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.     

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.     

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.     

Impact of cycloheximide addition on adenylates in soil

Cycloheximide inhibits specifically the ribosomal protein synthesis of eukaryotic cells, i.e. the metabolism of soil fungi. We measured cycloheximide effects on adenylates in 20 different soils (0-10 cm depth) from arable, grass and forest land with a large variety of soil properties. The aims were (1) to assess the interactions between cycloheximide effects and soil properties and (2) to prove the relationship between cycloheximide effects on ATP and the ergosterol-to-microbial biomass C ratio, which is an indicator for the fungal proportion of the total microbial biomass. The adenylates ATP, ADP and AMP were measured 6 h after adding either 10 mg cycloheximide per gram soil in combination with 24 mg talcum per gram soil or 24 mg talcum per gram soil solely. The medians of the relative increases in AMP and ADP were 45 and 25% and the medians of the relative decreases in ATP and adenylates were −36 and −12%. These changes in adenylate composition lead to a cycloheximide-induced relative decrease in the adenylate energy charge level of 15%. The relative decrease in ATP content after cycloheximide addition was significantly correlated with the ATP-to-microbial biomass C ratio, but not with the ergosterol-to-microbial biomass C ratio. The absolute increase in ADP and the absolute decrease in ATP were affected by the clay content according to principal component analysis. The reduction of the ATP-to-microbial biomass C ratio indicates that this ratio had the potential of being an important ecotoxicological indicator of direct toxic effects of organic pollutants on soil microorganisms.     

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.     

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.     

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.     

Estimation of the adenylate energy charge in unamended and amended agricultural soils

To determine relatively low concentrations of adenine nucleotides in agricultural soils a NaHCO₃-based extradant was developed and compared with the trichloroacetic acid-paraquat-phosphate extradant. The new medium, consisting of chloroform, sodium bicarbonate, phosphate and adenosine (pH 8.0) gave soil extracts which could be investigated without further neutralization and dilution. ATP was measured directly in the soil extracts by the luciferin-luciferase system. ADP and AMP were estimated after their enzymatic conversion to ATP by standard methods. The quantities of nucleotides corrected for recovery of standards were used to calculate the adenylate energy charge (AEC) from the formula AEC = [ATP] + 1/2[ADP]/[ATP] + [ADP] + [AMP], The AEC was estimated in six unplanted soils from agricultural fields. A very similar energy charge of 0.3-0.4 was found in all soils sampled which indicates a low metabolic activity of the soil population. Two other soils with a pronounced difference in biomass-C content were used to investigate the influence of different amendments on the AEC. In an experiment with low glucose supplements up to 500 μg C g^?1 soil, the soil with the low biomass-C (a cambisol) showed a distinct increase of the AEC from 0.34 to 0.50, whereas the soil with the high biomass-C content (a phaeozem) increased its AEC only slightly from 0.32 to 0.37. In another experiment with high glucose supplements the phaeozem reached its maximum AEC value of 0.56 after the addition of 4000 μg Cg^?1 soil. An amendment with 8000 μg C g^?1 soil gave no further increase. In the combisol the addition of 1000 μg C g^?1 soil increased the AEC to 0.61. Higher supplements gave only a slight further increase to a maximum value of 0.67 after the addition of 8000 μg C g^?1 soil. The same AEC value was reached when the cambisol was amended with a mixture of organic substrates at a concentration of 10,000 μg C g^?1 soil.     

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.     

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.     

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.     

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.     

Decomposition of Zn-rich Arabidopsis halleri Litter in Low and High Metal Soil in the Presence and Absence of EDTA

Hyperaccumulating plants are increasingly investigated in combination with EDTA addition to soil for phytoremediation of heavy metal contaminated soils. A 60-day incubation experiment was carried out to investigate the effects of heavy metal release during the decomposition of Zn-rich (15.7 mg g^?1 dry weight) Arabidopsis halleri litter on C mineralization, microbial biomass C, biomass N, ATP, and adenylate energy charge (AEC). These effects were investigated in two soils with different Zn, Cu, and Pb levels, with and without EDTA addition to soil. The sole addition of Zn-rich A. halleri litter to the two soils did not increase the contents of NH₄NO₃ extractable Zn, only with the combined additions of EDTA and litter was there a considerable increase, being equivalent to three times the added amount in the low metal soil and to 50% in the high metal soil. Litter amendment increased the CO₂ evolved; being equivalent to 44% of the added C in the two soils, but EDTA addition had no significant effect on CO₂ evolution. Litter amendment resulted also in an 18% increase in microbial biomass C, 27% increase in ATP and 6% increase in AEC in the two soils, but EDTA had again no effect on these indices at both metal levels. In contrast, the sole addition of litter had no effect on microbial biomass N, but EDTA addition increased microbial biomass N on average by 49%. The application of EDTA for chelate-assisted phytoextraction should in the future consider the risk of groundwater pollution, which is intensified by resistance of EDTA to microbial decomposition.     

Relation between respiration,ATP content,and Adenylate Energy Charge (AEC) after incubation at different temperatures and after drying and rewetting

Temperature, drying, and rewetting are important climatic factors that control microbial properties. In the present study we looked at the respiration rates, adenosine 5′‐triphosphate (ATP) content, and adenylate energy charge (AEC) as a measure for energy status of microbial biomass in the upper 5 cm of mineral soils of three beech forests at different temperatures and after rewetting. The soils differed widely in pH (4.0 to 6.0), microbial biomass C (92 to 916 μg (g DW)^—1) and ATP content (2.17 to 7.29 nmol ATP (g DW)^—1). The soils were incubated for three weeks at 7 °C, 14 °C, and 21 °C. After three weeks the microbial properties were determined, retaining temperature conditions. The temperature treatment did not significantly affect AEC or ATP content, but respiration rates increased significantly with increasing temperature. In a second experiment the soils were dried for 12 hours at 40 °C. Afterwards the soils were rewetted and microbial properties were monitored for 72 hours. After the drying, respiration rates dropped below the detection limit, but within one hour after rewetting respiration rates increased above control level. Drying reduced AEC by 16 % to 44 % and ATP content by 47 % to 78 %, respectively. Rewetting increased AEC and ATP content significantly as compared to dry soil, but after 72 hours the level of the controls was still not reached. The level of AEC values indicated dormant cells, but ATP content increased. These results indicate that the microbial carbon turnover was not directly linked to microbial growth or microbial energy status. Furthermore our results indicate that AEC may describe an average energy status but does not reflect phases of growing, dormant, or dying cells in the complex microbial populations of soils.     

Microbial reaction in activity, biomass, and community structure after long-term continuous mixing of a grassland soil

Long-term continuous mixing at 40% water holding capacity (WHC) or as slurry at 400% WHC should result in increased soil organic matter decomposition rates in comparison to a control treatment at 40% WHC, but may have strong impacts on soil microbial indices for activity, biomass, and community structure. The amount of extractable inorganic N (NO₃-N+NH₄-N) accumulated in the soil solution after 40 weeks of incubation at 25 °C was 3% of total N in the control treatment and 4% in the two continuous mixing treatments. However, in the treatment mixing at 40% WHC, this 33% increase compared to the control treatment might be explained solely by the decrease in microbial biomass N. In the control treatment, microbial indices decreased in the order microbial biomass C (−10%), microbial biomass N (−40%), ergosterol (−45%) and ATP (−60%). In the treatment mixing at 40% WHC, all four microbial biomass indices were significantly lower than the respective index in the control treatment. This was especially true for microbial biomass N. In the treatment mixing as slurry, only the contents of microbial biomass C and ATP were significantly lower in comparison to the control treatment. The correspondence analysis ordination biplot of the phospholipid fatty acid (PLFA) profiles showed distinct clusters for the three treatments at the end of the incubation. The strongest relative decline of 64% was observed for the fungi-specific PLFA 18:3ω6 in the treatment mixing as slurry in comparison to the control treatment. The content of total bacterial PLFA decreased only by 23%. The differences between the control treatment and the treatment mixing at 40% WHC were less apparent. Fungi represent on average 21% of total microbial biomass C at the end of the incubation if the ergosterol content is recalculated into fungal biomass C. In accordance with this percentage, 22% of the group-specific PLFA could be attributed to fungi.     

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.     

Adenosine triphosphate and microbial biomass in soil

Two methods for measuring adenosine 5'-triphosphate (ATP) in soil were compared, one based on extraction with NaHCO₃-CHCl₃ and thel other on extraction by a trichloracetic acid-phosphate-paraquat reagent. Recoveries of added ATP were greater with the NaHCO₃-CHCl₃ reagent but the extraction of “native” soil ATP by NaHCO₃-CHCl₃ was only about a third of that by TCA-phosphate-paraquat.Microbial biomass C and ATP were measured in 8 contrasting English soils, using the fumigation method to measure biomass C and the TCA-phosphate-paraquat method to measure ATP. Except in one acid woodland soil, the ratio (ATP content of the soil)/(biomass C content of the soil) was relatively constant, with a mean of 7.3 mg ATP g^?1 biomass C for the different soils. This value is very similar to that obtained earlier in a range of 11 grassland and arable soils from Australia. Taking the English and Australian grassland and arable soils together, there is a close (r = 0.975) linear relationship between ATP and microbial biomass C that holds over a wide range of soils and climates. From this relationship, the soil biomass contains 7.25 mg ATP g^?1 biomass C, equivalent to an ATP-to-C ratio of 138, or to 6.04 μmoles ATP g^?1 dry biomass.The acid woodland soil (pH 3.9) contained much less biomass C, as measured by the fumigation method, than would have been expected from this relationship. This, and other evidence, suggests that the fumigation method for measuring microbial biomass C breaks down in strongly acid soils.The ATP content of the biomass did not depend on the P status of the soil, as indicated by NaHCO₃-extractable P.     

Adenosine triphosphate (ATP) and microbial biomass in soil: Effects of storage at different temperatures and at different moisture levels

Abstract

On air‐drying, the ATP contents of two moist soils fell to about one quarter of their original values. When a freshly‐sampled soil (field temperature 5.5°C) was stored moist (43% water holding capacity) for 7 days at 25°C the ATP content increased from 4.54 to 7.84 μg ATP g^‐1 soil. Storage at 10°C caused a smaller increase; to 5.39 μg g^‐1 soil. Microbial biomass C also increased on storage but the relative increase was less than that of ATP. Thus the biomass C/ATP ratio fell from 234 in the freshly sampled soil to 168 in the soil stored moist for 7 days at 25°C. The ATP content declined to less than half its starting value if storage was under waterlogged conditions.

The ATP method for determining microbial biomass in soil depends on the use of a constant factor (5.85 mg ATP g^‐1 biomass C) for converting ATP content to biomass C. This factor came from work on soils that had been stored moist at 25°C for several days before biomass C and ATP measurements were made: it is only applicable to soils that have been stored in this way.     

Microbial biomass growth,following incorporation of biochars produced at 350 °C or 700 °C,in a silty-clay loam soil of high and low pH

Biochar has been widely proposed as a soil amendment, with reports of benefits to soil physical, chemical and biological properties. To quantify the changes in soil microbial biomass and to understand the mechanisms involved, two biochars were prepared at 350 °C (BC350) and 700 °C (BC700) from Miscanthus giganteus, a C4 plant, naturally enriched with ¹³C. The biochars were added to soils of about pH 4 and 8, which were both sampled from a soil pH gradient of the same soil type. Isotopic (¹³C) techniques were used to investigate biochar C availability to the biomass. Scanning Electron Microscopy (SEM) was used to observe the microbial colonization, and Attenuated Total Reflectance (ATR) to highlight structural changes at the surface of the biochars. After 90 days incubation, BC350 significantly increased the biomass C concentration relative to the controls in both the low (p < 0.05) and high pH soil (p < 0.01). It declined between day 90 and 180. The same trend occurred with soil microbial ATP. Overall, biomass C and ATP concentrations were closely correlated over all treatments (R² = 0.87). This indicates that neither the biomass C, nor ATP analyses were affected by the biochars, unless, of course, they were both affected in the same way, which is highly unlikely. About 20% of microbial biomass ¹³C was derived from BC350 after 90 days of incubation in both low and high pH soils. However, less than 2% of biomass ¹³C was derived from BC700 in the high pH soil, showing very low biological availability of BC700. After 90 days of incubation, microbial colonization in the charsphere (defined here as the interface between soil and biochar) was more pronounced with the BC350 in the low pH soil. This was consistent with the biomass C and ATP results. The microbial colonization following biochar addition in our study was mainly attributed to biochar C availability and its large surface area. There was a close linear relationship between ¹³CO₂ evolved and biomass ¹³C, suggesting that biochar mineralization is essentially a biological process. The interactions between non-living and living organic C forms, which are vital in terms of soil fertility and the global C cycle, may be favoured in the charsphere, which has unique properties, distinct from both the internal biochar and the bulk soil.