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.     

Adenylates as an estimate of microbial biomass C in different soil groups

Adenylate (i.e. adenosine tri- (ATP), di- (ADP) and monophosphates (AMP)) and microbial biomass C data were collected over a wide range of sites including forest floor layers and forest, grassland and arable soils. Microbial biomass C was measured by fumigation extraction and adenylates after alkaline Na₃PO₄/DMSO/EDTA extraction and HPLC detection. Our aims were (1) to test whether the sum of adenylates is a better estimate for microbial biomass than the determination of ATP, (2) to compare our conversion values with those proposed by others, and (3) to analyse whether soil properties or land use form affect the relationships between ATP, adenylates and microbial biomass C. A close relationship was found between microbial biomass C and ATP (r=0.96), but also with the sum of adenylates (r=0.96) within all appropriately conditioned soil samples (n=112). In the mineral soil (n=98), the geometric means of the ATP-to-microbial biomass C ratio and the adenylates-to-microbial biomass C ratio were 7.4 and 11.4 μmol g⁻¹, respectively. The mean ratios did not differ significantly between the different texture classes and land use forms. In the forest floor, the ATP-to-microbial biomass C ratio and the adenylates-to-microbial biomass C ratio were both roughly two-thirds of those of the mineral soil. The average adenylate energy charge (AEC) of all soil samples was 0.79 and showed a strong negative relationship with the soil pH (r=−0.69). However, the AEC is presumably only indirectly affected by the soil pH.     

Adenylate energy charge and ATP-to-microbial biomass C ratio in soils differing in the intensity of disturbance

We carried out an 8-days' incubation experiment with three different intensities of soil disturbance to analyse the effects on the ATP-to-microbial biomass C ratio and on the adenylate energy charge (AEC=(ATP+0.5×ADP)/(AMP+ADP+ATP). Single mixing of soil at 50% water holding capacity with a spatula during weighing of the samples into extraction jars at the end of the 8-days' incubation or 8-times repeated daily mixing for 2 min triggered the immediate formation of ATP, increasing both AEC and the ATP-to-microbial biomass C ratio. The energy for this extra ATP produced seems to be mainly derived from an accelerated turnover of C within the microbial biomass. In contrast, 8-days' continuous mixing led to a significant decrease in AEC and ATP-to-microbial biomass C ratio.     

Microbial reaction of secondary tropical forest soils to the addition of leaf litter

Two soils from a secondary tropical forest at La Union, Philippines, predominantly vegetated with Swietenia marcrophylla and Gmelina arborea were amended with different leaf litter types (Eucalyptus camaldulensis, S. macrophylla, G. arborea, and Calliandra calothyrsus) and incubated in the laboratory for 49 days at 25 °C. The experiment was carried out to elucidate the reasons for a low ATP-to-microbial biomass C ratio and a high microbial biomass C-to-N ratio. This has been measured repeatedly in tropical forest soils. In the non-amended soils, the microbial biomass C-to-N ratio of 12.1 exceeded the soil organic C-to-total N ratio of 11, while the ergosterol-to-microbial biomass C ratio of 0.14% and the ATP-to-microbial biomass C ratio of 4.1 μmol g⁻¹ were both low. At the end of the incubation, the addition of the different leaf litter types led generally to a decrease in the microbial biomass C-to-N ratio and to an increase in the ATP-to-microbial biomass C ratio, adenylate energy charge (AEC) and especially to an increase in the ergosterol-to-microbial biomass C ratio. The increase in the ATP-to-microbial biomass C ratio and the decrease in the microbial biomass C-to-N ratio were positively related to the N concentration in the leaf litter, the increase in the ergosterol-to-microbial biomass ratio negatively. The reasons for a low ATP-to-microbial biomass C ratio and a high microbial biomass C-to-N ratio are P deficiency and probably a reduced access of soil microorganisms to N containing organic components at low soil organic C levels.     

Long-term effects of organic farming on fungal and bacterial residues in relation to microbial energy metabolism

Samples from the bio-dynamic, bio-organic, and conventional trial, Therwil, Switzerland, were analyzed with the aim of determining the effects of organic land use management on the energy metabolism of the soil microbial biomass and on the fraction of microbial residues. The contents of adenylates, adenosine triphosphate (ATP), glucosamine, muramic acid, and galactosamine were significantly largest in the biodynamic organic farming (BYODIN) treatment and significantly lowest in the conventional farming treatment with inorganic fertilization (CONMIN). In contrast, the ergosterol-to-ATP ratio and fungal C-to-bacterial C ratios were significantly lowest in the BYODIN treatment and significantly largest in the CONMIN treatment. No clear treatment effects were observed for the ergosterol content and the adenylate energy charge (AEC), the ATP-to-microbial biomass C ratio and the ergosterol-to-fungal C ratio. Ergosterol, an indicator for saprotrophic fungal biomass, and fungal residues were significantly correlated. The microbial biomass carbon-to-nitrogen ratio showed a negative relationship with the AEC and strong positive relationships with the ratios ergosterol-to-microbial biomass C, ergosterol-to-ATP and fungal C-to-bacterial C. In conclusion, the long-term application of farmyard manure in combination with organic farming practices led to an increased accumulation of bacterial residues.     

Soil microbial properties down the profile of a black earth burie by colluvium

The activity and biomass of soil microorganisms were determined in samples at 0—140 cm depth taken from an arable site, where the soil has been developed by erosion and colluvial deposition overlaying a black earth at 70—110 cm depth. The central aim was to get an insight into the breakdown of increasingly old and thus recalcitrant soil organic matter down the profile, effects on the availability of C to microorganisms and the microbial community structure. From 0 to 140 cm depth, microbial biomass C decreased by 96%, biomass N by 97%, the adenylates ATP, ADP, and AMP as well as the basal respiration rate by 89%. No ergosterol was measured at 120—140 cm depth. All soil biological properties decreased in distinct steps after 30 cm and 50 cm depth. At 30—90 cm depth, the amounts of soil organic C and microbial biomass C per hectare of the present colluvium exceeded nearly three‐fold those in undisturbed aeolian loess sediments. The cation exchange significantly affected the relationships between microbial biomass C, biomass N, and the adenylates. As a consequence, none of the ratios between the soil microbial biomass properties revealed constant gradients throughout the profile. The adenylate energy charge (AEC) varied between the different soil layers insignificantly around a mean of 0.71. It was the most stable ratio down the profile showing absolutely no depth gradient, the lowest depth‐to‐depth variation, and also the lowest within depth variability. The other ratios between soil organic C, basal respiration, ergosterol, microbial biomass C and biomass N also did not reveal any marked changes in the microbial community structure.     

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.     

The adenylate energy charge of the soil microbial biomass

A method was developed for measuring adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphate (ADP) and adenosine 5'-monophosphate (AMP) in soil. All three adenine nucleotides were extracted from soil with a solution of trichloroacetic acid, paraquat and phosphate. ATP was measured in the neutralised (pH 7.4) soil extracts by the fire-fly luciferin-luciferase system. ADP was measured as ATP after incubating the neutralised extracts with pyruvate kinase (PK) and phosphoenolpyruvate (PEP) to convert ADP to ATP. AMP was converted to ATP by incubation with the coupled PK-PEP-myokinase system and measured as ATP. The quantities of nucleotides present in the extracts were corrected for incomplete extraction from soil by measuring the percentage recovery of added ATP, ADP and AMP. The adenylate energy charge (AEC) was calculated from the formula AEC = [[ATP] + 0.5[ADP]]/[[ATP] + [ADP] + [AMP]]. Measurements were made on (1) fresh soil, extracted as soon as possible after field sampling (2) soil stored air-dry at 5°C for 18 days and (3) soil stored air-dry at 5°C for 57 days and then rewetted to the original field moisture content and incubated aerobically for 2.5 h at 10°C before extraction.In moist soil the biomass maintains both ATP and AEC at levels close to those of activity growing cells, even though little of the biomass in soil can be in active growth at any given time. ATP accounted for 77% of the total adenine nucleotides (A_T) in the fresh soil, with an AEC of 0.85 (a value comparable to that found in microorganisms undergoing active growth in vitro. In contrast, ATP only accounted for 28% of A_T in the air-dried soil, with an AEC of 0.46. When the air-dried soil was rewetted, ATP increased to 66% of A_T and the AEC increased to 0.76. However, A_T in the air-dried soil (7.65 nmol g^?1 soil) was of the same order as that in rewetted soil (6.70 nmol g^?1) even though the AEC's were very different.These results show that the soil microbial biomass does not maintain a high AEC when air-dried. Once remoistened, the population tends to restore its AEC to the original value. This restoration occurs so rapidly that it cannot be due to the formation of a new biomass.     

Long‐term effects of leguminous cover crops on microbial indices and their relationships in soils of a coconut plantation of a humid tropical region

In this study, leguminous crops like Atylosia scarabaeoides, Centrosema pubescens, Calopogonium mucunoides, and Pueraria phaseoloides. grown as soil cover individually in the interspaces of a 19‐yr‐old coconut plantation in S. Andaman (India) were assessed for their influence on various microbial indices (microbial biomass C, biomass N, basal respiration, ergosterol, levels of ATP, AMP, ADP) in soils (0–50 cm) collected from these plots after 10 years. The effects of these cover crops on . CO₂ (metabolic quotient), adenylate energy charge (AEC), and the ratios of various soil microbial properties viz., biomass C : soil organic C, biomass C : N, biomass N : total N, ergosterol : biomass C, and ATP : biomass C were also examined. Cover cropping markedly enhanced the levels of organic matter and microbial activity in soils after the 10‐yr‐period. Microbial biomass C and N, basal respiration, . CO₂, ergosterol and levels of ATP, AMP, ADP in the cover‐cropped plots significantly exceeded the corresponding values in the control plot. While the biomass C : N ratio tended to decrease, the ratios of biomass N : total N, ergosterol : biomass C, and ATP : biomass C increased significantly due to cover cropping. Greater ergosterol : biomass C ratio in the cover‐cropped plots indicated a decomposition pathway dominated by fungi, and high . CO₂ levels in these plots indicated a decrease in substrate use efficiency probably due to the dominance of fungi. The AEC levels ranged from 0.80 to 0.83 in the cover‐cropped plots, thereby reflecting greater microbial proliferation and activity. The ratios of various microbial and chemical properties could be assigned to three different factors by principal components analysis. The first factor (PC1) with strong loadings of ATP : biomass C ratio, AEC, and . CO₂ reflected the specific metabolic activity of soil microbes. The ratios of ergosterol : biomass C, soil organic C : total N, and biomass N : total N formed the second factor (PC2) indicating a decomposition pathway dominated by fungi. The biomass C : N and biomass C : soil organic C ratios formed the third principal component (PC3), reflecting soil organic matter availability in relation to nutrient availability. Overall, the study suggested that Pueraria phaseoloides. or Atylosia scarabaeoides were better suited as cover crops for the humid tropics due to their positive contribution to soil organic C, N, and microbial activity.     

Adenylate energy charge measurements in soil

Adenosine 5'-triphosphate (ATP), adenosine 5'-diphosphatc (ADP) and adenosine 5'-monophosphate (AMP) were extracted from soil with either a solution of trichloroacetic acid, paraquat and phosphate (TCA reagent) or a mixture of chloroform, sodium hydrogen carbonate, phosphate and adenosine (NaHCO₃ reagent). Standard enzymic procedures were used to convert ADP and AMP to ATP, which was measured by the fire-fly luciferin-luciferase system. The measured quantities of nucleotides were corrected for incomplete extraction using the percentage recoveries of added ATP, ADP and AMP. The adenylate energy charge ratio (AEC) was calculated from the formula AEC = ([ATP] + 0.5 [ADP])/([ATP] + [ADP] + [AMP]).Measurements were made on a grassland soil, following a conditioning incubation at 15°C and 50% WHC for 7 days. Additional measurements were made on the same soil after a further 50- or 100-day incubation at 25°C and 50% WHC, with or without an amendment of 1100 μg ryegrass Cg⁻¹ soil, added at the end of the conditioning incubation. Biomass-ATP concentration, measured in TCA extracts, changed little, even on prolonged incubation, and was maintained at a level comparable to that observed in earlier work (about 10 p mol ATP g⁻¹ biomass C). AEC values in TCA soil extracts were high (0.8–0.9) for all soil treatments and independent of substrate addition or length of incubation.In contrast, AEC was low (0.4) in fresh soil extracted with NaHCO₃ reagent, but increased to 0.6 when ryegrass was incubated with the soil for 50 days. Although the total adenine nucleotide pool (i.e. [ATP] + [ADP] + [AMP]) was similar as measured in NaHCO₃ and in TCA soil extracts, both energy charge and ATP content were lower in the NaHCO₃ extracts. It was therefore concluded that the main reason for the lower AECs observed with the NaHCO₃ reagent was that microbial ATPases were still active during extraction and caused appreciable hydrolysis of microbial ATP to ADP and AMP. In contrast, the TCA reagent rapidly inactivates ATPases and is therefore preferable for extracting adenine nucleotides from soil.The results indicate that the soil microbial biomass, although a mainly dormant population, maintains both AEC and ATP at levels characteristic of exponentially growing organisms in vitro, even during prolonged incubation without fresh substrate. It was also concluded that roots make a negligible contribution to total ATP extracted from fresh sieved soil.     

Phosphatase activity does not limit the microbial use of low molecular weight organic-P substrates in soil

Plant roots and soil microorganisms contain significant quantities of low molecular weight (MW) phosphorylated nucleosides and sugars. Consequently, upon death these can represent a significant input of organic-P to the soil. Some of these organic-P substrates must first be dephosphorylated by phosphatases before being assimilated by the soil microbial community while others can be taken up directly from soil solution. To determine whether sorption or phosphatase activity was limiting the bioavailability of low MW organic-P in soil we compared the microbial uptake and C mineralization of a range of ¹⁴C-labeled organic-P substrates [glucose-6-phosphate, adenosine monophosphate (AMP), adenosine diphosphate (ADP) and adenosine triphosphate (ATP)] to that of the parent compounds (adenosine and glucose). In a fertile grassland soil we showed that at low organic-P substrate concentrations (<0.5 mM) phosphatase activity did not limit microbial uptake or mineralization in comparison to their non-phosphorylated counterparts. However, at high substrate concentrations (1-10 mM) the mineralization of the organic-P compounds was significantly lower than that of the non-phosphorylated compounds suggesting that phosphatase activity or microbial transporter capacity limited bioavailability. Sorption to the solid phase followed the series glucose<adenosine<G-6-P<AMP<ADP=ATP. However, sorption of the organic-P compounds to the solid phase did not appear to greatly affect bioavailability. The high adenosine mineralization capacity of the microbial biomass suggests that nucleosides may represent a significant source of C and N to the soil microbial biomass. We conclude that at low organic-P substrate concentrations typical of those in soil, neither phosphatase activity nor sorption greatly limits their bioavailability.     

Specific respiration rates, adenylates, and energy budgets of soil microorganisms after addition of transgenic Bt-maize straw

An arable soil was incubated with straw (stem+leaves) of two transgenic Bt-maize varieties (Novelis: event MON810 and Valmont: event Bt176) and the two corresponding near-isogenic varieties (Nobilis and Prelude). The aim was to evaluate the use of these substrates for microbial growth and maintenance in soil during early decomposition. The addition of Bt-maize straw increased CO₂ production rates and the specific respiration rates CO₂-C/microbial biomass C and CO₂-C/ATP significantly compared with the addition of non-Bt maize straw. This extra energy in the Bt-maize straw could not be used for microbial biomass or ATP and ADP production, and was lost for maintenance. In addition, increased death rates of microbial biomass occurred in the soils treated with the Bt-maize straw from day 3 to 21. Generally, most of the energy was stored in microbial biomass, whereas only 10% of energy was stored in ATP, and only 1-2% in ADP. The AEC (adenylate energy charge: (ATP+0.5×ADP)/(AMP+ADP+ATP)) was not affected by any treatment. The reasons for the lower efficiency of microbial substrate use after adding Bt-maize straw cannot be fully explained by the present experiment. However, a risk assessment has to look at the impact of transgenic plant material on soil microorganisms at different maturity stages.     

Microbial biomass and activity indices after organic substrate addition to a selenium-contaminated soil

High concentrations of Se in soil might have negative effects on microorganisms. For this reason, the effect of organic substrate addition (glucose + maize straw) on Se volatilisation in relation to changes in microbial biomass and activity indices was investigated using an artificially Se-contaminated soil. Microbial biomass N was reduced on average by more than 50% after substrate addition, but adenylate energy charge (AEC) and metabolic quotient qCO₂ were both increased. The Se content decreased by nearly 30% only with the addition of the organic substrate at 25°C. No significant Se loss occurred without substrate at 25°C or with substrate at 5°C. In the two treatments with substrate addition, the substrate-derived CO₂ evolution was about 30% lower with Se addition than without. In contrast, Se had no effect on any of the other soil microbial indices analysed, i.e. microbial biomass C, microbial biomass N, adenosine triphosphate (ATP), AEC, ATP-to-microbial biomass C, and qCO₂.     

Soil biochemical and microbial indices in wet tropical forests: Effects of deforestation and cultivation

Little information is available about the long‐term effects of deforestation and cultivation on biochemical and microbial properties in wet tropical forest soils. In this study, we evaluated the general and specific biochemical properties of soils under evergreen, semi‐evergreen, and moist deciduous forests and adjacent plantations of coconut, arecanut, and rubber, established by clear felling portions of these forests. We also examined the effects of change in land use on microbial indices and their interrelationships in soils. Significant differences between the sites occurred for the biochemical properties reflecting soil microbial activity. Microbial biomass C, biomass N, soil respiration, N mineralization capacity, ergosterol, levels of adenylates (ATP, AMP, ADP), and activities of dehydrogenase and catalase were, in general, significantly higher under the forests than under the plantations. Likewise, the activities of various hydrolytic enzymes such as acid phosphomonoesterase, phosphodiesterase, casein‐protease, BAA‐protease, β‐glucosidase, CM‐cellulase, invertase, urease, and arylsulfatase were significantly higher in the forest soils which suggested that deforestation and cultivation markedly reduced microbial activity, enzyme synthesis and accumulation due to decreased C turnover and nutrient availability. While the ratios of microbial biomass C : N and microbial biomass C : organic C did not vary significantly between the sites, the ratios of ergosterol : biomass C and ATP : biomass C, qCO2 and AEC (Adenylate Energy Charge) levels were significantly higher in the forest sites indicating high energy requirements of soil microbes at these sites.     

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.     

Development of ergosterol, microbial biomass C, N, and P after steaming as a result of sucrose addition, and Sinapis alba cultivation

A pot experiment was carried out to monitor the recovery of a steaming-reduced microbial biomass (C, N, and P) and fungal ergosterol by sucrose addition. The second objective was to investigate the recovery of a steaming-reduced microbial biomass by white mustard (Sinapis alba) cultivation and its interactions with microbial residues, freshly formed from sucrose addition. Thirty days after steaming, the soil microbial biomass C and N was still significantly reduced by 80%, leading to a rather constant microbial biomass C/N ratio around 7 throughout the experiment. The steaming-induced decreases of microbial biomass P and ergosterol were only roughly 50%, leading to a decrease in the microbial biomass C/P ratio and an increase in the ergosterol-to-microbial biomass C ratio. Sucrose addition led to a 25% reduction in the ergosterol-to-microbial biomass C ratio. Mustard cultivation had significant positive effects on microbial biomass C, N, P, and ergosterol, but the effects were smaller than those of sucrose addition. Cultivating mustard had no significant effects on the C loss or on the incorporation of sucrose C into the microbial biomass. In contrast, the application of sucrose led to a significant decrease in the mustard shoot biomass and especially in the mustard root biomass.     

The proportional mineralisation of microbial biomass and organic matter caused by air-drying and rewetting of a grassland soil

During the first few days after rewetting of an air-dried soil (AD-RW), microbial activity increases compared to that in the original moist soil, causing increased mineralisation (a flush) of soil organic carbon (C) and other nutrients. The AD-RW flush is believed to be derived from the enhanced mineralisation of both non-biomass soil organic matter (due to its physical release and enhanced availability) and microbial biomass killed during drying and rewetting. Our aim was to determine the effects of AD-RW on the mineralisation of soil organic matter and microbial biomass during and after repeated AD-RW cycles and to quantify their proportions in the CO₂-C flushes that resulted. To do this, a UK grassland soil was amended with ¹⁴C-labelled glucose to label the biomass and then given five AD-RW cycles, each followed by 7 d incubation at 25 °C and 50% water holding capacity. Each AD-RW cycle increased the amount of CO₂-C evolved (varying from 83 to 240 μg g⁻¹ soil), compared to the control with, overall, less CO₂-C being evolved as the number of AD-RW cycles increased. In the first cycle, the amount of biomass C decreased by 44% and microbial ATP by 70% while concentrations of extractable C nearly doubled. However, all rapidly recovered and within 1.3 d after rewetting, biomass C was 87% and ATP was 78% of the initial concentrations measured prior to air-drying. Similarly, by 2 d, extractable organic C had decreased to a similar concentration to the original. After the five AD-RW cycles, the amounts of total and ¹⁴C-labelled biomass C remaining in the soil accounted for 60 and 40% of those in the similarly incubated control soil, respectively. Soil biomass ATP concentrations following the first AD-RW cycle remained remarkably constant (ranging from about 10 to 14 μmol ATP g⁻¹ biomass C) and very similar to the concentration in the fresh soil prior to air-drying. We developed a simple mathematical procedure to estimate the proportion of CO₂-C derived from biomass C and non-biomass C during AD-RW. From it, we estimate that, over the five AD-RW cycles, about 60% of the CO₂-C evolved came from mineralisation of non-biomass organic C and the remainder from the biomass C itself.     

Microbial biomass dynamics in recently air-dried and rewetted soils compared to others stored air-dry for up to 103 years

Our aim was to compare the soil microbial biomass concentration and its activity (measured as CO₂-C evolved) following the rewetting and aerobic incubation of soils which have previously been stored air-dry for different periods. Some of the soils have been stored in the Rothamsted sample archive for 103 years, others were comparable freshly sampled soils following air-drying and rewetting and other soils were stored air-dry for 2 years then rewetted for the work described here. Following air-drying, soil ATP concentrations were variable in recently air-dried soil, comprising about 10-35% of the initial ATP concentrations in fresh soil. Following rewetting, the percentage recovery of ATP increased in all soils by 7 days, then declined to between 73% and 87% of the original ATP concentration in the air-dried soils by day 12. Storage of air-dried soils decreased the ability of the microbial biomass to restore its ATP concentrations. For example, the ATP concentration in a soil sampled from stubbed (i.e. tree seedling, saplings and bushes cut frequently to ground level) grassland of the Broadbalk continuous wheat experiment at Rothamsted then air-dried for 2 years was only about 14% of that in the fresh soil at 2 days after rewetting. In other soils from the Hoosfield Barley Experiment, also at Rothamsted, previously given NPK or FYM since 1852, and sampled then stored air-dry for between 13 and 83 years, from 52% to 57% of the ATP in the comparable fresh soils was measured at two days after rewetting. The soil ATP concentration then changed little more up to 12 days. One of the most interesting findings was that while the microbial biomass ATP concentration in the above NPK soils only ranged from about 2 to 4 μmol ATP g⁻¹ biomass C, in the FYM soil the microbial biomass ATP concentrations (range 11.5-13.6 μmol ATP g⁻¹ biomass C) were the same as we repeatedly measure in fresh moist aerobic soil. We do not yet know the reasons for this. More than twice as much CO₂-C was evolved from the long-term stored soils than from freshly sampled ones. However, the specific respiration of the microbial biomass did not change much after the first 12 years of storage, indicating that loss of viability mainly occurred in the earlier years.     

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.     

Simultaneous measurement of S, macronutrients, and heavy metals in the soil microbial biomass with CHCl3 fumigation and NH4NO3 extraction

The study was carried out to investigate whether 1 M NH₄NO₃ extraction is a useful alternative to 10 mM CaCl₂ extraction for estimating soil microbial biomass S and whether the data of CHCl₃-labile NH₄NO₃-extractable macronutrients and heavy metals are useful and in agreement with the available data on element concentrations in soil microorganisms. Microbial biomass C was followed by microbial biomass S after CaCl₂ extraction with an average C/S ratio of 82, and by microbial biomass S after NH₄NO₃ extraction with an average C/S ratio of 57. The mean contribution of CHCl₃-labile metals in relation to the NH₄NO₃-extractable fraction from non-fumigated soils ranged from 0.1 to 112% in the order potassium < magnesium < cadmium < sodium < zinc + nickel < manganese < copper. The mean contribution of CHCl₃-labile metals in relation to the microbial biomass C ranged from 0.03 to 22‰ in the order cadmium < nickel < zinc < manganese < magnesium < copper < sodium < potassium. These relative contributions varied within the different metals from a 4-fold (Na⁺) to a more than 200-fold range (Cu²⁺). Significant positive correlations with microbial biomass C were observed for CHCl₃-labile zinc, sodium and especially potassium. The concentration of all elements except copper in relation to microbial biomass C were in the range known from the limited literature on fungi grown on heavy metal contaminated soils.