Warming effects on grassland soil microbial communities are amplified in cool months.

Global warming modulates soil respiration (RS) via microbial decomposition, which is seasonally dependent. Yet, the magnitude and direction of this modulation remain unclear, partly owing to the lack of knowledge on how microorganisms respond to seasonal changes. Here, we investigated the temporal dynamics of soil microbial communities over 12 consecutive months under experimental warming in a tallgrass prairie ecosystem. The interplay between warming and time altered (P < 0.05) the taxonomic and functional compositions of microbial communities. During the cool months (January to February and October to December), warming induced a soil microbiome with a higher genomic potential for carbon decomposition, community-level ribosomal RNA operon (rrn) copy numbers, and microbial metabolic quotients, suggesting that warming stimulated fast-growing microorganisms that enhanced carbon decomposition. Modeling analyses further showed that warming reduced the temperature sensitivity of microbial carbon use efficiency (CUE) by 28.7% when monthly average temperature was low, resulting in lower microbial CUE and higher heterotrophic respiration (Rh) potentials. Structural equation modeling showed that warming modulated both Rh and RS directly by altering soil temperature and indirectly by influencing microbial community traits, soil moisture, nitrate content, soil pH, and gross primary productivity. The modulation of Rh by warming was more pronounced in cooler months compared to warmer ones. Together, our findings reveal distinct warming-induced effects on microbial functional traits in cool months, challenging the norm of soil sampling only in the peak growing season, and advancing our mechanistic understanding of the seasonal pattern of RS and Rh sensitivity to warming.

Experimental warming accelerates positive soil priming in a temperate grassland ecosystem.

Unravelling biosphere feedback mechanisms is crucial for predicting the impacts of global warming. Soil priming, an effect of fresh plant-derived carbon (C) on native soil organic carbon (SOC) decomposition, is a key feedback mechanism that could release large amounts of soil C into the atmosphere. However, the impacts of climate warming on soil priming remain elusive. Here, we show that experimental warming accelerates soil priming by 12.7% in a temperate grassland. Warming alters bacterial communities, with 38% of unique active phylotypes detected under warming. The functional genes essential for soil C decomposition are also stimulated, which could be linked to priming effects. We incorporate lab-derived information into an ecosystem model showing that model parameter uncertainty can be reduced by 32-37%. Model simulations from 2010 to 2016 indicate an increase in soil C decomposition under warming, with a 9.1% rise in priming-induced CO2 emissions. If our findings can be generalized to other ecosystems over an extended period of time, soil priming could play an important role in terrestrial C cycle feedbacks and climate change.

Simulating Temperature in a Soil Incubation Experiment.

The study of warming impact on soils requires a realistic and accurate representation of temperature. In laboratory incubation studies, a widely adopted method has been to render constant temperatures in multiple chambers, and via comparisons of soil responses between low- and high-temperature chambers, to derive the warming impact on soil changes. However, this commonly used method failed to imitate both the magnitude and amplitude of actual temperatures as observed in field conditions, thus potentially undermining the validity of such studies. With sophisticated environmental chambers becoming increasingly available, it is imperative to examine alternative methods of temperature control for soil incubation research. This protocol will introduce a state-of-the-art environmental chamber and demonstrate both conventional and new methods of temperature control to improve the experimental design of soil incubation. The protocol mainly comprises four steps: temperature monitoring and programming, soil collection, laboratory incubation, and warming effect comparison. One example will be presented to demonstrate different methods of temperature control and the resultant contrasting warming scenarios; that is, a constant temperature design referred to as stepwise warming (SW) and simulated in situ temperature design as gradual warming (GW), as well as their effects on soil respiration, microbial biomass, and extracellular enzyme activities. In addition, we present a strategy to diversify temperature change scenarios to meet specific climate change research needs (e.g., extreme heat). The temperature control protocol and the recommended well-tailored and diversified temperature change scenarios will assist researchers in establishing reliable and realistic soil incubation experiments in the laboratory.

Reduction of microbial diversity in grassland soil is driven by long-term climate warming.

Anthropogenic climate change threatens ecosystem functioning. Soil biodiversity is essential for maintaining the health of terrestrial systems, but how climate change affects the richness and abundance of soil microbial communities remains unresolved. We examined the effects of warming, altered precipitation and annual biomass removal on grassland soil bacterial, fungal and protistan communities over 7 years to determine how these representative climate changes impact microbial biodiversity and ecosystem functioning. We show that experimental warming and the concomitant reductions in soil moisture play a predominant role in shaping microbial biodiversity by decreasing the richness of bacteria (9.6%), fungi (14.5%) and protists (7.5%). Our results also show positive associations between microbial biodiversity and ecosystem functional processes, such as gross primary productivity and microbial biomass. We conclude that the detrimental effects of biodiversity loss might be more severe in a warmer world.

Multi-year incubation experiments boost confidence in model projections of long-term soil carbon dynamics.

Global soil organic carbon (SOC) stocks may decline with a warmer climate. However, model projections of changes in SOC due to climate warming depend on microbially-driven processes that are usually parameterized based on laboratory incubations. To assess how lab-scale incubation datasets inform model projections over decades, we optimized five microbially-relevant parameters in the Microbial-ENzyme Decomposition (MEND) model using 16 short-term glucose (6-day), 16 short-term cellulose (30-day) and 16 long-term cellulose (729-day) incubation datasets with soils from forests and grasslands across contrasting soil types. Our analysis identified consistently higher parameter estimates given the short-term versus long-term datasets. Implementing the short-term and long-term parameters, respectively, resulted in SOC loss (-8.2 ± 5.1% or -3.9 ± 2.8%), and minor SOC gain (1.8 ± 1.0%) in response to 5 °C warming, while only the latter is consistent with a meta-analysis of 149 field warming observations (1.6 ± 4.0%). Comparing multiple subsets of cellulose incubations (i.e., 6, 30, 90, 180, 360, 480 and 729-day) revealed comparable projections to the observed long-term SOC changes under warming only on 480- and 729-day. Integrating multi-year datasets of soil incubations (e.g., > 1.5 years) with microbial models can thus achieve more reasonable parameterization of key microbial processes and subsequently boost the accuracy and confidence of long-term SOC projections.

Effects of nitrogen fertilization and bioenergy crop species on central tendency and spatial heterogeneity of soil glycosidase activities.

Extracellular glycosidases in soil, produced by microorganisms, act as major agents for decomposing labile soil organic carbon (e.g., cellulose). Soil extracellular glycosidases are significantly affected by nitrogen (N) fertilization but fertilization effects on spatial distributions of soil glycosidases have not been well addressed. Whether the effects of N fertilization vary with bioenergy crop species also remains unclear. Based on a 3-year fertilization experiment in Middle Tennessee, USA, a total of 288 soil samples in topsoil (0-15 cm) were collected from two 15 m2 plots under three fertilization treatments in switchgrass (SG: Panicum virgatum L.) and gamagrass (GG: Tripsacum dactyloides L.) using a spatially explicit design. Four glycosidases, α-glucosidase (AG), ß-glucosidase (BG), ß-xylosidase (BX), cellobiohydrolase (CBH), and their sum associated with C acquisition (Cacq) were quantified. The three fertilization treatments were no N input (NN), low N input (LN: 84 kg N ha-1 year-1 in urea) and high N input (HN: 168 kg N ha-1 year-1 in urea). The descriptive and geostatistical approaches were used to evaluate their central tendency and spatial heterogeneity. Results showed significant interactive effects of N fertilization and crop type on BX such that LN and HN significantly enhanced BX by 14% and 44% in SG, respectively. The significant effect of crop type was identified and glycosidase activities were 15-39% higher in GG than those in SG except AG. Within-plot variances of glycosidases appeared higher in SG than GG but little differed with N fertilization due to large plot-plot variation. Spatial patterns were generally more evident in LN or HN plots than NN plots for BG in SG and CBH in GG. This study suggested that N fertilization elevated central tendency and spatial heterogeneity of glycosidase activities in surficial soil horizons and these effects however varied with crop and enzyme types. Future studies need to focus on specific enzyme in certain bioenergy cropland soil when N fertilization effect is evaluated.

Effects of nitrogen fertilization and bioenergy crop type on topsoil organic carbon and total Nitrogen contents in middle Tennessee USA.

Nitrogen (N) fertilization affects bioenergy crop growth and productivity and consequently carbon (C) and N contents in soil, it however remains unclear whether N fertilization and crop type individually or interactively influence soil organic carbon (SOC) and total N (TN). In a three-year long fertilization experiment in switchgrass (SG: Panicum virgatum L.) and gamagrass (GG: Tripsacum dactyloides L.) croplands in Middle Tennessee USA, soil samples (0-15cm) were collected in plots with no N input (NN), low N input (LN: 84 kg N ha-1 yr-1 in urea) and high N input (HN: 168 kg N ha-1 yr-1 in urea). Besides SOC and TN, the aboveground plant biomass was also quantified. In addition to a summary of published root morphology data based on a separated mesocosm experiment, the root leachable dissolved organic matter (DOM) of both crops was also measured using archived samples. Results showed no significant interaction of N fertilization and crop type on SOC, TN or plant aboveground biomass (ABG). Relative to NN, HN (not LN) significantly increased SOC and TN in both crops. Though SG showed a 15-68% significantly higher ABG than GG, GG showed a 9.3-12% significantly higher SOC and TN than SG. The positive linear relationships of SOC or TN with ABG were identified for SG. However, GG showed structurally more complex and less readily decomposed root DOM, a larger root volume, total root length and surface area than SG. Collectively, these suggested that intensive N fertilization could increase C and N stocks in bioenergy cropland soils but these effects may be more likely mediated by the aboveground biomass in SG and root chemistry and morphology in GG. Future studies are expected to examine the root characteristics in different bioenergy croplands under the field fertilization experiment.

Nitrogen Fertilization Restructured Spatial Patterns of Soil Organic Carbon and Total Nitrogen in Switchgrass and Gamagrass Croplands in Tennessee USA.

Nitrogen (N) fertilizers can potentially alter spatial distribution of soil organic carbon (SOC) and total nitrogen (TN)  concentrations in croplands such as switchgrass (SG: Panicum virgatum L.) and gamagrass (GG: Tripsacum dactyloides L.), but it remains unclear whether these effects are the same between crops and under different rates of fertilization. 13C and 15N are  two important proxy measures of soil biogeochemistry, but they were rarely examined as to their spatial distributions in soil. Based on a three-year long fertilization experiment in Middle Tennessee, USA, the top mineral horizon soils (0-15 cm) were collected using a spatially explicit design within two 15-m2 plots under three fertilization treatments in SG and GG croplands. A total of 288 samples were collected based on 12 plots and 24 samples in each plot. The fertilization treatments were no N input (NN), low N input (LN: 84 kg N ha-1 in urea) and high N input (HN: 168 kg N ha-1 in urea). The SOC, TN, SOC/TN (C: N), δ13C and δ15N were quantified and their within-plot variations and spatial distributions were achieved via descriptive and geostatistical methods. Results showed that SG generally displayed 10~120% higher plot-level variations in all variables than GG, and the plot-level variations were 20~77% higher in NN plots than LN and HN plots in SG but they were comparable in unfertilized and fertilized plots in GG. Relative to NN, LN and HN showed more significant surface trends and spatial structures in SOC and TN in both croplands, and the fertilization effect appeared more pronounced in SG. Spatial patterns in C: N, δ13C and δ15N were comparable among different fertilization treatments in both croplands. The descending within-plot variations were also identified among variables (SOC > TN > δ15N > C: N > δ13C). This study demonstrated that N fertilizations generally reduced the plot-level variance and simultaneously re-established spatial structures of SOC and TN in bioenergy croplands, which little varied with fertilization rate but was more responsive in switchgrass cropland.

Effects of fly ash application on plant biomass and element accumulations: a meta-analysis.

Fly ash generated from coal-fired power plants is a source of potential pollutants, but can be used as a soil ameliorant to increase plant biomass and yield in agriculture. However, the effects of fly ash soil application on plant biomass and the accumulation of both nutrient and toxic elements in plants remain unclear. Based on 85 articles, we conducted a comprehensive meta-analysis to evaluate changes in plant biomass and concentrations of 21 elements in plants in response to fly ash application. These elements included macro-nutrients (N, P, K, Ca, and S), micro-nutrients (B, Co, Cu, Fe, Mn, Mo, Ni, and Zn), and metal(loid)s (Al, As, Cd, Cr, Pb, and Se). Overall, fly ash application decreased plant biomass by 15.2%. However, plant biomass was enhanced by fly ash application by 11.6-29.2% at lower application rates (i.e. <25% of soil mass), and decreased by 45.8% at higher application rates (i.e. 50-100%). Belowground biomass was significantly reduced while yield was enhanced by fly ash application. Most of the element concentrations in plants were enhanced by fly ash application, and followed a descending order with metal(loid)s > micro-nutrients > macro-nutrients. Concentrations of elements tended to increase with an increase in fly ash application rate. Our syntheses indicated that fly ash should be applied at less than 25% in order to enhance plant biomass and yield but avoid high accumulations of metal(loid)s.

Nitrogen Fertilization Elevated Spatial Heterogeneity of Soil Microbial Biomass Carbon and Nitrogen in Switchgrass and Gamagrass Croplands.

The effects of intensive nitrogen (N) fertilizations on spatial distributions of soil microbes in bioenergy croplands remain unknown. To quantify N fertilization effect on spatial heterogeneity of soil microbial biomass carbon (MBC) and N (MBN), we sampled top mineral horizon soils (0-15 cm) using a spatially explicit design within two 15-m2 plots under three fertilization treatments in two bioenergy croplands in a three-year long fertilization experiment in Middle Tennessee, USA. The three fertilization treatments were no N input (NN), low N input (LN: 84 kg N ha-1 in urea) and high N input (HN: 168 kg N ha-1 in urea). The two crops were switchgrass (SG: Panicum virgatum L.) and gamagrass (GG: Tripsacum dactyloides L.). Results showed that N fertilizations little altered central tendencies of microbial variables but relative to LN, HN significantly increased MBC and MBC:MBN (GG only). HN possessed the greatest within-plot variances except for MBN (GG only). Spatial patterns were generally evident under HN and LN plots and much less so under NN plots. Substantially contrasting spatial variations were also identified between croplands (GG > SG) and among variables (MBN, MBC:MBN > MBC). This study demonstrated that spatial heterogeneity is elevated in microbial biomass of fertilized soils likely by uneven fertilizer application in bioenergy crops.

Costimulation of soil glycosidase activity and soil respiration by nitrogen addition.

Unprecedented levels of nitrogen (N) have been deposited in ecosystems over the past century, which is expected to have cascading effects on microbially mediated soil respiration (SR). Extracellular enzymes play critical roles on the degradation of soil organic matter, and measurements of their activities are potentially useful indicators of SR. The links between soil extracellular enzymatic activities (EEAs) and SR under N addition, however, have not been established. We therefore conducted a meta-analysis from 62 publications to synthesize the responses of soil EEAs and SR to elevated N. Nitrogen addition significantly increased glycosidase activity (GA) by 13.0%, α-1,4-glucosidase (AG) by 19.6%, ß-1,4-glucosidase (BG) by 11.1%, ß-1,4-xylosidase (BX) by 21.9% and ß-D-cellobiosidase (CBH) by 12.6%. Increases in GA were more evident for long duration, high rate, organic and mixed N addition (combination of organic and inorganic N addition), as well as for studies from farmland. The response ratios (RRs) of GA were positively correlated with the SR-RRs, even when evaluated individually for AG, BG, BX and CBH. This positive correlation between GA-RR and SR-RR was maintained for most types of vegetation and soil as well as for different methods of N addition. Our results provide the first evidence that GA is linked to SR under N addition over a range of ecosystems and highlight the need for further studies on the response of other soil EEAs to various global change factors and their implications for ecosystem functions.