A simple method for quantitative determination of ATP in soil

A simple method to measure soil ATP by the luciferin-luciferase system is described. The ATP is extracted from the soil by vigorous shaking with a sulfuric acid-phosphate solution for 15 min. An aliquot of the soil suspension is neutralized with a Tris-EDTA solution and mixed with a special ATP releasing reagent (NRB). ATP is measured after a 10 s exposure to the NRB reagent, followed by addition of luciferin-luciferase and integration over 10 s in a Lumacounter M 2080. The ATP content in soils which had been stored at 5°C for 90 days and then incubated at 25°C for 5 days, ranged from 0.37 to 7.52 μg ATP g^?1 dry wt, with standard deviations less than 10%. There was a close (r = 0.96) linear relationship between ATP content and biomass C determinated by fumigation for this group of soils. The soil biomass contained 4.2–7.1 μg ATP mg^?1 biomass C. The ATP content of the biomass declined during storage at 5°C for 210 days.     

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.     

Estimation of microbial biomass and activity in soil using microcalorimetry

Relationships between the rate of heat output from soil, the rate of respiration and the soil microbial biomass were investigated for 25 soils from northern Britain. The rate of heat output, measured in a Calvet microcalorimeter at 22°C, correlated well with the rate of carbon dioxide respiration. The average amount of heat evolved per cm³ of gas respired. 21.1 J cm^?3, suggests that the biomass metabolism was largely aerobic. The rate of heat output per unit of total microbial biomass was remarkably uniform over a wide range of soils, but showed differences depending upon whether the soil had been stored or amended. Mineral soils that had been stored at 4°C had the lowest heat output, 12.0 mW g^?1 biomass C, compared with a mean of 20.4 mW g^?1 biomass C for freshly-collected soils. Amendment with glucose (0.5% w/w) caused an immediate increase in respiration and heat output, up to 59.4 mW g^?1 biomass C for stored soils and 188.2 mW g^?1 biomass C for freshly collected soils. There was a consistent relationship between the biomass and the rate of heat output from freshly collected and amended mineral and organic soils which gave a linear fit using log transformed data: y= 0.6970+ 1.025x (r= 0.98, P < 0.001) (y=log₁₀ biomass C, μgC g^?1; x=log₁₀ rate of heat output at 22°C, μW g^?1). The overall relationship between biomass and the rate of heat output for all the amended samples was: 1 g biomass C= 180.05 ± 34.61 mW.     

The reaction between phosphate and dry soil. I. The effect of time, temperature and dryness

Phosphate (P) was added to soil in solution. The soil was air-dried or freeze-dried and then incubated at a range of temperatures for periods of up to 110 d. The rate of the continuing reaction between the P and soil was measured using the null-point method, and by measuring the amount of desorption induced by filter paper impregnated with iron oxide (Pi test). The reaction between soil and P continued in both air-dried and freeze-dried soil, albeit more slowly than in moist soil. Freezing the soil, whether moist or dry, virtually stopped the reaction. These results are consistent with the hypothesis that the continuing reaction between P and soil involves a solid-state diffusive penetration of the soil particles by the sorbed P ions. They also indicate that the common practice of storing soil air-dry, even for short periods at low temperature, will not preserve the P status of the soil as at sampling. It was estimated that for a sample of soil which remained moist at 25°C for 100d after the addition of 335 μg P g⁻¹ soil, before being sampled and stored air-dry at 4°C for 16 years, the measured P_i test value would be about 15 μg P g⁻¹. This compares with 46 μg P g⁻¹ which is the estimated Pⁱ test value measured on the same day as sampling. When samples cannot be analysed for P status immediately following sampling, they should be stored at the lowest convenient temperature, preferably below 0°C.     

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.     

Relationship between rate of carbon dioxide evolution,microbial biomass carbon,and amount of dissolved organic carbon as affected by temperature and water content of a forest and an arable soil

Abstract

Using an Ochrept soil of a forest at climax stage or of an arable site at Kita‐Ibaraki, a city in central Japan, the rates of carbon dioxide (CO₂)‐carbon (C) evolution, the amounts of microbial biomass carbon (MBC) and the amounts of dissolved organic carbon (DOC) were measured in a laboratory with special reference to the incubation temperature and the soil water content. The rates of CO₂‐C evolution increased exponentially with increase in the incubation temperature in the range of 4–40°C. The temperature coefficients (Q₁₀) were 2.0 for the forest and 1.9 for the arable soil. The amounts of MBC were almost constant of 980 μg g^‐1 soil in the incubation temperature up to 25°C for the forest, and 340 μg g^‐1 soil in the incubation temperature up to 31 °C for the arable soil. The amounts of DOC in soil solutions were almost constant at 3.1 μg g^‐1 soil in the incubation temperature up to 25°C for the forest, and 3.8 μg g^‐1 soil in the incubation temperature up to 31°C for the arable soil. The rates of CO₂‐C evolution and the amounts of DOC increased with increase in soil water content (% of soil dry weight) up to 91% for the forest or up to 26% for the arable soil. However, the rates of CO₂‐C evolution and the amounts of DOC were almost constant within soil water content in the range of 91–160% or 26–53%, respectively. The amounts of MBC of the forest or arable soil were almost constant over a wide range of soil water content in the range of 41–220% or 8–73%, respectively. The rates of CO₂‐C evolution of both the forest and the arable soils were highly correlated with the amounts of DOC, but not with the amounts of MBC, under laboratory conditions in the case that the amounts of DOC were changed by various treatments. The regression equation,     

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.     

Microbial biomass and activity in soils amended with glucose

Four contrasting soils were amended with glucose at concentrations up to 10 mg g^?1 soil. The soils were incubated at 22°C for 14 days and the biomass determined at various times by chloroform fumigation or substrate-induced respiration. The adenosine triphosphate (ATP) content or the amylase and dehydrogenase activities were also determined. The size of the increases in biomass, ATP content and the enzyme activities was generally related to the amount of glucose added. The initially higher ATP levels quickly declined, and apparent substrate conversion figures up to 84% indicated that substrate-induced respiration overestimated the biomass. There were generally no significant correlations between ATP, biomass or enzyme activities.     

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.     

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.     

Interference from plant roots in the estimation of soil microbial ATP,C, N and P

Excised, solution-grown roots of maize or ryegrass added to two pasture soils at the rate of 6.0mg g^?1 and 13.8 mg g^?, respectively, increased the flush (fumigated minus control values) of CO₂-C by up to 1.89-fold, KCl extractable N by up to 1.88-fold, and NaHCO₃ extractable P by 3.28-fold. The ATP content of the soil was increased by up to 1.42-fold. Because of high variability the effect of the roots on the C and N flushes was not significant at P < 0.05.Incubation of the root-amended soils for 7 days at 25°C prior to fumigation much decreased the contribution from the roots to the C and N flush, and to the ATP content. There was, however, still a large significant effect of the roots on the P-flush, this being up to 3 times greater than the equivalent soil without roots.In soil samples with a high viable root density (> 6mg g^?1) such as may occur in dense pastures, greenhouse pot experiments or rhizosphere soil samples, it is recommended that they be incubated for 7 days prior to fumigation and analyses. Without such prior incubation there is the risk that root material may be included in the “microbial” biomass estimations.     

Persistence of effects of nitrification inhibitors added to soils

Abstract

The persistence of the effects of four nitrification inhibitors (2‐ethynylpyridine, nitrapyrin, etridiazole, 3‐methylpyrazole‐l‐carboxamide) on nitrification in soil was assessed by measuring the ability of two soils to nitrify NH₄ ⁺ [added as (NH₄)₂SO₄] after they had been treated with 5 μg inhibitor g^‐1 soil and incubated at 10, 20, or 30°C for 0, 21, 42, 84, 126, or 168 days. The soils used differed markedly in organic‐matter content (1.2 and 4.2% organic C). The data obtained showed that the persistence of the effects of the inhibitors studied decreased markedly with increase in soil temperature from 10 to 30°C and that, whereas the initial inhibitory effects of the test compounds on nitrification were greatest with the soil having the lower organic‐matter content, the persistence of their effects at 20 or 30°C was greatest with the soil having the higher organic‐matter content. The inhibitory effects of 2‐ethynylpyridine and etridiazole on nitrification were considerably more persistent than those of nitrapyrin or 3‐methylpyrazole‐l‐carboxamide and were significant even after incubation of inhibitor‐treated soil at 20°C for 168 days.     

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.     

Activity and biomass of soil microorganisms at different depths

We measured microbial biomass C and soil organic C in soils from one grassland and two arable sites at depths of between 0 and 90 cm. The microbial biomass C content decreased from a maximum of 1147 (0–10 cm layer) to 24 g g^-1 soil (70–90 cm layer) at the grassland site, from 178 (acidic site) and 264 g g^-1 soil (neutral site) at 10–20 cm to values of between 13 and 12 g g^-1 soil (70–90 cm layer) at the two arable sites. No significant depth gradient was observed within the plough layer (0–30 cm depth) for biomass C and soil organic C contents. In general, the microbial biomass C to soil organic C ratio decreased with depth from a maximum of between 1.4 and 2.6% to a minimum of between 0.5 and 0.7% at 70–90 cm in the three soils. Over a 24-week incubation period at 25°C, we examined the survival of microbial biomass in our three soils at depths of between 0 and 90 cm without external substrate. At the end of the incubation experiment, the contents of microbial biomass C at 0–30 cm were significantly lower than the initial values. At depths of between 30 and 90 cm, the microbial biomass C content showed no significant decline in any of the four soils and remained constant up to the end of the experiment. On average, 5.8% of soil organic C was mineralized at 0–30 cm in the three soils and 4.8% at 30–90 cm. Generally, the metabolic quotient qCO₂ values increased with depth and were especially large at 70–90 cm in depth.     

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.     

Microbial biomass estimations in soils from tussock grasslands by three biochemical procedures

Microbial biomass was determined by three biochemical procedures in nine topsoils from a climosequence in tussock grasslands. The pH values of the samples ranged from 4.4 to 6.2 and organic C contents from 2.5 to 20.0%. When determined by a chloroform-fumigation procedure, contents of biomass C and mineral-N (Min-N) flush ranged from 530–2780 and 59–167 μgg^?1 dry soil respectively. Adenosine 5'-triphosphate (ATP) content ranged from 2.2 to 10.7 μg g^?1 dry soil. All three estimates were significantly correlated with each other and with several soil properties, including organic C and total N contents and CO₂ production. They were not significantly correlated with any climatic factor.In spite of these significant correlations, the ratios of the biomass estimates varied appreciably in the different soils. The ratios of biomass C/Min-N flush ranged from 7.8 to 22.8 (average 12.5), biomass C/ATP from 163 to 423 (average 248) and Min-N flush/ATP from 12 to 35 (average 22). These ratios were mostly higher than those found elsewhere for Australian and English soils. The high biomass C/ATP and Min-N flush/ATP ratios did not appear to originate from inefficient extraction of “native” ATP or from the soils' P status. Based on these results, care in the use of factors for obtaining soil microbial biomass content from Min-N flush or ATP values is indicated.     

Enzyme activities in Appalachian soils: 3. Pyro‐phosphatase

Abstract

The pyrophosphatase (PPi) enzyme catalyzes the hydrolysis of pyrophosphate to orthophosphate. The PPi activity was determined in 14 hill‐land soils of the Appalachian Region. The top two horizons from each soil were sampled and stored in a field‐moist state at 4°C (4°C) and in an air‐dry (AD) state prior to determination of PPi activity. Each soil type had its own level of PPi activity. The mean PPi activities of surface and subsurface horizons stored at 4°C were 1.3 and 1.7 times higher than those of AD samples. The average PPi activities of surface horizons were 6.2 and 4.6 times higher than subsurface horizons stored at AD and 4°C. respectively. The PPi activities were positively correlated with organic C, N, forms of S and P. exchangeable cations, original moisture by weight and volume, and percent air‐filled porosity. In both surface and subsurface horizons, significant positive correlationships were observed between PPi activities and Mn content, indicating the need for Mn in the activation of PPi. High PPi activities in surface horizons might induce a greater rate of hydrolysis of applied pyrophosphate (polyphosphate) fertilizers to orthophosphate and lead to undesirable fixation of P in acid soils.     

Influence of storage on soil microbial biomass estimated by three biochemical procedures

The effects of 28 and 56 days' storage at 25°, 4° and ?20°C on the microbial biomass content of four soils from tussock grasslands were studied by three biochemical procedures. Two of the procedures involved measurement of CO₂ and mineral-N (Min-N) production by chloroform-fumigated and unfumigated soil, and consequent estimation of biomass C and Min-N flush respectively. In the third, adenosine 5'-triphosphate (ATP) content was determined.Patterns of CO₂ production were often influenced by storage treatment. The use of fixed incubation periods for estimating the CO₂ flush of fumigated soil and the steady rate of CO₂ production by unfumigated soil did, however, give biomass C estimates that were generally similar to those calculated from individually determined incubation periods for each treatment and soil.Biomass C values could change significantly at all storage temperatures, but generally least at ?20°C. Storage at ?20°C was also the most suitable for retaining ATP contents, whereas 4°C was best for values of Min-N flush. Values of Min-N flush after storage of soil at ?20°C decreased significantly in two of the soils but increased in another. No storage temperature was thus satisfactory for all three indices of microbial biomass. Generally, however, 4°C was adequate for short periods, and 25°C the least suitable.     

Soil microbial activity in opencast coal mine restorations

Abstract. A number of restored areas, a soil store and undisturbed areas on opencast coal mine sites, all of similar soil type, were sampled. The microbiological activity (dehydrogenase assay), nitrogen mineralization and nitrifying potentials and physico-chemical characteristics of the soils were determined. Dehydrogenase activities ranged from 140 to 580 μg TPF g^-1 24 h^-1 in undisturbed control soils, whereas the disturbed soils had activities of 10 to 220 μg g^-1 24 h^-1, with the smallest activities being recorded in the stored and most recently reinstated soil, indicating that disturbance has depressed microbial activity. Nitrogen mineralization potential was significantly affected by disturbance, with a value of 18 to 26 μg inorganic N g^-1 21 d^-1 in the soil store and 38 μg^-1 21 d^-1 in a soil reinstated for six months, although the values were less than this in soils reinstated for up to six years. Nitrifying potential was not significantly less in the stored soils, being within the range of 60 to 135 μg nitrate N formed g^-1 soil 21 d^-1, which was similar to that found in the undisturbed control soil. The water-holding capacity was less in the stored soil than the undisturbed controls, and was significantly less in soil reinstated for 1.5 to 2.5 years, being only 65% of the undisturbed value (0.66 g water g^-1 soil). Ammonium content of the soil store was one hundred fold larger in the soil store than in the controls (0.6 to 1.7 μg ammonium N g^-1). The carbon contents in the control soils ranged from 4.5 to 7.2%, whereas the stored and reinstated soils had generally less amounts ranging from 1.6 to 5.8%. There was a significant and positive correlation between water-holding capacity and nitrifying potential. The implications for long-term restoration are discussed.     

Some factors affecting survival of sclerotia of Macrophomina phaseolina in soil

At least 75% of the sclerotia of Macrophomina phaseolina survived for 1 yr in most natural soils kept at 26°C and at 50–55% of the soil moisture holding capacity (m.h.c.). Although survivability was reduced in a very acid soil (pH 4.5) collected under a pine stand, 33% of the sclerotia survived for 1 yr. Soil pH had very little or no effect on sclerotial survivability. Of three organic amendments tested (alfalfa hay, chitin, pine needles) only ground alfalfa hay at 0.8% (w/w) reduced survivability of sclerotia in soil by about 75% in a year. Alfalfa hay at 0.4% reduced survivability by 36%. Various N sources added at 200 μg Ng^?1 soil had no effect on survival. Of 13 fungicides tested, only benomyl and captan at 20 μg a.i. g^?1 soil appreciably reduced populations of sclerotia in soil.Soil temperature and moisture content were the two most important factors affecting survivability of sclerotia. At ?5 or 5°C the biggest drop in sclerotial survivability occurred when the soil was incubated moist (at 50% m.h.c. or more). At 26°C the biggest drop occurred in air-dried soil (2–3% m.h.c.) and survivability was decreased to some extent at 15 and 30% m.h.c. Survivability also dropped rapidly in moist soil (50–55% m.h.c.) exposed to four cycles each having 3-week freezing (?5°C) and 1 week thawing (26°C). Sclerotia in air-dried soil (2–3% m.h.c.) continuously kept at ?5°C maintained nearly complete survivability after 16 weeks. Sclerotia survived almost 80–90% in moist soil (50–55% m.h.c.) kept for 16 weeks at 26°C or in moist soil exposed to four cycles each having 3-week thawing (26°C) and 1-week freezing (?5°C).