Soil Protists Research Articles

Elevated Atmospheric Co2 Levels Impact Soil Protist Functional Core Community Compositions

Protists, known as microeukaryotes, are a significant portion of soil microbial communities. They are crucial predators of bacteria and depend on bacterial community dynamics for the growth and evolution of protistan communities. In parallel, increased levels of atmospheric CO2 significantly impact bacterial metabolic activity in rhizosphere soils. In this study, we investigated the effect of elevated atmospheric CO2 levels on the metabolically active protist community composition and function and their co-occurrences with bacteria from bulk and rhizosphere soils from the Giessen Free-Air CO2 enrichment grassland experiment. Metabarcoding sequencing data analyses of partial 18S rRNA from total soil RNA showed that elevated CO2 concentrations stimulated only a few ASVs of phagotrophic predators of bacteria and other microeukaryotes, affecting protist community composition (P = 0.006, PERMANOVA). In parallel, phagotrophic and parasitic lineages appeared slightly favoured under ambient CO2 conditions, results that were corroborated by microbial signature analyses. Cross-comparisons of protist-bacteria co-occurrences showed mostly negative relations between prokaryotes and microeukaryotes, indicating that the ongoing increase in atmospheric CO2 will lead to changes in microbial soil communities and their interactions, potentially cascading to higher trophic levels in soil systems.

Functional traits data for testate amoebae of Northern Holarctic realm

The functional traits of soil protists have been employed in ecological research to enhance comprehension of the underlying mechanisms of ecological processes. Among the numerous soil protists, testate amoebae emerge as a prominent and abundant group, playing a pivotal role in soil micro-food webs. Furthermore, they are regarded as valuable bioindicators for environmental monitoring and palaeoecological studies due to their sensitivity to environmental changes. We screened 372 testate amoebae species widely distributed across Northern Holarctic realm and collected trait data, representing the morphological and feeding characteristics of testate amoebae. The dataset would provide valuable basis for investigation of the functional diversity and ecological roles of testate amoebae, thus facilitating further research on soil protist communities and ecosystem dynamics.

Impact of fallen wood on soil protists in the subtropics

Impact of fallen wood on soil protists in the subtropics

Investigating protistan predators and bacteria within soil microbiomes in agricultural ecosystems under organic and chemical fertilizer applications

Organic farming can enhance biodiversity and soil health and is a sustainable alternative to conventional farming. Yet, soil protists especially protistan predators, have received inadequate attention, and their contributions to the sustainability of organic farming remained underexplored. In this study, we examined soil microbial communities from 379 samples, including both organic and chemically fertilized soils from China. Our findings revealed higher bacterial diversity and increases in plant-beneficial bacteria in organically farmed soils. Notably, organic farming systems facilitated dynamic predator-prey interactions, which may be disrupted by the application of chemical fertilizers. Additionally, organic farming enriched protistan predators, enhancing the relative abundance of functional PGPR, thus improving soil health. We further conducted a case study highlighting the critical role of organic matter in sustaining protistan predator populations and their interactions with bacteria. We propose the crucial contributions of organic inputs for supporting protistan predators and the interplay of predator-prey, ultimately enhancing soil functions and promoting agricultural sustainability.

Colonization ability and uniformity of resources and environmental factors determine biological homogenization of soil protists in human land-use systems.

Humans have substantially transformed the global land surface, resulting in the decline in variation in biotic communities across scales, a phenomenon known as "biological homogenization." However, different biota are affected by biological homogenization to varying degrees, but this variation and the underlying mechanisms remain little studied, particularly in soil systems. To address this topic, we used metabarcoding to investigate the biogeography of soil protists and their prey/hosts (prokaryotes, fungi, and meso- and macrofauna) in three human land-use ecosystem types (farmlands, residential areas, and parks) and natural forest ecosystems across subtropical and temperate regions in China. Our results showed that the degree of community homogenization largely differed between taxa and functional groups of soil protists, and was strongly and positively linked to their colonization ability of human land-use systems. Removal analysis showed that the introduction of widespread, generalist taxa (OTUs, operational taxonomic units) rather than the loss of narrow-ranged, specialist OTUs was the major cause of biological homogenization. This increase in generalist OTUs seemingly alleviated the negative impact of land use on specialist taxa, but carried the risk of losing functional diversity. Finally, homogenization of prey/host biota and environmental conditions were also important drivers of biological homogenization in human land-use systems, with their importance being more pronounced in phagotrophic than parasitic and phototrophic protists. Overall, our study showed that the variation in biological homogenization strongly depends on the colonization ability of taxa in human land-use systems, but is also affected by the homogenization of resources and environmental conditions. Importantly, biological homogenization is not the major cause of the decline in the diversity of soil protists, and conservation and study efforts should target at taxa highly sensitive to local extinction, such as parasites.

The impact of fungi on soil protist communities in European cereal croplands.

Protists, a crucial part of the soil food web, are increasingly acknowledged as significant influencers of nutrient cycling and plant performance in farmlands. While topographical and climatic factors are often considered to drive microbial communities on a continental scale, higher trophic levels like heterotrophic protists also rely on their food sources. In this context, bacterivores have received more attention than fungivores. Our study explored the connection between the community composition of protists (specifically Rhizaria and Cercozoa) and fungi across 156 cereal fields in Europe, spanning a latitudinal gradient of 3000 km. We employed a machine-learning approach to measure the significance of fungal communities in comparison to bacterial communities, soil abiotic factors, and climate as determinants of the Cercozoa community composition. Our findings indicate that climatic variables and fungal communities are the primary drivers of cercozoan communities, accounting for 70% of their community composition. Structural equation modelling (SEM) unveiled indirect climatic effects on the cercozoan communities through a change in the composition of the fungal communities. Our data also imply that fungivory might be more prevalent among protists than generally believed. This study uncovers a hidden facet of the soil food web, suggesting that the benefits of microbial diversity could be more effectively integrated into sustainable agriculture practices.

Measurements of soil protist richness and community composition are influenced by primer pair, annealing temperature, and bioinformatics choices.

Protists are a diverse and understudied group of microbial eukaryotic organisms especially in terrestrial environments. Advances in molecular methods are increasing our understanding of the distribution and functions of these creatures; however, there is a vast array of choices researchers make including barcoding genes, primer pairs, PCR settings, and bioinformatic options that can impact the outcome of protist community surveys. Here, we tested four commonly used primer pairs targeting the V4 and V9 regions of the 18S rRNA gene using different PCR annealing temperatures and processed the sequences with different bioinformatic parameters in 10 diverse soils to evaluate how primer pair, amplification parameters, and bioinformatic choices influence the composition and richness of protist and non-protist taxa using Illumina sequencing. Our results showed that annealing temperature influenced sequencing depth and protist taxon richness for most primer pairs, and that merging forward and reverse sequencing reads for the V4 primer pairs dramatically reduced the number of sequences and taxon richness of protists. The data sets of primers that targeted the same 18S rRNA gene region (e.g., V4 or V9) had similar protist community compositions; however, data sets from primers targeting the V4 18S rRNA gene region detected a greater number of protist taxa compared to those prepared with primers targeting the V9 18S rRNA region. There was limited overlap of protist taxa between data sets targeting the two different gene regions (80/549 taxa). Together, we show that laboratory and bioinformatic choices can substantially affect the results and conclusions about protist diversity and community composition using metabarcoding.IMPORTANCEEcosystem functioning is driven by the activity and interactions of the microbial community, in both aquatic and terrestrial environments. Protists are a group of highly diverse, mostly unicellular microbes whose identity and roles in terrestrial ecosystem ecology have been largely ignored until recently. This study highlights the importance of choices researchers make, such as primer pair, on the results and conclusions about protist diversity and community composition in soils. In order to better understand the roles protist taxa play in terrestrial ecosystems, biases in methodological and analytical choices should be understood and acknowledged.

Species-specific predation determines the feeding impacts of six soil protist species on bacterial and eukaryotic prey

Predatory protists play a central role in nutrient cycling and are involved in other ecosystem functions by predating the microbiome. While most soil predatory protist species arguably are bacterivorous, some protist species can prey on eukaryotes. However, studies about soil protist feeding mainly focused on bacteria as prey and rarely tested both bacteria and eukaryotes as potential prey. In this study, we aimed to decipher soil predator–prey interactions of three amoebozoan and three heterolobosean soil protists and potential bacterial (Escherichia coli; 0.5–1.5 µm), fungal (Saccharomyces cerevisiae; 5–7 µm) and protist (Plasmodiophora brassicae; 3–5 µm) prey, either as individual prey or in all their combinations. We related protist performance (relative abundance) and prey consumption (qPCR) to the protist phylogenetic group and volume. We showed that for the six soil protist predators, the most suitable prey was E. coli, but some species also grew on P. brassicae or S. cerevisiae. While protist relative abundances and growth rates depended on prey type in a protist species-specific manner, phylogenetic groups and volume affected prey consumption. Yet we conclude that protist feeding patterns are mainly species-specific and that some known bacterivores might be more generalist than expected, even preying on eukaryotic plant pathogens such as P. brassicae.

Land use intensification homogenizes soil protist communities and alters their diversity across Europe

Soil protists have vital roles as major microbiome predators in soil functioning and plant performance. Protists are also suggested to be the most responsive microbial group to external changes, such as anthropogenic land use types. While protists were long used as models for biogeography such as to investigate if ‘everything is everywhere’ among microbes, their biogeography at the taxonomic level has never been explored in depth at the continental scale and linked to anthropogenic drivers. Here we evaluated how land-use types affect the diversity and structure of soil protist communities across 885 locations in Europe based on the European Commission's Land Use and Coverage Area frame Survey (LUCAS). We observed higher α-diversity of soil protists but lower community structure dissimilarity (β-diversity) in croplands compared with woodlands, with grasslands in an intermediate position. The diversity of protist groups with a broader spatial niche was higher in croplands, whereas taxa with a narrower niche increased in woodlands. The importance of climate factors on α-diversity variations reduced as land use intensity increased, but the opposite trend was observed for the effect of soil properties. Our study suggests that there is an interaction between land use type, environmental effects, and spatial niche attributes of soil protist groups, highlighting the importance of land-use type on the dynamics of protist communities.

Enhanced transport of bacteria along root systems by protists can impact plant health.

Soil protists have been shown to contribute to the structure and function of the rhizosphere in a variety of ways. Protists are key contributors to nutrient cycling through the microbial loop, where biomass is digested by protists and otherwise stored nutrients are returned to the environment. Protists have also been shown to feed on plant pathogenic bacteria and alter root microbiomes in ways that may benefit plants. Recently, a mechanism involving bacterial transport, facilitated by protists, has been hypothesized to contribute to the spatial distribution of bacteria in the rhizosphere. Here, we observe the differential abilities of three soil protists: a ciliate (Colpoda sp.), a flagellate (Cercomonas sp.), and a naked amoeba (Acanthamoeba castellanii) to transport nitrogen-fixing Sinorhizobium meliloti to infectible root tips. Co-inoculation of protists plus S. meliloti resulted in the movement of bacteria, as measured by the presence of nitrogen-fixing nodules, up to 15 cm farther down the root systems when compared to plants inoculated with S. meliloti alone. Co-inoculation of the ciliate, Colpoda sp., with S. meliloti, resulted in shoot weights that were similar to plants that grew in nitrogen-replete potting mix. Colpoda sp.-feeding style and motility likely contributed to their success at transporting bacteria through the rhizosphere. We observed that the addition of protists alone without the co-inoculum of S. meliloti resulted in plants with larger shoot weights than control plants. Follow-up experiments showed that protists plus their associated microbiomes were aiding in plant health, likely through means of nutrient cycling.IMPORTANCEProtists represent a significant portion of the rhizosphere microbiome and have been shown to contribute to plant health, yet they are understudied compared to their bacterial and fungal counterparts. This study elucidates their role in the rhizosphere community and suggests a mechanism by which protists can be used to move bacteria along plant roots. We found that the co-inoculation of protists with nitrogen-fixing beneficial bacteria, Sinorhizobium meliloti, resulted in nodules farther down the roots when compared to plants inoculated with S. meliloti alone, and shoot weights similar to plants that received nitrogen fertilizer. These data illustrate the ability of protists to transport viable bacteria to uninhabited regions of the root system.

The influence of soil factors on protist community dynamics during plant succession in subalpine natural and planted forests

Protists, as the core component of the soil microbiome, may play an important role in forest succession by altering soil nutrient cycling and plant performance. However, knowledge of how protists change with plant succession is extremely limited. To illustrate protistan community dynamics following plant succession in subalpine natural and planted ecosystems, we used high-throughput sequencing to identify protists in rhizosphere and bulk soils at different successional stages, and the effects of soil abiotic and biotic factors on protists were analysed. Overall, the α-diversity and community composition of protists differed significantly between natural and planted ecosystems, but not between rhizosphere and bulk soils. Moving along the successional gradient, the richness and diversity of protists in rhizosphere and bulk soils of natural ecosystems, as well as the richness of functional taxa (i.e., consumers, parasites and phototrophs), gradually increased. In contrast, the richness of protists and functional taxa in the plantations was generally lowest in the old stands (75 years). The community composition of protists in rhizosphere and bulk soils of these two ecosystems varied significantly among successional stages, and their community composition variation was positively correlated with the time interval of successional stages, indicating that community dynamics exhibited directionality along successional gradients. Soil abiotic and biotic factors jointly regulate protistan α-diversity and community composition, and forest succession regulates functional taxa through multiple pathways. However, the key influencing factors and specific regulatory pathways are not consistent across different ecosystems, soil types and functional taxa. Therefore, our study has revealed that protistan community assembly was distinct along two separate successional gradients, indicating potential functional differences. These results provide a basis for the development of future forest management measures.

Protist predation promotes antimicrobial resistance spread through antagonistic microbiome interactions.

Antibiotic resistance has grown into a major public health threat. In this study, we reveal predation by protists as an overlooked driver of antibiotic resistance dissemination in the soil microbiome. While previous studies have primarily focused on the distribution of antibiotic resistance genes, our work sheds light on the pivotal role of soil protists in shaping antibiotic resistance dynamics. Using a combination of metagenomics and controlled experiments in this study, we demonstrate that protists cause an increase in antibiotic resistance. We mechanistically link this increase to a fostering of antimicrobial activity in the microbiome. Protist predation gives a competitive edge to bacteria capable of producing antagonistic secondary metabolites, which secondary metabolites promote in turn antibiotic-resistant bacteria. This study provides insights into the complex interplay between protists and soil microbiomes in regulating antibiotic resistance dynamics. This study highlights the importance of top-down control on the spread of antibiotic resistance and directly connects it to cross-kingdom interactions within the microbiome. Managing protist communities may become an important tool to control outbreaks of antibiotic resistance in the environment.

Promotion of the symbiotic interaction between phagotrophic protists and beneficial bacteria, mediated via a balancing of soil nutrients, reduces the incidence of watermelon Fusarium wilt

Elucidating the microbial mechanisms that trigger Fusarium wilt represents a key step in addressing the barriers to sustainable cropping. However, from the perspective of the complete microbiome, the integrated role of soil nutrients and the microbial community in fields with different rate of wilt disease remains unclear. In this study, we examined the potential interrelationships among rhizosphere soil nutrients, bacteria, fungi, and protists in soils collected from fields in which watermelon had been cultivated for 8 years, and which were characterized by high (HW: 81.25%) or low (LW: 6.25%) incidence of watermelon Fusarium wilt. Soil from HW fields was found to contain a higher density of F. oxysporum (1.30-fold higher than that in the LW soil), along with higher contents of available phosphorus (1.31-fold higher), and available potassium (2.39-fold higher). In addition, the inter-kingdom correlation between protists and bacteria in the HW soil was 2.08-fold higher than that in LW soil. Furthermore, structural equation modeling revealed that an excess of soil available potassium enhances predation by potentially detrimental phagotrophic protists on potentially beneficial bacteria. Collectively, our findings indicate that a balanced nutrient input and the interaction between protists and beneficial bacteria play important roles in controlling the incidence of watermelon Fusarium wilt. These findings have implications for controlling the incidence of watermelon Fusarium wilt based on modifying the nutrient balance of soil and increasing the flora of certain beneficial protists (Cercomonas and Colpoda) that interact symbiotically with species of Bacillus.

Seven-year N and P inputs regulate soil microbial communities via bottom-up effects on carbon and nutrient supply and top-down effects on protist relative abundance

Information about the interactions between microbial carbon (C), nitrogen (N), and phosphorus (P) mineralization and nutrient inputs will improve our understanding of the effects of global change on soil biogeochemical cycling and ecosystem function. In this study, we explored how N, P, and NP inputs, when applied at low rates (50 kg ha–1 year–1) for seven years, affected the microbial community in an acidic, strongly weathered, red clay soil in a subtropical conifer plantation. N and P inputs regulate microbial communities holistically through influencing soil resources (bottom-up), such as the C and nutrient supply, and predators, such as protists (top-down). We observed that N, P, and NP inputs caused changes in the protist community, which then affected bacterial diversity rather than fungal diversity. The increases in the protist diversity that resulted from the P and NP inputs were associated with increases in the relative abundance of Calditrichaeota, which is involved in C-mineralization, and decreases in the relative abundance of Marinimicrobia, which is involved in N-mineralization. The N and NP inputs caused the soil to become more acidic, exacerbated the microbial C limitation, and reduced the overall microbial functional diversity to carry out C, N, and P mineralisation. We conclude that the nutrient supply directly impacted the C-, N-, and P-mineralising microbial communities (bottom-up), and directly and indirectly affected soil protists (top-down).

Environmental factors influencing the diversity and distribution of dictyostelid cellular slime molds in forest and farmland soils of western China.

Soil protists are an essential yet seriously understudied component of the soil microbiome. In this study, 11 new records of dictyostelids belonging to 2 orders, 3 families, and 4 genera were identified from 99 soil samples collected from different elevations and habitats in central Gansu and the southeastern and southcentral portions of Guizhou Province, China. We found that dictyostelid communities were significantly different between Gansu and Guizhou Provinces, apparently in response to different environmental factors. Moreover, dictyostelids were found to have the highest species diversity in mixed forests. Soil pH, temperature, and elevation were determined to be the primary factors that affect the distribution and occurrence of dictyostelids in Guizhou and Gansu Provinces. This work supplements the survey data available for dictyostelids elsewhere in China. These new findings have significant implications for our understanding of the diversity of soil microorganisms.

Urban greenspaces shape soil protist communities in a location-specific manner

The impacts of urbanization on aboveground biodiversity are well studied, and its impact on soil microorganisms are also receiving increased attention. However, the impact of urbanization on the soil protists are hardly investigated. Here, we studied how urbanization and distinct urban greenspaces affect protist communities. We used amplicon sequencing of the18 S rRNA gene of samples from five types of urban greenspaces (parks, greenbelts, industrial areas, residential areas and hospital lawns), neighboring natural forests and agricultural ecosystems in Ningbo, China. We found that urban greenspaces harbored higher protist α-diversity than forests, while protist β-diversity increased from agricultural systems to urban greenspaces to forests. Among the studied driving factors, soil bacterial α- and β-diversity best predicted phagotrophic protist α- and β-diversity in urban greenspaces, while differences in α- and β-diversity of phototrophic protists were best explained by soil carbon-to-nitrogen ratio and fungal β-diversity, respectively. Abiotic factors i.e., total phosphorus and carbon-to-nitrogen ratio, best predicted the α- and β-diversity of protist parasites in urban greenspaces, respectively. The results revealed that the composition and drivers of protist communities vary between functional groups and urban ecosystems. Overall, our findings contribute to a better understanding of drivers of soil protist communities and indicate that soil protist communities and associated soil functions could be managed in predictable ways in urban greenspaces.

Different long-term fertilization regimes affect soil protists and their top-down control on bacterial and fungal communities in Mollisols

Soil protists represent a vastly diverse component of soil microbial communities and significantly contribute to biogeochemical cycling. However, how different fertilization regimes impact the protistan communities and their top-down control on bacteria and fungi remain largely unknown. Here, using high-throughput sequencing, we investigated the differences in protist communities and their relationships with bacterial and fungal communities in Mollisols of Northeast China that were subjected to chemical and organic fertilization over 30 years. The results showed that manure addition rather than chemical fertilization significantly increased protistan alpha diversity and changed protistan community structure. Manure amendments markedly increased the relative abundances of protistan consumers (such as Cercozoa) and reduced the proportion of phototrophic protists (such as Chlorophyta). Soil pH was the most influential factor driving microbial communities, and protists were less sensitive to environmental disturbances than bacteria and fungi. Protistan communities exhibited more stronger relationships with bacterial communities than fungal communities, and Chlorococcum was the most important contributor in regulation of microbial taxa and functional genes. Furthermore, manure addition slightly simplified the microbial network, and chemical plus manure fertilization improved network stability with the highest robustness. Manure addition specifically mitigated the negative interactions between protists and bacteria while reinforced the positive interactions between protists and fungi. This study advanced our knowledge about the roles of protistan groups in regulating microbial communities and ecosystem functions associated with chemical and organic fertilization.

Water rather than nitrogen availability predominantly modulates soil microbial beta-diversity and co-occurrence networks in a secondary forest

Atmospheric nitrogen (N) deposition and changing precipitation regimes greatly affect the structure and functions of terrestrial ecosystems. However, their impacts on the diversity and assembly of soil microbial communities including bacteria, fungi and protists, remain largely unclear. As part of a six-year field experiment in a secondary forest in a warm temperate and subtropical climate transitional zone in China, we aimed to investigate the responses of soil microbial communities to N addition, increased and decreased precipitation. The results showed that N addition had no effect on soil microbial α- or β-diversity, but reduced the complexity of microbial network. Neither increased nor decreased precipitation influenced soil microbial α-diversity, but decreased precipitation rather than increased precipitation elevated bacterial and protistan community dissimilarities (β-diversity), which could have been largely attributed to species replacement processes through reducing soil water availability. In addition, decreased precipitation weakened microbial complexity and stability, but enhanced the node proportion of protists in the co-occurrence network. Our observations suggest the asymmetric responses of soil microbial β-diversity to increased and decreased precipitation, and underscore that water rather than N availability, especially drought condition, plays a predominant role in modulating soil microbial β-diversity. Moreover, the findings imply that global change can strengthen the importance of soil protists and then reshape microbial assembly in forests.

Toward a common set of functional traits for soil protists

Protists are major actors of soil communities and play key roles in shaping food webs, community assembly, and ecosystem processes, yet their functional diversity is understudied. High-throughput sequencing data have revealed their ubiquity and diversity, but lack of standardized traits has hampered the integration of functional information, limiting our understanding of soil ecosystems. Here, we propose a functional framework for soil protists, identify a set of common traits to characterize their functional diversity, and apply the framework on a broad-scale, real-world dataset. We reviewed studies on soil protists to identify the traits used in the literature, and define a framework based on 10 key traits that satisfy two criteria: availability of information, and applicability to most taxa. The framework was tested on a dataset of environmental DNA metabarcoding data from 1123 soil samples collected in 48 glacier forelands worldwide. Traits were assigned to the 570 Molecular Operational Taxonomic Units (MOTUs) detected in our dataset, leading to the production of a global trait-based dataset from glacier forelands. We estimated the functional space of protist communities and evaluated if the selected traits were effective in describing protist diversity. The functional space of protist communities showed that the MOTUs are clustered in three regions, mainly reflecting different nutritional and habitat preferences. The proposed framework is appropriate for multiple applications, including estimation of functional diversity and food web analyses, and provides a basis for ecological studies on soil protists, enabling the functional characterization of this essential but often neglected component of soil biodiversity.

Keystone predatory protists are associated closely with ammonia‐oxidizing microorganisms in an acidic Ultisol

AbstractPredatory protists are widely recognized as critical biotic forces driving soil microbial communities, but their top‐down controls on ammonia‐oxidizing microorganisms (AOMs), the major players in nitrification, are largely unresolved. Here, we investigated the communities of predatory protists and their associations with AOMs using high‐throughput sequencing and network analysis in soil aggregates following various long‐term organic substitutions. We found that organic substitutions increased while soil aggregation decreased the alpha diversity of predatory protists. Predatory protistan communities were significantly associated with AOMs. Variosea, an important group of Amoebozoa, were the keystone predatory protists associated with the AOMs. Collectively, our findings highlight the importance of predatory protists, especially Variosea, in regulating the communities of AOMs in an acidic Ultsisol, with implications for managing nitrification by predatory protists in agricultural soils.