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The soil microbiome is recognized as an essential component of healthy soils. Viruses are also diverse and abundant in soils, but their roles in soil systems remain unclear. Here we argue for the consideration of viruses in soil microbial food webs and describe the impact of viruses on soil biogeochemistry. The soil food web is an intricate series of trophic levels that span from autotrophic microorganisms to plants and animals. Each soil system encompasses contrasting and dynamic physicochemical conditions, with labyrinthine habitats composed of particles. Conditions are prone to shifts in space and time, and this variability can obstruct or facilitate interactions of microorganisms and viruses. Because viruses can infect all domains of life, they must be considered as key regulators of soil food web dynamics and biogeochemical cycling. We highlight future research avenues that will enable a more robust understanding of the roles of viruses in soil function and health., (© 2024. Springer Nature Limited.)

Soil biodiversity is threatened by intensification of land use. The consequences of different land use on belowground biodiversity remain insufficiently explored for soil protists. Alongside being abundant and extremely diverse in soil, protists provide many ecosystem services: key players in the microbial loop, turnover of organic matter and stimulation of plant growth-promoting rhizobacteria. However, we lack knowledge of effects of site, land use intensity and management on diversity of soil protists. Here we assessed protist communities in four European arable sites with contrasting land use intensities at each site: Lusignan, France; Moskanjci, Slovenia; Castro Verde, Portugal and Scheyern, Germany as well as two grassland sites: Hainich, Germany and Lancaster, UK. Each site has consistent agricultural management history of low and high land use intensities quantified in terms of land use index (LUI). We employed high-throughput sequencing of environmental DNA, targeting the V4 region of the 18S rRNA gene. By assigning the protist composition to trophic groups, we inspected for effects of management, and other biotic and abiotic variables. While overall protist richness was unaffected by LUI within sites, specific trophic groups such as plant pathogens and saprotrophs were affected. Effects on protist biome across land uses and sites were also observed. LUI sensitive taxa were taxonomically diverse in each plot, and their trophic groups responded in specific patterns to specific practices. The most abundant trophic group was phagotrophs (73%), followed by photoautotrophs (16%), plant pathogens (4%), animal parasites (2%) and saprotrophs (1%). Community compositions and factors affecting the structure of individual trophic groups differed between land uses and management systems. The agricultural management selected for distinct protist populations as well as specific functional traits, and the protist community and diversity were indeed affected by site, LUI and management, which indicates the ecological significance of protists in the soil food web., Competing Interests: Declaration of competing interest We declare that all authors approved the manuscript submission and we have no conflict of interest., (Copyright © 2020 Elsevier B.V. All rights reserved.)

This study linked growth inhibition of soil bacteria and protists to the chemical activity (a) of polycyclic aromatic hydrocarbons (PAHs) and compared the sensitivities of bacteria and protists. Passive dosing from pre-loaded silicone provided well-defined and constant a of PAHs in independent tests. Single-species growth inhibition with two bacterial (Pseuodomonas fluorescens DR54 and Sinorhizobium meliloti) and two protist (Cercomonas longicauda and Acanthamoeba castellanii) strains at maximum a (amax) of nine and four PAHs, respectively, showed no inhibition of PAHs with amax below 0.1 (pyrene and anthracene), while growth inhibition was observed for PAHs with amax above 0.1 (e.g. fluorene, fluoranthene and naphthalene). The bacteria were less sensitive than the protists. Soil bacterial community-level growth inhibition by naphthalene was in good agreement with single-species data, but also indicated the presence of sensitive bacteria that were inhibited by a below 0.05 and increasing pre-exposure time giving higher inhibition. The a of 50% inhibition (Ea50) was 0.434 and 0.329 for 0.5 and 4 h pre-exposure time, respectively. Invertebrates tended to be more sensitive than single-celled organisms tested here. This suggests that PAH exposure leads to differential toxicity in soil biota, which may affect soil food web structure and cycling of organic matter., (© FEMS 2019.)

Advances in sequencing technologies allow deeper studies of the soil protist diversity and function. However, little attention has been given to the impact of the chosen soil DNA extraction procedure to the overall results. We examined the effect of three acknowledged DNA recovery methods, two manual methods (ISOm-11063, GnS-GII) and one commercial kit (MoBio), on soil protist community structures obtained from different sites with different land uses. Results from 18S rRNA gene amplicon sequencing suggest that DNA extraction method significantly affect the replicate homogeneity, the total number of operational taxonomic units (OTUs) recovered and the overall taxonomic structure and diversity of soil protist communities. However, DNA extraction effects did not overwhelm the natural variation among samples, as the community data still strongly grouped by geographical location. The commercial DNA extraction kit was associated with the highest diversity estimates and with a corresponding higher retrieval of Excavata, Cercozoa and Amoebozoa-related taxa. Overall, our findings indicate that this extraction offers a compromise between rare and dominant taxa representation, while providing high replication reproducibility. A comprehensive understanding of the DNA extraction techniques impact on soil protist diversity can enable more accurate diversity assays., (Copyright © 2017 Elsevier GmbH. All rights reserved.)

Soil organisms have an important role in aboveground community dynamics and ecosystem functioning in terrestrial ecosystems. However, most studies have considered soil biota as a black box or focussed on specific groups, whereas little is known about entire soil networks. Here we show that during the course of nature restoration on abandoned arable land a compositional shift in soil biota, preceded by tightening of the belowground networks, corresponds with enhanced efficiency of carbon uptake. In mid- and long-term abandoned field soil, carbon uptake by fungi increases without an increase in fungal biomass or shift in bacterial-to-fungal ratio. The implication of our findings is that during nature restoration the efficiency of nutrient cycling and carbon uptake can increase by a shift in fungal composition and/or fungal activity. Therefore, we propose that relationships between soil food web structure and carbon cycling in soils need to be reconsidered.

The use of molecular methods to investigate protist communities in soil is in rapid development this decade. Molecular analysis of soil protist communities is usually dependant on direct genomic DNA extraction from soil and inefficient or differential DNA extraction of protist DNA can lead to bias in downstream community analysis. Three commonly used soil DNA extraction methods have been tested on soil samples from three European Long-Term Observatories (LTOs) with different land-use and three protist cultures belonging to different phylogenetic groups in different growth stages. The methods tested were: ISOm-11063 (a version of the ISO-11063 method modified to include a FastPrep ®-24 mechanical lysis step), GnS-GII (developed by the GenoSol platform to extract soil DNA in large-scale soil surveys) and a commercial DNA extraction kit - Power Lyzer™ PowerSoil® DNA Isolation Kit (MoBio). DNA yield and quality were evaluated along with DNA suitability for amplification of 18S rDNA fragments by PCR. On soil samples, ISOm-11063 yields significantly higher DNA for two of the three soil samples, however, MoBio extraction favors DNA quality. This method was also more effective to recover copies of 18S rDNA numbers from all soil types. In addition and despite the lower yields, higher DNA quality was observed with DNA extracted from protist cultures with the MoBio method. Likewise, a bead-beating step shows to be a good solution for DNA extraction of soil protists, since the recovery of DNA from protist cultures and from the different soil samples with the ISOm method proved to be efficient in recovering PCR-amplifiable DNA. This study showed that soil DNA extraction methods provide biased results towards the cyst stages of protist organism., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Sorption of PAHs to carbonaceous soil amendments reduces their dissolved concentrations, limiting toxicity but also potentially biodegradation. Therefore, the maximum abiotic desorption of freshly sorbed phenanthrene (≤5 mg kg(-1)) was measured in three soils amended with activated carbon (AC), biochar or compost. Total amounts of phenanthrene desorbed were similar between the different soils, but the amendment type had a large influence. Complete desorption was observed in the unamended and compost amended soils, but this reduced for biochar (41% desorbed) and AC (8% desorbed). Cumulative amounts mineralized were 28% for the unamended control, 19% for compost, 13% for biochar and 4% for AC. Therefore, the effects of the amendments in soil in reducing desorption were also reflected in the extents of mineralization. Modeling was used to analyze key processes, indicating that for the AC and charcoal treatments bacterial activity did not limit mineralization, but rather desorption into the dissolved phase., (Copyright © 2013 Elsevier Ltd. All rights reserved.)

Carbonaceous soil amendments are applied to contaminated soils and sediments to strongly sorb hydrophobic organic contaminants (HOCs) and reduce their freely dissolved concentrations. This limits biouptake and toxicity, but also biodegradation. To investigate whether HOCs sorbed to such amendments can be degraded at all, the desorption and biodegradation of low concentrations of (14)C-labelled phenanthrene (≤5 μg L(-1)) freshly sorbed to suspensions of the pure soil amendments activated carbon (AC), biochar (charcoal) and compost were compared. Firstly, the maximum abiotic desorption of phenanthrene from soil amendment suspensions in water, minimal salts medium (MSM) or tryptic soy broth (TSB) into a dominating silicone sink were measured. Highest fractions remained sorbed to AC (84±2.3%, 87±4.1%, and 53±1.2% for water, MSM and TSB, respectively), followed by charcoal (35±2.2%, 32±1.7%, and 12±0.3%, respectively) and compost (1.3±0.21%, similar for all media). Secondly, the mineralization of phenanthrene sorbed to AC, charcoal and compost by Sphingomonas sp. 10-1 (DSM 12247) was determined. In contrast to the amounts desorbed, phenanthrene mineralization was similar for all the soil amendments at about 56±11% of the initially applied radioactivity. Furthermore, HPLC analyses showed only minor amounts (<5%) of residual phenanthrene remaining in the suspensions, indicating almost complete biodegradation. Fitting the data to a coupled desorption and biodegradation model revealed that desorption did not limit biodegradation for any of the amendments, and that degradation could proceed due to the high numbers of bacteria and/or the production of biosurfactants or biofilms. Therefore, reduced desorption of phenanthrene from AC or charcoal did not inhibit its biodegradation, which implies that under the experimental conditions these amendments can reduce freely dissolved concentration without hindering biodegradation. In contrast, phenanthrene sorbed to compost was fully desorbed and biodegraded., (Copyright © 2012 Elsevier Ltd. All rights reserved.)

Microbial communities are integral parts of soil and their activity is very important to the functioning of soil. Therefore, microorganisms should be included in soil quality classification and assessment concepts. The challenges of using microbial indicators are to identify the best choice among the many techniques available to assess soil quality and to convert the information obtained from the microbial indicator into a form relevant for policy makers. In this article, we present a wide range of possible microbial indicators, some of them standardized; each provides slightly different information on soil quality. Experience with the use of indicators for assessment of microbial communities and soil quality is discussed. At present, as many microbial indicators as possible should be included to gain experience. At a minimum, measures of microbial biomass, respiration, and N mineralization and a community profiling method (e.g., DGGE, PLFA, or CLPP) should be included.

Soil viral ecology is a growing research field; however, the state of knowledge still lags behind that of aquatic systems. Therefore, to facilitate progress, the first Soil Viral Workshop was held to encourage international scientific discussion and collaboration, suggest guidelines for future research, and establish soil viral research as a concrete research area. The workshop took place at Søminestationen, Denmark, between 15 and 17th of June 2022. The meeting was primarily held in person, but the sessions were also streamed online. The workshop was attended by 23 researchers from ten different countries and from a wide range of subfields and career stages. Eleven talks were presented, followed by discussions revolving around three major topics: viral genomics, virus-host interactions, and viruses in the soil food web. The main take-home messages and suggestions from the discussions are summarized in this report.

Bacteria producing secondary metabolites with antagonistic effects on fungal pathogens have received attention during the last decades as an alternative to chemical pesticides. They, however, might also have effects on indigenous soil organisms like bacteria and protozoa, the latter ones being among the most important grazers of bacteria in soil. The present study reports on the effect of the potential biocontrol agent Pseudomonas fl uorescens CHA0 and its genetically modifi ed derivative CHA0/pME3424 on indigenous soil bacteria and protozoa in a soil system. CHA0/pME3424 overproduces two of the secondary metabolites produced by CHA0: the polyketide antibiotics pyoluteorin (Plt) and 2,4-diacetylphloroglucinol (DAPG). P. fl uorescens CHA0/gfp1 and CHA0/pME3424 both negatively affected the abundance of soil bacteria and protozoa and the genetic community structure of Kinetoplastida studied by PCR-DGGE. The negative effects were detectable after 14 days but were decreasing and are expected to be temporary. The overproducer of secondary metabolites did not differ in effect from the wild type. The soil respiration and bacterial genetic community structure were not signifi cantly affected. The study shows the soil bacteria and protozoa to be temporary affected by bacteria producing secondary metabolites, which can have implications for nutrient-cycling in soil and environmental risks of biocontrol agents. [ABSTRACT FROM AUTHOR]

Conventional identification based on morphology is today's tool of choice for monitoring earthworm diversity. However, it requires skills to distinguish specific features, which can become the bottleneck in soil ecological studies. DNA-based metabarcoding is increasingly used to monitor diversity of soil organisms. Few studies have compared this method with conventional methods. The aim of the study was to compare metabarcoding based on the mitochondrial 16S rRNA region with conventional morphology-based identification, where metabarcoding captured more species (eight) than identified by conventional method (five). Additionally, we tested the effects of two commercial DNA extraction kits: PowerSoil based on 0.25 g of soil and PowerMax based on 10 g of soil (both from Qiagen), where the PowerMax covered higher richness than PowerSoil. • DNA-based metabarcoding captured higher earthworm diversity than conventional morphology-based method. • Soil DNA extraction methods significantly affected earthworm diversity estimation. • Location drives earthworm community structure in DNA-based and conventional methods. [ABSTRACT FROM AUTHOR]

Despite the critical ecological roles of microeukaryotes in terrestrial ecosystems, most descriptive studies of soil microbes published so far focused only on specific groups. Meanwhile, the fast development of metagenome sequencing leads to considerable data accumulation in public repositories, providing microbiologists with substantial amounts of accessible information. We took advantage of public metagenomes in order to investigate microeukaryote communities in a well characterized grassland soil. The data gathered allowed the evaluation of several factors impacting the community structure, including the DNA extraction method, the database choice and also the annotation procedure. While most studies on soil microeukaryotes are based on sequencing of PCR-amplified taxonomic markers (18S rRNA genes, ITS regions), this work represents, to our knowledge, the first report based solely on metagenomic microeukaryote DNA. Choosing the correct annotation procedure and reference database has proven to be crucial, as it considerably limits the risk of wrong assignments. In addition, a significant and pronounced effect of the DNA extraction method on the taxonomical structure of soil microeukaryotes has been identified. Our analyses suggest that publicly available metagenome data can provide valuable information on soil microeukaryotes for comparative purposes when handled appropriately, complementing the current view provided by ribosomal amplicon sequencing methods. [ABSTRACT FROM AUTHOR]

Soil samples were analyzed with community-level physiological profiles (CLPP) using Biolog™ ECO-plates in the Netherlands Soil Monitoring Network (NSMN; 704 samples) and in a European-wide transect (73 samples). The selection of sites was based on a representative sample of major soil texture types, land uses and climate zones. This method for determination of CLPP was specifically designed to be applied in large and long-term monitoring activities. It encompassed a solution for the inoculum-density dependence of colour formation patterns in ECO-plates, through stepwise dilution to extinction of the bacterial inoculum. The CLPP in Dutch and European soil samples appeared to be reproducible and sensitive to land use and/or soil texture. Although the method is selective, CLPP based parameters correlated well with other microbial parameters and soil characteristics. Consistent patterns in CLPP and soil habitat characteristics are emerging, as brought about by environmental disturbances, land management and soil texture. The applicability of CLPP analysis in monitoring systems is discussed. [ABSTRACT FROM AUTHOR]