Accounting for variability in soil microbial communities of temperate upland grassland ecosystems

This study aimed to determine the factors which regulate soil microbial community organisation and function in temperate upland grassland ecosystems. Soil microbial biomass (C_mic), activity (respiration and potential carbon utilisation) and community structure (phospholipid fatty acid (PLFA) analysis, culturing and community level physiological profiles (CLPP) (Biolog^®)) were measured across a gradient of three upland grassland types; Festuca–Agrostis–Galium grassland (unimproved grassland, National Vegetation Classification (NVC) — U4a); Festuca–Agrostis–Galium grassland, Holcus–Trifolium sub-community (semi-improved grassland, NVC — U4b); Lolium–Cynosurus grassland (improved grassland, NVC — MG6) at three sites in different biogeographic areas of the UK over a period of 1 year. Variation in C_mic was mainly due to grassland type and site (accounting for 55% variance, v, in the data). C_mic was significantly (P<0.001) high in the unimproved grassland at Torridon (237.4 g C m⁻² cf. 81.2 g C m⁻² in semi- and 63.8 g C m⁻² in improved grasslands) and Sourhope (114.6 g C m⁻² cf. in 44.8 g C m⁻² semi- and 68.3 g C m⁻² in improved grasslands) and semi-improved grassland at Abergwyngregyn (76.0 g C m⁻² cf. 41.7 g C m⁻² in un- and 58.3 g C m⁻² in improved grasslands). C_mic showed little temporal variation (v=3.7%). Soil microbial activity, measured as basal respiration was also mainly affected by grassland type and site (n=32%). In contrast to C_mic, respiration was significantly (P<0.001) high in the improved grassland at Sourhope (263.4 l h⁻¹m⁻² cf. 79.6 l h⁻¹m⁻² in semi- and 203.9 l h⁻¹m⁻² unimproved grasslands) and Abergwyngregyn (198.8 l h⁻¹m⁻² cf. 173.7 l h⁻¹m⁻² in semi- and 88.2 l h⁻¹m⁻² unimproved grasslands). Microbial activity, measured as potential carbon utilisation, agreed with the respiration measurements and was significantly (P<0.001) high in the improved grassland at all three sites (A₅₉₀ 0.14 cf. 0.09 in semi- and 0.07 in unimproved grassland). However, date of sampling also had a significant (P<0.001) impact on C utilisation potential (v=24.7%) with samples from April 1997 having highest activity at all three sites. Variation in microbial community structure was due, predominantly, to grassland type (average v=23.6% for bacterial and fungal numbers and PLFA) and date of sampling (average v=39.7% for bacterial and fungal numbers and PLFA). Numbers of culturable bacteria and bacterial PLFA were significantly (P<0.001) high in the improved grassland at all three sites. Fungal populations were significantly (P<0.01) high in the unimproved grassland at Sourhope and Abergwyngregyn. The results demonstrate a shift in soil microbial community structure from one favouring fungi to one favouring bacteria as grassland improvement increased. Numbers of bacteria and fungi were also significantly (P<0.001) higher in August than any other sampling date. Canonical variate analysis (CVA) of the carbon utilisation data significantly (P<0.05) differentiated microbial communities from the three grassland types, mainly due to greater utilisation of sugars and citric acid in the improved grasslands compared to greater utilisation of carboxylic acids, phenolics and neutral amino acids in the unimproved grasslands, possibly reflecting substrate availability in these grasslands. Differences in C_mic, activity and community structure between grassland types were robust over time. In addition, broad scale measures of microbial growth and activity (C_mic and respiration) showed little temporal variation compared to measures of soil microbial community structure, which varied quantitatively with respect to environmental variables (temperature, moisture) and plant productivity, hence substrate supply.     

Interrelationships between microbial and soil properties in young volcanic ash soils of Nicaragua

The activity and biomass of soil microorganisms were measured in soils from 25 different arable sites in the Pacific region of Nicaragua with the objective of elucidating their interrelationship with soil textural and soil chemical properties. All soils developed from recent volcanic deposits but differ in their particle size distribution. Short-term phosphorus fixation capacity varied widely and was, on average, 11% of added P. In contrast, long-term P fixation capacity varied within a small range of around 55%. Mean basal respiration was 8.6 μg CO₂–C d⁻¹ g⁻¹ soil, average contents of biomass C, biomass P, and ergosterol as an indicator of fungal biomass were 116, 1.95, and 0.34 μg g⁻¹ soil, respectively. They were all, except biomass P, significantly lower in the sandy than in the loamy soils. The mean biomass C-to-soil C ratio was 0.69%, the mean metabolic quotient 95 mg CO₂–C d⁻¹ g⁻¹ biomass C, the mean ergosterol-to-biomass C ratio 0.31% and the mean biomass C-to-P ratio 107. The very low ergosterol-to-biomass C ratio indicates that fungi contribute only a relatively small percentage to the microbial biomass. The biomass C-to-P ratio exceeded considerably the soil C-to-total P ratio. Metabolic quotient qCO₂ and ergosterol-to-biomass C were both negatively correlated with biomass C-to-soil C ratio and clay content, indicating positive correlations between qCO₂ and ergosterol-to-biomass C ratio and between biomass C-to-soil C ratio and clay content. Key problems of soil fertility and soil quality in Nicaragua are low availability of soil organic matter and phosphorus to soil microorganisms, which are magnified by a low percentage of fungi, probably reducing the ability of soil to provide nutrients for plant growth.     

Animal manure and anhydrous ammonia amendment alter microbial carbon use efficiency,microbial biomass,and activities of dehydrogenase and amidohydrolases in semiarid agroecosystems

The potential negative impact of agricultural practices on soil and water quality is of environmental concern. The associated nutrient transformations and movements that lead to environmental concerns are inseparable from microbial and biochemical activities. Therefore, biochemical and microbiological parameters directing nitrogen (N) transformations in soils amended with different animal manures or inorganic N fertilizers were investigated. Soils under continuous corn cultivation were treated with N annually for 5 years at 56, 168, and 504 kg N ha⁻¹ in the form of swine effluent, beef manure, or anhydrous ammonia. Animal manure treatments increased dehydrogenase activity, microbial biomass carbon (C_mic) and N (N_mic) contents, and activities of amidohydrolases, including l-asparaginase, urease, l-glutaminase, amidase, and β-glucosaminidase. Soils receiving anhydrous ammonia demonstrated increased nitrate contents, but reduced microbiological and biochemical activities. All treatments decreased C_mic:organic C (C_org) ratios compared with the control, indicating reduced microbial C use efficiency and disturbance of C equilibrium in these soil environments. Activities of all enzymes tested were significantly correlated with soil C_org contents (P < 0.001, n = 108), but little correlation (r = 0.03, n = 36) was detected between C_mic and C_org. Activities of amidase and β-glucosaminidase were dominated by accumulated enzymes that were free of microbial cells, while activities of asparaginase and glutaminase were originated predominately from intracellular enzymes. Results indicated that soil microbial and biochemical activities are sensitive indicators of processes involved in N flow and C use efficiency in semiarid agroecosystems.     

Responses of soil microbial biomass and activity for practices of organic and conventional farming systems in Piauí state,Brazil

The aim of this work was to investigate the response of soil microbial biomass and activity to practices in organic and conventional farming systems. The study was carried out at the Irrigation District of Piauí, Brazil. Five different plots planted with “acerola” orchard (Malpighia glaba) and established at the following management were evaluated: (1) under 12 months of soil conventional management (CNV); (2) under six months of soil organic management (ORG6); (3) under 12 months of soil organic management (ORG12); (4) under 18 months of soil organic management (ORG18); and (5) under 24 months of soil organic management (ORG24). Soil microbial biomass C (C_mic), basal respiration, organic carbon (C_org), C_mic-to-C_org ratio and metabolic quotient (qCO₂) were evaluated in soil samples collected at 0–10 cm depth. The highest C_org and C_mic levels occurred in organic system plots ORG18 and ORG24 compared to the conventional system. Soil respiration and C_mic-to-C_org ratio were significantly enhanced by the organic system plots. The qCO₂ was greater in conventional than in organic system. These results indicate that the organic practices rapidly improved soil microbial characteristics and slowly increase soil organic C.     

Microbial respiration activities of soils from different climatic regions of European Russia

The aim of this study was to survey and evaluate the microbial respiration of main soil types (gleyic Cryosols, umbric Albeluvisols, albic Luvisols, luvic Chernozems, Kastanozems) across European Russia, from semiarid to polar climatic zones. Soil was sampled from 0–5 and 5–10 cm layers at natural (forest, grassland, fallow) and corresponding sites under agricultural land use. Soil microbial biomass carbon (C_mic) determined by the substrate-induced respiration method and basal respiration (BR) were measured under standardized laboratory conditions (22 °C, 60% WHC). The ratios of BR/C_mic and C_mic/C_org were also calculated. C_mic and BR were highest in polar (gleyic Cryosols) and temperate (albic Luvisols, luvic Chernozems) climatic zones, the lowest were in boreal (umbric Luvisols) and semiarid (Kastanozems). C_mic, BR and C_mic/C_org ratios were higher in 0–5 cm layers compared to the corresponding 5–10 cm and in natural sites versus in arable. Principal component analysis yielded a clear separation of the vegetation zones with respect to the several principal components (PC). PC 1 was composed of C_mic, BR, soil chemical (C_org, N_tot) and texture parameters. PC 2 was composed of climatic (MAT, MAP) and soil pH variables. Three-way ANOVA indicated that “soil type”, “ecosystem” and “layer” factors, and their interactions accounted for almost 98 and 99% of the total variance in C_mic and BR, respectively.     

Effects of chemical conditions in re-wetted peats on temporal variation in microbial biomass and acid phosphatase activity within the growing season

Possible effects of chemical alterations in peat following re-wetting on their microbial characteristics are insufficiently known. Microbial biomass carbon (C_mic), nitrogen (N_mic), phosphorus (P_mic) and acid phosphatase activity were investigated in re-wetted virtually undisturbed and differently degraded peatlands (Histosols) in northeast Germany to assess re-wetting effects on microbial biomass production and phosphorus (P) cycling in one growing season. The virtually undisturbed Eutri-Ombric Histosol had the largest content of microbial biomass (C_mic: 2132 mg/kg, N_mic: 309 mg/kg and P_mic: 48 mg/kg; means of six sampling dates, upper 10 cm). Increasingly lower contents of microbial biomass were observed in the more strongly degraded peats of two Ombri-Sapric Histosols. Furthermore, the proportions of P_mic as a percent of total P (P_t) were smallest in the strongly degraded Ombric-Sapric Histosol (1.6% of P_t) and gradually larger with better peat conservation (2.6% of P_t in the moderately degraded Ombri-Sapric Histosol and 3.0% of P_t in the virtually undisturbed Eutri-Ombric Histosol). The acid phosphatase activity was always greatest in May, irrespective of peat degradation. This maximum was lower for the Eutri-Ombric Histosol (2633 μg nitrophenol/(g h)) than for the two Ombri-Sapric Histosols (3963 and 3212 μg nitrophenol/(g h)). In the two degraded peats, the temporal variation in phosphatase activity was also more pronounced. Our results, in particular the higher peak phosphatase activity combined with an incorporation of P into microbial biomass, indicate that peat degradation may enhance the phosphate input to soil solution. Thus, it is concluded that modified biological P cycling could contribute to increased risks of P losses to adjacent surface water after re-wetting of degraded peats.     

Comparison of the eddy covariance and automated closed chamber methods for evaluating nocturnal CO2 exchange in a Japanese cypress forest

Underestimation of nocturnal CO₂ respiration using the eddy covariance method under calm conditions remains an unsolved problem at many flux observation sites in forests. To evaluate nocturnal CO₂ exchange in a Japanese cypress forest, we observed CO₂ flux above the canopy (F_c), changes in CO₂ storage in the canopy (S_t) and soil, and trunk and foliar respiration for 2 years (2003–2004). We scaled these chamber data to the soil, trunk, and foliar respiration per unit of ground area (F_s, F_t, F_f, respectively) and used the relationships of F_s, F_t, and F_f with air or soil temperature for comparison with canopy-scale CO₂ exchange measurements (=F_c + S_t). The annual average F_s, F_t, and F_f were 714 g C m⁻² year⁻¹, 170 g C m⁻² year⁻¹, and 575 g C m⁻² year⁻¹, respectively. At small friction velocity (u_*), nocturnal F_c + S_t was smaller than F_s + F_t + F_f estimated using the chamber method, whereas the two values were almost the same at large u_*. We replaced F_c + S_t measured during calm nocturnal periods with a value simulated using a temperature response function derived during well-mixed nocturnal periods. With this correction, the estimated net ecosystem exchange (NEE) from F_c + S_t data ranged from −713 g C m⁻² year⁻¹ to −412 g C m⁻² year⁻¹ in 2003 and from −883 g C m⁻² year⁻¹ to −603 g C m⁻² year⁻¹ in 2004, depending on the u_* threshold. When we replaced all nocturnal F_c + S_t data with F_s + F_t + F_f estimated using the chamber method, NEE was −506 g C m⁻² year⁻¹ and −682 g C m⁻² year⁻¹ for 2003 and 2004, respectively.     

Partitioning soil surface CO2 efflux into autotrophic and heterotrophic components,using natural gradients in soil δ13C in an undisturbed savannah soil

We used natural gradients in soil and vegetation δ¹³C signatures in a savannah ecosystem in Texas to partition soil respiration into the autotrophic (Ra) and heterotrophic (Rh) components. We measured soil respiration along short transects from under clusters of C₃ trees into the C₄ dominated grassland. The site chosen for the study was experiencing a prolonged drought, so an irrigation treatment was applied at two positions of each transect. Soil surface CO₂ efflux was measured along transects and CO₂ collected for analysis of the δ¹³C signature in order to: (i) determine how soil respiration rates varied along transects and were affected by localised change in soil moisture and (ii) partition the soil surface CO₂ efflux into Ra and Rh, which required measurement of the δ¹³C signature of root- and soil-derived CO₂ for use in a mass balance model.The soil at the site was unusually dry, with mean volumetric soil water content of 8.2%. Soil respiration rates were fastest in the centre of the tree cluster (1.5 ± 0.18 μmol m^?2 s^?1; mean ± SE) and slowest at the cluster–grassland transition (0.6 ± 0.12 μmol m^?2 s^?1). Irrigation produced a 7–11 fold increase in the soil respiration rate. There were no significant differences (p > 0.5) between the δ¹³C signature of root biomass and respired CO₂, but differences (p < 0.01) were observed between the respired CO₂ and soil when sampled at the edge of the clusters and in the grassland. Therefore, end member values were measured by root and soil incubations, with times kept constant at 30 min for roots and 2 h for soils. The δ¹³C signature of the soil surface CO₂ efflux and the two end member values were used to calculate that, in the irrigated soils, Rh comprised 51 ± 13.5% of the soil surface CO₂ efflux at the mid canopy position and 57 ± 7.4% at the drip line. In non-irrigated soil it was not possible to partition soil respiration, because the δ¹³C signature of the soil surface CO₂ efflux was enriched compared to both the end member values. This was probably due to a combination of the very dry porous soils at our study site (which may have been particularly susceptible to ingress of atmospheric CO₂) and the very slow respiration rates of the non-irrigated soils.     

Climatic influences on active fractions of soil organic matter

Biologically active fractions of soil organic matter are important in understanding decomposition potential of organic materials, nutrient cycling dynamics, and biophysical manipulation of soil structure. We evaluated the quantitative relationships among potential C and net N mineralization, soil microbial biomass C (SMBC), and soil organic C (SOC) under four contrasting climatic conditions. Mean SOC values were 28±11 mg g⁻¹ (n=24) in a frigid–dry region (Alberta/British Columbia), 25±5 mg g⁻¹ (n=12) in a frigid–wet region (Maine), 11±4 mg g⁻¹ (n=117) in a thermic–dry region (Texas), and 12±5 mg g⁻¹ (n=131) in a thermic–wet region (Georgia). Higher mean annual temperature resulted in consistently greater basal soil respiration (1.7 vs 0.8 mg CO₂–C g⁻¹ SOC d⁻¹ in the thermic compared with the frigid regions, P<0.001), greater net N mineralization (2.8 vs 1.3 mg inorganic N g⁻¹ SOC 24 d⁻¹, P<0.001), and greater SMBC (53 vs 21 mg SMBC g⁻¹ SOC, P<0.001). Specific respiratory activity of SMBC was, however, consistently lower in the thermic than in the frigid regions (29 vs 34 mg CO₂–C g⁻¹ SMBC d⁻¹, P<0.01). Higher mean annual precipitation resulted in consistently lower basal soil respiration (1.1 vs 1.3 mg CO₂–C g⁻¹ SOC d⁻¹ in the wet compared with the dry regions, P<0.01) and lower SMBC (31 vs 43 mg SMBC g⁻¹ SOC, P<0.001), but had inconsistent effects on net N mineralization that depended upon temperature regime. Specific respiratory activity of SMBC was consistently greater in the wet than the dry regions (≈33 vs 29 mg CO₂–C g⁻¹ SMBC d⁻¹, P<0.01). Although the thermic regions were not able to retain as high a level of SOC as the frigid regions, due likely to high annual decomposition rates, biologically active soil fractions were as high per mass of soil and even 2–3-times greater per unit of SOC in the thermic compared with the frigid regions. These results suggest that macroclimate has a large impact on the portion of soil organic matter that is potentially active, but a relatively small impact on the specific respiratory activity of SMBC.     

Formation and use of microbial residues after adding sugarcane sucrose to a heated soil devoid of soil organic matter

A 67-day incubation experiment was carried out with a soil initially devoid of any organic matter due to heating, which was amended with sugarcane sucrose (C₄-sucrose with a δ¹³C value of ?10.5‰), inorganic N and an inoculum for recolonisation and subsequently at day 33 with C₃-cellulose (δ¹³C value of ?23.4‰). In this soil, all organic matter is in the microbial biomass or in freshly formed residues, which makes it possible to analyse more clearly the role of microbial residues for decomposition of N-poor substrates. The average δ¹³C value over the whole incubation period was ?10.7‰ in soil total C in the treatments without C₃-cellulose addition. In the CO₂ evolved, the δ¹³C values decreased from ?13.4‰ to ?15.4‰ during incubation. In the microbial biomass, the δ¹³C values increased from ?11.5‰ to ?10.1‰ at days 33 and 38. At day 67, 36% of the C₄-sucrose was left in the treatment without a second amendment. The addition of C₃-cellulose resulted in a further 7% decrease, but 4% of the C₃-cellulose was lost during the second incubation period. Total microbial biomass C declined from 200 μg g^?1 soil at day 5 to 70 μg g^?1 soil at day 67. Fungal ergosterol increased to 1.5 μg g^?1 soil at day 12 and declined more or less linearly to 0.4 μg g^?1 soil at day 67. Bacterial muramic acid declined from a maximum of 35 μg g^?1 soil at day 5 to a constant level of around 16 μg g^?1 soil. Glucosamine showed a peak value at day 12. Galactosamine remained constant throughout the incubation. The fungal C/bacterial C ratio increased more or less linearly from 0.38 at day 5 to 1.1 at day 67 indicating a shift in the microbial community from bacteria to fungi during the incubation. The addition of C₃-cellulose led to a small increase in C₃-derived microbial biomass C, but to a strong increase in C₄-derived microbial biomass C. At days 45 and 67, the addition of N-free C₃-cellulose significantly decreased the C/N ratio of the microbial residues, suggesting that this fraction did not serve as an N-source, but as an energy source.     

Rapid estimation of microbial biomass in grassland soils by ultra-violet absorbance

A reliable and simple technique for estimating soil microbial biomass (SMB) is essential if the role of microbes in many soil processes is to be quantified. Conventional techniques are notoriously time-consuming and unreproducible. A technique was investigated that uses the UV absorbance at 280 nm of 0.5 M K₂SO₄ extracts of fumigated and unfumigated soils to estimate the concentrations of carbon, nitrogen and phosphorus in the SMB. The procedure is based on the fact that compounds released after chloroform fumigation from lysed microbial cells absorb in the near UV region. Using 29 UK permanent grassland soils, with a wide range of organic matter (2.9–8.0%) and clay contents (22–68%), it was demonstrated that the increase in UV absorbance at 280 nm after soil fumigation was strongly correlated with the SMB C (r=0.92), SMB N (r=0.90) and SMB P (r=0.89), as determined by conventional methods. The soils contained a wide range of SMB C (412–3412 μg g⁻¹ dry soil), N (57–346 μg g⁻¹ dry soil) and P (31–239 μg g⁻¹ dry soil) concentrations. It was thus confirmed that the UV absorbance technique described was a rapid, simple, precise and relatively inexpensive method of estimating soil microbial biomass.     

Microbial biomass in a semi arid soil of the central highlands of Mexico cultivated with maize or under natural vegetation

Microbial biomass (MB) is the key factor in nutrient dynamics in soil, but no information exists how clearing of vegetation to cultivate maize in the central highlands of Mexico might affect it. Soil MB was measured with the chloroform fumigation incubation (CFI) and fumigation extraction (CFE) techniques and the substrate-induced respiration (SIR) method in soil sampled under or outside the canopy of mesquite (Prosopis laevigata) and huisache (Acacia tortuoso), N₂ fixing shrubs, and from fields cultivated with maize. Microbial biomass C as measured with the CFI technique ranged from 122 mg C kg⁻¹ in agricultural soil to 373 mg C kg⁻¹ in soil sampled under mesquite shrubs. Microbial biomass N as measured with the CFI technique ranged from 11 mg N kg⁻¹ in agricultural soil to 116 mg N kg⁻¹ in soil sampled under mesquite shrub. The ratio of microbial biomass C as measured with CFI related to the ninhydrin-positive compounds (NPC) was 12.23 after 1 day and 8.43 after 10 days while the relationship with extractable C was 3.15 and 2.96, respectively. The metabolic quotient (qCO₂) decreased in the order OUTSIDE > MESQUITE > HUIZACHE > AGRICULTURE, and the microbial biomass:soil organic C ratio decreased in the order MESQUITE > HUIZACHE > OUTSIDE > AGRICULTURE using SIR to determine the microbial biomass. It was found that converting soil under natural vegetation to arable soil was not only detrimental for soil quality, but might be unsustainable as organic matter input is limited.     

Microbial properties and soil respiration in submontane forests of Venezuelian Guyana: characteristics and response to fertilizer treatments

The distribution of vegetation types in Venezuelan Guyana (in the ‘Canaima’ National Park) represents a transitional stage in a long term process of savannization, a process considered to be conditioned by a combined chemical and intermittent drought stress. All types of woody vegetation in this environment accumulate large amounts of litter and soil organic carbon (SOC). We hypothesized that this accumulation is caused by low microbial activity. During 1 year we measured microbial biomass carbon (Cmic), microbial respiration and soil respiration of stony Oxisols (Acrohumox) at a tall, a medium and a low forest and with three chemical modifications of site conditions by the addition of NO₃⁻, Ca²⁺ and PO₄³⁻ as possible limiting elements. Due to high SOC contents, mean Cmic was 1 mg g soil⁻¹ in the mineral topsoil and 3 mg g soil⁻¹ in the forest floor. Mean microbial respiration in the mineral topsoil and the forest floor were 165 and 192 μg CO₂-C g soil⁻¹ d⁻¹, respectively. We calculated high mean metabolic quotients (qCO₂) of 200 mg CO₂-C g Cmic⁻¹ d⁻¹ in the litter layer and 166 mg CO₂-C g Cmic⁻¹ d⁻¹ in the mineral topsoil, while the Cmic-to-SOC ratios were as low as 1.0% in the litter layer and 0.8% in the mineral topsoil. Annual soil respiration was 9, 12 and 10 Mg CO₂-C ha⁻¹ yr⁻¹ in the tall, medium and low forest, respectively. CO₂ production was significantly increased by CaHPO₄ fertilization, but no consistent effects were caused by Ca²⁺ and NO₃⁻, fertilization. Our findings indicate that Cmic and microbial respiration are reduced by low nutrient concentrations and low litter and SOC quality. Reduced microbial decomposition may have contributed to SOC accumulation in these forests.     

Microbial utilization and mineralization of [14C]glucose added in six orders of concentration to soil

The substrate availability for microbial biomass (MB) in soil is crucial for microbial biomass activity. Due to the fast microbial decomposition and the permanent production of easily available substrates in the rooted top soil mainly by plants during photosynthesis, easily available substrates make a very important contribution to many soil processes including soil organic matter turnover, microbial growth and maintenance, aggregate stabilization, CO₂ efflux, etc. Naturally occurring concentrations of easily available substances are low, ranging from 0.1 μM in soils free of roots and plant residues to 80 mM in root cells. We investigated the effect of adding ¹⁴C-labelled glucose at concentrations spanning the 6 orders of magnitude naturally occurring concentrations on glucose uptake and mineralization by microbial biomass. A positive correlation between the amount of added glucose and its portion mineralized to CO₂ was observed: After 22 days, from 26% to 44% of the added 0.0009 to 257 μg glucose C g^?1 soil was mineralized. The dependence of glucose mineralization on its amount can be described with two functions. Up to 2.6 μg glucose C g^?1 soil (corresponds to 0.78% of initial microbial biomass C), glucose mineralization increased with the slope of 1.8% more mineralized glucose C per 1 μg C added, accompanied by an increasing incorporation of glucose C into MB. An increased spatial contact between micro-organisms and glucose molecules with increasing concentration may be responsible for this fast increase in mineralization rates (at glucose additions <2.6 μg C g^?1). At glucose additions higher than 2.6 μg C g^?1 soil, however, the increase of the glucose mineralization per 1 μg added glucose was much smaller as at additions below 2.6 μg C g^?1 soil and was accompanied by decreasing portions of glucose ¹⁴C incorporated into microbial biomass. This supports the hypothesis of decreasing efficiency of glucose utilization by MB in response to increased substrate availability in the range 2.6–257 μg C g^?1 (=0.78–78% of microbial biomass C). At low glucose amounts, it was mainly stored in a chloroform-labile microbial pool, but not readily mineralized to CO₂. The addition of 257 μg glucose C g^?1 soil (0.78 μg C glucose μg^?1 C micro-organisms) caused a lag phase in mineralization of 19 h, indicating that glucose mineralization was not limited by the substrate availability but by the amount of MB which is typical for 2nd order kinetics.     

Applying an oxygen-based respiratory assay to assess soil microbial responses to substrate and N availability

Documented approaches for measuring soil microbial activities and their controlling factors under field conditions are needed to advance understanding of soil microbial processes for numerous applications. We manipulated field plots with carbon (C) and nitrogen (N) additions to test the capability of a respiratory assay to: (1) measure respiration of endogenous soil C in comparison to field-measured CO₂ fluxes; (2) determine substrate-induced respiratory (SIR) activities that are consistent with substrate availability in the field; and, (3) report N availability in the field based on assay responses with and without added N. The respiratory assay utilizes a microplate containing an oxygen-sensitive fluorescent ruthenium dye. Respiratory activities measured with this approach have previously been shown to occur within short (6–8 h) incubation periods using low substrate concentrations that minimize enrichment during the assay. Field treatments were conducted in a randomized full-factorial design with C substrate (casamino acids, glucose, or none) and inorganic N (±) as the treatment factors. With one exception, we found that respiration of endogenous soil C in the assay responded to the field treatments in a similar manner to CO₂ fluxes measured in the field. Patterns of SIR with low concentrations of added amino acid or carbohydrate substrate (200 μg C g⁻¹ soil) were consistent with field treatments. The ratio (N_ratio) of carbohydrate respiration with added N (25 μg N g⁻¹ soil) to the same without N in the assay was significantly (P < 0.05) decreased by field N amendment. The carbohydrate N_ratio exhibited a logarithmic relationship (r = 0.64, P < 0.05) with extractable inorganic soil nitrate and ammonium concentrations. These data significantly extend and support the capability of this oxygen-based respiratory assay to evaluate in situ soil activities and examine factors that limit these activities.     

Energetic eco‐physiology of the soil microbiota in two landscapes of southern and northern Germany

Interactions between microbial communities and organic matter were analyzed for soils from the project regions ’︁Ecosystem Research in the Agricultural Landscape/FAM, Munich’ in southern Germany and ’︁Ecosystem Research in the Bornhöved Lake district’ from northern Germany using ratios between microbial biomass content (C_mic), microbial metabolic quotient (qCO₂) and organic carbon content (C_org). In the agricultural soils in southern Germany, the qCO₂/C_org ratio differed significantly with respect to agricultural management in contrast to ecophysiological C_mic/C_org ratio. In addition, C_mic/C_org ratio decreased from 39 to 21 mg C_mic g^—1 C_org and qCO₂/C_org ratio increased from 72 to 180 mg CO₂‐C g^—1 C_mic h^—1 (g C_org g^—1 soil)^—1 with increasing soil depth. For the upper soil horizons from the landscape in northern Germany the two quotients differed significantly with reference to land use showing highest microbial colonization under grassland and lowest under beech forest. In contrast, C use efficiency was lowest in arable field under maize monoculture and highest in a wet grassland having a high organic C content.     

Influence of reduced tillage on earthworm and microbial communities under organic arable farming

Although reduced tillage is an agricultural practice reported to decrease soil erosion and external inputs while enhancing soil fertility, it has still rarely been adopted by European organic farmers. The objective of this study was to assess the long-term interactive effects of tillage (conventional (CT) vs. reduced (RT)) and fertilization (slurry (S) vs. composted manure/slurry (MCS)) on earthworms and microbial communities in a clay soil under spelt in an organic 6-year crop rotation. Earthworm populations (species, density and biomass, cocoons) were investigated by handsorting the soil nine years after initial implementation of the treatments. Soil microbial carbon (C_mic) and nitrogen (N_mic) were measured by chloroform-fumigation extraction and a simplified phospholipid fatty acid (PLFA) analysis was used to separate for populations of bacteria, fungi and protozoa. Significantly increased total earthworm density in RT plots was mainly attributed to increased numbers of juveniles. Moreover, we found five times more cocoons with RT. Species richness was not affected by the treatments, but tillage treatments had differentially affected populations at the species-level. In addition, cluster analysis at the community level revealed two distinct groups of plots in relation to tillage treatments. In RT plots C_mic increased in the 0–10 cm and 10–20 cm soil layers, while PLFA concentrations indicative of Gram-negative bacteria, fungi and protozoa only increased in the topsoil. Lower bacteria-to-fungi ratios in the upper soil layer of RT plots indicated a shift to fungal-based decomposition of organic matter whereas a higher C_mic-to-C_org ratio pointed towards enhanced substrate availability. Slurry application decreased microbial biomass and enhanced density of juvenile anecic earthworms but overall fertilization effect was weak and no interactions with tillage were found. In conclusion, tillage is a major driver in altering communities of earthworms and microorganisms in arable soils. The use of reduced tillage provides an approach for eco-intensification by enhancing inherent soil biota functions under organic arable farming.     

Evaluation of an optimal extraction method for measuring d-ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) in agricultural soils and its association with soil microbial CO2 assimilation

Assimilating atmospheric carbon (C) into terrestrial ecosystems is recognized as a primary measure to mitigate global warming. Ribulose-1,5-bisphosphate carboxylase/oxygenase (RubisCO) is the dominant enzyme by which terrestrial autotrophic bacteria and plants fix CO₂. To investigate the possibility of using RubisCO activity as an indicator of microbial CO₂ fixation potential, a valid and efficient method for extracting soil proteins is needed. We examined three methods commonly used for total soil protein extraction. A simple sonication method for extracting soil protein was more efficient than bead beating or freeze–thaw methods. Total soil protein, RubisCO activity, and microbial fixation of CO₂ in different agricultural soils were quantified in an incubation experiment using ¹⁴C-CO₂ as a tracer. The soil samples showed significant differences in protein content and RubisCO activity, defined as nmol CO₂ fixed g⁻¹ soil min⁻¹. RubisCO activities ranged from 10.68 to 68.07 nmol CO₂ kg⁻¹ soil min⁻¹, which were closely related to the abundance of cbbL genes (r = 0.900, P = 0.0140) and the rates of microbial CO₂ assimilation (r = 0.949, P = 0.0038). This suggests that RubisCO activity can be used as an indicator of soil microbial assimilation of atmospheric CO₂.     

Effects of Zn enriched sewage sludge on microbial activities and biomass in soil

An incubation study in closed static microcosms was performed to elucidate Zn effects on N mineralisation in relation to other microbial activities and biomass in a sandy soil. Sewage sludge equivalent to 25 t ha⁻¹ was enriched with five different rates of Zn to add concentrations between 50 and 800 μg Zn g⁻¹ soil. All microbial indices were increasingly depressed with increasing Zn concentration of the sewage sludge, but they were affected with different intensity: Zn had especially large effects on CO₂ production and qCO₂, moderate effects on N mineralisation and relatively small effects on protease activity, biomass C and arginine ammonification.     

Emission of nitrous oxide from hydrocarbon contaminated soil amended with waste water sludge and earthworms

Soils in Mexico are often contaminated with hydrocarbons and addition of waste water sludge and earthworms accelerates their removal. However, little is known how contamination and subsequent bioremediation affects emissions of N₂O and CO₂. A laboratory study was done to investigate the effect of waste water sludge and the earthworm Eisenia fetida on emission of N₂O and CO₂ in a sandy loam soil contaminated with the polycyclic aromatic hydrocarbons (PAHs): phenanthrene, anthracene and benzo(a)pyrene. Emissions of N₂O and CO₂, and concentrations of inorganic N (ammonium (NH₄⁺), nitrite (NO₂^?) nitrate (NO₃^?)) were monitored after 0, 5, 24, 72 and 168 h. Adding E. fetida to the PAHs contaminated soil increased CO₂ production rate significantly 2.0 times independent of the addition of sludge. The N₂O emission rate from unamended soil expressed on a daily base was 5 μg N kg^?1 d^?1 for the first 2 h and increased to a maximum of 325 μg N kg^?1 d^?1 after 48 h and then decreased to 10 μg N kg^?1 d^?1 after 168 h. Addition of PAHs, E. fetida or PAHs + E. fetida had no significant effect on the N₂O emission rate. Adding sludge to the soil sharply increased the N₂O emission rate to >400 μg N kg^?1 d^?1 for the entire incubation with a maximum of 1134 μg N kg^?1 d^?1 after 48 h. Addition of E. fetida, PAHs or PAHs + E. fetida to the sludge-amended soil reduced the N₂O emission rate significantly compared to soil amended with sludge after 24 h. It was found that contaminating soil with PAHs and adding earthworms had no effect on emissions of N₂O. Emission of N₂O, however, increased in sludge-amended soil, but addition of earthworms to this soil and contamination reduced it.