Enzyme activity in soils at subzero temperatures

Urease activity, phosphatase activity, and sulfatase activity were detected in soils at ?10 and ?20°C. The occurrence of enzyme activity in soils at subzero temperatures is attributed to enzyme-substrate interaction in unfrozen water at the surfaces of soil particles. Support for this explanation was obtained from experiments showing that hydrolysis of urea by jackbean urease occurs at ?10 or ?20°C in the presence, but not in the absence, of clay minerals or autoclaved soils. No enzyme activity could be detected in soils at ?30°C.     

A comparison of the effects of air-drying and acetone dehydration on soil enzyme activities

The effects of air-drying or acetone dehydration on the activities of invertase, amylase, cellulase, xylanase, urease, protease, phosphatase and sulphatase in nine New Zealand soils from pastures were determined. Generally, air-drying of the soils reduced activities, with losses ranging from slight for sulphatase to very large for protease. For most of the enzymes, acetone dehydration generally resulted in smaller losses of activity than did air-drying; exceptions were invertase and all but two enzymes in the two sandy soils with lowest clay contents. In contrast to the other activities, the activity of urease was generally increased by both drying treatments and that of sulphatase by acetone dehydration. The effects of 6 weeks' storage at 4°C on the enzyme activities of field-moist, air-dried and acetone-dehydrated soils were also assessed. Generally, field-moist storage was best for preserving the original activity, primarily because of the initial losses caused by both drying processes.     

Volatilization of sulfur from unamended and sulfate-treated soils

Volatilization of sulfur from unamended and sulfate-treated soils was studied by sensitive gas chromatographic techniques using a flame photometric detector fitted with a sulfur filter. The soils employed were surface samples of 25 Iowa soils selected to obtain a wide range in properties. No release of volatile sulfur compounds was detected when 11 of these soils were incubated under aerobic or waterlogged conditions before or after treatment with sulfate (400 μg sulfate S/g soil). Fourteen soils released volatile sulfur compounds when incubated under waterlogged conditions before and after addition of sulfate, but only 4 of these soils released volatile sulfur compounds when incubated under aerobic conditions. Where volatilization of sulfur was observed, the volatile sulfur detected was identified as dimethyl sulfide or as dimethyl sulfide associated with smaller amounts of carbonyl sulfide, carbon disulfide, methyl mercaptan, and (or) dimethyl disulfide. No trace of hydrogen sulfide was detected. Where release of volatile sulfur was observed, the amount of sulfur volatilized at 30°C in 60 days under aerobic or waterlogged conditions was very small and did not account for more than 0–05% of the sulfur in the unamended or sulfate-treated soils studied. It is concluded that gaseous loss of sulfur from unamended or sulfate-treated soils is insignificant under conditions likely to be encountered in the field.     

Production and persistence of urease activity in soils

Addition of urea to Iowa soils did not induce urease activity, but production of urease activity was observed on addition of glucose and other organic materials that promote microbial activity. The persistence of the urease activity produced on addition of these materials varied with the soil, but, with each soil studied, the urease activity after addition of organic materials eventually was identical to that of the unamended soil. No increase or decrease in urease activity was observed when unamended or urea-treated soils were incubated under aerobic conditions for several months. It is concluded that soil constituents protect urease against microbial degradation and other processes leading to inactivation of enzymes and that every soil has a stable level of urease activity determined by the ability of its constituents to provide this protection.     

Effects of liming and selected heavy metals on ammonium release in waterlogged agricultural soils

Many farmlands are periodically flooded or ponded by excessive precipitation resulting in changes to soil chemical and biochemical properties. In this study, one set (eight treatments with four replications) of field-moist surface soils (0–15 cm) and their air-dried counterparts obtained from a long-term liming experiment were incubated at 30 °C under waterlogged conditions for 10 days, and the amounts of net NH₄ ⁺-N released (soluble and exchangeable) were determined after extraction with 4 M KCl. Another set of three surface soils were used to evaluate the effect of six heavy metals on the NH₄ ⁺-N release under waterlogged conditions. Results showed that increasing the liming rate from 0 to 17,930 kg ha^?1 effective calcium carbonate equivalent increased the average soil pH from 4.98 to 7.06, averages of the amounts of NH₄ ⁺-N released ranged from 1.6 to 5.2 mg N kg^?1 field-moist soil, and the corresponding amounts released in air-dried soils ranged from 18.9 to 32.9 mg N kg^?1 soil. This increase of the amount NH₄ ⁺-N released in air-dried soil samples is presumably due to a slaking effect. At 5 mmol kg^?1 soil, all six heavy metals inhibited the NH₄ ⁺-N released. The relative effectiveness of the heavy metals in inhibition of the NH₄ ⁺-N released varied among the three soils. Lead(II) was the most effective inhibitor of NH ₄ ⁺-N release in Clarion and Harps soils and Cd(II) in Harps soil. Cobalt(II), Cu(II), and Cd(II) were the least effective inhibitors of NH₄ ⁺-N release in Clarion, Harps, and Okoboji soils, respectively.     

Assessing air-drying and rewetting pre-treatment effect on some soil enzyme activities under Mediterranean conditions

Soil enzyme activities are useful indicators of soil quality as they are very sensitive to disturbance. Sample storage or pre-treatments could affect the results in these assays, which are normally determined in fresh samples, kept cold or frozen. The objectives of this study were to (i) evaluate the effect of air-drying or air-drying and rewetting on β-glucosidase, acid phosphatase and urease activities in soils from different locations, degradation status and sampling seasons, and (ii) assess if air-drying or air-drying and rewetting is an accurate sample storage and pre-treatment procedure for enzyme activities in soil quality evaluations under semiarid Mediterranean conditions. Our results showed that urease, phosphatase and β-glucosidase activities were hardly affected by air-drying of degraded and non-degraded soils from the two locations studied in all seasons. Short incubations (4, 8 and 12 d at 23 °C) of rewetted air-dried soil at 55% of water-holding capacity showed different patterns depending on the enzyme studied. Urease and β-glucosidase activities were relatively stable during incubation, with several significant (P<0.05) shifts up and down in some soils and samplings. However, acid phosphatase showed an increase in activity with incubation, of between 5% and 50% relative to air-dried samples. These increases followed no pattern and were unrelated to soil characteristics or sampling date. Hence, urease, phosphatase and β-glucosidase activities determined in air-dried soil samples seem to be representative of those obtained under field-moist conditions. In contrast, short incubations of rewetted soil samples can produce fluctuations in these enzyme activities, mainly of acid phosphatase, and estimations in these conditions are not so representative of field-moist soil values.     

Short-term assay of soil urease activity using colorimetric determination of ammonium

Summary A rapid assay for soil urease in the absence of bacteriostatic agents has been developed. The method comprises incubation of soil with an aqueous or buffered urea solution, extraction of ammonium with 1 N KCl and 0.01 NHCl and colorimetric NH₄ ⁺ determination by a modified indophenol reaction. The method is characterized by high sensitivity and stability of the coloured complex formed. Measurements obtained by this method showed that no change in urease activity occurred when field-moist samples of soils were stored at –20°C for as long as 5 months. Air-drying of field-moist soil samples may lead to an increase in urease activity.     

Factors affecting the availability of organic carbon for denitrification of nitrate in subsoils

Summary Recent work in our laboratory indicated that the slow rate of denitrification in Iowa subsoils is not due to a lack of denitrifying microorganisms, but to a lack of organic C that can be utilized by these microorganisms for reduction of nitrate. To identify factors affecting the availability of leachable organic C in surface soils capable of promoting denitrification in subsoils, we studied the effects of freezing and drying and of plants and plant residues on the amounts of water-soluble organic C in surface soils and the ability of this organic C to promote denitrification in subsoils. We found that aqueous extracts of field-moist, frozen, and air-dried surface soils promoted denitrification in subsoils and that their stimulatory effects on denitrification were highly correlated (r=0.93) with their organic C contents and decreased in the order air-dried soils frozen soils >field-moist soils. But a detailed study of the effect of drying a surface soil to different water tensions indicated that drying of soils under natural conditions is not likely to lead to a substantial increase in their content of water-soluble organic C. Amendment of surface soils with corn or soybean residues led to a marked increase in the amount of organic C in aqueous extracts of the soils and in the ability of these extracts to promote denitrification in subsoils. These effects of plant residues could not be detected after incubation of residue-treated soils for a few days under aerobic conditions, but they increased markedly with an increase in the time of incubation from 1 to 10 days when residue-treated soils were incubated under anaerobic conditions. Analyses for organic acids indicated that this increase was largely due to fermentative production of acetic, propionic, and butyric acids by soil microorganisms. Growth chamber studies showed that growth of corn, soybean, wheat, and sorghum plants on surface soil did not significantly increase the organic C content of leachates of the soil or the ability of these leachates to promote denitrification in subsois. We conclude that plant residues are a major source of the leachable organic C in surface soils that is capable of promoting denitrification in subsoils.     

Effect of soil moisture and temperature on the survival of phytophthora cinnamomi rands in soil

Soil moisture and temperature affect hyphal lysis and sporangial formation in Phytophthora cinnamomi. Hyphal lysis was most rapid in soils at, or below, field capacity when incubated at 25–27° C. Sporangia were not formed in soils below field capacity and only very slowly in waterlogged soil. Sporangia were not formed at temperatures below 15°C.     

Effects of soil properties and trace metals on urease activities of calcareous soils

A study was conducted to investigate the relationship between urease activity and some physical, chemical, and microbiological properties of soils from central Iran. Inhibitory effects of Cr, Cd, and Pb on urease activity were also studied. Results indicated that no significant difference was observed between urease activity of field-moist and air-dried soils. Soil organic C and total N correlated highly significantly with urease activity, with r values of 0.899*** and 0.797***, respectively. There was also a significant correlation between urease activity and the number of bacteria grown on urea-agar media, with r value of 0.470*. A significant negative correlation (r =–0.492*) was observed between urease activity and electrical conductivity of saturation paste extracts. There were no significant correlations between urease activity and soil textural properties, pH, calcium carbonate equivalent, cation exchange capacity, and populations of soil bacteria on nutrient agar and population of soil fungi on potato dextrose agar. Both Cd and Pb inhibited urease activity to a similar extent and to a greater extent than did Cr.     

Residual Soil Nitrate: A Comparison between Air-Dried and Field-Moist Soil Samples

In the framework of the European nitrate directive (91/676/EEG), losses of nitrate (NO₃)– nitrogen (N) to both surface and groundwater are limited to 50 mg/l. Because the residual NO₃-N in the soil profile after harvest is considered the main determinant of nitrate leaching during wintertime, the Flemish government imposed a limit value of 90 kg NO₃-N ha^?1 up to a soil depth of 90 cm between 1 October and 15 November. This study compared two different soil sample preparation methodologies. When samples were analyzed immediately upon arrival, no differences in NO₃-N concentration were observed. However, although field-moist samples are maintained at 4 °C, nitrification is not completely stopped, as indicated by the increased NO₃-N concentration in field-moist samples 10 days after storage at 4 °C . In contrast, nitrification in air-dried samples is stopped during the oven drying when 40 °C is reached. Moreover, the reproducibility was significantly greater in air-dried samples as compared to field-moist samples.     

Assessing the effects of air-drying and rewetting pre-treatment on soil microbial biomass,basal respiration,metabolic quotient and soluble carbon under Mediterranean conditions

Soil biochemical properties are useful indicators of soil quality as they are very sensitive to disturbance. Sample storage or pre-treatments could affect the results in these assays, which are normally determined on fresh samples, kept cold or frozen. The objectives of this study were to (i) evaluate the effect of air-drying or incubation of rewetted air-dried soil samples on microbial biomass carbon (MBC), basal soil respiration (BSR), qCO₂ and water soluble carbon (WSC), in soils from different locations, with different degradation status and sampling seasons, and (ii) assess if air-drying or incubation of rewetted air-dried soil samples is an accurate sample storage and pre-treatment procedure for these soil properties in soil quality evaluations under semiarid Mediterranean conditions. Our results showed that air-drying does not have the same effects on MBC, BSR, qCO₂ and WSC depending on the geographical situation and sampling date. It seems that the warmest and driest place and season show less variation when using air-dried soil samples, with values representative of those obtained under field-moist conditions. Short incubations (4, 8 and 12 days at 23 °C) provoked a general decrease in all properties, probably due to labile organic compounds depletion. Hence, air-dried soils can be used as part of soil quality analysis to estimate these biochemical properties in summer time in the semiarid region of South-East Spain, because they have not suffered severe affections. Moreover, MBC could also be determined using air-dried soil in the driest zones during all year. In contrast, estimations with incubated soil samples are not, in any case, representative of field-moist soil values.     

Die N-Mineralisierung von Böden im Laborbrutversuch

N mineralization in soils under laboratory incubation conditions The potential rate of release of nitrogen by the organic matter in agriculturally used soils was determined under laboratory conditions by means of incubation. Mineralization of the more resistant soil organic matter proceeded linearly with time during an incubation period of 2–3 weeks, when field-moist and air-dried samples were used and at the beginning of the incubation experiment sufficient water was added to bring them to saturation. Mineralization was taking an exponential course in soils with additions of easily decomposable organic matter or in soils with a higher proportion of organic residues from crops. For the 14 investigated arable and grassland soils great variations in the average daily rate of mineralization were found ranging from 5–60 μg N_min/10 g DM. The data correlated very well with the biomass (r = 0.96) and the cell-free protease activity (r = 0.98) of the soils. Different measures of soil management (preceding crops, application of sewage sludge, addition of heavy metals) had a more or less pronounced influence on the rate of mineralization. The optimum temperature was 50°C for N mineralization and 26°C for nitrification. Contrary to nitrification, the soil reaction had only little influence on mineralization and proved also independent of the N_min content of soils. The results indicate that ammonification of organic N compounds may largely proceed via the microbial biomass.     

Interactions between soil organic matter status, cropping history, method of quantification and sample pretreatment and their effects on measured aggregate stability

 The effects of sample pretreatment (field-moist, air-dried or tension rewetted) on aggregate stability measured by wet sieving or turbidimetry were compared for a group of soil samples ranging in organic C content from 20 to 40 g C kg^–1. Concentrations of total N, total and hot-water-extractable carbohydrate and microbial biomass C were linearly related to those of organic C. Aggregate stability measured by wet sieving using air-dried or field-moist samples and that measured by turbidimetry, regardless of sample pretreatment, increased curvilinearly with increasing soil organic C content. However, when tension-rewetted samples were used for wet sieving, aggregate stability was essentially unaffected by soil organic C content. Measurements of aggregate stability (apart from wet sieving using rewetted soils) were closely correlated with one another and with organic C, total and extractable carbohydrate and microbial biomass C content of the soils. The short-term effects of aggregate stability were also studied. Soils from under long-term arable management and those under long-term arable followed by 1 or 3 years under pasture had similar organic C contents, but aggregate stability measured by turbidimetry and by wet sieving using air-dried or field-moist samples increased with increasing years under pasture. Light fraction C, microbial biomass and hot-water-extractable carbohydrate concentrations also increased. It was concluded that both total and labile soil organic C content are important in relation to water-stable aggregation and that the use of tension-rewetted samples to measure stability by wet sieving is unsatisfactory since little separation of values is achieved. Received: 6 January 1999     

淹水厌氧培养对水稻土中酶活性的影响

An incubation experiment with soil water content treatments of 0.15 （W1）, 0.20 （W2）, and 0.40 （W3） g g^-1 soil was carried out for two months to investigate the activities of important enzymes involved in C, N, P, and S cycling in a paddy soil from the Taihu Lake region, China, under waterlogged and aerobic conditions. Compared with air-dried soil, waterlogging resulted in a significant decrease （P ≤ 0.05） of fluorescein diacetate （FDA） and /3-D-glucosidase activities, and this effect was enhanced with increasing waterlogging time. Waterlogging also significantly inhibited （P ≤ 0.05） arylsulfatase as well as alkaline and acid phosphatase activities, but did not decrease the activities with the increase in waterlogging time. Short-term waterlogging did not affect urease activity, but prolonged waterlogging decreased it markedly. In contrast, the aerobic incubation （W1 and W2 treatments） significantly increased （P ≤ 0.05） FDA, alkaline phosphatase, and /3-D-glucosidase activities. With aerobic treatments the activities of FDA and alkaline phosphatase increased with incubation time, whereas /3-D-glucosidase activity decreased. A significant difference （P ≤ 0.05） was usually observed between the W1 and W2 treatments for the activities of FDA as well as alkaline and acid phosphatase; however,/3-D-glucosidase and urease were usually not significant （P ≤ 0.05）. No activity differences were observed between waterlogging and aerobic incubation for arylsulfatase and urease.     

Kinetics of Soil Urease in Four Agricultural Soils Affected by Urease Inhibitor PPD at Contrasting Moisture Regimes

An incubation test was conducted on four agricultural soils to investigate effects of phenyl phosphorodiamidate (PPD) on soil urease kinetics at contrasting moisture regimes with contrasting fertility levels. The PPD made the Michaelis constant (Km) increase and the maximum enzyme reaction velocity (Vmax) decrease, behaving as a mixed inhibitor. With incubation time, Km first increased and then decreased under saturation condition, whereas it decreased under field-moist condition, the opposite of the changes of Vmax. Compared with black soil and albic soils, brown and cinnamon soils had larger Km and lower Vmax and Vmax/Km ratios, indicating that the effectiveness of PPD was greater in soils with low fertility. Compared with brown soil and cinnamon soil, black soil and albic soil showed more increases in Km and decreases in Vmax. To apply PPD under waterlogged, saturated, and low-fertility conditions could be a reasonable way to increase fertilizer nitrogen (N)–use efficiency.     

Estimation of Changes in Available Soil Phosphate under Submerged Conditions Associated with Temperature during the Tillering Stage of Rice Plant in the Cool Climate Region of Japan

This study was performed to investigate changes in available soil phosphate associated with temperature under submerged conditions and to explore the possibility for estimating those under submerged conditions during the early growth (tillering) stage of rice plants (Oryza sativa L.). An incubation experiment was conducted under submerged conditions at three temperatures (10°C, 17.5°C and 25°C) using paddy soils collected over a widespread area in Japan. In most soils, significant positive correlations were observed between available soil phosphate and cumulative temperature to 650°C, which corresponded to the tillering stage in Japan. Relationships between the regression formulae of the available phosphate against cumulative temperature to 650°C and soil chemical properties measured in air-dried soil were investigated. The results indicate that the available phosphate of paddy soil against cumulative temperature during tillering stage under submerged conditions can be estimated from the results of air-dried soil analyses which can be conducted before a crop season.     

Persistence of the inhibitory effects of phosphoroamides on urea hydrolysis in soils

Abstract

The persistence of the inhibitory effects of three phosphoroamides [N‐(n‐butyl) thiophosphoric triamide (NBPT), phenylphosphorodiamidate (PPD), and thiophosphoryl triamide (TPT)] on urea hydrolysis in soils was assessed by measuring the ability of four soils to hydrolyze urea after they had been treated with 5 μg phosphoroamide/g soil and incubated at 15°C or 30°C for 0, 3, 7, 14, or 28 days. The soils used differed markedly in pH, texture, and organic‐matter content. The data obtained showed that the persistence of the effects of the phosphoroamides studied decreased with increase in soil temperature from 15°C to 30°C and that whereas the effect of PPD decreased with increase in the time of incubation, the effects of NBPT and TPT sometimes increased before decreasing with increased time of incubation. These observations are in harmony with the recent findings that PPD is a potent inhibitor of urease activity, but decomposes in soils with formation of phenol, which is a relatively weak inhibitor of urease activity, whereas NBPT and TPT do not inhibit urease activity but decompose in soil with formation of their oxon analogs, which are potent inhibitors of urease activity. The inhibitory effect of NBPT on urea hydrolysis was considerably more persistent than that of PPD or TPT and was significant even after incubation of NBPT‐treated soil at 15°C or 30°C for 28 days.     

Sorption of dissolved organic carbon in soils: effects of soil sample storage,soil-to-solution ratio,and temperature

Experiments on the sorption of dissolved organic carbon (DOC) in soils were mainly conducted in batch approaches. Because varying setups were used in these studies, comparison of the results requires knowledge on the effects that different experimental conditions may have on the sorption of DOC. This investigation evaluated the DOC sorption of soils using differently pretreated soil samples (field-fresh (two sampling dates), air-dried, stored at 3°C and −18°C), at different soil-to-solution ratios (1:40, 1:20, 1:10 and 1:5 w/v) and different temperatures (5°C, 15°C, 25°C and 35°C). The sorption of DOC was analyzed using the initial mass (IM) approach, which regressed the initial amount of sorbate (normalized to soil mass) against the sorbed amount (normalized to soil mass). The DOC release — when a solution without DOC was added — strongly increased with temperature and soil-to-solution ratio. Among the different types of sample storage and preparation, air-drying resulted in the largest DOC release. The smallest release was from the field-fresh samples. Freezing and storage at 3°C resulted in intermediate DOC release with freezing having the greater effect. The release from air-dried samples exceeded that of field-fresh samples by a factor of four at maximum. In contrast, none of the experimental setups influenced the slope of the IM isotherms. Thus, it seems possible to compare directly the binding affinity of DOC to different soils as determined at varying experimental conditions.     

Formation of phosphoryl triamide by decomposition of thiophosphoryl triamide in soil

Summary Thiophosphoryl triamide inhibits soil urease activity when added to soil and is under consideration as a fertilizer amendment for retarding hydrolysis of urea fertilizer by soil urease. Work reported showed that this compound is a poor inhibitor of jackbean urease and that its ability to retard soil urease activity is due to its decomposition in soil with formation of phosphoryl triamide, which is a potent inhibitor of urease activity. This conclusion was supported by studies showing a close relationship between the peak area of the ³¹P-nuclear magnetic resonance (NMR) signal from phosphoryl triamide in thiophosphoryl triamide solutions that had been incubated with soil for various times and the ability of these solutions to inhibit jackbean urease activity.