Considerations on the use of the p-nitrophenyl phosphomonoesterase assay in the study of the phosphorus nutrition of soil borne fungi

The p-nitrophenyl phosphomonoesterase assay (p NPPase) is commonly used to measure cell-wall-associated and extracellular phosphatase activity of soil fungi. p NPPases are usually assayed in the context of fungal nutrition, where inorganic P supply might be enhanced by the mineralisation of monoester organic P sources in the soil. The importance of the assay to the P nutrition of soil fungi is considered based on the evidence currently available including the consistency of methodological approach. The nature of organic P in the soil and the relevance of the assay to some specific soil substrates is discussed, particularly the chemistry and bioavailability of myo-inositol hexakisphosphate and the lower inositol phosphates. The evidence for the long-term stability of p NPPases in the soil is examined in the light of the persistence of p NPPase in soils. The role of persistent extracellular fungal p NPPases in the soil P cycle is discussed. Conclusions from p NPPase based studies must be based upon an appreciation of the constraints of the assay and the complex chemistry of organic P and p NPPase in the soil.

Some potential inaccuracies of the p -nitrophenyl phosphomonoesterase assay in the study of the phosphorus nutrition of soil borne fungi

The p-nitrophenol phosphomonoesterase assay (pNPPase) is commonly used to measure cell-wall-associated and extracellular phosphatase activity of soil fungi. pNPPases are usually assayed in the context of fungal nutrition, where inorganic P supply might be enhanced by the mineralisation of organic P sources in the soil. We report here on a series of experiments with the ectomycorrhizal basidiomycete Hebeloma cylindrosporum that highlight components of accepted methodology that might impinge on the reliability of the assay. These include the loss of pNPPase after filtration, inaccuracies in measuring wall-associated enzyme and the ample pool of intracellular pNPPase can be mistakenly measured as external pNPPase if cells are accidentally damaged.

Soil bacterial phoD gene abundance and expression in response to applied phosphorus and long-term management

Bacterial transformation of phosphorus (P) compounds in soil is largely dependent on soil microbial community function, and is therefore sensitive to anthropogenic disturbances such as fertilization or cropping systems. However, the effect of soil management on the transcription of bacterial genes that encode phosphatases, such as phoD, is largely unknown. This greenhouse study examined the effect of long-term management and P amendment on potential alkaline phosphatase (ALP) activity and phoD gene (DNA) and transcript (RNA) abundance. Soil samples (0–15 cm) were collected from the Glenlea Long-term Rotation near Winnipeg, Manitoba, to compare organic, conventional and prairie management systems. In the greenhouse, pots of soil from each management system were amended with P as either soluble mineral fertilizer or cattle manure and then planted with Italian ryegrass (Lolium multiforum). Soils from each pot were sampled for analysis immediately and after 30 and 106 days. Significant differences among the soil/P treatments were detected for inorganic P, but not the organic P in NaHCO3-extracts. At day 0, ALP activity was similar among the soil/P treatments, but was higher after 30 days for all P amendments in soil from organically managed plots. In contrast, ALP activity in soils under conventional and prairie management responded to increasing rates of manure only, with significant effects from medium and high manure application rates at 30 and 106 days. Differences in ALP activity at 30 days corresponded to the abundance of bacterial phoD genes, which were also significantly higher in soils under organic management. However, this correlation was not significant for transcript abundance. Next-generation sequencing allowed the identification of 199 unique phoD operational taxonomic units (OTUs) from the metagenome (soil DNA) and 35 unique OTUs from the metatranscriptome (soil RNA), indicating that a subset of phoD genes was being transcribed in all soils.

Decomposition and phosphorus release from organic residues as affected by residue quality and added inorganic phosphorus

The combined use of organic residue and inorganic fertiliser-phosphorus (P) is appropriate in meeting both the short and long-term P requirement of crops. To assess the influence of added inorganic fertiliser-P on the processes of decomposition and P release from the residue and the relationships with quality, prunings of Gliricidia sepium, Leucaena leucocephela, Senna siamea, Acacia mangium and Paraserienthus falcataria were incubated without and with added inorganic fertiliser-P for 56 days. Soil was added only as inoculum. Decomposition rate and amounts of acid extractable-P (P release) were in the same order: G. sepium > S. siamea > L. leucocepheta > P falcataria > A. mangium. Unlike the other residues, A. mangium released no P despite the loss of half its mass during the 8 weeks of incubation. The residue P content correlated with P release. However, decomposition rate did not correlate with residue P content but with the lignin, polyphenol and cellulose content, and ratios to P. These ratios were negatively correlated with P release suggesting that lignin and polyphenol contents influence P release more when the residue-P content is low. Results suggest that rate of decomposition influences the release of P. The critical residue P content for P release was estimated to be 0.12% < P < 0.19%. Added P had no effect on decomposition and P release from the residues.

Utilisation by upland rice of plant residue- and fertiliser-phosphorus in two tropical acid soils

A dual isotopic technique was used to assess the effects of soil type, and residues of Gliricidia sepium, without and with added fertiliser-P on the utilisation of P. Upland rice (Oryza sativa) was grown for 70 days in two tropical acid soils of different P sorbing capacity and P status. Uniformly P-32-labelled soils were treated with inorganic fertiliser-P tagged with P-33, Gliricidia sepium residue applied at planting and 3 weeks earlier, and in a combination of fertiliser-P and Gliricidia applied at and 3 weeks before planting. There were significant responses of shoot and root weights, and total P uptake to Gliricidia- and/or fertiliser-P addition in the Ultisol (low P status) but not the Oxisol (high P status), suggesting that P in the latter soil was not yield limiting, despite the high standard P requirement. Similarly, incorporation of Gliricidia three weeks before planting further increased shoot weight only in the Ultisol. There were generally higher proportions, quantities and percent utilisations of the Gliricidia- P and fertiliser-P in the Ultisol than in the Oxisol. Gliricidia significantly increased the utilisation of fertiliser-P only in the Ultisol. However, early application of Gliricidia increased Gliricidia- P but not fertiliser-P utilisation in the Ultisol. Added fertiliser-P did not influence Gliricidia- P utilisation.

The Phosphorus Indicators Tool: a simple model of diffuse P loss from agricultural land to water

The Phosphorus Indicators Tool provides a catchment-scale estimation of diffuse phosphorus (P) loss from agricultural land to surface waters using the most appropriate indicators of P loss. The Tool provides a framework that may be applied across the UK to estimate P loss, which is sensitive not only to land use and management but also to environmental factors such as climate, soil type and topography. The model complexity incorporated in the P Indicators Tool has been adapted to the level of detail in the available data and the need to reflect the impact of changes in agriculture. Currently, the Tool runs on an annual timestep and at a 1 km(2) grid scale. We demonstrate that the P Indicators Tool works in principle and that its modular structure provides a means of accounting for P loss from one layer to the next, and ultimately to receiving waters. Trial runs of the Tool suggest that modelled P delivery to water approximates measured water quality records. The transparency of the structure of the P Indicators Tool means that identification of poorly performing coefficients is possible, and further refinements of the Tool can be made to ensure it is better calibrated and subsequently validated against empirical data, as it becomes available.

Zinc tolerance in wheat cultivars as affected by varying levels of phosphorus

The effect of zinc-phosphorus (Zn-P) interaction on Zn efficiency of six wheat cultivars was studied. The higher dry matter yields were observed when Zn was applied at 5 mu g g(-1) soil than with no Zn application. Phosphorus applications also increased dry matter yield up to the application of 25 mu g P g(-1) soil. The dry matter yield was significantly lower at the P rate of 250 mu g g(-1) soil. At the Zn-deficient level, the Zn-efficient cultivars had higher Zn concentrations in the shoots. Zinc concentrations in all cultivars increased when the P level in the soil was increased from 0 to 25 mu g P g(-1) soil except for the cv. Durati, in which Zn concentrations decreased with increases in P levels. However, when ZnxP interactions were investigated, it was observed that at a Zn-deficient level, Zn concentrations in the plant shoot decreased with each higher level of P, and more severe Zn deficiency was observed at P level of 250 mu g g(-1) soil.

Phosphorus dynamics and export in streams draining micro-catchments: Development of empirical models

Annual total phosphorus (TP) export data from 108 European micro-catchments were analyzed against descriptive catchment data on climate (runoff), soil types, catchment size, and land use. The best possible empirical model developed included runoff, proportion of agricultural land and catchment size as explanatory variables but with a low explanation of the variance in the dataset (R-2 = 0.37). Improved country specific empirical models could be developed in some cases. The best example was from Norway where an analysis of TP-export data from 12 predominantly agricultural micro-catchments revealed a relationship explaining 96% of the variance in TP-export. The explanatory variables were in this case soil-P status (P-AL), proportion of organic soil, and the export of suspended sediment. Another example is from Denmark where an empirical model was established for the basic annual average TP-export from 24 catchments with percentage sandy soils, percentage organic soils, runoff, and application of phosphorus in fertilizer and animal manure as explanatory variables (R-2 = 0.97).

Arsenate-induced phosphate release from soils and its effect on plant phosphorus

Adsorption of arsenic onto soil was investigated as a means of understanding arsenic-induced release of phosphate. In batch adsorption experiments As adsorption was accompanied by P desorption. At low As additions, the ratio As adsorbed: P desorbed remained constant. At higher As additions, P desorption reached a maximum while As adsorption continued to increase. The P desorption maximum coincided with an increase in pH. Barley plants were grown on soils spiked with arsenate (0-360 mg As kg(-1)) to investigate the effect on plant growth and P uptake. As arsenic concentration increased, above ground plant yield decreased and the plants showed symptoms typical of As toxicity and P deficiency. At low As additions to the soil, uptake of As and P by barley increased. At higher As additions P uptake decreased. It is argued that this was due to the change in As:P ratio in the soil solution. It is concluded that input of arsenic to the soil could mobilise phosphate. Crop yield is likely to be affected, either due to reduced phosphate availability at low arsenic additions or arsenic toxicity at higher additions.

Mitigation of diffuse phosphorus pollution during rewetting of fen peat soils: A trans-European case study

Intensive cultivation of fen peat soils (Eutric Histosols) for agricultural purposes, started in Europe about 250 years ago, resulting in decreased soil fertility, increased oxidation of peat and corresponding CO2-emissions to the atmosphere, nutrient transfer to aquatic ecosystems and losses in the total area of the former native wetlands. To prevent these negative environmental effects set-aside programs and rewetting measures were promoted in recent years. Literature results and practical experiences showed that large scale rewetting of intensively used agricultural Histosols may result in the mobilisation of phosphorus (P), its transport to adjacent surface waters and an accelerated eutrophication risk. The paper summarises results from an international European Community sponsored research project and demonstrates how results obtained at different scales and from different scientific disciplines were compiled to derive a strategy to carry out rewetting measures. A decision support system (DSS) for a hydrologically sensitive area in the Droemling catchment in north-eastern Germany was developed and is presented as a tool to regulate rewetting in order to control P release. It is demonstrated that additional laboratory experiments to identify essential processes of P release during rewetting and the site-specific management of the water table, the involvement of specific knowledge and experience of the stakeholders are necessary to develop an applicable DSS. The presented DSS is practically used to prevent freshwater resources from diffuse P pollution.

Phosphorus dynamics in the ditch system of a restored peat wetland

The restoration of wetlands as bird habitats often involves the maintenance of a fluctuating water regime by careful, localised ditch water management using pumps and sluices. However, there is evidence in the literature to Suggest that alternate flood/drainage cycles can accelerate nutrient cycling and transport within the soil and, therefore, pose a threat to water quality through the process of eutrophication. This study focused on the dynamics and losses of soil P in a recently re-wetted, eutrophic fen peat developed on alluvium ill South west England. During the 2-year Study (2001 and 2002), soil water tensiometry revealed that the field water table (fluctuating annually between +20 and 60 cm relative to ground level) was extensively influenced across most of the 8.4 ha field site by the management of the adjacent ditch water levels. This conservation-led, prescribed water balance was facilitated by the high hydraulic conductivity (1.1 x 10(-s) ms(-1)) of the lower (70-140 cm), degraded layer of peat. However, only during a 7-day period of water table drawdown by intermittent pump drainage, approximately 45 g ha(-1) of dissolved reactive P (DRP) entered the pumped ditch from the field via this degraded layer. Summer rainfall events >35 mm d(-1) also coincided with significant peaks ill ditch water P concentration (up to 200 mu g L-1 DRP). Even larger peaks (Up to 700 mu g L-1 DRP) Occurred With the annual onset of autumn reflooding. These episodic P loss events pose a serious potential threat to biological water quality. (C) 2009 Elsevier B.V. All rights reserved.

The Representative Soil Sampling Scheme of England and Wales: a statistical analysis of topsoil nutrient status and pH between 1971 and 2001

The Representative Soil Sampling Scheme of England and Wales has recorded information on the soil of agricultural land in England and Wales since 1969. It is a valuable source of information about the soil in the context of monitoring for sustainable agricultural development. Changes in soil nutrient status and pH were examined over the period 1971-2001. Several methods of statistical analysis were applied to data from the surveys during this period. The main focus here is on the data for 1971, 1981, 1991 and 2001. The results of examining change over time in general show that levels of potassium in the soil have increased, those of magnesium have remained fairly constant, those of phosphorus have declined and pH has changed little. Future sampling needs have been assessed in the context of monitoring, to determine the mean at a given level of confidence and tolerable error and to detect change in the mean over time at these same levels over periods of 5 and 10 years. The results of a non-hierarchical multivariate classification suggest that England and Wales could be stratified to optimize future sampling and analysis. To monitor soil quality and health more generally than for agriculture, more of the country should be sampled and a wider range of properties recorded.

Effect of ammonium and nitrate on indigenous mycorrhizal infection, rhizosphere pH change, and phosphorus uptake by sorghum

Sorghum (Sorghum bicolor L.) plants were grown in split pots in three Rothamsted soils with different soil pH values and phosphorus (P) contents. Ammonium addition resulted in higher plant dry weight and P content than comparable nitrate treatments. The pH of soils in the rhizosphere (0.51-mm average thickness) differed from the bulk soil depending on nitrogen (N) form and level. Ammonium application resulted in a pH decrease, but nitrate application slightly increased pH. To examine the effect of rhizosphere acidification on mobilization of phosphate, 0.5 M NaHCO3 extractable phosphate was measured. The lowering rhizosphere pH enhanced the solubility of P in the soil and maybe availability of P to plants. Rhizosphere-P depletion increased with increasing ammonium supply, but when N was supplied as nitrate, P depletion was not related to increasing nitrate supply. Low P status Hoosfield soils developed mycorrhizal infection., and as a result, P inflow was increased. Geescroft soil, which initially had a high P status, did not develop mycorrhizal infection, and P inflow was much smaller and was unaffected by N treatments. Therefore, plant growth and P uptake were influenced by both rhizosphere pH and indigenous mycorrhizal infection.

Phosphorus sorption and availability in soils amended with animal manures and sewage sludge

Soils that receive large applications of animal wastes and sewage sludge are vulnerable to releasing environmentally significant concentrations of dissolved P available to subsurface flow owing to the gradual saturation of the soil's P sorption capacity. This study evaluated P sorption (calculated from Langmuir isotherms) and availability of P (as CaCl2-P and resin P) in soils incubated for 20 d with poultry litter, poultry manure, cattle slurry, municipal sewage sludge, or KH2PO4, added on a P-equivalent basis (100 mg P kg(-1)). All the P sources had a marked negative effect on P sorption and a positive effect on P availability in all soils. In the cattle slurry- and KH2PO4- treated soils, the decreases in P sorption maximum (19-66%) and binding energy (25-89%) were consistently larger than the corresponding decreases (7-41% and 11-30%) in poultry litter-, poultry manure-, and sewage sludge-treated soils. The effects of cattle slurry and KH2PO4 on P availability were, in most cases, larger than those of the other P sources. In the poultry litter, poultry manure, and sewage sludge treatments, the increase in soil solution P was inversely related (R-2 = 0.75) to the input of Ca from these relatively high Ca (13.5-42 g kg(-1)) sources. Correlation analyses implied that the magnitude of the changes in P sorption and availability was not related to the water-extractable P content of the P sources. Future research on the sustainable application of organic wastes to agricultural soils needs to consider the non-P- as well as P-containing components of the waste.

Differences in phosphorus retention and release in soils amended with animal manures and sewage sludge

Excessive levels of P in agricultural soils pose a threat to local water quality. This study evaluated (i) time-dependent changes in soil P sorption (expressed as a phosphorus sorption index, PSI) and P availability (as resin P) during incubation (100 d) with poultry litter, cattle slurry, sewage sludge, or KH2PO4, added on a P-equivalent basis (100 mg P kg(-1)), and (ii) the subsequent kinetics of P release, measured by repeated extractions with a mixed cation-anion exchange resin. Soil exchangeable Ca and ammonium oxalate-extractable Fe and Al were also determined at 100 d of incubation. The small decrease in P sorption in the litter and sludge treatments (25%), compared with that in the slurry and KH2PO4 treatments (52%) between 20 and 100 d of incubation was attributed partly to the formation of new adsorption sites for P. Subsequent P release was described by a power equation: Resin P = a(extraction number)(b), where the constants a and b represent resin P obtained with a single extraction and the rate of P release per resin extraction, respectively. On average, the rate of P release decreased in the order: KH2PO4 and slurry > litter > sludge, and was inversely related to exchangeable Ca content of the incubated soils (R-2 = 0.57). The slower rates of P release in the litter and sludge treatments (P < 0.001) are attributed to the large values for exchangeable Ca (1050-2640 and 1070-2710 mg kg(-1), respectively) in these amended soils. Future research concerned with short-term declines in environmentally harmful levels of P in recently amended soils should consider the differential effects of the amendments on soil P dynamics.