FUELS↗

Soil Environmental Conditions in the East River Watershed, CO

This dataset contains depth-resolved soil environmental variables collected between June 2016 and October 2018 from the Bradley Meadow site in the East River watershed. Datalogger output from 7 Campbell CS655 12 cm Soil Moisture and Temperature Sensors including depth, temperature, volumetric water content, and electrical conductivity are included in ‘BM_Merged_T_VWC_0616_10_18.csv’. Sensors were installed in June 2016. Data have been processed to merge raw sensor data and to remove additional variables. No additional QA/QC was performed. Measured pCO2 and depth are contained in ‘BM_EGM_WELL_CO2.csv’. These data represent gas well samples measured via injection using a PP Systems EGM-5 infra-red gas analyzer as described in the methods section. Measured CO2 surface flux values are contained in 'BM_EGM_Surface_Flux.csv'. These were measured using the PP Systems EGM-5 with the Soil Respiration Chamber attachment. Multiple reps are reported for each measurement including CO2 flux rate calculations using built-in linear and quadratic function methods, and average and standard deviations are taken across all reps and calculation methods.

54 ENVIRONMENTAL SCIENCES↗

Biodegradation in Soil of PLA/PBAT Blends Compatibilized with Chain Extender

This paper presents a study of biodegradation, in soil, of samples of poly(butylene adipate-co-terephthalate)(PBAT), poly(lactic acid) (PLA) and blends of these materials prepared in torque rheometer with the addition of a chain extender. Film samples of these materials were buried in soil under controlled laboratory conditions. The degraded samples were regularly taken from soil and analyzed by visual inspection, size exclusion chromatography, differential scanning calorimetry and infrared spectroscopy. Respirometry biodegradation tests were conducted to assess samples mineralization degree. Blends showed higher degree of crystallinity compared to pure polymers. Crystallinity degree enhanced during the biodegradation process in all samples, being able to causing the samples to degrade slowly. The study showed the great complexity of the biodegradation process of PLA and PBAT blends when compatibilized with a chain extender. The biodegradation rate showed different results depending on the characteristic applied to evaluate it: visual, molecular weight or mineralization. The chain extender had strong influence in PBAT and blends degradation, slowing the process as observed by the variation of molecular weight and carbonyl index. Blends showed an intermediate behavior compared to the original polymers.

37 INORGANIC, ORGANIC, PHYSICAL, AND ANALYTICAL CH↗

Data for Genetic Variation in Zea mays Influences Microbial Nitrification and DeNitrification in Conventional Agroecosystems

Nitrogenous fertilizers provide a short-lived benefit to crops in agroecosystems, but stimulate nitrification and denitrification, processes that result in nitrate pollution, N2O production, and reduced soil fertility. Recent advances in plant microbiome science suggest that genetic variation in plants can modulate the composition and activity of rhizosphere N-cycling microorganisms. Here we attempted to determine whether genetic variation exists in Zea mays for the ability to influence the rhizosphere nitrifier and denitrifier microbiome under “real-world” conventional agricultural conditions. To capture an extensive amount of genetic diversity within maize we grew and sampled the rhizosphere microbiome of a diversity panel of germplasm that included ex-PVP inbreds ( Z. mays ssp. mays ), ex-PVP hybrids ( Z. mays ssp. may s), and teosinte ( Z. mays ssp. mexicana and Z. mays ssp. parviglumis ). From these samples, we characterized the microbiome, a suite of microbial genes involved in nitrification and denitrification and carried out N-cycling potential assays. Here we are showing that populations/genotypes of a single species can vary in their ecological interaction with denitrifers and nitrifers. Some hybrid and teosinte genotypes supported microbial communities with lower potential nitrification and potential denitrification activity in the rhizosphere, while inbred genotypes stimulated/did not inhibit these N-cycling activities. These potential differences translated to functional differences in N2O fluxes, with teosinte plots producing less GHG than maize plots. Taken together, these results suggest that Zea genetic variation can lead to changes in N-cycling processes that result in N leaching and N2O production, and thereby are selectable targets for crop improvement. Understanding the underlying genetic variation contributing to belowground microbiome N-cycling into our conventional agricultural system could be useful for sustainability.

Nitrogen↗

16S and ITS Amplicon Sequencing Fastq files and metadata from PARCHED Panama Tropical Forest soils, 2019-2020,

Model projections predict tropical forests will experience longer periods of drought and more intense precipitation cycles under a changing climate. Such transitions have implications for structure-function relationships within microbial communities. We examine how chronic drying might reshape prokaryotic and fungal communities across four lowland forests in Panama with a wide variation in mean annual precipitation and soil fertility. Four sites were established across a 1000 mm span in mean annual precipitation (2335 to 3300 mm). We expected microbial communities at sites with lower MAP to be less sensitive to chronic drying than sites with higher MAP; while fungal communities to be more resistant to disturbance than prokaryotes. At each location, partial throughfall exclusion structures were established over 10 x 10 m plots to reduce direct precipitation input. Raw demultiplexed sequences (bacteria, archaea, fungal) from soil samples taken from PARCHED Panama Tropical Forest throughfall exclusion experiments. Files that contain 16S are sequences from prokaryotes, ITS indicates sequences from fungi. Compressed fastq files are contained in Field_PARCHED_ITS_fastq_2020.zip, Field_PARCHED_ITS_fastq_2019.zip, Field_PARCHED_16S_fastq_2020.zip, Field_PARCHED_16S_fastq_2019.zip.There are four sites across the isthmus of Panama, each with 4 control and 4 exclusion plots. Throughfall exclusion shelters were built to intercept 50% of throughfall precipitation that hits the soil. Samples were taken on May 2019 and Jan 2020 from 0-10 cm and 10-20 cm depth approximately 9 and 18 months after shelter installation. Metadata and sample IDs for fastq files for 2019 sampling are within Field_PARCHED_Metadata_2019_ITS.csv and Field_PARCHED_Metadata_2019_16S.csv. The metadata for both 16S and ITS fastq files for January 2020 is included in Field_PARCHED_Metadata_2020.csv. No data processing or QA/QC was done on the raw data. Data processing example provided in R notebook file.

54 ENVIRONMENTAL SCIENCES↗

An initial exploration of Bayesian model calibration for estimating the composition of rocks and soils on Mars

The Mars Curiosity rover carries an instrument, ChemCam, designed to measure the composition of surface rocks and soil using laser-induced breakdown spectroscopy (LIBS). The measured spectra from this instrument must be analyzed to identify the component elements in the target sample, as well as their relative proportions. This process, which we call disaggregation, is complicated by so-called matrix effects, which describe nonlinear changes in the relative heights of emission lines as an unknown function of composition due to atomic interactions within the LIBS plasma. In this work, we explore the use of the plasma physics code ATOMIC, developed at Los Alamos National Laboratory, for the disaggregation task. ATOMIC has recently been used to model LIBS spectra and can robustly reproduce matrix effects from first principles. The ability of ATOMIC to predict LIBS spectra presents an exciting opportunity to perform disaggregation in a manner not yet tried in the LIBS community, namely via Bayesian model calibration. However, using it directly to solve our inverse problem is computationally intractable due to the large parameter space and the computation time required to produce a single output. Therefore, we also explore the use of emulators as a fast solution for this analysis. We discuss a proof of concept Gaussian process emulator for disaggregating two-element compounds of sodium and copper. The training and test datasets were simulated with ATOMIC using a Latin hypercube design. After testing the performance of the emulator, we successfully recover the composition of 25 test spectra with Bayesian model calibration.

97 MATHEMATICS AND COMPUTING↗

Interpreting mixing relationships in energetic melts to estimate vapor contribution and composition

Heterogeneous, glassy debris can be produced by energetic events such as near surface nuclear explosions and asteroid impacts. In these events, the resultant debris comprises a rapidly-cooled mixture of the projectile or bomb material with proximate materials such as soil and anthropogenic structures. Glasses formed in these events preserve inter and/or intra sample compositional variation reflecting complex mixtures of the precursor materials or their chemically fractionated products. Understanding formation mechanisms and chemical processes relating to energetic glass formation based on study of the end products requires an objective method for calculating the precursor components that mixed together to produce the final product. Here, we compare a set of multivariate linear-least squares approaches to address the unmixing problem of heterogeneous glasses including classical least squares (CLS), principal component analysis (PCA) and two multivariate curve resolution – alternating least squares (MCR-ALS) approaches. We generate a synthetic data set representing spatially resolved, compositional data in a set of heterogeneously mixed glasses to understand model sensitivity to precursor abundance and composition. We explore the impact of incomplete knowledge of the system prior to the formation of the melts on the resulting confidence and accuracy of the solution (i.e., the number, composition, and abundances of the precursor materials). We conclude that a closure and equality constrained MCR-ALS approach succeeds in identifying unrecognized precursor components and is a suitable alternative to CLS approaches when the investigator has incomplete knowledge of the starting precursor chemical compositions. Finally, we demonstrate application of this MCR-ALS approach to measurements of Trinitite.

38 RADIATION CHEMISTRY, RADIOCHEMISTRY, AND NUCLEA↗

Method Validation Summary for L16.2 AD-ISO-0015 for ISO-17025 / NFAC Applications

Methods for the analysis of semi-volatile organic compounds are employed at Savannah River National Laboratory (SRNL) for routine and non-routine samples from tank waste, process control, waste acceptance, and a wide array of other process and research samples. A method has been developed by SRNL, based on existing methods for semi-volatile analysis, for the analysis of nitroaromatic high explosives by gas chromatography / mass spectrometry in soils and sediments. Relative to past work performed at SRNL on nitroaromatic explosives, this developed, optimized, and validated method can achieve lower limits of detection and quantitation, greater precision, lower bias, higher linearity, and accuracy across a greater linear range.

12 MANAGEMENT OF RADIOACTIVE AND NON-RADIOACTIVE W↗

Geochemistry and metagenomics analyses of bacterial community structure in selected waste dumpsites in Lagos Metropolis, Nigeria

Dumpsites are reservoirs of persistent organic pollutants (POPs) and heavy metals (HMs), constituting environmental hazards to humanity. Autochthonous microorganisms in dumpsites exhibit various degrees of responses to contaminants. Unfortunately, there is a dearth of information on the types and concentration of pollutants and the array of microorganisms in these dumpsites which may play important roles in the metabolism of such pollutants or other community processes. Therefore, determining the microbial community structure in such contaminated sites across a municipality is essential for profiling the taxa that would serve as consensus degraders of the pollutants. In this study, soil samples from three dumpsites (Cele, CS; Solous, SS; and Computer Village, CVS) were characterized for geochemical properties using GC-MS, MP-AES, and other analytical protocols, while the dynamics of bacterial communities were evaluated based on their 16S rRNA gene barcodes. A significant difference in the bacterial communities was observed among the dumpsites in relation to the extent of pollution caused by POPs and HMs. CVS, with the highest HM contamination, was rich in Actinobacteria (41.7%) and Acidobacteria (10.2%), in contrast to CS and SS. Proteobacteria (34.1%) and Firmicutes (20%) were the dominant phyla in CS (highest POP contamination), while Bacteroidetes (45.5%) and Proteobacteria (39.9%) were dominant in SS soil.Bacilluswas the dominant genus in the most polluted dumpsite. Canonical correspondence analysis revealed that polycyclic aromatic hydrocarbons (PAHs) and HMs shaped the structure of the bacterial operational taxonomic units (OTUs) in the most polluted dumpsite. Out of a total of 706 OTUs, 628 OTUs exhibited a significant correlation (>50%) with benzo(b)fluoranthene, azobenzene, dibenzofurans, pyrene, dibenzo(a,l)pyrene, Cu, and Zn. In particular, Proteobacteria (Achromobactersp. andSerratiasp.), Bacteroidetes (Zhouiasp.), and Firmicutes (Bacillussp.) were suggested to be pivotal to the ecophysiology of dumpsite soils contaminated with POPs and HMs. The results generally underscored the importance of metagenomic and physicochemical analyses of polluted systems in enabling correlations for useful prediction of drivers of such ecosystems. This will further improve our understanding of the metabolic potential and adaptation of organisms in such systems.

Environmental Sciences & Ecology↗

Novel soil bacteria possess diverse genes for secondary metabolite biosynthesis

In soil ecosystems, microorganisms produce diverse secondary metabolites such as antibiotics, antifungals and siderophores that mediate communication, competition and interactions with other organisms and the environment. Most known antibiotics are derived from a few culturable microbial taxa3, and the biosynthetic potential of the vast majority of bacteria in soil has rarely been investigated4. Here we reconstruct hundreds of near-complete genomes from grassland soil metagenomes and identify microorganisms from previously understudied phyla that encode diverse polyketide and nonribosomal peptide biosynthetic gene clusters that are divergent from well-studied clusters. These biosynthetic loci are encoded by newly identified members of the Acidobacteria, Verrucomicobia and Gemmatimonadetes, and the candidate phylum Rokubacteria. Bacteria from these groups are highly abundant in soils, but have not previously been genomically linked to secondary metabolite production with confidence. In particular, large numbers of biosynthetic genes were characterized in newly identified members of the Acidobacteria, which is the most abundant bacterial phylum across soil biomes5. We identify two acidobacterial genomes from divergent lineages, each of which encodes an unusually large repertoire of biosynthetic genes with up to fifteen large polyketide and nonribosomal peptide biosynthetic loci per genome. To track gene expression of genes encoding polyketide synthases and nonribosomal peptide synthetases in the soil ecosystem that we studied, we sampled 120 time points in a microcosm manipulation experiment and, using metatranscriptomics, found that gene clusters were differentially co-expressed in response to environmental perturbations. Transcriptional co-expression networks for specific organisms associated biosynthetic genes with two-component systems, transcriptional activation, putative antimicrobial resistance and iron regulation, linking metabolite biosynthesis to processes of environmental sensing and ecological competition. We conclude that the biosynthetic potential of abundant and phylogenetically diverse soil microorganisms has previously been underestimated. These organisms may represent a source of natural products that can address needs for new antibiotics and other pharmaceutical compounds.

59 BASIC BIOLOGICAL SCIENCES↗

BONCAT-FACS-Seq reveals the active fraction of a biocrust community undergoing a wet-up event

Determining which microorganisms are active within soil communities remains a major technical endeavor in microbial ecology research. One promising method to accomplish this is coupling bioorthogonal non-canonical amino acid tagging (BONCAT) with fluorescence activated cell sorting (FACS) which sorts cells based on whether or not they are producing new proteins. Combined with shotgun metagenomic sequencing (Seq), we apply this method to profile the diversity and potential functional capabilities of both active and inactive microorganisms in a biocrust community after being resuscitated by a simulated rain event. We find that BONCAT-FACS-Seq is capable of discerning the pools of active and inactive microorganisms, especially within hours of applying the BONCAT probe. The active and inactive components of the biocrust community differed in species richness and composition at both 4 and 21 h after the wetting event. The active fraction of the biocrust community is marked by taxa commonly observed in other biocrust communities, many of which play important roles in species interactions and nutrient transformations. Among these, 11 families within the Firmicutes are enriched in the active fraction, supporting previous reports indicating that the Firmicutes are key early responders to biocrust wetting. We highlight the apparent inactivity of many Actinobacteria and Proteobacteria through 21 h after wetting, and note that members of the Chitinophagaceae, enriched in the active fraction, may play important ecological roles following wetting. Based on the enrichment of COGs in the active fraction, predation by phage and other bacterial members, as well as scavenging and recycling of labile nutrients, appear to be important ecological processes soon after wetting. To our knowledge, this is the first time BONCAT-FACS-Seq has been applied to biocrust samples, and therefore we discuss the potential advantages and shortcomings of coupling metagenomics to BONCAT to intact soil communities such as biocrust. In all, by pairing BONCAT-FACS and metagenomics, we are capable of highlighting the taxa and potential functions that typifies the microbes actively responding to a rain event.

59 BASIC BIOLOGICAL SCIENCES↗

Artificial Replication of Field Soiling Losses on PV Modules

In this paper, we experimentally demonstrate an improved replication of field soiling losses using an indoor artificial soiling chamber and tests on anti-soiling coated PV modules and coupons. The primary focus is to use site-specific soil collected from module surface and replicate the natural soiling processes including dust concentration in the air, slow and gradual dust accumulation and sedimentation on the module surface during the dominant soiling season of the site of interest. The experiments were conducted on two sample sets having different anti-soiling properties. The first set contains commercial modules with two different surface properties retrieved after three years of exposure from a single PV plant in a mid-Atlantic location; the other set contain glass coupons with three different coating materials that were installed and exposed over 4 months, at Lemoore, California. Major field-representative factors considered here for the close replication in the chamber include: the use of dust collected from modules surfaces at the outdoor sites to give the same dust chemistry; dust particle size distribution and concentration; the charge size of dust (< 0.15 g per injection); field humidity, and module temperature. The effectiveness of antisoiling coatings (or surface properties) for both sample sets were ranked in the artificial testing and were found to be closely matching with the field rank orders of the respective sites and sample sets. This paper provides the rank ordering results to objectively demonstrate the replication of field soiling losses in the artificial soiling chamber.

artificial soiling↗

Metatranscriptomic reconstruction reveals RNA viruses with the potential to shape carbon cycling in soil

Viruses impact nearly all organisms on Earth, with ripples of influence in agriculture, health, and biogeochemical processes. However, very little is known about RNA viruses in an environmental context, and even less is known about their diversity and ecology in soil, 1 of the most complex microbial systems. Here, we assembled 48 individual metatranscriptomes from 4 habitats within a planted soil sampled over a 22-d time series: Rhizosphere alone, detritosphere alone, rhizosphere with added root detritus, and unamended soil (4 time points and 3 biological replicates). We resolved the RNA viral community, uncovering a high diversity of viral sequences. We also investigated possible host organisms by analyzing metatranscriptome marker genes. Based on viral phylogeny, much of the diversity was Narnaviridae that may parasitize fungi or Leviviridae , which may infect Proteobacteria. Both host and viral communities appear to be highly dynamic, and rapidly diverged depending on experimental conditions. The viral and host communities were structured based on the presence of root litter. Clear temporal dynamics by Leviviridae and their hosts indicated that viruses were replicating. With this time-resolved analysis, we show that RNA viruses are diverse, abundant, and active in soil. When viral infection causes host cell death, it may mobilize cell carbon in a process that may represent an overlooked component of soil carbon cycling.

54 ENVIRONMENTAL SCIENCES↗