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.     

Role of soil drying in nitrogen mineralization and microbial community function in semi-arid grasslands of north-west Australia

We examined effects of wetting and then progressive drying on nitrogen (N) mineralization rates and microbial community composition, biomass and activity of soils from spinifex (Triodia R. Br.) grasslands of the semi-arid Pilbara region of northern Australia. We compared soils under and between spinifex hummocks and also examined impacts of fire history on soils over a 28 d laboratory incubation. Soil water potentials were initially adjusted to −100 kPa and monitored as soils dried. We estimated N mineralization by measuring changes in amounts of nitrate (NO₃⁻-N) and ammonium (NH₄⁺-N) over time and with change in soil water potential. Microbial activity was assessed by amounts of CO₂ respired. Phospholipid fatty acid (PLFA) analyses were used to characterize shifts in microbial community composition during soil drying. Net N mineralized under hummocks was twice that of open spaces between hummocks and mineralization rates followed first-order kinetics. An initial N mineralization flush following re-wetting accounted for more than 90% of the total amount of N mineralized during the incubation. Initial microbial biomass under hummocks was twice that of open areas between hummocks, but after 28 d microbial biomass was<2 μ g⁻¹ ninhydrin N regardless of position. Respiration of CO₂ from soils under hummocks was more than double that of soils from between hummocks. N mineralization, microbial biomass and microbial activity were negligible once soils had dried to −1000 kPa. Microbial community composition was also significantly different between 0 and 28 d of the incubation but was not influenced by burning treatment or position. Regression analysis showed that soil water potential, microbial biomass N, NO₃⁻-N, % C and δ¹⁵N all explained significant proportions of the variance in microbial community composition when modelled individually. However, sequential multiple regression analysis determined only microbial biomass was significant in explaining variance of microbial community compositions. Nitrogen mineralization rates and microbial biomass did not differ between burned and unburned sites suggesting that any effects of fire are mostly short-lived. We conclude that the highly labile nature of much of soil organic N in these semi-arid grasslands provides a ready substrate for N mineralization. However, process rates are likely to be primarily limited by the amount of substrate available as well as water availability and less so by substrate quality or microbial community composition.     

Soil microbial community composition as affected by restoration practices in California grassland

Agricultural practices have strong impacts on soil microbes including both the indices related to biomass and activity as well as those related to community composition. In a grassland restoration project in California, where native perennial bunchgrasses were introduced into non-native annual grassland after a period of intensive tillage, weeding, and herbicide use to reduce the annual seed bank, microbial community composition was investigated. Three treatments were compared: annual grassland, bare soil fallow, and restored perennial grassland. Soil profiles down to 80 cm in depth were investigated in four separate layers (0-15, 15-30, 30-60, and 60-80 cm) using both phospholipid ester-linked fatty acid (PLFAs) and ergosterol as biomarkers in addition to microbial biomass C by fumigation extraction. PLFA fingerprinting showed much stronger differences between the tilled bare fallow treatment vs. grasslands, compared to fewer differences between restored perennial grassland and annual grassland. The presence or absence of plants over several years clearly distinguished microbial communities. Microbial communities in lower soil layers were little affected by management practices. Regardless of treatment, soil depth caused a strong gradient of changing habitat conditions, which was reflected in Canonical Correspondence Analysis of PLFAs. Fungal organisms were associated with the presence of plants and/or litter since the total amount and the relative proportion of fungal markers were reduced in the tilled bare fallow and in lower layers of the grassland treatments. Total PLFA and soil microbial biomass were highly correlated, and fungal PLFA biomarkers showed strong correlations to ergosterol content. In conclusion, microbial communities are resilient to the grassland restoration process, but do not reflect the change in plant species composition that occurred after planting native bunchgrasses.     

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.     

Community composition and activities of denitrifying bacteria from adjacent agricultural soil, riparian soil, and creek sediment in Oregon, USA

We examined denitrifying bacteria from wet soils and creek sediment in an agroecosystem in Oregon, USA that received inputs of nitrogen (N) fertilizer. Our objective was to determine the variation in denitrifying community composition and activities across three adjacent habitats: a fertilized agricultural field planted to perennial ryegrass, a naturally vegetated riparian area, and creek sediment. Using C₂H₂ inhibition, denitrifying enzyme and N₂O-reductase activities were determined in short-term incubations of anaerobic slurries. A key gene in the denitrification pathway, N₂O reductase (nosZ), served as a marker for denitrifiers. Mean denitrifying enzyme activity (DEA) was similar among habitats, ranging from 0.5 to 1.8 μg N g⁻¹ dry soil h⁻¹. However, the ratio of N₂O production, without C₂H₂, to DEA was substantially higher in riparian soil (0.64±0.02; mean±standard error, n=12) than in agricultural soil (0.19±0.02) or creek sediment (0.32±0.03). Mean N₂O-reductase activity ranged from 0.5 to 3.2 μg N g⁻¹ dry soil h⁻¹, with greater activity in agricultural soil than in riparian soil. Denitrifying community composition differed significantly among habitats based on nosZ terminal-restriction fragment length polymorphisms. The creek sediment community was unique. Communities in the agricultural and riparian soil were more closely related but distinct. A number of unique nosZ genotypes were detected in creek sediment. Sequences of nosZ obtained from riparian soil were closely related to nosZ from Bradyrhizobium japonicum. Although nosZ distribution and N₂O-reductase activity differed among habitats, relationships between activity and community composition appeared uncoupled across the agroecosystem.     

Response of microbial communities to water stress in irrigated and drought-prone tallgrass prairie soils

To better understand how water stress and availability affect the structure of microbial communities in soil, I measured the change in phospholipid fatty acids (PLFA) and the incorporation of ¹³C-labeled glucose into the PLFA following exposure to water stress. Overlaid on the laboratory water stress treatment, samples were collected from drought-prone and irrigated (11 years) tallgrass prairie soil (0-10 cm depth). In the laboratory, soils were either incubated at −250 kPa or dried steadily over a 3-d period to −45 MPa. On the fourth day, the dried samples were brought up to −250 kPa and then all samples received 250 μg of glucose-C (+4000 δ¹³C-PDB) solution that brought them to −33 kPa matric water potential. Samples were then extracted for PLFA following 6 and 24 h of incubation (25 °C). Non-metric multidimensional scaling (NMS) techniques and multi-response permutation procedure (MRPP) showed that the largest effect on the mol% distribution of PLFA was related to the field scale water addition experiment. In response to irrigation, the PLFA 16:1ω5, 18:1+, and 18:2ω6,9 showed increases, and a15:0, a17:0, and cy19:0 showed decreases in their respective mol%. Effects related to the induction of laboratory water stress were predominantly associated with a decrease in the mol% distribution of the putative fungal biomarker (18:2ω6,9) with little to no change in the mol% distribution of the bacterial biomarkers. Interestingly, the flow of C to the microbial community was not strongly related to any single PLFA, and differences were rather subtle, but multivariate MRPP detected change to the community structure related to the laboratory water stress treatment but not related to the 11 years of field irrigation. Our results suggest that both the total and the actively metabolizing bacterial community in soil were generally resistant to the effects of water stress brought by rewetting of dry soil. However, more research is needed to understand the nature of the fungal response to drying and rewetting in soil.     

Do organic inputs alter resistance and resilience of soil microbial community to drying?

Grassland ecosystems in south-eastern Australia are important for dairy and livestock farming. Their productivity relies heavily on water availability, as well as the ecosystem services provided by soil microbial communities including carbon and nutrient cycling. Management practices such as compost application are being encouraged as a means to improve both soil water holding capacity and fertility, thereby buffering against the impacts of increasing climate variability. Such buffering consists of two complementary processes: resistance, which measures the ability of an ecosystem to maintain community structure and function during a period of stress (such as drying); and resilience, which measures the ability of an ecosystem to recover community structure and function post-stress. We investigated the effects of compost on the resistance and resilience of the grassland soil ecosystem under drying and drying with rewetting events, in a terrestrial model ecosystem. Overall, compost addition led to an increase in soil moisture, greater plant available P and higher plant δ¹⁵N. Soil C:nutrient ratios, mineral N content (NH₄⁺ and NO₃⁻) and soil microbial PLFA composition were similar between amended and unamended soils. Rainfall treatment led to differences in soil moisture, plant above-ground and below-ground biomass, plant δ¹⁵N, soil mineral N content (NH₄⁺ and NO₃⁻) and microbial biomass C, N and P composition but had no effects on soil C:nutrient ratios, plant available P and soil microbial PLFA composition. There was little interaction between rainfall and compost. Generally, the soil microbial community was resistant and resilient to fluctuations in rainfall regardless of compost amendment. However, these properties of the soil microbial community were translated to resilience and not resistance in soil functions. Overall, the results below-ground showed much greater response to rainfall than compost amendment. Water was the key factor shaping the soil microbial community, and nutrients were not strong co-limiting factors. Future projections of increasing rainfall variability will have important below-ground functional consequences in the grassland, including altered nutrient cycling.     

Microbial communities, biomass, and activities in soils as affected by freeze thaw cycles

Two Finnish agricultural soils (peat soil and loamy sand) were exposed to four freeze-thaw cycles (FTC), with a temperature change from −17.3±0.4 °C to +4.1±0.4 °C. Control cores from both soils were kept at constant temperature (+6.6±2.0 °C) without FTCs. Soil N₂O and CO₂ emissions were monitored during soil thawing, and the effects of FTCs on soil microbes were studied. N₂O emissions were extremely low in peat soil, possibly due to low soil water content. Loamy sand had high N₂O emission, with the highest emission after the second FTC. Soil freeze-thaw increased anaerobic respiration in both soil types during the first 3-4 FTCs, and this increase was higher in the peat soil. The microbial community structure and biomass analysed with lipid biomarkers (phospholipid fatty acids, 3- and 2- hydroxy fatty acids) were not affected by freezing-thawing cycles, nor was soil microbial biomass carbon (MIB-C). Molecular analysis of the microbial community structure with temperature gradient gel electrophoresis (TGGE) also showed no changes due the FTCs. These results show that freezing and thawing of boreal soils does not have a strong effect on microbial biomass or community structure.     

Identification of groups of metabolically-active rhizosphere microorganisms by stable isotope probing of PLFAs

We combined microbial community phospholipid fatty acid (PLFA) analyses with an in situ stable isotope ¹³CO₂ labelling approach to identify microbial groups actively involved in assimilation of root-derived C in limed grassland soils. We hypothesized that the application of lime would stimulate more rapid ¹³C assimilation and turnover in microbial PLFAs. Four and 8 d after label application, 18:1ω9, 18:2ω6,9 (fungal biomarkers) and 16:1ω7, 18:1ω7, 19:0cy (Gram-negative bacterial biomarkers) showed the most ¹³C enrichment and rapid turnover rates. This suggests that these microorganisms were assimilating recently-photosynthesized root C inputs to soils. Contrary to our hypothesis, liming did not affect assimilation or turnover rates of ¹³C-labelled C. ¹³C stable isotope pulse-labelling technique paired with analyses of PLFA microbial biomarkers shows promise for in situ investigations of microbial function in soils.     

Reaction of microorganisms to rewetting in continuous cereal and legume rotation soils of semi-arid Sub-Saharan Africa

A 20-day incubation experiment with continuous cereal (CC) versus cereal legume (CL) rotation soils of two semi-arid Sub-Saharan sites (Fada-Kouaré in Burkina Faso, F, and Koukombo in Togo, K) were carried out to investigate the effects of rewetting on soil microbial properties. Site- and system-specific reactions of soil microorganisms were observed on cumulative CO₂ production, adenylates (ATP, ADP, and AMP), microbial biomass C and N, ergosterol, muramic acid and glucosamine. Higher values of all parameters were found in the CL rotation soils and in both soils from Fada-Kouaré. While the inorganic N concentration showed only a system-specific response to rewetting, the adenylate energy charge (AEC) showed only a site-specific response. ATP recovered within 6 h after rewetting from ADP and AMP due to rehydration of microorganisms and not due to microbial growth. Consequently, no N seemed to be immobilized by microorganisms and all NO₃ in the soil was immediately available to the plants. The fungal cell-membrane component ergosterol was three (CC) and five (CL) times larger at Fada than in the respective soils at Koukombo. The concentrations of the bacterial cell-wall component muramic acid were by 20% and of mainly fungal glucosamine by 10% larger in the CL rotation soils than in the CC soils. This indicates long-shifts in the microbial community structure.     

Do plant clumps constitute microbial hotspots in semiarid Mediterranean patchy landscapes?

Three semiarid Mediterranean patchy landscapes were investigated to test the existence of a microsite effect (i.e. plant canopy vs. inter-canopy) on soil microbial communities. Surface soil samples were independently taken from both microsites under naturally changing conditions of humidity and temperature through the year. In gypsiferous soils covered with a shrub steppe, improved physical and chemical soil properties were registered underneath the plant canopy, where the densest and most active microbial communities were also detected (e.g. microbial biomass C averaged 531 and 202 mg kg⁻¹ in canopy and inter-canopy areas, respectively). In calcareous perennial tussock grasslands, either growing on soils over limestones or alluvial deposits, the microsite effect was not so marked. Soil humidity, temperature and total organic C were homogeneously distributed over the landscape conditioning their uniform microbial activity under field moisture conditions (ATP content averaged 853 and 885 nmol kg⁻¹ in canopy and intercanopy areas, respectively). However, readily mineralizable C and microbial biomass C were preferentially accumulated in soils underneath the tussocks determining their larger potential microbial activity (e.g. C hydrolysis capacity under optimal conditions). In conclusion, plant clumps either functioned as microbial hotspots where enhanced microbially driven ecosystem processes took place or as microbial banks capable of undergoing a burst of activity under favourable climatic conditions. Our results provide experimental evidence of a non-patchy distribution of certain soil microbial properties in semi-arid Mediterranean patchy ecosystems.     

Grazing effects on microbial community composition, growth and nutrient cycling in salt marsh and sand dune grasslands

The effect of grazing by large herbivores on the microbial community and the ecosystem functions they provide are relatively unknown in grassland systems. In this study, the impact of grazing upon the size, composition and activity of the soil microbial community was measured in field experiments in two coastal ecosystems: one salt marsh and one sand dune grassland. Bacterial, fungal and total microbial biomass were not systematically affected by grazing across ecosystems, although, within an ecosystem, differences could be detected. Fungal-to-bacterial ratio did not differ with grazing for either habitat. Redundancy analysis showed that soil moisture, bulk density and root biomass significantly explained the composition of phospholipid fatty acid (PLFA) markers, dominated by the distinction between the two grassland habitats, but where the grazing effect could also be resolved. PLFA markers for Gram-positive bacteria were more proportionally abundant in un-grazed, and markers for Gram-negative bacteria in grazed grasslands. Bacterial growth rate (leucine incorporation) was highest in un-grazed salt marsh but did not vary with grazing intensity in the sand dune grassland. We conclude that grazing consistently affects the composition of the soil microbial community in semi-natural grasslands but that its influence is small (7 % of the total variation in PLFA composition), compared with differences between grassland types (89 %). The relatively small effect of grazing translated to small effects on measurements of soil microbial functions, including N and C mineralisation. This study is an early step toward assessing consequences of land-use change for global nutrient cycles driven by the microbial community.     

A mass-balance approach to measuring microbial uptake and pools of phosphorus in nutrient-amended soils

Soil microbial biomass P is usually determined through fumigation-extraction (FE), in which partially extractable P from lysed biomass is converted to biomass P using a conversion factor (K_p). Estimation of K_p has been usually based on cultured microorganisms, which may not adequately represent the soil microbial community in either nutrient-poor or in altered carbon and nutrient conditions following fertilisation. We report an alternative approach in which changes in microbial P storage are determined as the residual in a mass balance of extractable P before and after incubation. This approach was applied in three low-fertility sandy soils of southwestern Australia, to determine microbial P immobilisation during 5-day incubations in response to the amendment by 2.323 mg C g⁻¹, 100 μg N g⁻¹ and 20 μg P g⁻¹. The net P immobilisation during the amended incubations determined to be 18.1, 14.1 and 16.3 μg P g⁻¹ in the three soils, accounting for 70.6-90.5% of P added through amendment. Such estimates do not rely on fumigation and K_p values, but for comparison with the FE method we estimated ‘nominal’ K_p values to be 0.20-0.31 for the soils under the amended conditions. Our results showed that microbial P immobilisation was a dominant process regulating P concentration in soil water following the CNP amendment. The mass-balance approach provides information not only about changes in the microbial P compartment, but also about other major P-pools and their fluxes in regulating soil-water P concentrations under substrate- and nutrient-amended conditions.     

Regional and local factors affecting diversity, abundance and activity of free-living, N2-fixing bacteria in Australian agricultural soils

N₂-fixation by free-living (diazotrophic) microorganism is a key process affecting ecosystem functioning in soils. Understanding drivers affecting diazotrophic community assemblages and activities may lead to management practices to increase primary production and/or environmental sustainability. We used PCR-DGGE to determine the fundamental relationships between diazotrophic community structure and in a wide range of soils across southern Australia. In addition qPCR, RT-qPCR and N₂-fixation (acetylene reduction) were used to investigate factors influencing gene abundance, expression and processes in similar soils with different agricultural inputs. Across 22 soils, the structural composition of the nifH community was significantly influenced by site (ANOSIM R = 0.876; P = 0.001). The effects of management practices were evident, and often larger than between-soil differences, but were only present at some sites. Differences in nifH communities between sites correlated to particulate organic carbon (POC; measured by mid-infrared spectroscopy) content of the soils (BIO-ENV test; ρ = 0.502; P = 0.001), but not other factors including total soil C. In 3 soils from the Murrumbidgee irrigation region of NSW, intensification of the farming systems was associated with increasing N₂-fixation (P < 0.05), except where rice was cultivated. N₂-fixation correlated either with nifH abundance or gene expression in soils, but not both. Our data shows that soil C is closely linked to diazotrophic ecology. Principally, the amount of C entering the soil system is directly related to the abundance and N₂-fixation activity of free-living bacteria. However, we also show that C in the POC pool has associative links to the genetic diversity of the soil diazotroph community. Given the importance of diversity and abundance of functional organisms in supporting ecosystem processes, we suggest that soil C inputs should be considered for both qualitative and quantitative properties when considering impacts on diazotrophic bacterial ecology.     

Soil microbial community composition and land use history in cultivated and grassland ecosystems of coastal California

Phospholipid ester-linked fatty acid (PLFA) profiles were used to evaluate soil microbial community composition for 9 land use types in two coastal valleys in California. These included irrigated and non-irrigated agricultural sites, non-native annual grasslands and relict, never-tilled or old field perennial grasslands. All 42 sites were on loams or sandy loams of similar soil taxa derived from granitic and alluvial material. We hypothesized that land use history and its associated management inputs and practices may produce a unique soil environment, for which microbes with specific environmental requirements may be selected and supported. We investigated the relationship between soil physical and chemical characteristics, management factors, and vegetation type with microbial community composition. Higher values of total soil C, N, and microbial biomass (total PLFA) and lower values of soil pH occurred in the grassland than cultivated soils. The correspondence analysis (CA) of the PLFA profiles and the canonical correspondence analysis (CCA) of PLFA profiles, soil characteristics, and site and management factors showed distinct groupings for land use types. A given land use type could thus be identified by soil microbial community composition as well as similar soil characteristics and management factors. Differences in soil microbial community composition were highly associated with total PLFA, a measure of soil microbial biomass, suggesting that labile soil organic matter affects microbial composition. Management inputs, such as fertilizer, herbicide, and irrigation, also were associated with the distinctive microbial community composition of the different cultivated land use types.     

Controls on soil nitrogen cycling and microbial community composition across land use and incubation temperature

We conducted a laboratory incubation of forest (Scots pine (Pinus sylvestris) or beech (Fagus sylvatica)), grassland (Trifolium repens/Lolium perenne) and arable (organic and conventional) soils at 5 and 25 °C. We aimed to clarify the mechanisms of short-term (2-weeks) nitrogen (N) cycling processes and microbial community composition in relation to dissolved organic carbon (DOC) and N (DON) availability and selected soil properties. N cycling was measured by ¹⁵N pool dilution and microbial community composition by denaturing gradient gel electrophoresis (DGGE), phospholipid fatty acid (PLFA) and community level physiological profiles (CLPP). Soil DOC increased in the order of arable<grassland<forest soil while DON and gross N fluxes increased in the order of forest<arable<grassland soil; land use had no affect on respiration rate. Soil DOC was lower, while respiration, DON and gross N fluxes were higher at 25 than 5 °C. Gross N fluxes, respiration and bacterial biomass were all positively correlated with each other. Gross N fluxes were positively correlated with pH and DON, and negatively correlated with organic matter, fungal biomass, DOC and DOC/DON ratio. Respiration rate was positively correlated with bacterial biomass, DON and DOC/DON ratio. Multiple linear modelling indicated that soil pH, organic matter, bacterial biomass, DON and DOC/DON ratio were important in predicting gross N mineralization. Incubation temperature, pH and total-C were important in predicting gross nitrification, while gross N mineralization, gross nitrification and pH were important in predicting gross N immobilization. Permutation multivariate analysis of variance indicated that DGGE, CLPP and PLFA profiles were all significantly (P<0.05) affected by land use and incubation temperature. Multivariate regressions indicated that incubation temperature, pH and organic matter content were important in predicting DGGE, CLPP and PLFA profiles. PLFA and CLPP were also related to DON, DOC, ammonium and nitrate contents. Canonical correlation analysis showed that PLFA and CLPP were related to differences in the rates of gross N mineralization, gross nitrification and soil respiration. Our study indicates that vegetation type and/or management practices which control soil pH and mediate dissolved organic matter availability were important predictors of gross N fluxes and microbial composition in this short-term experiment.     

Shifts in rhizosphere microbial communities and enzyme activity of Poa alpina across an alpine chronosequence

This study quantifies the influence of Poa alpina on the soil microbial community in primary succession of alpine ecosystems, and whether these effects are controlled by the successional stage. Four successional sites representative of four stages of grassland development (initial, 4 years (non-vegetated); pioneer, 20 years; transition, 75 years; mature, 9500 years old) on the Rotmoos glacier foreland, Austria, were sampled. The size, composition and activity of the microbial community in the rhizosphere and bulk soil were characterized using the chloroform-fumigation extraction procedure, phospholipid fatty acid (PLFA) analysis and measurements of the enzymes β-glucosidase, β-xylosidase, N-acetyl-β-glucosaminidase, leucine aminopeptidase, acid phosphatase and sulfatase. The interplay between the host plant and the successional stage was quantified using principal component (PCA) and multidimensional scaling analyses. Correlation analyses were applied to evaluate the relationship between soil factors (C_org, N_t, C/N ratio, pH, ammonium, phosphorus, potassium) and microbial properties in the bulk soil. In the pioneer stage microbial colonization of the rhizosphere of P. alpina was dependent on the reservoir of microbial species in the bulk soil. As a consequence, the rhizosphere and bulk soil were similar in microbial biomass (ninhydrin-reactive nitrogen (NHR-N)), community composition (PLFA), and enzyme activity. In the transition and mature grassland stage, more benign soil conditions stimulated microbial growth (NHR-N, total amount of PLFA, bacterial PLFA, Gram-positive bacteria, Gram-negative bacteria), and microbial diversity (Shannon index H) in the rhizosphere either directly or indirectly through enhanced carbon allocation. In the same period, the rhizosphere microflora shifted from a G⁻ to a more G⁺, and from a fungal to a more bacteria-dominated community. Rhizosphere β-xylosidase, N-acetyl-β-glucosaminidase, and sulfatase activity peaked in the mature grassland soil, whereas rhizosphere leucine aminopeptidase, β-glucosidase, and phosphatase activity were highest in the transition stage, probably because of enhanced carbon and nutrient allocation into the rhizosphere due to better growth conditions. Soil organic matter appeared to be the most important driver of microbial colonization in the bulk soil. The decrease in soil pH and soil C/N ratio mediated the shifts in the soil microbial community composition (bacPLFA, bacPLFA/fungPLFA, G⁻, G⁺/G⁻). The activities of β-glucosidase, β-xylosidase and phosphatase were related to soil ammonium and phosphorus, indicating that higher decomposition rates enhanced the nutrient availability in the bulk soil. We conclude that the major determinants of the microflora vary along the successional gradient: in the pioneer stage the rhizosphere microflora was primarily determined by the harsh soil environment; under more favourable environmental conditions, however, the host plant selected for a specific microbial community that was related to the dynamic interplay between soil properties and carbon supply.     

Soil CO2 efflux and extractable organic carbon fractions under simulated precipitation events in a Mediterranean Dehesa

The magnitude of CO₂ efflux pulses after rewetting a dry soil is highly variable and the factors regulating these pulses are poorly understood. In this field experiment, we aimed to study the C dynamics after simulated summer rainstorms in a Mediterranean open holm oak woodland (dehesa). We hypothesized that because the herbaceous cover is mostly dead during the summer in this ecosystem, the short-term CO₂ efflux (SR) after rewetting could mainly be explained by different measurable soil C fractions: i) K₂SO₄-extracted soil C (EOC); ii) microbial biomass C (MBC); or iii) chloroform-fumigated extracted C (CFE). On both grazed and abandoned dehesa sites, we simulated three summer rain events at two-week intervals and we measured SR discontinuously in three plots under tree canopy and in another three plots in open grassland. In each plot, C fractions and water content were estimated before (2 h) and after (36 h) each irrigation event. Following rewettings, SR increased up to ten times compared with non-irrigated plots. The CFE actually increased after rewetting in the first two irrigations but not in the third event, suggesting that the capacity of the soil to release labile organic C from soil aggregates or litter was reduced after each irrigation event. Overall, the C released as CO₂ in the first 24 h was related to the CFE existing before rewetting, which may help to explain the spatial variability in SR. However, the explained variability decreased after each irrigation, suggesting a change to a less labile composition of the CFE fraction as a consequence of multiple drying-rewetting cycles.     

Microbial response time to sugar and amino acid additions to soil

Soil microbial respiration is derived predominantly from the turnover of carbohydrates and proteins in soil. In most agricultural ecosystems, these C compounds enter soil mainly from rhizodeposition (root exudation and turnover). Our aim was to determine how long it takes for the microbial population to reach their maximum mineralization potential after the addition of low-molecular-weight (MW) rhizodeposits to the soil. We added sugar in the form of glucose and amino acids in the form of glycine to an arable, grazed grassland, Eucalyptus forest and boreal forest soil and monitored CO₂ efflux over a 6-h period. Artificial rainwater amended (zero C addition) or unamended soils were used as controls. The Michaelis-Menten substrate utilization profiles showed vastly different patterns of microbial mineralization capacity and substrate affinity between the soils. However, in all soils we showed that activation of the soil microbial community to C addition occurred almost instantaneously (?60 s) with the average time taken to reach half maximal CO₂ production being 14±8 min for glucose and 10±8 min for glycine. After reaching their maximal mineralization potential, the rate of CO₂ evolution remained constant for the remainder of the experiment. Our results showed that while substrate uptake and mineralization within the soil microbial biomass was activated quickly, subsequent adaptation and upregulation of its C processing capacity did not occur at least in the short term. The fast rate of microbial activation and substrate use we partially attribute to the large degree of functional redundancy that exists within the soil microbial community for processing rhizodeposits.     

Microbial biomass dynamics in recently air-dried and rewetted soils compared to others stored air-dry for up to 103 years

Our aim was to compare the soil microbial biomass concentration and its activity (measured as CO₂-C evolved) following the rewetting and aerobic incubation of soils which have previously been stored air-dry for different periods. Some of the soils have been stored in the Rothamsted sample archive for 103 years, others were comparable freshly sampled soils following air-drying and rewetting and other soils were stored air-dry for 2 years then rewetted for the work described here. Following air-drying, soil ATP concentrations were variable in recently air-dried soil, comprising about 10-35% of the initial ATP concentrations in fresh soil. Following rewetting, the percentage recovery of ATP increased in all soils by 7 days, then declined to between 73% and 87% of the original ATP concentration in the air-dried soils by day 12. Storage of air-dried soils decreased the ability of the microbial biomass to restore its ATP concentrations. For example, the ATP concentration in a soil sampled from stubbed (i.e. tree seedling, saplings and bushes cut frequently to ground level) grassland of the Broadbalk continuous wheat experiment at Rothamsted then air-dried for 2 years was only about 14% of that in the fresh soil at 2 days after rewetting. In other soils from the Hoosfield Barley Experiment, also at Rothamsted, previously given NPK or FYM since 1852, and sampled then stored air-dry for between 13 and 83 years, from 52% to 57% of the ATP in the comparable fresh soils was measured at two days after rewetting. The soil ATP concentration then changed little more up to 12 days. One of the most interesting findings was that while the microbial biomass ATP concentration in the above NPK soils only ranged from about 2 to 4 μmol ATP g⁻¹ biomass C, in the FYM soil the microbial biomass ATP concentrations (range 11.5-13.6 μmol ATP g⁻¹ biomass C) were the same as we repeatedly measure in fresh moist aerobic soil. We do not yet know the reasons for this. More than twice as much CO₂-C was evolved from the long-term stored soils than from freshly sampled ones. However, the specific respiration of the microbial biomass did not change much after the first 12 years of storage, indicating that loss of viability mainly occurred in the earlier years.