Nitrogen balance in a soil‐wheat system under plow‐ and no‐tillage in the argentine humid pampa

Abstract

Changing conventional tillage to conservation tillage systems affects nitrogen (N) cycling in agroecosystems. Our objective was to evaluate the role of soil organic pools, specially plant residues, as sources‐sinks of nitrogen in an humid and warm temperate environment cropped to wheat, under plow‐ and no‐tillage. The experimental site was in the Argentine Pampa on a Typic Hapludoll. A balance‐sheet method was used: Nupt+Nres=Nsow+Nmin, where Nupt=N uptake by the crop at harvest; Nsow=soil mineral N as NH₄ and NO₃ at 0–90 cm depth, one month before sowing, plus N added as fertilizer; Nres=residual soil mineral N as NH₄ and NO₃ at 0–90 cm depth, at harvest; Nmin=N mineralized from humus and plant residues during wheat growing period. Nupt did not differ between tillage systems. Nitrogen supply by the mineral N pool, estimated by the difference Nsow‐Nres, was ca. 150 kg N ha^‐1 in both tillage systems. Plant residues decomposed and released N under both treatments. This organic N pool decreased 77% along the crop cycle. Nmin, calculated using the balance equation was 83 kg N ha^‐1, and did not differ between tillage managements, representing 35% of Nupt. This results highlight the importance of the organic pools as sources of N for wheat in the Humid Pampa. They also brink our attention on the importance for evaluate residue decomposition and humus mineralization in warm‐temperate regions when fertilizer requirements are determined, in order to minimize environmental hazard and economic losses by overfertilization.     

Effects of seedbed properties on crop emergence. 4. Inhibitory effects of oxygen deficiency

Abstract

For studies of the effects of seedbed properties on crop emergence, experiments were carried out in shallow plastic boxes. In some experiments, it was examined whether rainfall after sowing could cause oxygen deficiency in the seedbed sufficiently severe to hamper emergence. Crops studied were barley (Hordeum vulgare L.), wheat (Triticum aestivum L.), oilseed rape (Brassica napus var. oleifera, L., Metzg.) and pea (Pisum sativum L.). For harmful oxygen deficiency to develop it appeared that rainfall would need to cause structural collapse of the surface layer followed by continuously wet weather accompanied by slow drainage and high oxygen consumption in the soil; in the experiments the latter was achieved by large amounts of easily decomposable organic matter. It was concluded that such conditions are rare in the field. Therefore, unless rainfall after sowing generates surface water for an extended period, the poor crop emergence often observed after such rainfall is nearly always caused not by oxygen deficiency, but by surface layer hardening when this layer dries.     

Modification of the Cataldo method for the determination of nitrate in soil extracts by potassium chloride

The simple and sensitive colorimetric method developed by Cataldo et al. for the determination of nitrate in solution was modified to apply it for 2 M KCI soil extracts. The color intensity was decreased markedly when the KCI concentration of the sample solution increased, in contrast, the sensitivity was improved when the ratio of mixing volume of the sample and salicylic acid-sulfuric acid reagent was changed, and when the sample volume was increased. The modified method showed almost the same accuracy as the steam distillation method with MgO and Devarda alloy.     

A sensitive method for detecting the depletion of nonexchangealble potassium in the rhizosphere using a sequential extraction with 0.01 molar hydrochloric acid

Twenty-one strains of fluorescent pigment-producing Pseudomonas (abbreviated to FPP-Pseudomonas) species were isolated from soil and roots of apple and peach trees using selective media. FPP-Pseudomonas strains were identified as Pseudomonas fluorescens. Moreover, on the basis of the utilization of several organic compounds, these strains were divided into three groups.

P. fluorescens strains isolated from the roots were assigned to mainly groups 1 and 2, and most of the isolates from the soil to group 3. All the strains of group 2 exhibited antifungal activity (in vitro) against three soilborne plant pathogenic fungi: Rhizoctonia solani, Verticillium dahliae, and Rosellinia necatrix. These results suggest that the strains of group 2 play an important role as antifungal rhizobacteria.     

Evaluation of readily available nonexchangeable potassium in soil by sequential extractions with 0.01 molar hydrochloric acid

Nonexchangeable potassium (K-ne), i.e. 1 M NH₄OAc-nonexchangeable K, often contributes significantly to plant nutrition. However conventional extraction methods often extract much more K-ne than plants even after intensive cropping, suggesting the difficulty in evaluating the amount of readily available soil K-ne. In this study, we used a milder extraction method (0.01 M HCl method) to examine its applicability to evaluate the amount of readily available K-ne in soil. In the first experiment, the concentration of K-ne in twenty surface soils sampled from agricultural fields in Japan and K-bearing minerals was determined by the 0.01 M HCl method, i.e. sequential extraction with 0.01 M HCl over a period of 10 d after removal of exchangeable K, and by conventional methods. The average percentage of the soil K-ne extracted by the 0.01 M HCl method amounted to 0.66% of the total K amount, and was much lower than that by a single extraction with 1 M HNO₃ (2.0%) or with 0.2 M sodium tetraphenylboron for 2 d (22%). In the second experiment, the amount of K-ne removed by chemical extractions was compared with that of K-ne removed by maize plants grown for 29 d in five of the above soils. The amount of the K-ne evaluated by the 0.01 M HCl method gave the highest correlation (p < 0.05) with that of the K-ne utilized by plants among the extraction methods applied. The amount of soil K-ne extracted by the 0.01 M Hel method could therefore become a suitable index of the amount of readily available K-ne in soil. Extraction of K-ne in soils after maize planting further indicated that plants had removed K-ne more intensively than the 0.01 < HCl method probably only from the rhizosphere, although a high correlation was observed between the amount of K-ne removed by the 0.01 M Hel method and that by plants. This implies that the estimation of the amount of K-ne utilized by plants requires not only soil chemical analysis but also the evaluation of the percentage of the soil volume where the plant-induced release of K-ne actually occurs.     

Prediction of potentially mineralizable N from amidohydrolase activities in a manure-applied, corn residue-amended soil

Prediction of potentially mineralizable N as an important N pool from soil amidohydrolases was investigated. Composite soil samples were collected from plots of a field experiment in which 0, 50 and 100 Mg cow manure ha⁻¹ year⁻¹ had been applied for five consecutive years. The soils were treated with corn shoots or roots or remained untreated in a factorial combination with the manure treatments, with three replications. The mineralized inorganic N was measured periodically in 20-week incubations and potentially mineralizable N (N₀) was calculated based on a first-order kinetic model. Urease, l-glutaminase and l-asparaginase activities were measured before and after incubation. The values of N₀ ranged from 208.6 in the controls to 388.4 in soils that had received 50 Mg ha⁻¹ year⁻¹ of cow manure and were amended with corn shoots. Corn residue amendment in the manure treated soils, increased the values of N₀ or changed the N mineralization kinetic pattern from a first-order to a zero-order model. According to a relative sensitivity index, l-asparaginase was the most sensitive enzyme to the treatments. Multiple regression analysis revealed that 92% of N₀ variations can be described by the activities of urease and l-asparaginase and therefore the soil amidohydrolase activities have the potential to evaluate potentially mineralizable N.     

Effect of potato plants expressing snowdrop lectin (GNA) on the performance and colonization behaviour of the peach-potato aphid Myzus persicae

Abstract

Transgenic potato plants expressing snowdrop lectin (GNA potatoes) are developed to increase resistance against sap-feeding insects. When expressing GNA at relatively high levels such potatoes may have a negative effect on the fecundity and development of the first generation of the important pest, the peach-potato aphid Myzus persicae (Sulzer) (Hemiptera: Aphididae). However, the effects on M. persicae over several generations, and how such plants affect the alate aphids’ colonization behaviour have not been reported. In this laboratory study, the performance of single M. persicae on potatoes with low GNA expression, measured as developmental time, fecundity, size and survival, was compared with the control, following two successive generations of single apterous aphids. Aphid population growth on the two plant lines was also studied. In addition, colonization behaviour was investigated in a choice experiment where the alate aphids could choose between the GNA and an isogenic control potato plant in a cage. The present study showed that the apterous aphid performance was not significantly different on the two potato lines, although the aphids tended to perform slightly poorer on the transgenic potato. However, the transgenic potato was less likely to be colonized by alate aphids. It is concluded that such transgenic potato plants expressing the lectin at a relatively low level, maximum 0.2% of the soluble protein, have no significant impact on the performance of apterous M. persicae once on the plant, but may have a potential in controlling the aphids by altering the colonization behaviour of alates.     

The impact of increasing heavy metal stress on the diversity and structure of the bacterial and actinobacterial communities of metallophytic grassland soil

A study was undertaken to investigate the bacterial community found in metallophytic grassland soil contaminated with Zn and Pb. We hypothesised that such communities would be tolerant of additional heavy metal stress due to phylogenetic and functional adaptation. In microcosm experiments, lasting 51 days, denaturing gradient gel electrophoresis (DGGE) analyses was used to compare the total bacterial and actinobacterial communities in non-amended soils and those to which additional Pb and Zn concentrations were added. There was a decrease in total bacterial diversity with each addition of Pb and Zn; in contrast, the actinobacterial community diversity remained unaffected. The community structures were analysed using multivariate analyses of the DGGE profiles. Total bacterial community profiles showed two distinct groups sharing less than 80% similarity, irrespective of Pb and Zn addition. The first contained profiles sampled during the first 7 days of the experiment; the second contained those sampled from day 10 onwards. Actinobacterial profiles from those that were non-amended showed a similar distribution to those of the total bacterial community. However, in soil amended with fivefold additional Pb and Zn, all the profiles shared more than 80% similarity. Raup and Crick analyses suggested that total bacterial soil communities were subject deterministic selection becoming significantly similar as the experiment progressed, but this was inhibited by the highest concentration of additional Pb and Zn. Actinobacterial communities showed a similar response but were less affected by elevated Pb and Zn concentrations. These data indicate that the diversity of the actinobacterial community was not negatively affected by additional heavy metal stress in contrast to total bacterial community diversity.     

Seasonal variations in enzyme activity and organic carbon in soil of a burned and unburned hardwood forest

This study examined variations in soil organic C content and the activity of acid phosphatase, α-glucosidase, phenol oxidase, chitinase, and l-glutaminase in ultisols of burned and unburned areas in Quercus-dominated forests in Ohio, USA. The low intensity, prescribed fires were conducted in April 2001, with temperature 10 cm above the forest floor averaging 160-240 °C. Sampling was conducted throughout the six month growing season (May-October) of 2003, two years after the fire. Organic C content in these ultisols varied between 20 and 30 g C/kg soil, and varied little through the growing season, except for a late season increase to ∼32 g C/kg soil in the burned areas. When enzyme activity was expressed per unit soil organic C, there was no statistically significant variation among sample dates in soil enzyme activity except l-glutaminase, which demonstrated a distinct maximum in activity in spring. Non-metric multidimensional scaling (NMS) ordination resulted in no clear separation of burned and unburned sample areas based on soil organic C and enzyme activity. When the growing season was divided into three segments (early spring, late spring/early summer, and late summer/early autumn), there was again a lack of separation between burned and unburned areas in the earlier two segments, whereas in the late summer/early autumn segment the burned and unburned areas were clearly separated on the basis of differences in soil organic C and l-glutaminase activity. As environmental factors (e.g. soil temperature, moisture) and substrate availability do not vary in parallel through the growing season in this region, seasonal patterns often differ among enzyme systems based on their predominant control mechanism. Sampling time during the growing season appears to have little effect on holistic judgments of fire effects based on soil enzymes, except under restrictive conditions.     

Changes in amino acid enantiomers and microbial performance in soils from a subtropical mountain oasis in Oman abandoned for different periods

An important feature of maintaining the agricultural stability in millennia-old mountain oases of northern Oman is the temporary abandonment of terraces. To analyse the effects of a fallow period on soil microbial performance, i.e. microbial activity and microbial biomass, samples of eight terrace soils abandoned for different periods were collected in situ, assigned to four fallow age classes and incubated for 30 days in the laboratory after rewetting. The younger fallow age classes of 1 and 5 years were based on the records of the farmers recollections, the two older fallow age classes of 10–20 and 25–60 years according to the increase in the d -to- l ratio of valine and leucine enantiomers. The increase in these two ratios was in agreement with that of the d -to- l ratio of lysine. The strongest relationship was observed between the increase in the d -to- l ratio of lysine and the decrease in soil microbial biomass C. However, the most stringent coherence between the increase in fallow age and soil properties was revealed by the decreases in cumulative respiration and net N mineralisation rates with decreasing availability of substrate to soil microorganisms. During the 30-day incubation following rewetting, relative changes in microbial activity (respiration and net N mineralisation) and microbial biomass (C and N) indices were similar in the eight terrace soils on a fallow age-class-specific level, indicating that the same basic processes occurred in all of the sandy terrace soils investigated.     

The influence of time, storage temperature, and substrate age on potential soil enzyme activity in acidic forest soils using MUB-linked substrates and l-DOPA

The purpose of this experiment was to evaluate whether soil storage and processing methods significantly influence measurements of potential in situ enzyme activity in acidic forest soils. More specifically, the objectives were to determine if: (1) duration and temperature of soil storage; (2) duration of soil slurry in buffer; and (3) age of model substrates significantly influence the activity of six commonly measured soil extracellular enzymes using methylumbelliferone (MUB)-linked substrates and l-dihydroxyphenylalanine (l-DOPA). Soil collected and analyzed for enzyme activity within 2 h was considered the best measure of potential in situ enzyme activity and the benchmark for all statistical comparisons. Sub-samples of the same soil were stored at either 4 °C or −20 °C. In addition to the temperature manipulation, soils experienced two more experimental treatments. First, enzyme activity was analyzed 2, 7, 14, and 21 days after collection. Second, MUB-linked substrate was added immediately (i.e. <20 min) or 2 h after mixing soil with buffer. Enzyme activity of soil stored at 4 °C was not significantly different from soil stored at −20 °C. The duration of soil storage was minimal for β-glucosidase, β-xylosidase, and peroxidase activity. N-acetyl-glucosaminidase (NAGase), phosphatase, and phenol oxidase activity appeared to change the most when compared to fresh soils, but the direction of change varied. Likewise, the activities of these enzymes were most sensitive to extended time in buffer. Fluorometric MUB and MUB-linked substrates generally had a 3-day shelf life before they start to significantly suppress reported activities when kept at 4 °C. These findings suggest that the manner in which acidic forest soils are stored and processed are site and enzyme specific and should not initially be trivialized when conducting enzyme assays focusing on NAGase, phosphatase, and phenol oxidase. The activities of β-glucosidase, β-xylosidase, and peroxidase are insensitive to storage and processing methods.     

Effect of acid deposition on forested catchment in the western Tokyo,Japan

Acid deposition and its effect on Andisols were investigated in the forest experimental station, the Rolling Land Laboratory (RLL), located in the Tama Hill region of Central Japan. The annual volume-weighted mean pH value of open bulk precipitation was 4.8 in the period 1990 to 1992. Nitrate deposition at RLL was larger than for sulfate, which was obviously different from results in Japan Environment Agency (JEA) stations or in study areas of Europe and North America. Abundant nitrate deposition was ascribed to the high emissions from a non-point source, mainly cars. Although sulfate concentration in throughfall fluctuated, its concentration in soil solution was kept at a low level. This was attributed to the high sulfate adsorption capacity of the Andisols.     

Selected Soil Enzyme Activities,Soil Microbial Biomass Carbon,and Root Yield as Influenced by Organic Production Systems in Sweet Potato

A field experiment was conducted on Typic Rhedustalfs to determine the effects of various organic production systems. Results revealed that the soil microbial biomass carbon (SMBC) content was greatest with the application of 100% N through farmyard manure. The ratio index value of biofertilizer along with 50% N through any one of the organic sources were greater than 100% N through green leaf manure?/?vermicompost, integrated use of manure and fertilizer (conventional production system), and control (traditional system of production). Soil enzymes varied with the production systems. The urease, phosphatase, and β-glucosidase activities were more with greater nitrogen, phosphorus, and organic-matter treatments, respectively. The SMBC, soil enzymes, and microbial activity were very responsive to organic production systems, but their levels and activities were not reflected in sweet potato root yield.     

Effects of Topographic Factors of the Site on the Essential Oil Compounds of Artemisia aucheri Aerial Parts Grown in a Mountainous Region

In this study, the chemical compositions of the essential oils from the aerial parts of Artemisia aucheri, grown in a mountainous region in Ghamsar Province, central Iran, have been analyzed by using gas chromatography (GC)–mass spectroscopy (MS) to determine how they are affected by topographic factors (site direction and elevation). Plants were sampled at random in full flowering stage in a completely randomized (CR) design with three replications. The essential oil was extracted by a modified Likens–Nickerson's simultaneous distillation–extraction (SDE) technique. Analysis of the plant oils on the sites [in two directions, north–south (N/S) and east–west (E/W)] and five elevations resulted in 30 identified compounds. Of the oil samples collected, the 99.94% of the components at the N/S site and 99.89% at the E/W site were identified. The main component in the two directions (N/S and E/W) was α-thujone with 19.5% and 18%, respectively. Other significant components in the two directions include α-pinene, davanone, camphor, and camphene. The percentages of all these main compounds on the N/S site were more than on the E/W site. Also, the main compounds increased with increasing altitude and at all five elevations. Among all the oil compounds, the percentages of α-thujone, α-pinene, and camphor were the greatest. Finally, at all the elevations, the percentage of the α-thujone was the greatest among the different oil compounds.     

氮素对不同大豆品种根系分泌物中有机酸的影响

采用室内溶液培养方法,分别研究了接种根瘤菌处理下,两种氮源和两种氮浓度对两个大豆品种根系分泌物中有机酸的影响。结果表明,合丰25号根系分泌的有机酸种类和数量无论苗期或花期,接种或不接种根瘤菌,均表现为硝态氮处理高于酰胺态氮处理,表明合丰25号大豆更喜硝态氮,硝态氮促进了有机酸的分泌。绥农10号在酰胺态氮下的有机酸种类和数量均高于硝态氮处理,表明其更喜酰胺态氮,酰胺态氮下根瘤菌的存在增加其根系分泌物中有机酸种类和数量。可见,大豆根系分泌物中有机酸的种类和数量因品种而异,因品种对氮源的喜好而变化;根瘤菌在不同程度上增强或减弱根系有机酸的分泌作用。柠檬酸受氮素供应浓度影响很大,当氮素供应浓度较低时,大豆根系分泌物中可检测到柠檬酸,供氮浓度升高则检测不到。     

Optimization of hydrolytic and oxidative enzyme methods for ecosystem studies

Microbial digestive enzymes in soil and litter have been studied for over a half century, yet the understanding of microbial enzymes as drivers of ecosystem processes remains hindered by methodological differences among researchers and laboratories. Modern techniques enable the comparison of enzyme activities from different sites and experiments, but most researchers do not optimize enzyme assay methods for their study sites, and thus may not properly assay potential enzyme activity. In this review, we characterize important procedural details of enzyme assays, and define the steps necessary to properly assay potential enzyme activities in environmental samples. We make the following recommendations to investigators measuring soil enzyme activities: 1) run enzyme assays at the environmental pH and temperature; 2) run proper standards, and if using fluorescent substrates with NaOH addition, use a standard time of 1 min between the addition of NaOH and reading in a fluorometer; 3) run enzyme assays under saturating substrate concentrations to ensure V_max is being measured; 4) confirm that product is produced linearly over the duration of the assay; 5) examine whether mixing during the reaction is necessary to properly measure enzyme activity; 6) find the balance between dilution of soil homogenate and assay variation; and 7) ensure that enzyme activity values are properly calculated. These steps should help develop a unified understanding of enzyme activities in ecosystem ecology.     

Effect of tillage and residue management on enzyme activities in soils

Recent interest in soil tillage and residue management has focused on low-input sustainable agriculture. In this study we investigated the effect of three tillage systems (no-till, chisel plow, and moldboard plow) and four residue placements (bare, normal, mulch, and double mulch) on the activities of four amidohydrolases (amidase, L-asparaginase, L-glutaminase, and urease) in soils from four replicated field-plots. Correlation coefficients (r) for linear regressions between the activities of each of the enzymes and organic C or pH and between all possible paired amidohydrolases were also calculated. The results showed that the effects of tillage and residue management on pH in the 28 surface soil (0–15 cm) samples were not significant. The organic C content, however, was affected significantly by the different tillage and residue-management practices studied, being the greatest in soils with notill/double mulch treatment, and the least with no-till/bare and moldboard/normal treatments. Within the same tillage system, mulch treatment resulted in greater organic C content compared with normal or bare treatment. The activities of the amidohydrolases studied were generally greater in mulch-treated plots than in non-treated plots, and were significantly correlated with organic C contents of soils, with r values ranging from 0.70^*** to 0.90^***. Linear regression analyses of enzyme activities on pH values (in 0.01 M CaCl₂) of the 28 surface soils showed significant correlations for L-asparaginase, L-glutaminase, and urease, with r values of 0.74^***, 0.77^***, and 0.72^***, respectively, but not for amidase (r=0.24). The activities of the four amidohydrolases studied in the 40 soil samples tested were significantly intercorrelated, with r values ranging from 0.72^*** to 0.92^***. The activities of the four amidohydrolases decreased with increasing soil depth of the plow layer, and were accompanied by a decrease in organic C content.     

Nine years of measurements of atmospheric nitrogen and sulphur deposition to Danish forest

Since 1985 measurements of gasses, aerosols, precipitation and throughfall have been carried out at three forest sites in Denmark with equal aged Norway Spruce plantations. The times series show a downward trend in the concentration of sulphur dioxide. Particulate sulphate, ammonia and particulate ammonium and the total nitrate seem to have a more constant concentration level. The wet deposition measurements show a decreasing trend in the content of acid (protons), sulphate, ammonium and nitrate, though for the nitrogen compounds it is only a slight fall. A decrease in concentrations of protons and sulphate is also seen in the throughfall measurements, in throughfall the nitrogen compounds hardly seem to decrease.     

Development of an appropriate procedure for estimation of RUSLE EI30 index and preparation of erosivity maps for Pulau Penang in Peninsular Malaysia

Pluviographic data at 15 min interval from 6 stations in Pulau Penang of Peninsular Malaysia were used to compute rainfall erosivity factor (R) for the revised universal soil loss equation (RUSLE). Three different modelling procedures were applied for the estimation of monthly rainfall erosivity (EI₃₀) values. While storm rainfall (P) and duration (D) data were used in the first approach, the second approach used monthly rainfall for days with rainfall ≥ 10 mm (rain₁₀) and monthly number of days with rainfall ≥ 10 (days₁₀). The third approach however used the Fournier index as the independent variable. Based on the root mean squared error (RMSE) and the percentage error (PE) criteria, models developed using the Fournier index approach was adjudged the best with an average PE value of 0.92 and an average RMSE value of 164.6. Further, this approach was extended to the development of a regional model. Using data from additional sixteen stations and the Fournier index based regional model, EI₃₀ values were computed for each month. ArcView GIS was used to generate monthly maps of EI₃₀ values and also annual rainfall erosivity (R). The rainfall erosivity factor (R) in the region was estimated to vary from 9000 to 14,000 MJ mm ha^− 1 h^− 1 year^− 1.     

Stability of hydrolytic enzyme activity and microbial phosphorus during storage of tropical rain forest soils

Storage can markedly influence microbial and biochemical properties in soils, yet recommendations for sample storage are based on studies of temperate soils that regularly experience extended cold periods. We assessed the influence of storage conditions on microbial phosphorus and the activity of four hydrolytic enzymes (phosphomonoesterase, phosphodiesterase, β-glucosidase, and N-acetyl-β-d-glucosaminidase) in three lowland tropical forest soils from the Republic of Panama that experience a constant warm temperature. The soils spanned a strong rainfall gradient and contained contrasting physical and chemical properties (pH 3.6-5.9; total carbon 26-50 g C kg⁻¹; clay 33-62%; total phosphorus 0.30-0.60 g P kg⁻¹). Storage treatments were: (i) room temperature (22 °C in the dark), (ii) refrigerated (4 °C in the dark), (iii) air-dried (10 d, 22 °C), and (iv) frozen (−35 °C). There were significant changes in enzyme activities and microbial phosphorus during refrigerated and room temperature storage, although changes were relatively small during the first two weeks. An initial marked decline in enzyme activities for one soil analyzed within 2 h of sampling was attributed to a flush of activity caused by sampling and soil preparation (sieving, etc.). For longer-term storage (>2 weeks), ambient laboratory temperature appeared preferable to freezing and cold storage, because one month of storage caused a marked decline in enzyme activities and microbial phosphorus in one soil. Freezing preserved the activities of some enzymes in some soils at rates comparable to cold or room temperature storage, but caused a marked decline in microbial phosphorus in two soils. Air-drying caused a marked decline in microbial phosphorus and the activity of all enzymes. We therefore conclude that enzyme assays and microbial phosphorus should be determined in tropical forest soils after no more than two weeks storage in the dark at ambient laboratory temperature.