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.     

Estimation of conversion factors for fungal biomass determination in compost using ergosterol and PLFA 18:2ω6,9

Eleven species of common fungi from compost were analysed for their content of ergosterol and phospholipid fatty acids (PLFAs) after growth on agar media. Mean content of ergosterol was 3.1 mg g⁻¹ dw of fungal mycelium (range 1-24 mg g⁻¹ dw). Total amount of PLFAs varied between 2.6 and 43.5 μmol g⁻¹ dw of fungi (mean 14.9 μmol g⁻¹ dw). The most common PLFAs were 16:0, 18:2ω6,9 and 18:1ω9, comprising between 79 and 97 mol% of the total amount of PLFAs. The PLFA 18:2ω6,9, suggested as a marker molecule for fungi, comprised between 36 and 61 mol% of the total PLFAs in the Ascomycetes, between 45 and 57 mol% in the Basidiomycetes and 12-22 mol% in the Zygomycetes. There was a good correlation between the content of the two fungal marker molecules, ergosterol and the PLFA 18:2ω6,9, with a mean content of 1 mg ergosterol being equivalent to 2.1 μmol of 18:2ω6,9. Based on results from the fungal isolates, conversion factors were calculated (5.4 mg ergosterol g⁻¹ biomass C and 11.8 μmol 18:2ω6,9 g⁻¹ biomass C) and applied to compost samples in which both the ergosterol and the PLFA 18:2ω6,9 content had been measured. This resulted in similar estimates of fungal biomass C using the two marker molecules, but was three to five times higher than total microbial biomass C calculated using ATP content in the compost. This could partly be explained by the fact that both of the markers used for fungal biomass are cell membrane constituents. Thus, the ergosterol and the PLFA content were related to the hyphal diameter of the fungi, where fungi with thinner hyphae had higher ergosterol concentrations than fungi with thicker hyphae. This could also partly explain the large interspecific variation in content of the two marker molecules.     

Relationships between microbial indices in roots and silt loam soils forming a gradient in soil organic matter

Perennial rye grass (Lolium perenne) was grown in a greenhouse pot experiment on seven soils to answer the question whether the microbial colonisation of roots is related to existing differences in soil microbial indices. The soils were similar in texture, but differed considerably in soil organic matter, microbial biomass, and microbial community structure. Ergosterol and fungal glucosamine were significantly interrelated in the root material. This ergosterol was also significantly correlated with the average ergosterol content of bulk and rhizosphere soil. In addition, the sum of fungal C and bacterial C in the root material revealed a significant linear relationship with microbial biomass C in soil. The colonisation of roots with microorganisms increased apparently with an increase in soil microbial biomass. In the root material, microbial tissue consisted of 77% fungi and 23% bacteria. In soil, the fungal dominance was slightly, but significantly lower, with 70% fungi and 30% bacteria. Fungal glucosamine in the root material was significantly correlated with that in soil (r=0.65). This indicates a close relationship between the composition of dead microbial remains in soil and the living fraction in soil and root material for unknown reasons.     

Does soil ergosterol concentration provide a reliable estimate of soil fungal biomass?

Our aim was to determine if soil ergosterol concentration provides a quantitative estimate of the soil fungal biomass concentration, as is usually assumed. This was done by comparing soil ergosterol measurements with soil fungal biomass (fungal biomass C) concentrations estimated by microscopic measurements and by the selective inhibition technique linked to substrate-induced respiration (SIR). The measurements were compared in a silty-clay loam soil given a range of previous treatments designed to increase or decrease the soil fungal biomass and so also to change the soil ergosterol concentration. The treatments used were ryegrass amendment, to increase the total and fungal biomass, and CHCl₃-fumigation and the addition of the biocides, captan, bronopol and dinoseb, to decrease both ergosterol and fungal biomass C concentrations. The mineralization of ergosterol following addition to sand innoculated with soil extract, and to a sandy loam soil, was also determined. The added ergosterol was little, if at all, degraded following addition to either sand or the unfumigated or fumigated soil during a 10 d aerobic incubation. Similarly, pesticide addition did not significantly change soil ergosterol concentrations yet the soil fungal biomass C concentration decreased significantly. Thus, the ratio: (soil ergosterol concentration/soil fungal biomass C concentration) was much higher in the pesticide-treated soils than the control soil. Following ryegrass amendment, soil ergosterol concentration increased from about 6-12 μg⁻¹ soil within 5 d and then decreased gradually to about 7 μg g⁻¹ soil by 20 d incubation. Changes in fungal biomass C (measured by direct microscopy) closely mirrored changes in soil ergosterol over this period. However, when the amended soil was fumigated and then incubated for a further 5 d, the initial ergosterol concentration declined from 7 to 5 μg g⁻¹ soil by 20 d incubation (a decline of about 0.4). The comparable decline in fungal biomass C was about eight-fold. Thus the ratio of ergosterol to fungal biomass C increased from 0.005 to about 0.01. There was a significant correlation (r>0.84, P<0.001) between soil ergosterol concentration and fungal biomass measured by either SIR or microscopy. However, three data points played a vital role in the correlation. When these points were excluded the relationship was very poor (r<0.4). Our results therefore suggest that substantial amounts of ergosterol may exist, other than in living cells, for considerable periods, with little, if any mineralization. Thus, these results indicate that ergosterol and fungal biomass C concentrations are not always closely correlated, due to the slow metabolism of ergosterol in recently dead fugal biomass and/or the existence of exocellular ergosterol in soil.     

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.     

Respiration pattern and microbial use of field-grown transgenic Bt-maize residues

A 49-day incubation experiment was carried out with the addition of field-grown maize stem and leaf residues to soil at three different temperatures (5, 15, and 25 °C). The aim was to study the effects of two transgenic Bt-maize varieties in comparison to their two parental non-Bt varieties on the mineralization of the residues, on their incorporation into the microbial biomass and on changes in the microbial community structure. The stem and leaf residues of Novelis-Bt contained 3.9 μg g⁻¹ dry weight of the Bt toxin Cry1Ab and those of Valmont-Bt only 0.8 μg g⁻¹. The residues of the two parental non-Bt varieties Nobilis and Prelude contained higher concentrations of ergosterol (+220%) and glucosamine (+190%) and had a larger fungal C-to-bacterial C ratio (+240%) than the two Bt varieties. After adding the Bt residues, an initial peak in respiration of an extra 700 μg CO₂-C g⁻¹ soil or 4% of the added amount was observed in comparison to the two non-Bt varieties at all three temperatures. On average of the four varieties, 19-38% of the maize C added was mineralized during the 49-day incubation at the three different temperatures. The overall mean increase in total maize-derived CO₂ evolution corresponded to a Q₁₀ value of 1.4 for both temperature steps, i.e. from 5 to 15 °C and from 15 to 25 °C. The addition of maize residues led to a strong increase in all microbial properties analyzed. The highest contents were always measured at 5 °C and the lowest at 25 °C. The variety-specific contents of microbial biomass C, biomass N, ATP and adenylates increased in the order Novelis-Bt ? Prelude<Valmont-Bt ? Nobilis. The mineralization of Novelis-Bt residues with the highest Bt concentration and lowest N concentration and their incorporation into the microbial biomass was significantly reduced compared to the parental non-Bt variety Nobilis. These negative effects increased considerably from 5 to 25 °C. The transgenic Bt variety Valmont did not show further significant effects except for the initial peak in respiration at any temperature.     

Ergosterol and microbial biomass relationship in soil

Ergosterol and microbial biomass C were measured in 26 arable, 16 grassland and 30 forest soils. The ergosterol content ranged from 0.75 to 12.94 g g^-1 soil. The geometric mean ergosterol content of grassland and forest soils was around 5.5 g g^-1, that of the arable soils 2.14 g g^-1. The ergosterol was significantly correlated with biomass C in the entire group of soils, but not in the subgroups of grassland and forest soils. The geometric mean of the ergosterol: microbial biomass C ratio was 6.0 mg g^-1, increasing in the order grassland (5.1), arable land (5.4) and woodland (7.2). The ergosterol:microbial biomass C ratio had a strong negative relationship with the decreasing cation exchange capacity and soil pH, indicating that the fungal part of the total microbial biomass in soils increased when the buffer capacity decreased. The average ergosterol concentration calculated from literature data was 5.1 mg g^-1 fungal dry weight. Assuming that fungi contain 46% C, the conversion factor from micrograms ergosterol to micrograms fungal biomass C is 90. For soil samples, neither saponification of the extract nor the more effective direct saponification during extraction seems to be really necessary.     

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

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

Specific response of fungal and bacterial residues to one-season tillage and repeated slurry application in a permanent grassland soil

The dynamics of fungal and bacterial residues to a one-season tillage event in combination with manure application in a grassland soil are unknown. The objectives of this study were (1) to assess the effects of one-season tillage event in two field trials on the stocks of microbial biomass, fungal biomass, microbial residues, soil organic C (SOC) and total N in comparison with permanent grassland; (2) to determine the effects of repeated manure application to restore negative tillage effects on soil microbial biomass and residues. One trial was started 2 years before sampling and the other 5 years before sampling. Mouldboard ploughing decreased the stocks of SOC, total N, microbial biomass C, and microbial residues (muramic acid and glucosamine), but increased those of the fungal biomarker ergosterol in both trials. Slurry application increased stocks of SOC and total N only in the short-term, whereas the stocks of microbial biomass C, ergosterol and microbial residues were generally increased in both trials, especially in combination with tillage. The ergosterol to microbial biomass C ratio was increased by tillage, and decreased by slurry application in both trials. The fungal C to bacterial C ratio was generally decreased by these two treatments. The metabolic quotient qCO₂ showed a significant negative linear relationship with the microbial biomass C to SOC ratio and a significant positive relationship with the soil C/N ratio. The ergosterol to microbial biomass C ratio revealed a significant positive linear relationship with the fungal C to bacterial C ratio, but a negative one with the SOC content. Our results suggest that slurry application in grassland soil may promote SOC storage without increasing the role of saprotrophic fungi in soil organic matter dynamics relative to that of bacteria.     

Soil microorganisms and the growth of Lupinus albus on a high metal soil in the presence of EDTA

A pot experiment was designed with the objective of determining whether the presence of ethylenediaminetetraacetic acid (EDTA) and the resulting mobilization of heavy metals have any affect on: (i) soil microorganisms, (ii) growth of L. albus, and (iii) microbial colonization of roots. There was no effect of the different treatments on the contents of soil microbial biomass C and microbial biomass N. Increasing addition of EDTA to soil led to proportionate increases in extractable C and N, being roughly equivalent to the added amount. Increasing EDTA addition to soil led also to a proportionate increase in mobile heavy metals. Plant height, total amount of shoot and root C were not affected by EDTA addition. Fungal ergosterol in the lupine roots showed a 5- to 8-fold increase in the 0.1 and 0.2% EDTA treatments in comparison with the control. In contrast, EDTA addition did not affect fungal glucosamine or bacterial muramic acid concentrations in the lupine roots. Increasing EDTA addition to soil led also to a proportionate increase in the metal concentrations in the lupine shoots. The concentrations of heavy metals in the lupine shoots and in the NH₄NO₃ soil extracts were all highly significantly correlated.     

Partitioning root and microbial contributions to soil respiration in Leymus chinensis populations

Based on the enclosed chamber method, soil respiration measurements of Leymus chinensis populations with four planting densities (30, 60, 90 and 120 plants/0.25 m²) and blank control were made from July 31 to November 24, 2003. In terms of soil respiration rates of L. chinensis populations with four planting densities and their corresponding root biomass, linear regressive equations between soil respiration rates and dry root weights were obtained at different observation times. Thus, soil respiration rates attributed to soil microbial activity could be estimated by extrapolating the regressive equations to zero root biomass. The soil microbial respiration rates of L. chinensis populations during the growing season ranged from 52.08 to 256.35 mg CO₂ m⁻² h⁻¹. Soil microbial respiration rates in blank control plots were also observed directly, ranging from 65.00 to 267.40 mg CO₂ m⁻² h⁻¹. The difference of soil microbial respiration rates between the inferred and the observed methods ranged from −26.09 to 9.35 mg CO₂ m⁻² h⁻¹. Some assumptions associated with these two approaches were not completely valid, which might result in this discrepancy. However, these two methods' application could provide new insights into separating root respiration from soil microbial respiration. The root respiration rates of L. chinensis populations with four planting densities could be estimated based on measured soil respiration rates, soil microbial respiration rates and corresponding mean dry root weight, and the highest values appeared at the early stage, then dropped off rapidly and tended to be constant after September 10. The mean proportions of soil respiration rates of L. chinensis populations attributable to the inferred and the observed root respiration rates were 36.8% (ranging from 9.7 to 52.9%) and 30.0% (ranging from 5.8 to 41.2%), respectively. Although root respiration rates of L. chinensis populations declined rapidly, the proportion of root respiration to soil respiration still increased gradually with the increase of root biomass.     

Elevated enzyme activities in soils under the invasive nitrogen-fixing tree Falcataria moluccana

Like other N-fixing invasive species in Hawaii, Falcataria moluccana dramatically alters forest structure, litterfall quality and quantity, and nutrient dynamics. We hypothesized that these biogeochemical changes would also affect the soil microbial community and the extracellular enzymes responsible for carbon and nutrient mineralization. Across three sites differing in substrate texture and age (50-300 years old), we measured soil enzyme activities and microbial community parameters in native-dominated and Falcataria-invaded plots. Falcataria invasion increased acid phosphatase (AP) activities to >90 μmol g⁻¹ soil h⁻¹ compared to 30-60 μmol g⁻¹ soil h⁻¹ in native-dominated stands. Extracellular enzymes that mineralize carbon and nitrogen also increased significantly under Falcataria on the younger substrates. By contrast, total microbial biomass and mycorrhizal abundance changed little with invasion or substrate. However, fungal:bacterial ratios declined dramatically with invasion, from 2.69 and 1.35 to <0.89 on the 50- and 200-year-old substrates, respectively. These results suggest that Falcataria invasion alters the composition and function of belowground soil communities in addition to forest structure and biogeochemistry. The increased activities of AP and other enzymes that we observed are consistent with a shift toward phosphorus limitation and rapid microbial processing of litterfall C and N following Falcataria invasion.     

Identification of ergosterol in vesicular-arbuscular mycorrhizae

Summary Ergosta-5,7,22-tri-3-enol (ergosterol) was identified by gas chromatography-mass spectrometry in roots of berseem (Trifolium alexandrinum L., cv. Landsorte) and sweet corn (Zea mays L., cv. Honeycomb-F1) infected with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus intraradices. The fungal-derived compound ergosterol was determined quantitatively in root extracts using reverse-phase high performance liquid chromatography. The concentrations of ergosterol in VAM-infected roots reached 72 g^-1 dry material in berseem and 52 g^-1 in sweet corn after 80 days of growth, whereas concentrations in non-infected roots remained below 8 g^-1 dry weight. Additionally, phytosterols such as -sitosterol, campesterol, and stigmasterol were detected in both infected and non-infected roots. Ergosterol, as a characteristic fungal substance, is proposed as an indicator of fungal biomass in the early stages of VAM infection.     

Volatile organic compounds in the roots and rhizosphere of Pinus spp.

Plant roots normally release a complex mixture of chemicals which have important effects in the rhizosphere. Among these different root-emitted compounds, volatile isoprenoids have received very little attention, yet they may play important and diverse roles in the rhizosphere, contributing to the regulation of microbial activity and nutrient availability. It is therefore important to estimate their abundance in the rhizosphere, but so far, there is no reliable sampling method that can be used to measure realistic rates of root emissions from plants growing in field conditions, or even in pots. Here, we measured root content of volatile isoprenoids (specifically monoterpenes) for Pinus pinea, and explored the feasibility of using a dynamic bag enclosure method to measure emissions from roots of intact pot-grown plants with different degrees of root cleaning. We also investigated a passive diffusion method for exploring monoterpenes in soil at incremental distances from mature Pinus sylvestris trees growing in field conditions. Total monoterpene content of P. pinea roots was 415±50 μg g⁻¹ fresh wt in an initial screening study, and between 688±103 and 1144±208 μg g⁻¹ dry wt in subsequent investigations. Emissions from shaken-clean roots of intact plants and roots of intact plants washed to remove remaining soil after shaken-clean experiments were 119±14 and 26±5 μg g⁻¹ dry wt h⁻¹, respectively. Emissions from intact roots in soil-balls were an order of magnitude lower than from shaken-clean roots, and probably reflected the amount of emitted compounds taken up by physical, chemical or biological processes in the soil matrix surrounding the roots. Although monoterpene content was not significantly different in droughted roots, emission rates from droughted roots were generally significantly lower than from well-watered roots. Finally, passive sampling of monoterpenes in the soil at different distances from mature P. sylvestris trees in field conditions showed significantly decreasing sampling rates with increasing distance from the trunk. We conclude that it is feasible to measure volatile isoprenoid emissions from roots but the method of root preparation affects magnitude of measured emissions and therefore must be decided according to the application. We also conclude that the rhizosphere of Pinus species is a strong and previously un-characterized source of volatile isoprenoid emissions and these are likely to impact significantly on rhizosphere function.     

Impact of activated charcoal and tannin amendments on microbial biomass and residues in an irrigated sandy soil under arid subtropical conditions

Effects of goat manure application combined with charcoal and tannins, added as feed additives or mixed directly, on microbial biomass, microbial residues and soil organic matter were tested in a 2-year field trial on a sandy soil under Omani irrigated subtropical conditions. Soil microbial biomass C revealed the fastest response to manure application, followed by microbial residue C, estimated on the basis of fungal glucosamine and bacterial muramic acid, and finally soil organic C (SOC), showing the slowest, but still significant response. At the end of the trial, microbial biomass C reached 220 μg g^?1 soil, i.e. contents similar to sandy soils in temperate humid climate, and showed a relatively high contribution of saprotrophic fungi, as indicated by an average ergosterol to microbial biomass C ratio of 0.35 % in the manure treatments. The mean fungal C to bacterial C ratio was 0.55, indicating bacterial dominance of microbial residues. This fraction contributed relatively low concentrations of between 20 and 35 % to SOC. Charcoal added to manure increased the SOC content and the soil C/N ratio, but did not affect any of the soil microbial properties analysed. Tannins added to manure reduce the 0.5 M K₂SO₄-extractable N to N total ratio compared to manure control. These effects occurred regardless of whether charcoal or tannins were supplied as feed additive or directly mixed to the manure.     

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.     

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.     

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.     

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.     

Fungal biomass production and turnover in soil estimated using the acetate-in-ergosterol technique

We report the first attempt to estimate fungal biomass production in soil by correlating relative fungal growth rates (i.e., acetate incorporation into ergosterol) with fungal biomass increase (i.e., ergosterol) following amendments with dried alfalfa or barley straw in soil. The conversion factor obtained was then used in unamended soil, resulting in fungal biomass productions of 10-12 μg C g⁻¹ soil, yielding fungal turnover times between 130 and 150 days. Using a conversion factor from alfalfa-treated soil only resulted in two times higher estimates for biomass production and consequently lower turnover times. Comparing fungal biomass production with basal respiration indicated that these calculations overestimated the former. Still, the turnover times of fungal biomass in soil were in the same range as turnover times estimated in aquatic systems. The slow turnover of fungal biomass contrasts with the short turnover times found for bacteria. The study thus presents empirical data substantiating the theoretical division of bacteria and fungi into a fast and a slow energy channel, respectively, in the soil food web.