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.     

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.     

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.     

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.     

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.     

Immobilization and mineralization of nitrogen in a saline and alkaline soil during microbial use of sugarcane filter cake amended with glucose

A 42-day incubation was conducted to study the effect of glucose and ammonium addition adjusted to a C/N ratio of 12.5 on sugarcane filter cake decomposition and on the release of inorganic N from microbial residues formed initially. The CO₂ evolved increased in comparison with the non-amended control from 35% of the added C with pure +5 mg g⁻¹ soil filter cake amendment to 41% with +5 mg g⁻¹ soil filter cake +2.5 mg g⁻¹ soil glucose amendment to 48% with 5 mg g⁻¹ soil filter cake +5 mg g⁻¹ soil glucose amendment. The different amendments increased microbial biomass C and microbial biomass N within 6 h and such an increase persisted. The fungal cell-membrane component ergosterol initially showed a disproportionate increase in relation to microbial biomass C, which completely disappeared by the end of the incubation. The cellulase activity showed a 5-fold increase after filter cake addition, which was not further increased by the additional glucose amendment. The cellulase activity showed an exponential decline to values around 4% of the initial value in all treatments. The amount of inorganic N immobilized from day 0 to day 14 increased with increasing amount of C added, in contrast to the control treatment. After day 14, the immobilized N was re-mineralized at rates between 1.3 and 1.5 μg N g⁻¹ soil d⁻¹ in the treatments being more than twice as high as in the control treatment. This means that the re-mineralization rate is independent of the actual size of the microbial residues pool and also independent of the size of the soil microbial biomass.     

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.     

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.     

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.     

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.     

Gross sulphur mineralisation-immobilisation turnover in soil amended with plant residues

The rates of sulphur (S) released to and removed from the soil inorganic pool were estimated using the isotopic dilution technique. In an initial study fresh soil was mixed with combinations of two inorganic S levels (0 and 10 μg S g⁻¹ soil) and three plant residues (wheat straw, perennial ryegrass and oilseed rape) and followed over 32 days of incubation. As ³⁵S recovery was inadequate prior to day 2 and re-mineralisation of immobilised ³⁵S occurred after day 8 thereby invalidating the method, estimates of gross S transformation rates should be based on data sampled between days 2 and 8. In the main experiment 16 plant residues with ranges in S contents of 0.08-0.81%, C/S ratios of 50-604 and lignin content of 0.9-10.8 were mixed with soil and carrier-free ³⁵S label. Net turnover rates varied from 58% of S in Persian clover being immobilised to 76% of S in winter cress being mineralised within 5 days of incubation. Gross S mineralisation varied from 0.9-14.9 μg S g⁻¹ soil d⁻¹, whereas gross immobilisation only varied from 0.5 to 3.1 μg S g⁻¹ d⁻¹. Gross S immobilisation was strongly correlated to the C/S ratio of the plant material (P<0.001), whereas gross S mineralisation showed a weaker, but still significant, correlation with lignin content (P<0.05). The results indicate that immobilisation may predominantly have been a biological process in response to carbon addition while early mineralisation may have been dominated by the biochemical hydrolysis of organic sulphates in the residues. If attention is paid to the various constraints and limitations, isotopic pool dilution using ³⁵S offers a tool that may prove valuable in understanding and modelling soil S turnover.     

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.     

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.     

Decomposition of wheat straw differing in nitrogen content in soils under conventional and organic farming management

An incubation experiment was carried out to investigate the interactions of two straw qualities differing in N content and two soils differently accustomed to straw additions. One soil under conventional farming management (CFM) regularly received straw, the other soil under organic farming management (OFM) only farmyard manure. The soils of the two sites were similar in texture, pH, cation‐exchange capacity, and glucosamine content. The soil from the OFM site had higher contents of organic C, total N, muramic acid, microbial biomass C and N (C_mic and N_mic), but a lower ergosterol content and lower ratios ergosterol to C_mic and fungal C to bacterial C. The straw from the CFM had threefold higher contents of total N, twofold higher contents of ergosterol and glucosamine, a 50% higher content of muramic acid, and a 30% higher fungal C–to–bacterial C ratio. The straw amendments led to significant net increases in C_mic, N_mic, and ergosterol. Microbial biomass C showed on average a 50% higher net increase in the organic than in the CFM soil. In contrast, the net increases in N_mic and ergosterol differed only slightly between the two soils after straw amendment. The CO₂ evolution from the CFM soil always exceeded that from the OFM, by 50% or 200 µg (g soil)^–1 in the nonamended control soil and by 55% or additional 600 µg (g soil)^–1 in the two straw treatments. In both soils, 180 µg g^–1 less was evolved as CO₂‐C from the OFM straw. The metabolic quotient qCO₂ was nearly twice as high in the control and in the straw treatments of the CFM soil compared with that of the OFM. In contrast, the difference in qCO₂ was insignificant between the two straw qualities. Differences in the fungal‐community structure may explain to a large extent the difference in the microbial use of straw in the two soils under different managements.     

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.     

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

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

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.     

Food choice and reproductive success of Folsomia candida feeding on copper-contaminated mycelium of the soil fungus Alternaria alternata

We examined collembolan food preference for fungal mycelium grown on copper-contaminated medium, and the relationship between copper content, food selectivity and collembolan fitness when fed contaminated mycelium.To clarify whether collembolan food selectivity is related to fitness parameters, Folsomia candida were fed mycelium of the dark-pigmented fungus Alternaria alternata grown on medium with different copper concentrations. Copper-contaminated food (fungus grown on 50, 125, 250 and 500 μg Cu g⁻¹ medium, fresh wt.) was offered together with untreated food for 4 weeks. F. candida fed selectively on the provided mycelium and discriminated clearly between mycelium grown on high and low levels of contamination, distinctly preferring fungus grown on medium with a total copper concentration of 50 and 125 μg g⁻¹. In contrast, fungus grown on highly contaminated medium (250 and 500 μg g⁻¹) was avoided. Collembolan food preference generally matched fitness parameters. Reproduction was significantly affected by the total copper concentration of the fungal growth medium. When fed their preferred mycelium, collembolan reproduction was enhanced, whereas a diet of highly contaminated mycelium (250 or 500 μg g⁻¹) resulted in a strong decrease in reproduction. Adult survival was affected only marginally. Even though heavy metal contamination is a potential stress factor for many soil microarthropods, F. candida is able to discriminate between high and low quality food sources, and even benefits from moderately elevated copper concentrations.     

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.