Microbial community response to nitrogen deposition in northern forest ecosystems

The productivity of temperate forests is often limited by soil N availability, suggesting that elevated atmospheric N deposition could increase ecosystem C storage. However, the magnitude of this increase is dependent on rates of soil organic matter formation as well as rates of plant production. Nonetheless, we have a limited understanding of the potential for atmospheric N deposition to alter microbial activity in soil, and hence rates of soil organic matter formation. Because high levels of inorganic N suppress lignin oxidation by white rot basidiomycetes and generally enhance cellulose hydrolysis, we hypothesized that atmospheric N deposition would alter microbial decomposition in a manner that was consistent with changes in enzyme activity and shift decomposition from fungi to less efficient bacteria. To test our idea, we experimentally manipulated atmospheric N deposition (0, 30 and 80 kg NO₃⁻-N) in three northern temperate forests (black oak/white oak (BOWO), sugar maple/red oak (SMRO), and sugar maple/basswood (SMBW)). After one year, we measured the activity of ligninolytic and cellulolytic soil enzymes, and traced the fate of lignin and cellulose breakdown products (¹³C-vanillin, catechol and cellobiose).In the BOWO ecosystem, the highest level of N deposition tended to reduce phenol oxidase activity (131±13 versus 104±5 μmol h⁻¹ g⁻¹) and peroxidase activity (210±26 versus 190±21 μmol h⁻¹ g⁻¹) and it reduced ¹³C-vanillin and ¹³C-catechol degradation and the incorporation of ¹³C into fungal phospholipids (p<0.05). Conversely, in the SMRO and SMBW ecosystems, N deposition tended to increase phenol oxidase and peroxidase activities and increased vanillin and catechol degradation and the incorporation of isotope into fungal phospholipids (p<0.05). We observed no effect of experimental N deposition on the degradation of ¹³C-cellulose, although cellulase activity showed a small and marginally significant increase (p<0.10). The ecosystem-specific response of microbial activity and soil C cycling to experimental N addition indicates that accurate prediction of soil C storage requires a better understanding of the physiological response of microbial communities to atmospheric N deposition.     

Performance of para-nitrophenyl phosphate and 4-methylumbelliferyl phosphate as substrate analogues for phosphomonoesterase in soils with different organic matter content

Phosphomonoesterase (PMEase) activity plays a key role in nutrient cycling and is a potential indicator of soil condition and ecosystem stress. We compared para-nitrophenyl phosphate (pNPP) and 4-methylumbelliferyl phosphate (MUP) as substrate analogues for PMEase in 7 natural ecosystem soils and 8 agricultural top soils with contrasting C contents (8.0-414 g kg⁻¹ C) and pH (3.0-7.5). PMEase activities obtained with pNPP (0.05-5 μmol g⁻¹ h⁻¹) were significantly less than activities obtained with MUP (0.9-13 μmol g⁻¹ h⁻¹), especially in soils with a high organic matter content (>130 g kg⁻¹). Only PMEase activities assayed with MUP correlated significantly with total C and total N (r=0.7, P<0.01 all), and pH (r=−0.71, P<0.01). PMEase activities obtained with the two substrate analogues were correlated when expressed on a C-content basis (r=0.8, P<0.001), but not when expressed on an oven-dry soil weight basis. This indicated that interference by organic matter is related to the quantity rather than to the quality of organic matter. Overall, assaying with MUP was more sensitive compared to assaying with pNPP, particularly in the case of high organic and acid soils.     

Changes in soil biological and biochemical characteristics in a long-term field trial on a sub-tropical inceptisol

Soil organic carbon (SOC), microbial biomass carbon (MBC), their ratio (MBC/SOC) which is also known as microbial quotient, soil respiration, dehydrogenase and phosphatase activities were evaluated in a long-term (31 years) field experiment involving fertility treatments (manure and inorganic fertilizers) and a maize (Zea mays L.)-wheat (Triticum aestivum L.)-cowpea (Vigna unguiculata L.) rotation at the Indian Agricultural Research Institute near New Delhi, India. Applying farmyard manure (FYM) plus NPK fertilizer significantly increased SOC (4.5-7.5 g kg⁻¹), microbial biomass (124-291 mg kg⁻¹) and microbial quotient from 2.88 to 3.87. Soil respiration, dehydrogenase and phosphatase activities were also increased by FYM applications. The MBC response to FYM+100% NPK compared to 100% NPK (193 vs. 291 mg kg⁻¹) was much greater than that for soil respiration (6.24 vs. 6.93 μl O₂ g⁻¹ h⁻¹) indicating a considerable portion of MBC in FYM plots was inactive. Dehydrogenase activity increased slightly as NPK rates were increased from 50% to 100%, but excessive fertilization (150% NPK) decreased it. Acid phosphatase activity (31.1 vs. 51.8 μg PNP g⁻¹ h⁻¹) was much lower than alkali phosphatase activity (289 vs. 366 μg PNP g⁻¹ h⁻¹) in all treatments. Phosphatase activity was influenced more by season or crop (e.g. tilling wheat residue) than fertilizer treatment, although both MBC and phosphatase activity were increased with optimum or balanced fertilization. SOC, MBC, soil respiration and acid phosphatase activity in control (no NPK, no manure) treatment was lower than uncultivated reference soil, and soil respiration was limiting at N alone or NP alone treatments.     

Adenylates in the soil microbial biomass at different temperatures

Five soils from temperate sites (Germany; 2 arable and 3 grassland) were incubated aerobically at 5, 10, 15, 20, 25, 35, and 40 °C for 8 days. Soils were analysed for soil microbial biomass C, biomass N, AMP, ADP, and ATP to determine whether the increase in the ATP-to-microbial biomass C ratio with increasing temperature was either due to an increase in the adenylate energy charge (AEC) or de novo synthesis of ATP, or both. Around 80% of the variance in microbial biomass C and biomass N was explained by differences in soil properties, only 7% by the temperature treatments. Averaging the data of all 5 soils for each incubation temperature, the microbial biomass C content decreased with increasing temperature from 15 to 40 °C continuously by 2.5 μg g⁻¹ soil °C⁻¹ after 8-days' incubation. However, this decrease was not accompanied by a similar decrease in microbial biomass N. The average microbial biomass C/N ratio was 6.8. Between 54 and 76% of the variance in AMP, ADP, ATP and the sum of adenylates was explained by differences in soil properties and between 14 (ADP) and 27% (ATP) by the temperature treatments. However, temperature effects on AMP and ADP were variable and inconsistent. In contrast, ATP and consequently also the sum of adenylates increased continuously from 5 to 30 °C followed by a decline to 40 °C. The AEC showed similarly a small, but significant increase with increasing temperature from 0.73 to 0.85 at 30 °C. Consequently, the majority of the variance, i.e. roughly 60% in AEC values, but also in ATP-to-microbial biomass C ratios was explained by the incubation temperature. The mean ATP-to-microbial biomass C ratio increased from 4.7 μmol g⁻¹ at 5 °C to a 2.5 fold maximum of 12.0 μmol g⁻¹ at 35 °C. This increase was linear with a rate of 0.26 μmol ATP g⁻¹ microbial biomass C °C⁻¹. The energy for the extra ATP produced during temperature increase is probably derived from an accelerated turnover of endocellular C reserves in the microbial biomass.     

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.     

Microbial activity in pig slurry-amended soils under semiarid conditions

Soil amendment with manures from intensive animal industries is nowadays a common practice that may favorably or adversely affect several soil properties, including soil microbial activity. In this work, the effect of consecutive annual additions of pig slurry (PS) at rates of 30, 60, 90, 120 and 150 m³ ha⁻¹ y⁻¹ over a 4-year period on soil chemical properties and microbial activity was investigated and compared to that of an inorganic fertilization and a control (without amendment). Field plot experiment conducted under a continuous barley monoculture and semiarid conditions were used. Eight months after the fourth yearly PS and mineral fertilizer application (i.e. soon after the fourth barley harvest), surface soil samples (Ap horizon, 0-15 cm depth) from control and amended soils were collected and analysed for pH, electrical conductivity (EC), contents of total organic C, total N, available P and K, microbial biomass C, basal respiration and different enzymatic activities. The control soil had a slightly acidic pH (6.0), a small EC (0.07 dS m⁻¹), adequate levels of total N (1.2 g kg⁻¹) and available K (483 mg kg⁻¹) for barley growth, and small contents of total organic C (13.2 g kg⁻¹) and available P (52 mg kg⁻¹). With respect to the control and mineral fertilized soils, the PS-amended soils had greater pH values (around neutrality or slightly alkaline), electrical conductivities (still low) and contents of available P and K, and slightly larger total N contents. A significant decrease of total organic C was observed in soils amended at high slurry rate (12.3 g kg⁻¹). Compared with the control and mineral treatments, which produced almost similar results, the PS-amended soils were characterized by a higher microbial biomass C content (from 311 to 442 g kg⁻¹), microbial biomass C/total organic C ratio (from 2.3 to 3.6%) and dehydrogenase (from 35 to 173 μg INTF g⁻¹), catalase (from 5 to 24 μmol O₂ g⁻¹ min⁻¹), BAA-protease (from 0.7 to 1.9 μmol  g⁻¹ h⁻¹) and β-glucosidase (from 117 to 269 μmol PNP g⁻¹ h⁻¹) activities, similar basal respirations (from 48 to 77 μg C-CO₂ g⁻¹ d⁻¹) and urease activities (from 1.5 to 2.2 μmol  g⁻¹ h⁻¹), and smaller metabolic quotients (from 6.4 to 7.7 ng C-CO₂ μg⁻¹ biomass C h⁻¹) and phosphatese activities (from 374 to 159 μmol PNP g⁻¹ h⁻¹). For example, statistical analysis of experimental data showed that, with the exception of metabolic quotient and total organic C content, these effects generally increased with increasing cumulative amount of PS. In conclusion, cumulative PS application to soil over time under semiarid conditions may produce not only beneficial effects but also adverse effects on soil properties, such us the partial mineralization of soil organic C through extended microbial oxidation. Thus, PS should not be considered as a mature organic amendment and should be treated appropriately before it is applied to soil, so as to enhance its potential as a soil organic fertilizer.     

Enzyme activities and microbial biomass in coastal soils of India

Soil salinity is a serious problem for agriculture in coastal regions, wherein salinity is temporal in nature. We studied the effect of salinity, in summer, monsoon and winter seasons, on microbial biomass carbon (MBC) and enzyme activities (EAs) of the salt-affected soils of the coastal region of the Bay of Bengal, Sundarbans, India. The average pH of soils collected from different sites, during different seasons varied from 4.8 to 7.8. The average organic C (OC) and total N (TN) content of the soils ranged between 5.2-14.1 and 0.6-1.4 g kg⁻¹, respectively. The electrical conductivity of the saturation extract (ECe) of soils, averaged over season, varied from 2.2 to 16.3 dSm⁻¹. The ECe of the soils increased five fold during the summer season (13.8 dSm⁻¹) than the monsoon season (2.7 dSm⁻¹). The major cation and anion detected were Na⁺ and Cl⁻, respectively. Seasonality exerted considerable effects on MBC and soil EAs, with the lowest values recorded during the summer season. The activities of β-glucosidase, urease, acid phosphatase and alkaline phosphatase were similar during the winter and monsoon season. The dehydrogenase activity of soils was higher in monsoon than in winter. Average MBC, dehydrogenase, β-glucosidase, urease, acid phosphatase and alkaline phosphatase activities of the saline soils ranged from 125 to 346 mg kg⁻¹ oven dry soil, 6-9.9 mg triphenyl formazan (TPF) kg⁻¹ oven dry soil h⁻¹, 18-53 mg p-nitro phenol (PNP) kg⁻¹ oven dry soil h⁻¹, 38-86 mg urea hydrolyzed kg⁻¹ oven dry soil h⁻¹, 213-584 mg PNP kg⁻¹ oven dry soil h⁻¹ and 176-362 mg PNP g⁻¹ oven dry soil h⁻¹, respectively. The same for the non-saline soils were 274-446 mg kg⁻¹ oven dry soil, 8.8-14.4 mg TPF kg⁻¹ oven dry soil h⁻¹, 41-80 mg PNP kg⁻¹ oven dry soil h⁻¹, 89-134 mg urea hydrolyzed kg⁻¹ oven dry soil h⁻¹, 219-287 mg PNP kg⁻¹ oven dry soil h⁻¹ and 407-417 mg PNP kg⁻¹ oven dry soil h⁻¹, respectively. About 48%, 82%, 48%, 63%, 40% and 48% variation in MBC, dehydrogenase activity, β-glucosidase activity, urease activity, acid phosphatase activity and alkaline phosphatase activity, respectively, could be explained by the variation in ECe of saline soils. Suppression of EAs of the coastal soils during summer due to salinity rise is of immense agronomic significance and needs suitable interventions for sustainable crop production.     

Microbial enzyme activities in leaf litter, humus and mineral soil layers of European forests

The rationale of the study was to investigate microbial activity in different soil horizons in European forests. Hence, activities of chitinase and cellulase, microbial biomass carbon (C_mic) and basal respiration were measured in litter, fragmentation, humus and mineral soil layers collected several times from various beech and spruce forests. Sites were selected to form a gradient in N availability. Analyses were also performed on beech litter from a litterbag transplant experiment. Furthermore, microbiological parameters were measured in horizons of beech and spruce chronosequence sites with different stand age in order to investigate the influence of forest rotation, and hence changes in soil organic matter (SOM) dynamics, on microbial activity. Finally in horizons of one beech forest, the seasonal variation of selected microbiological parameters was measured more intensively. β-Glucosaminidase and cellobiohydrolase activities were measured using fluorogenic 4-methylumbelliferyl substrates to estimate chitinase and cellulase activities, respectively. On a spatial scale, chitinase and cellulase activities, C_mic determined by substrate induced respiration, and basal respiration ranged from 144 to 1924 and 6-177 nmol 4-MU g⁻¹ org-C h⁻¹, 8-48 mg C g⁻¹ org-C and 11-149 μg CO₂-C g⁻¹ org-C h⁻¹, respectively; in general values were significantly lower in layers of humus and mineral soil than of litter. Chitinase activity, C_mic and basal respiration from humus and mineral soil layers, together, correlated positively, while none correlated with cellulase activity. Similarly in the litter layer, no correlations were found between the microbiological parameters. On a seasonal scale, a time lag between a burst in basal respiration rate and activities of both enzymes were observed. In general, activities of cellulase and chitinase, C_mic and basal respiration, did not change with stand age, except in the humus layer in the spruce chronosequence, where C_mic decreased with stand age. In the litter layer, cellulase activity was significantly and positively related to the C:N ratio, while only a tendency for chitinase activity was shown, indicating that enzyme activities decreased with increasing N availability. In accordance, the enzyme activities and C_mic decreased significantly with increasing chronic N deposition in the humus layer, while basal respiration only tended to decrease with increasing N deposition. In contrast, enzyme activities in beech litter from litterbags after 2 years of incubation were generally higher at sites with higher N deposition. The results show different layer-specific responses of enzyme activities to changes in N availability, indicating different impacts of N availability on decomposition of SOM and stage of litter decomposition.     

Soil biochemical activity and phosphorus transformations and losses from acidified forest soils

Three Bohemian Forest catchments, Plešné, ?erné and ?ertovo, were studied. These catchments have similar climatic conditions, relief and vegetation, but differ in their bedrock composition. The granitic bedrock in the Plešné catchment was more susceptible to phosphorus (P) leaching under acid conditions than was the mica schist bedrock in the other catchments. The goal of this study was to determine if higher P leaching from the Plešné catchment was associated with differences in microbial P transformations and enzymatic P hydrolysis. Phosphorus and nitrogen contents in soil microbial biomass (P_MB, N_MB; chloroform fumigation), C mineralisation rate (C_min; CO₂ production by GC) and phosphatase activity (MUF-phosphate), were measured in three successive years. Phosphatase activity, P_MB, and C_min were used to characterise the enzymatic hydrolysis of organic P, microbial P accumulation, and microbial mineralisation rates of organic compounds, respectively. Soil chemical properties were characterised by C, N and P content, pH, and by oxalate-extractable P, Fe and Al. Spatial variability in N_MB, P_MB, C_min and phosphatase activity within the catchment was higher (coefficient of variation, CV<50%) than their temporal variability (CV<30%). Multivariate analysis revealed a significant soil layer effect but not that of catchment. When soil layers were evaluated separately, a difference between the Plešné and ?erné or ?ertovo catchments was found in litter and mineral layers, even though the variability within one catchment was high. Within soil profile, phosphatase activity was positively correlated with C_tot, N_MB and C_min (r²=0.89-0.92) being very correlated with P_MB (r²=0.99). Phosphatase activity was higher in the litter (14.0 nmol g⁻¹ h⁻¹) and humus (8.65 nmol g⁻¹ h⁻¹) layers of Plešné than in the same layers of the ?erné (9.65 and 6.40 nmol g⁻¹ h⁻¹) and ?ertovo (12.8 and 6.0 nmol g⁻¹ h⁻¹) soils. Similarly, P_MB in the litter and humus layers of Plešné soil (161 and 93 μg g⁻¹) was higher than P_MB of the same layers of the ?erné (120 and 66 μg g⁻¹) and ?ertovo (148 and 89 μg g⁻¹) soils. High MUFP hydrolysis rate: C_min molar ratio (0.16-1.17 M of P per 1 M of respired C) indicated that potential enzymatic P hydrolysis exceeded estimated microbial P demand (0.034 M of P per 1 M of respired C) in all catchments. The results suggest that higher microbial P transformations and enzymatic P hydrolysis could contribute to enhanced P leaching from the Plešné catchment, which could be enhanced by the lower Fe content in the soil of this catchment as compared to the ?erné and ?ertovo catchments.     

Microbial activity and functional composition among northern peatland ecosystems

The extent to which complex interrelationships between plants and microorganisms influence organic matter dynamics is critical to our understanding of global C cycles in changing environments. We examined the hypothesis that patterns of soil microbial activity and functional composition differ among vegetation types in northern peatland ecosystems. Microbial characteristics were compared among peatlands differing in plant growth form (tree, shrub/moss, sedge) in two regions (New York State and West Virginia). Microbial activity (basal respiration) was greater in surface (0-15 cm) than subsurface (15-30 cm) peat and from sites dominated by shrubs and Sphagnum moss (3.9±0.65 μg C g⁻¹ h⁻¹) compared to forested (1.8±0.20 μg C g⁻¹ h⁻¹) or sedge-dominated sites (1.9±0.38 μg C g⁻¹ h⁻¹). Microbial activity was not related to decomposability of peat organic matter among vegetation types, and activity was unexpectedly higher in sites with lower peat pH and higher water table level. Substrate-induced respiration (SIR) did not show a clear pattern among vegetation types, but was greater in surface than subsurface peat. Microbial responsiveness to added glucose was very low. The ratio of basal respiration to SIR varied between 0.39 and 0.72 and, like activity, was highest in shrub/Sphagnum sites. Microbial substrate utilization patterns (assayed with BIOLOG^® GN plates) also differed between shrub/Sphagnum sites and forest or sedge sites, suggesting that C fluxes were mediated by different assemblages of microorganisms in shrub/Sphagnum peatlands. Principal component (PC) scores indicated more utilization of N-containing compounds and carboxylic acids, and less utilization of carbohydrates by microbial communities in shrub/Sphagnum sites. PC scores were much more variable both within and among vegetation types for sites in West Virginia than in New York State, and a greater diversity of C sources were utilized in WV (57±3) than NYS (47±2) peat. Our results suggest a link between microbial respiratory activity and microbial functional composition as they vary among these peatland vegetation types.