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.     

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.     

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.     

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.     

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.     

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 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.     

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.     

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.     

Heterotrophic and autotrophic nitrification in two acid pasture soils

Laboratory incubation experiments, using ¹⁵N-labeling techniques and simple analytical models, were conducted to measure heterotrophic and autotrophic nitrification rates in two acid soils (pH 4.8-5.3; 1/5 in H₂O) with high organic carbon contents (6.2-6.8% in top 5 cm soil). The soils were from pastures located near Maindample and Ruffy in the Northeast Victoria, Australia. Gross rates of N mineralization, nitrification and immobilization were measured. The gross rates of autotrophic nitrification were 0.157 and 0.119 μg N g⁻¹ h⁻¹ and heterotrophic nitrification rates were 0.036 and 0.009 μg N g⁻¹ h⁻¹ for the Maindample and Ruffy soils, respectively. Heterotrophic nitrification accounted for 19% and 7% of the total nitrification in the Maindample and Ruffy soils, respectively. The heterotrophic nitrifiers used organic N compounds and no as the substrate for nitrification.     

Biochemical characterization of urban soil profiles from Stuttgart, Germany

The knowledge of biochemical properties of urban soils can help to understand nutrient cycling in urban areas and provide a database for urban soil management. Soil samples were taken from 10 soil profiles in the city of Stuttgart, Germany, differing in land use—from an essentially undisturbed garden area to highly disturbed high-density and railway areas. A variety of soil biotic (microbial biomass, enzyme activities) and abiotic properties (total organic C, elemental C, total N) were measured up to 1.9 m depth. Soil organic matter was frequently enriched in the subsoil. Microbial biomass in the top horizons ranged from 0.17 to 1.64 g C kg⁻¹, and from 0.01 to 0.30 g N kg⁻¹, respectively. The deepest soil horizon at 170-190 cm, however, contained 0.12 g C kg⁻¹ and 0.05 kg N kg⁻¹ in the microbial biomass. In general, arylsulphatase and urease activity decreased with depth but in three profiles potentially mineralizable N in the deepest horizons was higher than in soil layers directly overlying. In deeply modified urban soils, subsoil beside topsoil properties have to be included in the evaluation of soil quality. This knowledge is essential because consumption of natural soils for housing and traffic has to be reduced by promoting inner city densification.     

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.     

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.     

Measuring rates of gross and net mineralisation of organic phosphorus in soils

The isotopic dilution method developed by Oehl et al. [2001b. Organic phosphorus mineralisation studies using isotopic dilution techniques. Soil Science Society of America Journal 65, 780-787] to measure gross mineralisation of soil organic phosphorus (P) was tested on a range of low-P sorbing soils. This isotopic dilution method relies on accurate prediction of radiolabel behaviour due to soil physicochemical processes. Based on experimental validation of the extrapolation for isotopic dilution due to physicochemical processes using autoclaved soils, a simple power function was used for extrapolation rather than the more complex equation used in the original method. For several soils, however, a potential overestimation of gross mineralisation by 0.1-2.0 mg P kg⁻¹ d⁻¹ was revealed. In addition, the detection limit of P mineralisation ranged between 0.6 and 2.6 mg P kg⁻¹ d⁻¹. The method is likely to be at the detection limit for soils that are high in available P and low in biological activity. The method was modified with respect to the extrapolation and successfully applied to a soil with relatively high microbial P (18 mg P kg⁻¹) and soil respiration rates (29 mg C kg⁻¹ d⁻¹), revealing gross mineralisation rates of organic P of 0.9-1.2 mg P kg⁻¹ d⁻¹. Measurement of uptake of ³²P by the microbial biomass allowed derivation of a net organic P mineralisation rate of 0.5-0.9 mg P kg⁻¹ d⁻¹.     

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.     

Organic compounds that reach subsoil may threaten groundwater quality; effect of benzotriazole on degradation kinetics and microbial community composition

Toxic compounds in soils threaten groundwater quality in two ways: as potential contaminants themselves, and by retarding the microbial degradation of other organic compounds, thus enhancing their deep penetration. Benzotriazole (BTA) is a chemical with versatile industrial applications, used in large quantities worldwide, and represents a potential threat to the environment due to its apparent toxicity and recalcitrance. When used as an additive in aircraft deicing/antiicing fluid on airports, substantial spills of these mixtures and jet fuel will inevitably reach the soil. We have investigated the subsoil (1-2 m depth) microbial degradation and growth on four relevant organic substrates found in airport run-off (acetate, formate, glycol and toluene) in the presence of concentrations of BTA which can be found in airport run-off. Monitoring CO₂ evolution showed growth-dependent degradation rates for all substrates (sigmoid CO₂ accumulation curves), which were significantly affected by BTA. The mineralization of acetate was only moderately retarded and only by the highest BTA concentration used (400 mg l⁻¹ in soil solution); formate and glycol mineralization was substantially retarded at 200 mg l⁻¹, and toluene mineralization already at 10 mg l⁻¹ BTA. Mass balances (fraction of added C recovered as CO₂) suggested that the microbial growth yield (g biomass-C formed per g substrate C) was severely reduced with increasing concentrations of BTA. The analysis of phospholipid fatty acids (PLFA) demonstrated that Gram-negative bacteria were dominating among the organisms growing on all four substrates. The total amount of PLFA increased with approximately 1000 pmol PLFA g⁻¹ soil in response to a dose of 0.93 μmol glycol-C g⁻¹ soil, but this increase was gradually reduced with increasing BTA concentrations. This was in agreement with C mass balances based on CO₂ measurements, verifying that BTA severely reduced the growth yields. The response of individual PLFA's to BTA and substrates demonstrated that non-growing organisms were largely unaffected (i.e. the PLFA's of which the absolute amounts did not increase in response to substrates were not affected by BTA), whereas those which were growing on the added substrates were uniformly reduced by BTA (all the PLFA's which increased in response to the substrates were negatively affected by BTA). The results suggest that BTA functions as an uncoupler, i.e. a substance that reduces the yield of ATP per mole of substrate used, or that the defence mechanisms represent a large energy burden to all microbial cells.     

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.     

Filtration increases the correlation between water extractable organic carbon and soil microbial activity

A study was carried out in order to establish the relationship between the water extractable organic carbon (WEOC) content of soils and soil microbial activity, and to determine how variations in the extraction procedure might influence the quantity of WEOC recovered. Concentrations of WEOC were determined in soils taken from 12 different sites in the south east of Scotland, using a procedure in which samples were shaken with distilled water, centrifuged at 5000g and then filtered through 0.45 μm Millipore filters. Filtration resulted in between 30 and 400 μg C g⁻¹ being extracted using this procedure and the concentration of WEOC in the resultant extracts correlated with soil microbial production of CO₂ and dehydrogenase activity (P<0.001). Without filtration, although more WEOC was extracted (between 31 and 716 μg C g⁻¹), there was no significant correlation with biological activity. There was also no correlation between WEOC and nitrous oxide release during the incubations. Centrifugation at 20,000g for at least 10 min prior to filtration was required to remove particulate organic materials. Storage of samples at 4 °C or for up to 1 week or freezing for up to 3 months was not found to have a large influence on the concentration of WEOC in extracts, although amounts increased with soil:extractant ratio and increasing extraction time (from 15 to 60 min).     

Amino sugars and muramic acid—biomarkers for soil microbial community structure analysis

Characterizing functional and phylogenetic microbial community structure in soil is important for understanding the fate of microbially-derived compounds during the decomposition and turn-over of soil organic matter. This study was conducted to test whether amino sugars and muramic acid are suitable biomarkers to trace bacterial, fungal, and actinomycetal residues in soil. For this aim, we investigated the pattern, amounts, and dynamics of three amino sugars (glucosamine, mannosamine and galactosamine) and muramic acid in the total microbial biomass and selectively cultivated bacteria, fungi, and actinomycetes of five different soils amended with and without glucose. Our results revealed that total amino sugar and muramic acid concentrations in microbial biomass, extracted from soil after chloroform fumigation varied between 1 and 27 mg kg⁻¹ soil. In all soils investigated, glucose addition resulted in a 50-360% increase of these values. In reference to soil microbial biomass-C, the total amino sugar- and muramic acid-C concentrations ranged from 1-71 g C kg⁻¹ biomass-C. After an initial lag phase, the cultivated microbes revealed similar amino sugar concentrations of about 35, 27 and 17 g glucosamine-C kg⁻¹ TOC in bacteria, fungi, and actinomycetes, respectively. Mannosamine and galactosamine concentrations were lower than those for glucosamine. Mannosamine was not found in actinomycete cultures. The highest muramic acid concentrations were found in bacteria, but small amounts were also found in actinomycete cultures. The concentrations of the three amino sugars studied and muramic acid differed significantly between bacteria and the other phylogenetic microbial groups under investigation (fungi and actinomycetes). Comparison between the amino sugar and muramic acid concentrations in soil microbial biomass, extracted after chloroform fumigation, and total concentrations in the soil showed that living microbial biomass contributed negligible amounts to total amino sugar contents in the soil, being at least two orders of magnitude greater in the soils than in the soil inherent microbial biomass. Thus, amino sugars are significantly stabilized in soil.     

Microbial biomass and activity in alkalized magnesic soils under arid conditions

The effects of salinity and Mg²⁺ alkalinity on the size and activity of the soil microbial communities were investigated. The study was conducted along the border area of the alluvial fan of the Taolai River. Thirty soil samples were taken which had an electrical conductivity (EC) gradient of 0.93-29.60 mS cm⁻¹. Soil pH ranged from 8.60 to 9.33 and correlated positively with Mg²⁺/Ca²⁺ ratio, exchangeable Mg²⁺ percentage and HCO₃⁻+CO₃²⁻. Mg²⁺/Ca²⁺ varied considerably from 3.04 to 61.31, with an average of 23.03. Exchangeable Mg²⁺ percentage generally exceeded 60% and had a positive correlation with Mg²⁺/Ca²⁺. HCO₃⁻+CO₃²⁻ averaged 1.63 cmol kg⁻¹ and usually did not exceed 2.0 cmol kg⁻¹.Microbial biomass, indices of microbial activity and the activities of the hydrolases negatively correlated with Mg²⁺/Ca²⁺ or exchangeable Mg²⁺ percentage. Biomass C, biomass N, microbial quotient (the percentage of soil organic C present as biomass C), biomass N as a percentage of total N, potentially mineralizable N, FDA hydrolysis rate and arginine ammonification rate decreased exponentially with increasing EC. The biomass C/N tended to be lower in soils with higher salinity and Mg²⁺ alkalinity, probably reflecting the bacterial dominance in microbial biomass in alkalized magnesic soils. The metabolic quotient (qCO₂) positively correlated with salinity and Mg²⁺ alkalinity, and showed a quadratic relationship with EC, indicating that increasing salinity and Mg²⁺ alkalinity resulted in a progressively smaller, more stressed microbial communities which was less metabolically efficient. Consequently, our data suggest that salinity and Mg²⁺ alkalinity are stressful environments for soil microorganisms.