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Summary: Rising atmospheric carbon dioxide concentrations (CO2) and atmospheric nitrogen (N) deposition have contrasting effects on ectomycorrhizal (EM) and arbuscular mycorrhizal (AM) symbioses, potentially mediating forest responses to environmental change.In this study, we evaluated the cumulative effects of historical environmental change on N concentrations and δ15N values in AM plants, EM plants, EM fungi, and saprotrophic fungi using herbarium specimens collected in Minnesota, USA from 1871 to 2016. To better understand mycorrhizal mediation of foliar δ15N, we also analyzed a subset of previously published foliar δ15N values from across the United States to parse the effects of N deposition and CO2 rise.Over the last century in Minnesota, N concentrations declined among all groups except saprotrophic fungi. δ15N also declined among all groups of plants and fungi; however, foliar δ15N declined less in EM plants than in AM plants. In the analysis of previously published foliar δ15N values, this slope difference between EM and AM plants was better explained by nitrogen deposition than by CO2 rise.Mycorrhizal type did not explain trajectories of plant N concentrations. Instead, plants and EM fungi exhibited similar declines in N concentrations, consistent with declining forest N status despite moderate levels of N deposition. [ABSTRACT FROM AUTHOR]

Global climate models predict that the frequency and intensity of precipitation events will increase in many regions across the world. However, the biosphere‐climate feedback to elevated precipitation (eP) remains elusive. Here, we report a study on one of the longest field experiments assessing the effects of eP, alone or in combination with other climate change drivers such as elevated CO2 (eCO2), warming and nitrogen deposition. Soil total carbon (C) decreased after a decade of eP treatment, while plant root production decreased after 2 years. To explain this asynchrony, we found that the relative abundances of fungal genes associated with chitin and protein degradation increased and were positively correlated with bacteriophage genes, suggesting a potential viral shunt in C degradation. In addition, eP increased the relative abundances of microbial stress tolerance genes, which are essential for coping with environmental stressors. Microbial responses to eP were phylogenetically conserved. The effects of eP on soil total C, root production, and microbes were interactively affected by eCO2. Collectively, we demonstrate that long‐term eP induces soil C loss, owing to changes in microbial community composition, functional traits, root production, and soil moisture. Our study unveils an important, previously unknown biosphere‐climate feedback in Mediterranean‐type water‐limited ecosystems, namely how eP induces soil C loss via microbe‐plant–soil interplay. [ABSTRACT FROM AUTHOR]

Intermediate wheatgrass (Thinopyrum intermedium; IWG) is a perennial grass under development as a grain and forage crop. Although IWG is known for its ability to take up nitrate and improve water quality, seasonal nitrogen (N) demand and uptake by IWG is not well known. We measured IWG shoot, root, and grain production, tissue N concentration, and soil mineral N at multiple plant growth stages in 1- and 2-year-old IWG stands fertilized with various rates of N: (1) 80 kg N ha−1 applied at spring regrowth (spring), (2) 40–40 kg N ha−1 applied at spring regrowth and anthesis (split), and (3) an unfertilized control. We also calculated nitrogen use efficiency and biomass N yield. Soil mineral N, N-mineralization rates, and plant N concentration increased with fertilization, and lodging increased with spring fertilization, while root physiological N use efficiency (PNUE) declined with fertilization. Seasonally, shoot and root N concentration declined at physiological maturity, while shoot PNUE was highest at maturity, suggesting either that surplus N was allocated to grain or that more biomass was being produced per unit N taken up. In the 1-year-old stand, during fall regrowth, soil mineral N levels were among the lowest; however, the total soil N was highest compared with other sampling times, suggesting a large influx of organic N between physiological maturity and fall regrowth. Based on our results, IWG is well suited to use nitrogen inputs and avoid excess N leaching into groundwater, but it is also clear that IWG has strong seasonal N allocation patterns that should be taken into consideration with N recommendations and best practices. [ABSTRACT FROM AUTHOR]

Fire is a crucial event regulating the structure and functioning of many ecosystems. Yet few studies have focused on how fire affects taxonomic and functional diversities of soil microbial communities, along with changes in plant communities and soil carbon (C) and nitrogen (N) dynamics. Here, we analyze these effects in a grassland ecosystem 9 months after an experimental fire at the Jasper Ridge Global Change Experiment site in California, USA. Fire altered soil microbial communities considerably, with community assembly process analysis showing that environmental selection pressure was higher in burned sites. However, a small subset of highly connected taxa was able to withstand the disturbance. In addition, fire decreased the relative abundances of most functional genes associated with C degradation and N cycling, implicating a slowdown of microbial processes linked to soil C and N dynamics. In contrast, fire stimulated above‐ and belowground plant growth, likely enhancing plant–microbe competition for soil inorganic N, which was reduced by a factor of about 2. To synthesize those findings, we performed structural equation modeling, which showed that plants but not microbial communities were responsible for significantly higher soil respiration rates in burned sites. Together, our results demonstrate that fire ‘reboots’ the grassland ecosystem by differentially regulating plant and soil microbial communities, leading to significant changes in soil C and N dynamics.Fire significantly increased environmental selection pressure on soil microbial community, where a small subset of highly connected taxa was able to withstand the disturbance. Fire decreased the relative abundances of most functional genes associated with C degradation and N cycling, but stimulated above‐ and belowground plant growth, likely enhancing plant–microbe competition for soil inorganic N. Plants but not microbial communities were responsible for significantly higher soil respiration rates in burned sites. [ABSTRACT FROM AUTHOR]

Soil microbial communities play a critical role in nutrient transformation and storage in all ecosystems. Quantifying the seasonal and long-term temporal extent of genetic and functional variation of soil microorganisms in response to biotic and abiotic changes within and across ecosystems will inform our understanding of the effect of climate change on these processes. We examined spatial and seasonal variation in microbial communities based on 16S rRNA gene sequencing and phospholipid fatty acid (PLFA) composition across four biomes: a tropical broadleaf forest (Hawaii), taiga (Alaska), semiarid grassland-shrubland (Utah), and a subtropical coniferous forest (Florida). In this study, we used a team-based instructional approach leveraging the iPlant Collaborative to examine publicly available National Ecological Observatory Network (NEON) 16S gene and PLFA measurements that quantify microbial diversity, composition, and growth. Both profiling techniques revealed that microbial communities grouped strongly by ecosystem and were predominately influenced by three edaphic factors: pH, soil water content, and cation exchange capacity. Temporal variability of microbial communities differed by profiling technique; 16S-based community measurements showed significant temporal variability only in the subtropical coniferous forest communities, specifically through changes within subgroups of Acidobacteria. Conversely, PLFA-based community measurements showed seasonal shifts in taiga and tropical broadleaf forest systems. These differences may be due to the premise that 16S-based measurements are predominantly influenced by large shifts in the abiotic soil environment, while PLFA-based analyses reflect the metabolically active fraction of the microbial community, which is more sensitive to local disturbances and biotic interactions. To address the technical issue of the response of soil microbial communities to sample storage temperature, we compared 16S-based community structure in soils stored at -80°C and -20°C and found no significant differences in community composition based on storage temperature. Free, open access datasets and data sharing platforms are powerful tools for integrating research and teaching in undergraduate and graduate student classrooms. They are a valuable resource for fostering interdisciplinary collaborations, testing ecological theory, model development and validation, and generating novel hypotheses. Training in data analysis and interpretation of large datasets in university classrooms through project-based learning improves the learning experience for students and enables their use of these significant resources throughout their careers. [ABSTRACT FROM AUTHOR]

Nests of the yellow meadow ant (Lasius flavus) occur at high densities in grasslands worldwide. Although many studies have shown that L. flavus nests influence soil nutrient contents, little is known about their effect on soil nutrient cycling rates. The aim of this study was to examine the role of nest-mounds inhabited by L. flavus as potential 'hot spots' for soil nutrient cycling. Six pairs of nest-mounds and control soils were selected at a grassland site at the plateau of the Alter Gleisberg (Thuringia, Central Germany). L. flavus significantly modified the soil environment within the nest. In comparison to the control soils, nest-mounds were characterized by slightly higher soil temperatures during the summer months. In addition, we found that nests were related to decreased potential C mineralization rates and increased potential net N mineralization rates. Nest-mound soil exhibited lower amounts of SOC, hot-water extractable DOC and DN, and higher concentrations of leachable DOC and DN. Moreover, ants promoted the enrichment of base cations in the nest. Differences in the soil environment between nests and control soils were possibly a result of the burrowing activity of ants, soil mixing, accumulation of aphid honeydew, and decreased plant-derived nutrient inputs into the nest-mound soil. In conclusion, L. flavus nest-mounds had a significant but element dependent effect on the soil nutrient cycling and may represent cold spots for C cycling and hot spots for N cycling. Thus, L. flavus nests increase the spatial heterogeneity of soil properties and create unique micro-sites within grassland ecosystems. [ABSTRACT FROM AUTHOR]

The widespread paradigm in ecology that community structure determines function has recently been challenged by the high complexity of microbial communities. Here, we investigate the patterns of and connections between microbial community structure and microbially-mediated ecological function across different forest management practices and temporal changes in leaf litter across beech forest ecosystems in Central Europe. Our results clearly indicate distinct pattern of microbial community structure in response to forest management and time. However, those patterns were not reflected when potential enzymatic activities of microbes were measured. We postulate that in our forest ecosystems, a disconnect between microbial community structure and function may be present due to differences between the drivers of microbial growth and those of microbial function. [ABSTRACT FROM AUTHOR]

Living roots and their rhizodeposits affect microbial activity and soil carbon (C) and nitrogen (N) mineralization. This so-called rhizosphere priming effect (RPE) has been increasingly recognized recently. However, the magnitude of the RPE and its driving mechanisms remain elusive. Here we investigated the RPE of two plant species (soybean and sunflower) grown in two soil types (a farm or a prairie soil) and sampled at two phenological stages (vegetative and mature stages) over an 88-day period in a greenhouse experiment. We measured soil C mineralization using a continuous 13C-labeling method, and quantified gross N mineralization with a 15N-pool dilution technique. We found that living roots significantly enhanced soil C mineralization, by 27-245%. This positive RPE on soil C mineralization did not vary between the two soils or the two phenological stages, but was significantly greater in sunflower compared to soybean. The magnitude of the RPE was positively correlated with rhizosphere respiration rate across all treatments, suggesting the variation of RPE among treatments was likely caused by variations in root activity and rhizodeposit quantity. Moreover, living roots stimulated gross N mineralization rate by 36-62% in five treatments, while they had no significant impact in the other three treatments. We also quantified soil microbial biomass and extracellular enzyme activity when plants were at the vegetative stage. Generally, living roots increased microbial biomass carbon by 0-28%, β-glucosidase activity by 19-56%, and oxidative enzyme activity by 0-46%. These results are consistent with the positive rhizosphere effect on soil C (45-79%) and N (10-52%) mineralization measured at the same period. We also found significant positive relationships between β-glucosidase activity and soil C mineralization rates and between oxidative enzyme activity and gross N mineralization rates across treatments. These relationships provide clear evidence for the microbial activation hypothesis of RPE. Our results demonstrate that root-soil-microbial interactions can stimulate soil C and N mineralization through rhizosphere effects. The relationships between the RPE and rhizosphere respiration rate and soil enzyme activity can be used for explicit representations of RPE in soil organic matter models. [ABSTRACT FROM AUTHOR]

Biodiversity-ecosystem functioning ( BEF) experiments address ecosystem-level consequences of species loss by comparing communities of high species richness with communities from which species have been gradually eliminated. BEF experiments originally started with microcosms in the laboratory and with grassland ecosystems. A new frontier in experimental BEF research is manipulating tree diversity in forest ecosystems, compelling researchers to think big and comprehensively., We present and discuss some of the major issues to be considered in the design of BEF experiments with trees and illustrate these with a new forest biodiversity experiment established in subtropical China ( Xingangshan, Jiangxi Province) in 2009/2010. Using a pool of 40 tree species, extinction scenarios were simulated with tree richness levels of 1, 2, 4, 8 and 16 species on a total of 566 plots of 25·8 × 25·8 m each., The goal of this experiment is to estimate effects of tree and shrub species richness on carbon storage and soil erosion; therefore, the experiment was established on sloped terrain. The following important design choices were made: (i) establishing many small rather than fewer larger plots, (ii) using high planting density and random mixing of species rather than lower planting density and patchwise mixing of species, (iii) establishing a map of the initial 'ecoscape' to characterize site heterogeneity before the onset of biodiversity effects and (iv) manipulating tree species richness not only in random but also in trait-oriented extinction scenarios., Data management and analysis are particularly challenging in BEF experiments with their hierarchical designs nesting individuals within-species populations within plots within-species compositions. Statistical analysis best proceeds by partitioning these random terms into fixed-term contrasts, for example, species composition into contrasts for species richness and the presence of particular functional groups, which can then be tested against the remaining random variation among compositions., We conclude that forest BEF experiments provide exciting and timely research options. They especially require careful thinking to allow multiple disciplines to measure and analyse data jointly and effectively. Achieving specific research goals and synergy with previous experiments involves trade-offs between different designs and requires manifold design decisions. [ABSTRACT FROM AUTHOR]

We used microbial lipid analysis to analyze microbial biomass and community structure during 6 years of experimental treatment at the Jasper Ridge Global Change Experiment ( JRGCE), a long-term multi-factor global change experiment in a California annual grassland. The microbial community fingerprint and specific biomarkers varied substantially from year to year, in both control and experimental treatment plots. Possible drivers of the variability included plant growth, soil moisture, and ambient temperature. Surprisingly, background variation in the microbial community was of a larger magnitude than even very significant treatment effects, and this variation appeared to constrain responses to treatment. Microbial communities were mostly not responsive or not consistently responsive to the experimental treatments. Both arbuscular mycorrhizal fungi biomarker abundance (16 : 1 ω5c) and the fungal to bacterial ratio were lower under nitrogen addition in most years. Bacterial lipid biomarker abundances (15 : 0 iso and 16 : 1 ω7c) were higher under nitrogen addition in 2002, the year of largest microbial biomass, suggesting that bacteria could respond more to nitrogen addition in years of better growth conditions. Nitrogen addition and warming led to an interactive effect on the Gram-positive bacterial biomarker and the fungal to bacterial ratio. These patterns indicate that in California grassland ecosystems, microbial communities may not respond substantially to future changes in climate and that nitrogen deposition may be a determinant of the soil response to global change. Further, year-to-year variation in microbial growth or community composition may be important determinants of ecosystem response to global change. [ABSTRACT FROM AUTHOR]

It is increasingly recognized that a detailed understanding of the impacts of land use on soil carbon pools and microbially mediated carbon dynamics is required in order to accurately describe terrestrial carbon budgets and improve soil carbon retention. Toward this understanding, we analyzed the levels of biomarkers including phospholipid fatty acids, amino acids, monosaccharides, amino sugars, and several indicators of labile and stabilized carbon in soil samples from a long-term agricultural field experiment. Our results imply that the composition of soil organic carbon (SOC) strongly depends on both the applied fertilization regime and the cultivated crop. In addition, our approach allowed us to identify possible mechanisms of microbial growth and contributions to soil carbon storage under different long-term agricultural management regimes. Amino acids and monosaccharides were quantitatively the most dominant biomarkers and their levels correlated strongly positively with microbial biomass. The relative contributions of the studied biomarkers to the total SOC varied only slightly among the treatments except in cases of extreme fertilization and without any fertilizer. In case of extreme fertilization and with alfalfa as crop type, we found evidence for accumulation of microbially derived monosaccharides and amino acids within the labile OC pool, probably resulting from soil C saturation. Interestingly, we also found an accumulation of microbially derived monosaccharides and amino acids in completely unfertilized plots, which we assumed to be caused by the smaller pore space volume and subsequent oxygen limitation for microbial growth. Mineral fertilization also had substantial effects on soil organic N when applied to plots containing alfalfa, a leguminous plant. Our results demonstrate that over-fertilization, fertilizer type, and the cultivated crop type can have major impacts on the turnover and composition of soil organic carbon, and should be considered when assessing management effects on soil C dynamics. [ABSTRACT FROM AUTHOR]

Fires transform soil microbial communities directly via heat-induced mortality and indirectly by altering plant and soil characteristics. Emerging evidence suggests the magnitude of changes to some plant and soil properties increases with burn severity, but the persistence of changes varies among plant and soil characteristics, ranging from months to years post-fire. Thus, which environmental attributes shape microbial communities at intermediate time points during ecosystem recovery, and how these characteristics vary with severity, remains poorly understood. We identified the network of properties that influence microbial communities three years after fire, along a burn severity gradient in Sierra Nevada mixed-conifer forest. We used phospholipid fatty acid (PLFA) analysis and bacterial 16S-rDNA amplicon sequencing to characterize the microbial community in mineral soil. Using structural equation modelling, we applied a systems approach to identifying the interconnected relationships among severity, vegetation, soil, and microbial communities. Dead tree basal area, soil pH, and extractable phosphorus increased with severity, whereas live tree basal area, forest floor mass, and the proportion of the ≥53 μm soil fraction decreased. Forest floor loss was associated with decreased soil moisture across the severity gradient, decreased live tree basal area was associated with increased shrub coverage, and increased dead tree basal area was associated with increases in total and inorganic soil nitrogen. Soil fungal abundance decreased across the severity gradient, despite a slightly positive response of fungi to lower soil moisture in high severity areas. Bacterial phylogenetic diversity was negatively related to severity and was driven by differences in nutrients and soil texture. The abundance of Bacteroidetes increased and the abundance of Acidobacteria decreased across the severity gradient due to differences in soil pH. Overall, we found that the effects of burn severity on vegetation and soil physicochemical characteristics interact to shape microbial communities at an intermediate time point in ecosystem recovery., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Microbial communities are important drivers and regulators of ecosystem processes. To understand how management of ecosystems may affect microbial communities, a relatively precise but effort-intensive technique to assay microbial community composition is phospholipid fatty acid (PLFA) analysis. PLFA was developed to analyze phospholipid biomarkers, which can be used as indicators of microbial biomass and the composition of broad functional groups of fungi and bacteria. It has commonly been used to compare soils under alternative plant communities, ecology, and management regimes. The PLFA method has been shown to be sensitive to detecting shifts in microbial community composition. An alternative method, fatty acid methyl ester extraction and analysis (MIDI-FA) was developed for rapid extraction of total lipids, without separation of the phospholipid fraction, from pure cultures as a microbial identification technique. This method is rapid but is less suited for soil samples because it lacks an initial step separating soil particles and begins instead with a saponification reaction that likely produces artifacts from the background organic matter in the soil. This article describes a method that increases throughput while balancing effort and accuracy for extraction of lipids from the cell membranes of microorganisms for use in characterizing both total lipids and the relative abundance of indicator lipids to determine soil microbial community structure in studies with many samples. The method combines the accuracy achieved through PLFA profiling by extracting and concentrating soil lipids as a first step, and a reduction in effort by saponifying the organic material extracted and processing with the MIDI-FA method as a second step.

Global environmental change is predicted to have major consequences for carbon cycling and the functioning of soil ecosystems. However, we have limited knowledge about its impacts on the microorganisms, which act as a "valve" between carbon sequestered in soils versus released into the atmosphere. In this study we examined microbial response to continuous 9-years manipulation of three global change factors (elevated CO2, warming, and nitrogen deposition), singly and in combination using two methods: lipid and amino sugar biomarkers at the Jasper Ridge Global Change Experiment (JRGCE). The two methods yielded important distinctions. There were limited microbial lipid differences, but many significant effects for microbial amino sugars. We found that CO2 was not a direct factor influencing soil carbon and major amino sugar pools, but had a positive impact on bacterial-derived muramic acid. Likewise, warming and nitrogen deposition appeared to enrich residues specific to bacteria despite an overall depletion in total amino sugars. The results indicate that elevated CO2, warming, and nitrogen deposition all appeared to increase bacterial-derived residues, but this accumulation effect was far offset by a corresponding decline in fungal residues. The sensitivity of microbial residue biomarker amino sugars to warming and nitrogen deposition may have implications for our predictions of global change impacts on soil stored carbon.