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.     

Measuring microbial uptake of nitrogen in nutrient-amended sandy soils—A mass-balance based approach

Soil microbial biomass N is commonly determined through fumigation-extraction (FE), and a conversion factor (K_EN) is necessary to convert extractable N to actual soil biomass N. Estimation of K_EN has been constrained by various uncertainties including potential microbial immobilisation. We developed a mass-balance approach to quantify changes in microbial N storage during nutrient-amended incubation, in which microbial uptake is determined as the residual in a ‘mass-balance’ based on soil-water N before and after amended incubation. The approach was applied to three sandy soils of southwestern Australia, to determine microbial N immobilisation during 5-day incubation in response to supply of 2.323 mg C g⁻¹, 100 μg N g⁻¹ and 20 μg P g⁻¹. The net N immobilisation was estimated to be 95-114 μg N g⁻¹ in the three soils, equivalent to 82.7-85.1% of soil-water N following the amendment. Such estimation for microbial uptake does not depend on fumigation and K_EN conversion, but for comparison purposes we estimated ‘nominal’ K_EN values (0.11-0.14) for the three soils, which were comparable to previously reported K_EN from soils receiving C and N amendment. The accuracy of our approach depends on the mass-balance equation and the integrated measurement errors of the multiple N pools, and was assessed practically through recoveries of added-N when microbial uptake can be minimised. Near-satisfactory recoveries were achieved under such conditions. Our mass-balance approach provides information not only about changes in the microbial biomass nitrogen storage, but also major N-pools and their fluxes in regulating soil N concentrations under substrate and nutrient amended conditions.     

Initial recovery of soil carbon and nitrogen pools and dynamics following disturbance in jack pine forests: A comparison of wildfire and clearcut harvesting

Forests naturally maintained by stand-replacing wildfires are often managed with clearcut harvesting, yet we know little about how replacing wildfire with clearcutting affects soil processes and properties. We compared the initial recovery of carbon (C) and nitrogen (N) pools and dynamics following disturbance in jack pine (Pinus banksiana) stands in northern Lower Michigan, USA, by sampling soils (Oa+A horizons) from three “treatments”: 3-6-year-old harvest-regenerated stands, 3-6-year-old wildfire-regenerated stands and 40-55-year-old intact, mature stands (n=4 stands per treatment). We measured total C and N; microbial biomass and potentially mineralizable C and N; net nitrification; and gross rates of N mineralization and nitrification. Burned stands exhibited reduced soil N but not C, whereas clearcut and mature stands had similar quantities of soil organic matter. Both disturbance types reduced microbial biomass C compared to mature stands; however, microbial biomass N was reduced in burned stands but not in clearcut stands. The experimental C and N mineralization values were fit to a first-order rate equation to estimate potentially mineralizable pool size (C₀ and N₀) and rate parameters. Values for C₀ in burned and clearcut stands were approximately half that of the mature treatment, with no difference between disturbance types. In contrast, N₀ was lowest in the wildfire stands (170.2 μg N g⁻¹), intermediate in the clearcuts (215.4 μg N g⁻¹) and highest in the mature stands (244.6 μg N g⁻¹). The most pronounced difference between disturbance types was for net nitrification. These data were fit to a sigmoidal growth equation to estimate potential NO₃⁻ accumulation (Nit_max) and kinetic parameters. Values of Nit_max in clearcut soils exceeded that of wildfire and mature soils (149.2 vs. 83.5 vs. 96.5 μg NO₃⁻-N g⁻¹, respectively). Moreover, the clearcut treatment exhibited no lag period for net NO₃⁻ production, whereas the burned and mature treatments exhibited an approximate 8-week lag period before producing appreciable quantities of NO₃⁻. There were no differences between disturbances in gross rates of mineralization or nitrification; rather, lower NO₃⁻ immobilization rates in the clearcut soils, 0.20 μg NO₃⁻ g⁻¹ d⁻¹ compared to 0.65 in the burned soils, explained the difference in net nitrification. Because the mobility of NO₃⁻ and NH₄⁺ differs markedly in soil, our results suggest that differences in nitrification between wildfire and clearcutting could have important consequences for plant nutrition and leaching losses following disturbance.     

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.     

Humus accumulation and microbial activities in calcari-epigleyic fluvisols under grassland and forest diked in for 30 years

The accumulation and transformation of organic matter during soil development is rarely investigated although such processes are relevant when discussing about carbon sequestration in soil. Here, we investigated soils under grassland and forest close to the North Sea that began its genesis under terrestrial conditions 30 years ago after dikes were closed. Organic C contents of up to 99 mg g⁻¹ soil were found until 6 cm soil depth. The humus consisted mainly of the fraction lighter than 1.6 g cm⁻³ which refers to poorly degraded organic carbon. High microbial respiratory activity was determined with values between 1.57 and 1.17 μg CO₂-C g⁻¹ soil h⁻¹ at 22 °C and 40 to 70% water-holding capacity for the grassland and forest topsoils, respectively. The microbial C to organic C ratio showed values up to 20 mg C_mic g⁻¹ C_org. Although up to 2.69 kg C m⁻² were estimated to be sequestered during 30 years, the microbial indicators showed intensive colonisation and high transformation rates under both forest and grassland which were higher than those determined in agricultural and forest topsoils in Northern Germany.     

Taxon-specific responses of soil bacteria to the addition of low level C inputs

The addition of small or trace amounts of carbon to soils can result in the release of 2-5 times more C as CO₂ than was added in the original solution. The identity of the microorganisms responsible for these so-called trigger effects remains largely unknown. This paper reports on the response of individual bacterial taxa to the addition of a range of ¹⁴C-glucose concentrations (150, 50 and 15 and 0 μg C g⁻¹ soil) similar to the low levels of labile C found in soil. Taxon-specific responses were identified using a modification of the stable isotope probing (SIP) protocol and the recovery of [¹⁴C] labelled ribosomal RNA using equilibrium density gradient centrifugation. This provided good resolution of the ‘heavy’ fractions ([¹⁴C] labelled RNA) from the ‘light’ fractions ([¹²C] unlabelled RNA). The extent of the separation was verified using autoradiography. The addition of [¹⁴C] glucose at all concentrations was characterised by changes in the relative intensity of particular bands. Canonical correspondence analysis (CCA) showed that the rRNA response in both the ‘heavy’ and ‘light’ fractions differed according to the concentration of glucose added but was most pronounced in soils amended with 150 μg C g⁻¹ soil. In the ‘heavy RNA’ fractions there was a clear separation between soils amended with 150 μg C g⁻¹ soil and those receiving 50 and 15 μg C g⁻¹ soil indicating that at low C inputs the microbial community response is quite distinct from that seen at higher concentrations. To investigate these differences further, bands that changed in relative intensity following amendment were excised from the DGGE gels, reamplified and sequenced. Sequence analysis identified 8 taxa that responded to glucose amendment (Bacillus, Pseudomonas, Burkholderia, Bradyrhizobium, Actinobacteria, Nitrosomonas, Acidobacteria and an uncultured β-proteobacteria). These results show that radioisotope probing (RNA-RIP) can be used successfully to study the fate of labile C substrates, such as glucose, in soil.     

Dynamics of microbial biomass and nitrogen supply during primary succession on blastfurnace slag dumps in dry tropics

This paper reports the role of microbial biomass in the establishment of N pools in the substratum during primary succession (till 40-year age) in Blastfurnace Slag Dumps, an anthropogenically created land form in the tropics. Initially in the depressions in the slag dumps fine soil particles (silt+clay) accumulate, retaining moisture therein, and providing microsites for the accumulation of microbial biomass. In all sites microbial biomass showed distinct seasonality, with summer-peak and rainy season-low standing crops. During the summer season microbial biomass C ranged from 18.6 μg g⁻¹ in the 1-year old site to ca. 235 μg g⁻¹ in the 40-year old site; correspondingly, microbial biomass N ranged from 1.22 to 40 μg g⁻¹. On sites 2.5-years of age and younger, the microbial biomass N content accounted for more than 50% of the organic N in the soil, whereas the proportion of microbial biomass N was ca. 7% of organic N in 40-year old site. The strong correlation between microbial biomass and total N in soil indicated a significant role of microbes in the build-up of nitrogen during the initial stages of succession in the slag dumps. Though the organic N pool in the soil was low (594 mg kg⁻¹) even after 40 years of succession, the available N (NH₄-N and NO₃-N) contents in the soil were generally high through the entire age series (ca. 16-32 μg g⁻¹) during the rainy season (which supports active growth of the herbaceous community). The high mineral-N status on the slag dump was related with high N-mineralization rates, particularly in the young sites (20.6 and 13.9 μg g⁻¹ month⁻¹ at 1 and 2.5-year age). We suggest that along with the abiotic factors having strong effect on ecosystem functioning, the microbial biomass, an important biotic factor, shows considerable influence on soil nutrient build-up during early stages of primary succession on the slag dumps. The microbial biomass dynamics initiates biotic control in developing slag dumps ecosystem through its effect on nitrogen pools and availability.     

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.     

Controls over pathways of carbon efflux from soils along climate and black spruce productivity gradients in interior Alaska

Small changes in C cycling in boreal forests can change the sign of their C balance, so it is important to gain an understanding of the factors controlling small exports like water-soluble organic carbon (WSOC) fluxes from the soils in these systems. To examine this, we estimated WSOC fluxes based on measured concentrations along four replicate gradients in upland black spruce (Picea mariana [Mill.] BSP) productivity and soil temperature in interior Alaska and compared them to concurrent rates of soil CO₂ efflux. Concentrations of WSOC in organic and mineral horizons ranged from 4.9 to 22.7 g C m⁻² and from 1.4 to 8.4 g C m⁻², respectively. Annual WSOC fluxes (4.5-12.0 g C m⁻² y⁻¹) increased with annual soil CO₂ effluxes (365-739 g C m⁻² y⁻¹) across all sites (R²=0.55, p=0.02), with higher fluxes occurring in warmer, more productive stands. Although annual WSOC flux was relatively small compared to total soil CO₂ efflux across all sites (<3%), its relative contribution was highest in warmer, more productive stands which harbored less soil organic carbon. The proportions of relatively bioavailable organic fractions (hydrophilic organic matter and low molecular weight acids) were highest in WSOC in colder, low-productivity stands whereas the more degraded products of microbial activity (fulvic acids) were highest in warmer, more productive stands. These data suggest that WSOC mineralization may be a mechanism for increased soil C loss if the climate warms and therefore should be accounted for in order to accurately determine the sensitivity of boreal soil organic C balance to climate change.     

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.     

Cyanobacterial inoculation of heated soils: effect on microorganisms of C and N cycles and on chemical composition in soil surface

Physiological groups of soil microorganisms, total C and N and available nutrients were investigated in four heated (350 °C, 1 h) soils (one Ortic Podsol over sandstone and three Humic Cambisol over granite, schist or limestone) inoculated (1.5 μg chlorophyll a g⁻¹ soil or 3.0 μg chlorophyll a g⁻¹ soil) with four cyanobacterial strains of the genus Oscillatoria, Nostoc or Scytonema and a mixture of them.Cyanobacterial inoculation promoted the formation of microbiotic crusts which contained a relatively high number of NH₄⁺-producers (7.4×10⁹ g⁻¹ crust), starch-mineralizing microbes (1.7×10⁸ g⁻¹ crust), cellulose-mineralizing microbes (1.4×10⁶ g⁻¹ crust) and NO₂⁻ and NO₃⁻ producers (6.9×10⁴ and 7.3×10³ g⁻¹ crust, respectively). These crusts showed a wide range of C and N contents with an average of 293 g C kg⁻¹ crust and 50 g N kg⁻¹ crust, respectively. In general, Ca was the most abundant available nutrient (804 mg kg⁻¹ crust), followed by Mg (269 mg kg⁻¹ crust), K (173 mg kg⁻¹ crust), Na (164 mg kg⁻¹ crust) and P (129 mg kg⁻¹ crust). There were close positive correlations among all the biotic and abiotic components of the crusts.Biofertilization with cyanobacteria induced great microbial proliferation as well as high increases in organic matter and nutrients in the surface of the heated soils. In general, cellulolytics were increased by four logarithmic units, amylolytics and ammonifiers by three logarithmic units and nitrifiers by more than two logarithmic units. C and N contents rose an average of 275 g C kg⁻¹ soil and 50 g N kg⁻¹ soil while the C:N ratio decreased up to 7 units. Among the available nutrients the highest increase was for Ca (315 mg kg⁻¹ soil) followed by Mg (189 mg kg⁻¹ soil), K (111 mg kg⁻¹ soil), Na (109 mg kg⁻¹ soil) and P (89 mg kg⁻¹ soil). Fluctuations of the microbial groups as well as those of organic matter and nutrients were positively correlated.The efficacy of inoculation depended on both the type of soil and the class of inoculum. The best treatment was the mixture of the four strains and, whatever the inoculum used, the soil over lime showed the most developed crust followed by the soils over schist, granite and sandstone. In the medium term there were not significant differences between the two inocula amounts tested.These results showed that inoculation of burned soils with alien N₂-fixing cyanobacteria may be a biotechnological means of promoting microbiotic crust formation, enhancing C and N cycling microorganisms and increasing organic matter and nutrient contents in heated soils.     

Shifts in amino sugar and ergosterol contents after addition of sucrose and cellulose to soil

An incubation experiment with organic soil amendments was carried out with the aim to determine whether formation and use of microbial tissue (biomass and residues) could be monitored by measuring glucosamine and muramic acid. Living fungal tissue was additionally determined by the cell-membrane component ergosterol. The organic amendments were fibrous maize cellulose and sugarcane sucrose adjusted to the same C/N ratio of 15. In a subsequent step, spherical cellulose was added without N to determine whether the microbial residues formed initially were preferentially decomposed. In the non-amended control treatment, ergosterol remained constant at 0.44 μg g⁻¹ soil throughout the 67-day incubation. It increased to a highest value of 1.9 μg g⁻¹ soil at day 5 in the sucrose treatment and to 5.0 μg g⁻¹ soil at day 33 in the fibrous cellulose treatment. Then, the ergosterol content declined again. The addition of spherical cellulose had no further significant effects on the ergosterol content in these two treatments. The non-amended control treatment contained 48 μg muramic acid and 650 μg glucosamine g⁻¹ soil at day 5. During incubation, these contents decreased by 17% and 19%, respectively. A 33% increase in muramic acid and an 8% increase in glucosamine were observed after adding sucrose. Consequently, the ratio of fungal C to bacterial C based on bacterial muramic acid and fungal glucosamine was lowered in comparison with the other two treatments. No effect on the two amino sugars was observed after adding cellulose initially or subsequently during the second incubation period. This indicates that the differences in quality between sucrose and cellulose had a strong impact on the formation of microbial residues. However, the amino sugars did not indicate a preferential decomposition of microbial residues as N sources.     

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.     

High specific activity in low microbial biomass soils across a no-till evapotranspiration gradient in Colorado

The need to identify microbial community parameters that predict microbial activity is becoming more urgent, due to the desire to manage microbial communities for ecosystem services as well as the desire to incorporate microbial community parameters within ecosystem models. In dryland agroecosystems, microbial biomass C (MBC) can be increased by adopting alternative management strategies that increase crop residue retention, nutrient reserves, improve soil structure and result in greater water retention. Changes in MBC could subsequently affect microbial activities related to decomposition, C stabilization and sequestration. We hypothesized that MBC and potential microbial activities that broadly relate to decomposition (basal and substrate-induced respiration, N mineralization, and β-glucosidase and arylsulfatase enzyme activities) would be similarly affected by no-till, dryland winter wheat rotations distributed along a potential evapotranspiration (PET) gradient in eastern Colorado. Microbial biomass was smaller in March 2004 than in November 2003 (417 vs. 231 μg g⁻¹ soil), and consistently smaller in soils from the high PET soil (191 μg g⁻¹) than in the medium and low PET soils (379 and 398 μg g⁻¹, respectively). Among treatments, MBC was largest under perennial grass (398 μg g⁻¹). Potential microbial activities did not consistently follow the same trends as MBC, and the only activities significantly correlated with MBC were β-glucosidase (r = 0.61) and substrate-induced respiration (r = 0.27). In contrast to MBC, specific microbial activities (expressed on a per MBC basis) were greatest in the high PET soils. Specific but not total activities were correlated with microbial community structure, which was determined in a previous study. High specific activity in low biomass, high PET soils may be due to higher microbial maintenance requirements, as well as to the unique microbial community structure (lower bacterial-to-fungal fatty acid ratio and lower 17:0 cy-to-16:1ω7c stress ratio) associated with these soils. In conclusion, microbial biomass should not be utilized as the sole predictor of microbial activity when comparing soils with different community structures and levels of physiological stress, due to the influence of these factors on specific activity.     

Microbial colonisation of roots as a function of plant species

Fifteen plants species were grown in the greenhouse on the same soil and sampled at flowering to obtain rhizosphere soil and root material. In both fractions, the data on fungal and bacterial tissue obtained by amino sugar analysis were compared with the total microbial biomass based on fumigation-extraction and ergosterol data. The available literature on glucosamine concentrations in fungi and on muramic acid concentrations in bacteria was reviewed to prove the possibility of generating conversion values for general use in root material. All microbial properties analysed revealed strong species-specific differences in microbial colonisation of plant roots. The root material contained considerable amounts of microbial biomass C and biomass N, reaching mean levels of 10.9 and 1.4 mg g⁻¹ dry weight, respectively. However, the majority of CHCl₃ labile C and N, i.e. 89 and 55% was root derived. The average amount of ergosterol was 13 μg g⁻¹ dry weight and varied between 0.0 for Phacelia roots and 45.5 μg g⁻¹ dry weight for Vicia roots. The ergosterol content in root material of mycorrhizal and non-mycorrhizal plant species did not differ significantly. Fungal glucosamine was converted to fungal C by multiplication by 9 giving a range of 7.1-25.9 mg g⁻¹ dry weight in the root material. Fungal C and ergosterol were significantly correlated. Bacterial C was calculated by multiplying muramic acid by 45 giving a range from 1.7 to 21.6 mg g⁻¹ dry weight in the root material. In the root material of the 15 plant species, the ratio of fungal C-to-bacterial C ranged from 1.0 in mycorrhizal Trifolium roots to 9.5 in non-mycorrhizal Lupinus roots and it was on average 3.1. These figures mean that the microbial tissue in the root material consists on average of 76% fungal C and 24% bacterial C. The differences in microbial colonisation of the roots were reflected by differences in microbial indices found in the rhizosphere soil, most strongly for microbial biomass C and ergosterol, but to some extent also for glucosamine and muramic acid.     

Long-term organic phosphorus mineralization in Spodosols under forests and its relation to carbon and nitrogen mineralization

In forest soils where a large fraction of total phosphorus (P) is in organic forms, soil micro-organisms play a major role in the P cycle and plant availability since they mediate organic P transformations. However, the correct assessment of organic P mineralization is usually a challenging task because mineralized P is rapidly sorbed and most mineralization fluxes are very weak. The objectives of the present work were to quantify in five forest Spodosols at soil depths of 0-15 cm net mineralization of total organic P and the resulting increase in plant available inorganic P and to verify whether net or gross P mineralization could be estimated using the C or N mineralization rates. Net mineralization of total organic P was derived from the net changes in microbial P and gross mineralization of P in dead soil organic matter. We studied very low P-sorbing soils enabling us to use lower extractants to assess the change in total inorganic P as a result of gross mineralization of P in dead soil organic matter. In addition, to enable detection of gross mineralization of P in dead soil organic matter, a long-term incubation (517 days) experiment was carried out. At the beginning of the experiment, total P contents of the soils were very low (19-51 μg g⁻¹) and were essentially present as organic P (17-44 μg g⁻¹, 85-91%) or microbial P (6-14 μg g⁻¹; 24-39%). Conversely, the initial contents of inorganic P were low (2-7 μg g⁻¹; 9-15%). The net changes in the pool size of microbial P during the 517 days of incubation (4-8 μg g⁻¹) and the amounts of P resulting from gross mineralization of dead soil organic matter (0.001-0.018 μg g⁻¹ day⁻¹; 0.4-9.5 μg g⁻¹ for the entire incubation period) were considerable compared to the initial amounts of organic P and also when compared to the initial diffusive iP fraction (<0.3 μg g⁻¹). Diffusive iP corresponds to the phosphate ions that can be transferred from the solid constituents to the soil solution under a gradient of concentration. Net mineralization of organic P induced an important increase in iP in soil solution (0.6-10 μg g⁻¹; 600-5000% increase) and lower increases in diffusive iP fractions (0.3-5 μg g⁻¹; 300-2000% increase), soil solid constituents having an extremely low reactivity relative to iP. Therefore, soil micro-organisms and organic P transformations play a major role in the bioavailability of P in these forest soils. In our study, the dead soil organic matter was defined as a recalcitrant organic fraction. Probably because gross mineralization of P from this recalcitrant organic fraction was mainly driven by the micro-organisms’ needs for energy, the rates of gross mineralization of C, N and P in the recalcitrant organic fraction were similar. Indirect estimation of gross mineralization of P in dead soil organic matter using the gross C mineralization rate seems thus an alternative method for the studied soils. However, additional studies are needed to verify this alternative method in other soils. No relationships were found between microbial P release and microbial C and N releases.     

CO2 emission in an intensively cultivated loam as affected by long-term application of organic manure and nitrogen fertilizer

A long-term field experiment was conducted to examine the influence of mineral fertilizer and organic manure on the equilibrium dynamics of soil organic C in an intensively cultivated fluvo-aquic soil in the Fengqiu State Key Agro-Ecological Experimental Station (Fengqiu county, Henan province, China) since September 1989. Soil CO₂ flux was measured during the maize and wheat growing seasons in 2002-2003 and 2004 to evaluate the response of soil respiration to additions and/or alterations in mineral fertilizer, organic manure and various environmental factors. The study included seven treatments: organic manure (OM), half-organic manure plus half-fertilizer N (NOM), fertilizer NPK (NPK), fertilizer NP (NP), fertilizer NK (NK), fertilizer PK (PK) and control (CK). Organic C in soil and the soil heavy fraction (organo-mineral complex) was increased from 4.47 to 8.61 mg C g⁻¹ and from 3.32 to 5.68 mg C g⁻¹, respectively, after the 13 yr application of organic manure. In contrast, organic C and the soil heavy fraction increased in NPK soil to only 5.41 and 4.38 mg C g⁻¹, respectively. In the CK treatment, these parameters actually decreased from the initial C concentrations (4.47 and 3.32 mg C g⁻¹) to 3.77 and 3.11 mg C g⁻¹, respectively. Therefore, organic manure efficiently elevated soil organic C. However, only 66% of the increased soil organic C was combined with clay minerals in the OM treatment. Cumulative soil CO₂ emissions from inter-row soil in the OM and NPK treatments were 228 and 188 g C m⁻² during the 2002 maize growing season, 132 and 123 g C m⁻² during the 2002/2003 wheat growing season, and 401 and 346 g C m⁻² yr⁻¹ in 2002-2003, respectively. However, during the 2004 maize growing season, cumulative soil CO₂ emissions were as high as 617 and 556 g C m⁻², respectively, due to the contribution of rhizosphere respiration. The addition of organic manure contributed to a 16% increase in soil CO₂ emission in 2002-2003 (compared to NPK), where only 27%, 36% and 24% of applied organic C was released as CO₂ during the 2002 and 2004 maize growing seasons and in 2002-2003, respectively. During the 2002/2003 wheat growing season, soil CO₂ flux was significantly affected by soil temperature below 20 °C, but by soil moisture (WFPS) during the 2004 maize growing season at soil temperatures above 18 °C. Optimum soil WFPS for soil CO₂ flux was approximately 70%. When WFPS was below 50%, it no longer had a significant impact on soil CO₂ flux during the 2002 maize growing season. This study indicates the application of organic manure composted with wheat straw may be a preferred strategy for increasing soil organic C and sequestering C in soil.     

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.     

Long-term effects of metal-containing farmyard manure and sewage sludge on soil organic matter in a fluvisol

Our aim was to establish the long-term effects of repeated applications after 20 y of organic amendments (farmyard manure at 10 t ha⁻¹ y⁻¹, and urban sewage sludge at two different rates, 10 t ha⁻¹ y⁻¹ and 100 t ha⁻¹ every 2 y) on the quality of a sandy and poorly buffered soil (Fluvisol, pH 6). Chemical characteristics and biodegradability of the labile organic matter, which is mainly derived from microbial biomass and biodegradation products of organic residues, were chosen as indicators for soil quality. The organic C content had reached a maximal value (30.6 g C kg⁻¹ in the 100 t sludge-treated soil), i.e. about 2.5 times that in the control. Six years after the last application, the organic C content and the microbial biomass content remained higher in sludge-treated soils than in the control. In contrast, the proportion of labile organic matter was significantly lower in sludge-treated soils than in manure-treated and control soils. The labile organic matter of sludge extracts appeared less humified than that of manure-treated and control soils.     

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.