Protists are important key players in the microbial loop and influence their environment by grazing, which leads to the return of nutrients into the soil and reduces pathogen pressure on plants. Specifically, protists on and around plant roots are important for plants\' development and growth. For this study, the fourth most important crop in the world, Hordeum vulgare, was selected. Seeds of H. vulgare were inoculated with Acanthamoeba castellanii alone or with additional soil bacteria at the beginning and during the experiment. The germination of the seeds and the growth of the plants in pouches were monitored over 3 weeks. No differences were found in leaf growth, root growth, root and leaf nitrogen content or ammonia content of the liquid from the pouches. In contrast, the relative increase in root and leaf dry weight showed a small difference compared to the controls. The results of this experiment demonstrated that seed inoculation with A. castellanii alone or with additional unidentified soil bacteria did not have a major effect on the growth and development of barley. Nevertheless, small changes in plant development were detected, indicating that A. castellanii should be considered for further investigation of co-inoculations with plant growth-promoting bacteria and additional nutrients.

The swift proliferation of forests converted into monoculture plantations has profound impacts on soil nutrients, microbial communities, and many ecological processes and functions. Nematodes are soil microfauna that play a pivotal role in biogeochemical cycling and in soil food web, whereas the response of soil nematode communities and energy flows to forest conversion remains unknown. Here, we assessed the community composition and the energy flows of the nematode food webs as a function of soil chemistry after conversion from natural forests (Forest) to four plantations (8-year-old): Amygdalus persica (Peach), Myrica rubra (Berry), Camellia oleifera (Oil), and Cunninghamia lanceolata (Fir). After forest conversion, soil organic carbon (SOC) and total nitrogen (TN) contents decreased by 65 % and 55 %, respectively. Forest conversion strongly reduced the abundance (particularly large-bodied omnivorous-predatory nematodes), diversity, maturity, and stability of the soil nematode community. The shifts in composition and structure of nematode communities after forest conversion are reflected in changes in the abundance of predominant genera and trophic taxa, especially bacterivorous, fungivorous, and omnivorous-predatory nematodes. Acrobeloides notably increased, whereas Plectus, Prismatolaimus, Tylencholaimus, and Tripyla decreased. Accordingly, the abundances of r-strategy nematodes (cp value = 1-2) increased, but that of the K-strategists (cp value = 3-5) declined. Additionally, the energy flow across the soil nematode food web was reduced by 36 % and flow uniformity declined by 24 % after forest conversion. These changes in nematode diversity and abundance were triggered by diminishing soil C and N contents, thereby affecting the energy flows via the nematode food webs. Thus, forest conversion affects soil biotas and multi-functions from the perspective of nematode food web structure and energy flows, and underlines the interconnections between ecosystem and energy dynamics across multi-trophic levels, which is crucial for sustainable forest management.

Root-knot nematodes (RKNs) pose a serious threat to crop production. Flooding soil with biogas slurry, combined with soil heating before crop planting, has the potential for RKN disease suppression. However, the actual effect of this method has not been verified under field conditions. Here, we present the results of a two-year field experiment in a greenhouse demonstrating the control effect on RKN disease and plant growth using this method, as well as its influence on the soil nematode community. Four treatments were set: untreated control (CK), local control method for RKN (CC), soil flooded with 70 % biogas slurry (BS70), and soil flooded with undiluted biogas slurry (BS100). In the first year, all three RKN control treatments significantly reduced the root-knot index (p < 0.05). In the next year, only BS70 and BS100 still presented significantly suppressed effects (p < 0.05), and it was more obvious under BS70 with a relative control effect of 74.6 %. In the first year, BS70 and BS100 significantly inhibited the plant height of watermelon (p < 0.05). In the next year, however, all three RKN control treatments promoted the growth of watermelon, and their stem diameter was significantly greater than that of CK. The application of biogas slurry (BS70 and BS100) significantly increased nematode richness and the Shannon index in the second year (p < 0.05). However, the structure index showed no significant difference among treatments (p > 0.05), indicating that biogas slurry application did not increase the soil food web complex. Principal component analysis showed that the application of biogas slurry changed the nematode community, especially under BS70, which presented a more lasting influence. The high-level input of biogas slurry also caused soil NH4+-N and heavy-metal and arsenic accumulation in the first year, but these soil-pollution risks disappeared in the second year.

While it is acknowledged that alpine soil bacterial communities are primarily driven by season and elevation, there is no consensus on the factors influencing fungi and protists. Here we used a holistic approach of the microbiome to investigate the seasonal dynamics in alpine grasslands, focusing on soil food web interactions. We collected 158 soil samples along elevation transects from three mountains in the Alps, in spring during snowmelt and in the following summer. Using metatranscriptomics, we simultaneously assessed prokaryotic and eukaryotic communities, further classified into trophic guilds. Our findings reveal that the consumers\' pressure increases from spring to summer, leading to more diverse and evenly distributed prey communities. Consequently, consumers effectively maintain the diverse soil bacterial and fungal communities essential for ecosystem functioning. Our research highlights the significance of biotic interactions in understanding the distribution and dynamics of alpine microbial communities.

Arbuscular mycorrhizal fungi (AMF) transport substantial plant carbon (C) that serves as a substrate for soil organisms, a precursor of soil organic matter (SOM), and a driver of soil microbial dynamics. Using two-chamber microcosms where an air gap isolated AMF from roots, we 13CO2-labeled Avena barbata for 6 wk and measured the C Rhizophagus intraradices transferred to SOM and hyphosphere microorganisms. NanoSIMS imaging revealed hyphae and roots had similar 13C enrichment. SOM density fractionation, 13C NMR, and IRMS showed AMF transferred 0.77 mg C g-1 of soil (increasing total C by 2% relative to non-mycorrhizal controls); 33% was found in occluded or mineral-associated pools. In the AMF hyphosphere, there was no overall change in community diversity but 36 bacterial ASVs significantly changed in relative abundance. With stable isotope probing (SIP)-enabled shotgun sequencing, we found taxa from the Solibacterales, Sphingobacteriales, Myxococcales, and Nitrososphaerales (ammonium oxidizing archaea) were highly enriched in AMF-imported 13C (> 20 atom%). Mapping sequences from 13C-SIP metagenomes to total ASVs showed at least 92 bacteria and archaea were significantly 13C-enriched. Our results illustrate the quantitative and ecological impact of hyphal C transport on the formation of potentially protective SOM pools and microbial roles in the AMF hyphosphere soil food web.

Impacts of long-term fertilization and cultivation were evaluated on nematode communities associated with tall fescue turfgrass following 11 years of treatment applications. Fertilizer treatments of biosolid, synthetic, and plant-based fertilizers and cultivation treatments of 0×, 1×, and 2× aerification passes were applied to randomized and replicated tall fescue plots at the University of Maryland Paint Branch Turfgrass facility in College Park, Maryland. Free-living and plant-parasitic nematodes were identified, enumerated, and categorized into functional groups. Nematode count data were compared using generalized linear mixed modeling with negative binomial distribution and two-way ANOVA was used to compare nematode ecological indices. Biosolid treatments resulted in lower omnivore-predator densities than plant-based fertilizer treatments (p ≤ .001) and significantly greater Hoplolaimus densities than plant-based fertilizer plots (p ≤ .05). Synthetic fertilizer applications resulted in the greatest Eucephalobus (p ≤ .05) and total bacterivore densities (p ≤ .001) of all fertilizer treatments. Plant-based fertilizer-treated plots had the largest Maturity Index cp 2-5 and Structure Index (p ≤ .05). Cultivation of 1× resulted in fewer total bacterivore densities than 2× (p ≤ .01) while omnivore-predator densities were greater in 1× than 0× (p ≤ .001). Plant health, as measured by NDVI, was lowest in biosolid-treated turfgrass (p ≤ .05). These findings suggest that long-term turfgrass management practices can have variable impacts on nematode abundance and community structure in tall fescue and provide insights into ecological impacts of turfgrass management practices.

Climate change will likely increase habitat loss of endemic tree species and drives forest conversion in mountainous forests. Elevation gradients provide the opportunity to predict possible consequences of such changes. While species compositions of various taxa have been investigated along elevation gradients, data on trophic changes in soil-dwelling organisms are scarce. Here, we investigated trophic changes of the Collembola communities along the northern slope of Changbai Mountain, China. We sampled Collembola in primary forests at seven elevations (800-1700 m asl). We measured individual body lengths and bulk stable isotopes on species level. We further categorized Collembola species into life forms. The community-weighted means of Δ15N and Δ13C values as well as minimum Δ15N values and isotopic uniqueness of Collembola communities increased with increasing elevation, while the range of Δ15N values decreased. Maximum and minimum of Δ13C values differed between elevations but showed no linear trend. Further, Δ15N values of Collembola species occurring across all elevations increased with elevation. Changes in Δ15N values with elevation were most pronounced in hemiedaphic species, while Δ13C values increased strongest with elevation in euedaphic species. Δ15N values increased with decreasing body size in hemiedaphic and euedaphic species. Overall, the results suggest that Collembola species functioning as primary decomposers at lower elevations shift towards functioning as secondary decomposers or even predators or scavengers at higher elevation forests. The results further indicate that access to alternative food resources depends on Collembola life form as well as body size and varies between ecosystems.

Molecular gut content analysis via diagnostic PCR or high-throughput sequencing (metabarcoding) of consumers allows unravelling of feeding interactions in a wide range of animals. This is of particular advantage for analyzing the diet of small invertebrates living in opaque habitats such as the soil. Due to their small body size, which complicates dissection, microarthropods are subjected to whole-body DNA extraction-step before their gut content is screened for DNA of their food. This poses the problem that body surface contaminants, such as fungal spores may be incorrectly identified as ingested food particles for fungivorous species. We investigated the effectiveness of ten methods for body surface decontamination in litter-dwelling oribatid mites using Steganacarus magnus as model species. Furthermore, we tested for potential adverse effects of the decontamination techniques on the molecular detection of ingested prey organisms. Prior to decontamination, oribatid mites were fed with an oversupply of nematodes (Plectus sp.) and postmortem contaminated with fungal spores (Chaetomium globosum). We used diagnostic PCR with primers specific for C. globosum and Plectus sp. to detect contaminants and prey, respectively. The results suggest that chlorine bleach (sodium hypochloride, NaClO, 5%) is most efficient in removing fungal surface contamination without significantly affecting the detection of prey DNA in the gut. Based on these results, we provide a standard protocol for efficient body surface decontamination allowing to trace the prey spectrum of microarthropods using molecular gut content analysis.

Stable isotope technique is important for understanding the structure and function of soil food web, which is considered as a belowground black box. We reviewed typical application cases of stable isotope techniques in the research of soil food webs, including to determine food sources and feeding preferences of soil fauna by using isotopes, and to analyze the trophic structure of soil food webs through isotope fractionation effects during the process of feeding and nutrient sequestration by soil fauna. Additionally, stable isotope techniques could reveal the role of soil biota at different trophic levels within soil food web in ecosystem matter and energy flow, which favored to carry out accurate and efficient research on the contribution of soil food webs to soil carbon and nitrogen cycling process and the corresponding influence mechanism. We further put forward the limitations of current stable isotope techniques and the future development directions.
稳定同位素技术的发展为深入揭示地下黑箱-土壤食物网的结构和功能提供了重要的方法和手段。本文回顾了稳定同位素技术在土壤食物网研究中的应用,主要包括利用同位素确定土壤动物的食源和食性偏好;通过土壤动物取食与固持养分过程中的同位素分馏效应,分析土壤食物网的营养级结构;此外,稳定同位素技术还揭示了土壤食物网各营养级土壤生物在土壤生态系统物质能量流动中的作用,有助于准确高效地开展土壤食物网在土壤碳氮循环过程中的贡献及作用机理研究。本文同时指出了目前稳定同位素技术的局限性,展望了未来重点发展的方向。.

Selecting the appropriate indicators and measuring time point numbers is important for accurately examining the shift in soil gross decomposition channel structure. Through a selected case study on a natural forest vs. rainfed arable system over a two-month-long experiment, the utility of three commonly employed indicators (fungi to bacteria ratio (F:B), fungivore to bacterivore ratio (FF:BF), and glucosamine to muramic acid ratio (GlcN:MurN)) were compared to reflect the shift in soil gross decomposition channel structure. The requirement of measuring the time point numbers for the three indicators was also assessed, and we suggest a potential methodology. Our results revealed that the GlcN:MurN ratio was more reliable for assessing the shifts in gross decomposition channel structure for long-term land use changes, while it was less sensitive to short-term drought compared with the other two indicators. The F:B ratio was more applicable than the FF:BF ratio for reflecting both long- and short-term changes. Furthermore, the reliability of the GlcN:MurN ratio was the least dependent on measuring time point numbers. We suggest the use of multiple indicators and the adoption of multiple measuring time points for the overall methodology.