Using landscape and depth gradients to decouple the impact of correlated environmental variables on soil microbial community composition

Simultaneously assessing shifts in microbial community composition along landscape and depth gradients allows us to decouple correlations among environmental variables, thus revealing underlying controls on microbial community composition. We examined how soil microbial community composition changed with depth and along a successional gradient of native prairie restoration. We predicted that carbon would be the primary control on both microbial biomass and community composition, and that deeper, low-carbon soils would be more similar to low-carbon agricultural soils than to high carbon remnant prairie soils. Soil microbial community composition was characterized using phospholipid fatty acid (PLFA) analysis, and explicitly linked to environmental data using structural equations modeling (SEM). We found that total microbial biomass declined strongly with depth, and increased with restoration age, and that changes in microbial biomass were largely attributable to changes in soil C and/or N concentrations, together with both direct and indirect impacts of root biomass and magnesium. Community composition also shifted with depth and age: the relative abundance of sulfate-reducing bacteria increased with both depth and restoration age, while gram-negative bacteria declined with depth and age. In contrast to prediction, deeper, low-C soils were more similar to high-C remnant prairie soils than to low-C agricultural soils, suggesting that carbon is not the primary control on soil microbial community composition. Instead, the effects of depth and restoration age on microbial community composition were mediated via changes in available phosphorus, exchangeable calcium, and soil water, together with a large undetermined effect of depth. Only by examining soil microbial community composition shifts across sites and down the soil column simultaneously were we able to tease apart the impact of these correlates environmental variables.     

Soil texture affects soil microbial and structural recovery during grassland restoration

Many biotic and abiotic factors influence recovery of soil communities following prolonged disturbance. We investigated the role of soil texture in the recovery of soil microbial community structure and changes in microbial stress, as indexed by phospholipid fatty acid (PLFA) profiles, using two chronosequences of grasslands restored from 0 to 19 years on silty clay loam and loamy fine sand soils in Nebraska, USA. All restorations were formerly cultivated fields seeded to native warm-season grasses through the USDA’s Conservation Reserve Program. Increases in many PLFA concentrations occurred across the silty clay loam chronosequence including total PLFA biomass, richness, fungi, arbuscular mycorrhizal fungi, Gram-positive bacteria, Gram-negative bacteria, and actinomycetes. Ratios of saturated:monounsaturated and iso:anteiso PLFAs decreased across the silty clay loam chronosequence indicating reduction in nutrient stress of the microbial community as grassland established. Multivariate analysis of entire PLFA profiles across the silty clay loam chronosequence showed recovery of microbial community structure on the trajectory toward native prairie. Conversely, no microbial groups exhibited a directional change across the loamy fine sand chronosequence. Changes in soil structure were also only observed across the silty clay loam chronosequence. Aggregate mean weighted diameter (MWD) exhibited an exponential rise to maximum resulting from an exponential rise to maximum in the proportion of large macroaggregates (>2000 μm) and exponential decay in microaggregates (<250 μm and >53 μm) and the silt and clay fraction (<53 μm). Across both chronosequences, MWD was highly correlated with total PLFA biomass and the biomass of many microbial groups. Strong correlations between many PLFA groups and the MWD of aggregates underscore the interdependence between the recovery of soil microbial communities and soil structure that may explain more variation than time for some soils (i.e., loamy fine sand). This study demonstrates that soil microbial responses to grassland restoration are modulated by soil texture with implications for estimating the true capacity of restoration efforts to rehabilitate ecosystem functions.     

Comparison of soil fungal/bacterial ratios in a pH gradient using physiological and PLFA-based techniques

We have compared the total microbial biomass and the fungal/bacterial ratio estimated using substrate-induced respiration (SIR) in combination with the selective inhibition technique and using the phospholipid fatty acid (PLFA) technique in a pH gradient (3.0-7.2) consisting of 53 mature broad-leaved forest soils. A fungal/bacterial biomass index using the PLFA technique was calculated using the PLFA 18:2ω6,9 as an indicator of fungal biomass and the sum of 13 bacterial specific PLFAs as indicator of the bacterial biomass. Good linear correlation (p<0.001) was found between the total microbial biomass estimated with SIR and total PLFAs (totPLFA), indicating that 1 mg biomass-C was equivalent to 130 nmol totPLFA. Both biomass estimates were positively correlated to soil pH. The fungal/bacterial ratio measured using the selective inhibition technique decreased significantly with increasing pH from about 9 at pH 3 to approximately 2 at pH 7, while the fungal/bacterial biomass index using PLFA measurements tended to increase slightly with increasing soil pH. Good correlation between the soil content of ergosterol and of the PLFA 18:2ω6,9 indicated that the lack of congruency between the two methods in estimating fungal/bacterial ratios was not due to PLFA 18:2ω6,9-related non-fungal structures to any significant degree. Several PLFAs were strongly correlated to soil pH (R² values >0.8); for example the PLFAs 16:1ω5 and 16:1ω7c increased with increasing soil pH, while i16:0 and cy19:0 decreased. A principal component analysis of the total PLFA pattern gave a first component that was strongly correlated to soil pH (R²=0.85, p<0.001) indicating that the microbial community composition in these beech/beech-oak forest soils was to a large extent determined by soil pH.     

Microbial communities of a native perennial bunchgrass do not respond consistently across a gradient of land-use intensification

To test if native perennial bunchgrasses cultivate the same microbial community composition across a gradient in land-use intensification, soils were sampled in fall, winter and spring in areas under bunchgrasses (‘plant’) and in bare soils (‘removal’) in which plots were cleared of living plants adjacent to native perennial bunchgrasses (Nassella pulchra). The gradient in land-use intensification was represented by a relict perennial grassland, a restored perennial grassland, and a perennial grass agriculture site on the same soil type. An exotic annual grassland site was also included because perennial bunchgrasses often exist within a matrix of annual grasses in California. Differences in soil resource pools between ‘plant’ and ‘removal’ soils were observed mainly in the relict perennial grassland and perennial grass agriculture site. Seasonal responses occurred in all sites. Microbial biomass carbon (C) and dissolved organic C were greater under perennial bunchgrasses in the relict perennial grassland and perennial grass agriculture site when comparing treatment means of ‘plant’ vs. ‘removal’ soil. In general, soil moisture, microbial respiration, and nitrate decreased from fall to spring in ‘plant’ and ‘removal’ soils, while soil ammonium and net mineralizable nitrogen (N) increased only in ‘plant’ soils. A canonical correspondence analysis (CCA) of phospholipid fatty acid (PLFA) profiles from all sites showed that land-use history limits the similarity of microbial community composition as do soil C and N dynamics among sites. When PLFA profiles from individual sites were analyzed by CCA, different microbial PLFA markers were associated with N. pulchra in each site, indicating that the same plant species does not retain a unique microbial fingerprint across the gradient of land-use intensification.     

Microbial community structure in restored riparian soils of the Canadian prairie pothole region

In the Prairie Pothole Region (PPR) of Canada, wetlands once utilized for agricultural purposes are restored through the placement of a ditch plug to return them to their pre-existing hydrological state. The overall objective of this research was to assess differences in riparian soil microbial community structure between reference wetlands, those which had never been utilized for agricultural purposes, and restored wetlands, of varying times since restoration. Soil samples (0-6 cm) were taken from 15 reference and 28 restored wetlands. The soil microbial community was characterized using phospholipid fatty acid (PLFA) analysis. Data were analyzed using non-metric multidimensional scaling, multivariate regression trees (MRT) and indicator species analysis. The microbial community of younger restored soils (1-3 and 4-6 yrs) differed significantly from the reference soils, with reference soils having higher microbial biomass, evenness, and diversity. Richness showed an increasing trend with time since restoration. Results from the MRT underlined the importance of climatic factors, specifically precipitation - potential evapotranspiration (P-PE) in explaining the variation found in the microbial community. More specifically, drier sites had strong indicator species values associated with PLFAs of actinomycetal origin and fungal origin. Within the wetter sites, it was found that the older restored sites (7-11 yrs) and reference sites had strong indicator species values associated with PLFAs of Gram negative and fungal origin. The similarities in microbial community composition and biomass of the older restored sites (7-11 yrs) and the reference sites indicate that this component of the wetland ecosystems begins to recover within this time period.     

Multivariate approach to characterizing soil microbial communities in pristine and agricultural sites in Northwest Argentina

Land use effects on microbial communities may have profound impacts on agricultural productivity and ecosystem sustainability as they are critical in soil quality and health. The main aim of this study was to characterize the microbial communities of pristine and agricultural soils in the central Yungas region in Northwest Argentina. As a first step in the development of biological indicators of soil quality in this region, a comprehensive approach involving a structural and functional evaluation of microbial communities was used to detect changes in soil as consequence of land use. The sites selected included two pristine montane forest sites (MF1 and MF2), two plots under sugarcane monoculture for 40 and 100 years (SC40 and SC100), one plot under 20 years of soybean monoculture (SB20), a recently deforested and soybean cropped site (RC), and two reference sites of native forest adjacent to the sugarcane and soybean plots (PF1 and PF2). We used three microbial community profiling methods: denaturing gradient gel electrophoresis (DGGE) analysis of PCR amplified 16S rRNA genes, community-level physiological profiling (CLPP) using a BD oxygen biosensor system (BDOBS-CLPP) and phospholipid fatty acid (PLFA) analysis. Deforestation and agriculture caused expected increases in pH and decreases in organic carbon and microbial biomass. Additionally, shifts in the microbial community structure and physiology were detected with disturbance, including reduced diversity based on PLFA data. The higher respiratory response to several carbon substrates observed in agricultural soils suggested the presence of microbial communities with lower growth yield efficiency that could further reduce carbon storage in these soils.Using an integrated multivariate analysis of all data measured in this study we propose a minimum data set of variables (organic carbon, pH, sucrose and valeric acid utilizations, a17:0 and a15:0 PLFA biomarkers and the value of impact on microbial diversity) to be used for future studies of soil quality in Northwest Argentina.     

Re-establishment of suppressiveness to soil- and air-borne diseases by re-inoculation of soil microbial communities

The aim of this study was to investigate the potentials and limitations in restoring soil suppressiveness in disturbed soils. Soils from three sites in UK and Switzerland (STC, REC, THE) differing in their level of suppressiveness to soil-borne and air-borne diseases were γ-irradiated and this soil matrix was re-inoculated with 1% (w/w) of either parent native soil or native soil from the other sites (‘soil inoculum’). Suppressiveness to air-borne and soil-borne diseases was quantified by means of the host-pathogen systems Lepidium sativum (cress)-Pythium ultimum, an oomycete causing root rot and seedling damping-off, and Arabidopsis thaliana-Hyaloperonospora parasitica, an oomycete causing downy mildew. Soil microbial biomass, activity and community structure, as determined by phospholipid fatty acid (PLFA) profiles, were measured in native, γ-irradiated, and re-inoculated soils. Both, L. sativum and A. thaliana were highly susceptible to the pathogens if grown on γ-irradiated soils. Re-inoculation completely restored suppressiveness of soils to the foliar pathogen H. parasitica, independently of soil matrix or soil inoculum, whereas suppressiveness to P. ultimum depended on the soil matrix and, to a lesser extent, on the soil inoculum. However, the soil with the highest inherent suppressiveness did not reach the initial level of suppressiveness after re-inoculation. In addition, native microbial populations as defined by microbial biomass, activity and community structure, could not be fully restored in re-inoculated soils. As for suppressiveness to P. ultimum, the soil matrix, rather than the source of soil inoculum was identified as the key factor for re-establishing the microbial community structure. Our data show that soils do not or only slowly fully recover from sterilisation by γ-irradiation, indicating that agricultural soil management practices such as soil fumigation or heat treatments frequently used in vegetable cropping should be avoided.     

Microbial community composition on native and drastically disturbed serpentine soils

During construction of roads, entire hillsides can be cut away, dramatically disturbing the ecosystem. Microbial communities play important, but poorly understood roles in revegetating roadcuts because of the many functions they perform in nutrient cycling, plant symbioses, decomposition, and other ecosystem processes. Our objective was to determine relationships among microbial community composition, soil chemistry, and disturbance on a serpentine soil disturbed by a roadcut and then partially revegetated. We hypothesized that the adjacent undisturbed serpentine soil would have a different microbial community composition from barren and revegetated sections of the roadcut and that undisturbed soils would have the greatest microbial biomass and diversity. We measured phospholipid fatty acids (PLFA) and soil nutrient concentrations on barren and revegetated sections of the roadcut and on adjacent undisturbed serpentine and nonserpentine soils. Most roadcut samples had soil chemistry similar to the serpentine reference soil. The microbial biomass and diversity of barren sites was lower than that of revegetated or the serpentine reference site. The nonserpentine reference site had significantly (P≤0.05) greater microbial biomass than serpentine or disturbed sites but significantly lower relative proportions of actinomycetes, and slow growth biomarkers. The Barren site had the lowest microbial biomass and a significantly (P≤0.05) greater proportion of that biomass was fungi. Barren, revegetated, and serpentine sites all had dissimilar microbial community composition. The composition of the revegetated communities, however, was intermediate between the serpentine reference and barren soils, suggesting that community composition of revegetated soils is approaching that of an undisturbed site with similar soil chemistry.     

Development of soil microbial communities during tallgrass prairie restoration

Soil microbial communities were examined in a chronosequence of four different land-use treatments at the Konza Prairie Biological Station, Kansas. The time series comprised a conventionally tilled cropland (CTC) developed on former prairie soils, two restored grasslands that were initiated on former agricultural soils in 1998 (RG₉₈) and 1978 (RG₇₈), and an annually burned native tallgrass prairie (BNP), all on similar soil types. In addition, an unburned native tallgrass prairie (UNP) and another grassland restored in 2000 (RG₀₀) on a different soil type were studied to examine the effect of long-term fire exclusion vs. annual burning in native prairie and the influence of soil type on soil microbial communities in restored grasslands. Both 16S rRNA gene clone libraries and phospholipid fatty acid analyses indicated that the structure and composition of bacterial communities in the CTC soil were significantly different from those in prairie soils. Within the time series, soil physicochemical characteristics changed monotonically. However, changes in the microbial communities were not monotonic, and a transitional bacterial community formed during restoration that differed from communities in either the highly disturbed cropland or the undisturbed original prairie. The microbial communities of RG₉₈ and RG₀₀ grasslands were also significantly different even though they were restored at approximately the same time and were managed similarly; a result attributable to the differences in soil type and associated soil chemistry such as pH and Ca. Burning and seasonal effects on soil microbial communities were small. Similarly, changing plot size from 300 m² to 150 m² in area caused small differences in the estimates of microbial community structure. In conclusion, microbial community structure and biochemical properties of soil from the tallgrass prairie were strongly impacted by cultivation, and the microbial community was not fully restored even after 30 years.     

Response of microbial community composition and activity in agricultural and grassland soils after a simulated rainfall

Rainfall in Mediterranean climates may affect soil microbial processes and communities differently in agricultural vs. grassland soils. We explored the hypothesis that land use intensification decreases the resistance of microbial community composition and activity to perturbation. Soil carbon (C) and nitrogen (N) dynamics and microbial responses to a simulated Spring rainfall were measured in grassland and agricultural ecosystems. The California ecosystems consisted of two paired sets: annual vegetable crops and annual grassland in Salinas Valley, and perennial grass agriculture and native perennial grassland in Carmel Valley. Soil types of the respective ecosystem pairs were derived from granitic parent material and had sandy loam textures. Intact cores (30 cm deep) were collected in March 1999. After equilibration, dry soil cores (approx. −1 to −2 MPa) were exposed to a simulated Spring rainfall of 2.4 cm, and then were measured at 0, 6, 24, and 120 h after rewetting. Microbial biomass C (MBC) and inorganic N did not respond to rewetting. N₂O and CO₂ efflux and respiration increased after rewetting in all soils, with larger responses in the grassland than in the agricultural soils. Phospholipid fatty acid (PLFA) profiles indicated that changes in microbial community composition after rewetting were most pronounced in intensive vegetable production, followed by the relict perennial grassland. Changes in specific PLFA markers were not consistent across all sites. There were more similarities among microbial groups associated with PLFA markers in agricultural ecosystems than grassland ecosystems. Differences in responses of microbial communities may be related to the different plant species composition of the grasslands. Agricultural intensification appeared to decrease microbial diversity, as estimated from numbers of individual PLFA identified for each ecosystem, and reduce resistance to change in microbial community composition after rewetting. In the agricultural systems, reductions in both the measures of microbial diversity and the resistance of the microbial community composition to change after a perturbation were associated with lower ecosystem function, i.e. lower microbial responses to increased moisture availability.