Microbially Induced Soil Colloidal Phosphorus Mobilization Under Anoxic Conditions.

Understanding the behavior of colloidal phosphorus (Pcoll) under anoxic conditions is pivotal for addressing soil phosphorus (P) mobilization and transport and its impact on nutrient cycling. Our study investigated Pcoll dynamics in acidic floodplain soil during a 30-day flooding event. The sudden oxic-to-anoxic shift led to a significant rise in pore-water Pcoll levels, which exceeded soluble P levels by more than 2.7-fold. Colloidal fractions transitioned from dispersed forms (<220 nm) to colloid-associated microaggregates (>220 nm), as confirmed by electron microscopy. The observed increase in colloidal sizes was paralleled by their heightened ability to form aggregates. Compared to sterile control conditions, anoxia prompted the transformation of initially dispersed colloids into larger particles through microbial activity. Curiously, the 16S rRNA and ITS microbial diversity analysis indicated that fungi were more strongly associated with anoxia-induced colloidal release than bacteria. These microbially induced shifts in Pcoll lead to its higher mobility and transport, with direct implications for P release from soil into floodwaters.

Insoluble-Phytate Improves Plant Growth and Arsenic Accumulation in As-Hyperaccumulator Pteris vittata: Phytase Activity, Nutrient Uptake, and As-Metabolism.

Phytate, the principal P storage in plant seeds, is also an important organic P in soils, but it is unavailable for plant uptake. However, the As-hyperaccumulator Pteris vittata can effectively utilize soluble Na-phytate, while its ability to utilize insoluble Ca/Fe-phytate is unclear. Here, we investigated phytate uptake and the underlying mechanisms based on the phytase activity, nutrient uptake, and expression of genes involved in As metabolisms. P. vittata plants were cultivated hydroponically in 0.2-strength Hoagland nutrient solution containing 50 µM As and 0.2 mM Na/Ca/Fe-phytate, with 0.2 mM soluble-P as the control. As the sole P source, all three phytates supported P. vittata growth, with its biomass being 3.2-4.1 g plant-1 and Ca/Fe-phytate being 19-29% more effective than Na-phytate. Phytate supplied soluble P to P. vittata probably via phytase hydrolysis, which was supported by 0.4-0.7 nmol P min-1 g-1 root fresh weight day-1 phytase activity in its root exudates, with 29-545 µM phytate-P being released into the growth media. Besides, compared to Na-phytate, Ca/Fe-phytate enhanced the As contents by 102-140% to 657-781 mg kg-1 in P. vittata roots and by 43-86% to 1109-1447 mg kg-1 in the fronds, which was accompanied by 21-108% increase in Ca and Fe uptake. The increased plant As is probably attributed to 1.3-2.6 fold upregulation of P transporters PvPht1;3/4 for root As uptake, and 1.8-4.3 fold upregulation of arsenite antiporters PvACR3/3;1/3;3 for As translocation to and As sequestration into the fronds. This is the first report to show that, besides soluble Na-phytate, P. vittata can also effectively utilize insoluble Ca/Fe-phytate as the sole P source, which sheds light onto improving its application in phytoremediation of As-contaminated sites.

Insight into the availability and desorption kinetics of Se and Cd in naturally-rich soils using diffusive gradients in thin-films technique.

Cadmium (Cd) contamination of selenium (Se)-rich soils may jeopardize the nutritional benefits of Se-biofortified crops. This study employed diffusive gradients in thin-films (DGT) technique and DIFS (DGT-induced fluxes in soils) model to understand the interdependency and driving factors of Se and Cd distribution and desorption kinetics across 50 soils from south China with naturally elevated levels. DGT-labile Se was the highest (up to 2.66 µg L-1) in non-carbonate/shale-derived soils, while Cd was maximal (5.53 µg L-1) in carbonate-based soils, reflecting soil background concentrations and soil characteristics. Over one-third of the soils showed labile Se:Cd molar ratio below 0.7, suggesting Cd phytotoxicity risks. The DIFS-derived response times (Tc) and desorption rate constants (k-1) suggested that Se was resupplied to the soil solution faster than Cd in soils with higher pH and SOM level, but Se resupply was still restricted due to the rapid depletion of its labile pool. As the first study of Se and Cd release kinetics in soils, our results reveal dependence on soil parent materials, with low labile Se:Cd soils presenting greater Cd hazards. By elucidating Se and Cd lability and interactions in soils, our findings help to inform management strategies to balance reduced Cd risk with adequate Se availability.

Nano and fine colloids suspended in the soil solution regulate phosphorus desorption and lability in organic fertiliser-amended soils.

Mobile colloids impact phosphorus (P) binding and transport in agroecosystems. However, their relationship to P-lability and their relative importance to P-bioavailability is unclear. In soils amended with organic fertilisers, we investigated the effects of nano (NC; 1-20 nm), fine (FC; 20-220 nm), and medium (MC; 220-450 nm) colloids suspended in soil solution on soil P-desorption and lability. The underlying hypothesis is that mobile colloids of different sizes, i.e., NC, FC, and MC, may contribute differently to P-lability in soils enriched with organic fertiliser. NC- and FC-bound Pcoll were positively correlated with P-lability parameters from diffusive gradient in thin films (DGTA-labile P concentration, r ≥ 0.88; and DGTA-effective P concentration, r ≥ 0.87). The corresponding relations with MC-bound Pcoll are weaker (r values of 0.50 and 0.51). NC- and FC-bound Pcoll were also strongly correlated with soil P-resupply (r ≥ 0.64) and desorption (r ≥ 0.79) parameters during DGTA deployment, and the mobility of these colloids was corroborated by electron microscopy of DGTA gels. MC-bound Pcoll was negatively correlated with the solid-to-solution distribution coefficient (r = -0.42), indicating this fraction is unlikely to be the source of P-release from the solid phase after P-depletion from the soil solution. We conclude that NC and FC mainly contribute to regulating soil desorbable-P supply to the soil solution in the DGTA depletion zone (in vitro proxy for plant rhizosphere), and consequently may act as critical conditioners of P-bioavailability, whereas MC tends to form complexes that lead to P-occlusion rather than lability.

Simultaneous Quantification of Soil Phosphorus Labile Pool and Desorption Kinetics Using DGTs and 3D-DIFS.

The buffering of phosphorus concentrations in soil solution by the soil-solid phase is an important process for providing plant root access to nutrients. Accordingly, the size of labile solid phase-bound phosphorus pool and the rate at which it can resupply phosphorous into the dissolved phase can be important variables in determining when the plant availability of the nutrient may be limited. The phosphorus labile pool (Plabile) and its desorption kinetics were simultaneously evaluated in 10 agricultural UK soils using the diffusive gradients in thin-films (DGT) technique. The DGT-induced fluxes in the soil and sediments model (DIFS) was fitted to the time series of DGT deployments (1-240 h), which allowed the estimation of Plabile, and the system response time ( Tc). The Plabile concentration was then compared to that obtained by several soil P extracts including Olsen P, FeO-P, and water extractable P, in order to assess if the data from these analytical procedures can be used to represent the labile P across different soils. The Olsen P concentration, commonly used as a representation of the soil labile P pool, overestimated the desorbable P concentration by 6-fold. The use of this approach for the quantification of soil P desorption kinetic parameters found a wide range of equally valid solutions for Tc. Additionally, the performance of different DIFS model versions working in different dimensions (1D, 2D, and 3D) was compared. Although all models could provide a good fit to the experimental DGT time series data, the fitted parameters showed a poor agreement between different model versions. The limitations of the DIFS model family are associated with the assumptions taken in the modeling approach and the three-dimensional (3D) version is here considered to be the most precise among them.

Phosphorus availability and dynamics in soil affected by long-term ruzigrass cover crop.

The use of grasses as cover crops in the off-season of cash crops under no-till has been largely adopted. However, soil phosphorus (P) uptake was previously shown to be reduced when ruzigrass is introduced in the rotation, affecting the viability and sustainability of this cropping system. The objective of this study was to assess the effect of ruzigrass on soil P availability and desorption kinetics under different P fertilizer application rates. A long-term field experiment where soybean (Glycine max) has been grown in rotation with ruzigrass (Urochloa ruziziensis) or fallow for 10 years, with the application of 0, 13, and 26 kg ha-1 of P, was evaluated for two consecutive years. Soil P desorption kinetics was assessed using diffusive equilibrium (DET) and gradient in thin films (DGT) techniques, as well as the DGT-induced fluxes in soils model (DIFS). Microbial biomass P (MBP) was assessed to verify if soil solution P (PDET) was reduced due to immobilization by microorganisms. Ruzigrass reduced MBP and PDET especially when P fertilizer was applied. The concentration of labile P (PDGT) was also lower after ruzigrass than in fallow. The soil ability to resupply P to soil solution was lower after ruzigrass regardless of P rates due to a slower desorption in response to the perturbation imposed by DGT. Growing ruzigrass as cover crop in the soybean off-season decreases soil P availability regardless of P fertilizer application rates by fundamentally reducing P mobility and P resupply from soil solid phase into soil solution.

The effect of exogenous phytase supplementation on nutrient digestibility, ruminal fermentation and phosphorous bioavailability in Rambouillet sheep.

BACKGROUND: The effect of phytase supplementation with respect to a high sorghum grain diet on sheep voluntary feed intake, apparent nutrient digestibility, ruminal fermentation, phosphorus (P) excretion and blood serum P concentration was evaluated. RESULTS: Phytase supplementation significantly decreased fecal P excretion (P = 0.003), resulting in a 26% decrease in relation to the phytase free diet. Dry matter intake, nutrient digestibility, ruminal butyrate and serum P were not (P > 0.10) affected by the phytase level. Neutral detergent digestibility showed a tendency to increase linearly (P = 0.10) with increasing phytase levels. Ruminal pH was lower for phytase supplemented sheep, with a significant decrease (P = 0.007) at 9 h post feeding, whereas ruminal ammonia-N at 3 h post feeding was lower (P = 0.004) for the phytase treatment groups, resulting in a decreasing linear response (P = 0.001) with an increasing phytase dose. Duodenal pH was significantly reduced at 6 h post feeding. Propionate tended (P = 0.051) to be increased linearly as the phytase supplementation level increased. CONCLUSION: Exogenous phytase supplementation of high sorghum grain diets significantly decreased fecal P excretion in Rambouillet rams. Phytase supplementation appears to affect neutral detergent fiber digestibility, duodenal and ruminal pH, ammonia and propionate. © 2018 Society of Chemical Industry.

Phosphorus acquisition by citrate- and phytase-exuding Nicotiana tabacum plant mixtures depends on soil phosphorus availability and root intermingling.

Citrate and phytase root exudates contribute to improved phosphorus (P) acquisition efficiency in Nicotiana tabacum (tobacco) when both exudates are produced in a P deficient soil. To test the importance of root intermingling in the interaction of citrate and phytase exudates, Nicotiana tabacum plant-lines with constitutive expression of heterologous citrate (Cit) or fungal phytase (Phy) exudation traits were grown under two root treatments (roots separated or intermingled) and in two soils with contrasting soil P availability. Complementarity of plant mixtures varying in citrate efflux rate and mobility of the expressed phytase in soil was determined based on plant biomass and P accumulation. Soil P composition was evaluated using solution 31 P NMR spectroscopy. In the soil with limited available P, positive complementarity occurred in Cit+Phy mixtures with roots intermingled. Root separation eliminated positive interactions in mixtures expressing the less mobile phytase (Aspergillus niger PhyA) whereas positive complementarity persisted in mixtures that expressed the more mobile phytase (Peniophora lycii PhyA). Soils from Cit+Phy mixtures contained less inorganic P and more organic P compared to monocultures. Exudate-specific strategies for the acquisition of soil P were most effective in P-limited soil and depended on citrate efflux rate and the relative mobility of the expressed phytase in soil. Plant growth and soil P utilization in plant systems with complementary exudation strategies are expected to be greatest where exudates persist in soil and are expressed synchronously in space and time.

Screening and Characterization of Phytases from Bacteria Isolated from Chilean Hydrothermal Environments.

Phytases are enzymes involved in organic phosphorus cycling in nature and widely used as feed additives in animal diets. Thermal tolerance is a desired property of phytases. The objectives of this study were to screen and characterize bacterial phytases from Chilean hydrothermal environments. In this study, 60% (30 of 63) of screened thermophilic (60 °C) isolates showed phytase activity in crude protein extracts. The characterization of phytase from two selected isolates (9B and 15C) revealed that both isolates produce phytases with a pH optimum at 5.0. The temperature optimum for phytate dephosphorylation was determined to be 60 and 50 °C for the phytases from the isolates 9B and 15C, respectively. Interestingly, the phytase from the isolate 15C showed a residual activity of 46% after incubation at 90 °C for 20 min. The stepwise dephosphorylation of phytate by protein extracts of the isolates 9B and 15C was verified by HLPC analysis. Finally, the isolates 9B and 15C were identified by partial sequencing of the 16S rRNA gene as members of the genera Bacillus and Geobacillus, respectively.

Opportunities for mobilizing recalcitrant phosphorus from agricultural soils: a review.

BACKGROUND: Phosphorus (P) fertilizer is usually applied in excess of plant requirement and accumulates in soils due to its strong adsorption, rapid precipitation and immobilisation into unavailable forms including organic moieties. As soils are complex and diverse chemical, biochemical and biological systems, strategies to access recalcitrant soil P are often inefficient, case specific and inconsistently applicable in different soils. Finding a near-universal or at least widely applicable solution to the inefficiency in agricultural P use by plants is an important unsolved problem that has been under investigation for more than half a century. SCOPE: In this paper we critically review the strategies proposed for the remobilization of recalcitrant soil phosphorus for crops and pastures worldwide. We have additionally performed a meta-analysis of available soil 31P-NMR data to establish the potential agronomic value of different stored P forms in agricultural soils. CONCLUSIONS: Soil inorganic P stocks accounted on average for 1006 ± 115 kg ha-1 (57 ± 7%), while the monoester P pool accounted for 587 ± 32 kg ha-1 (33 ± 2%), indicating the huge potential for the future agronomic use of the soil legacy P. New impact driven research is needed in order to create solutions for the sustainable management of soil P stocks.

Inter- and intra-species intercropping of barley cultivars and legume species, as affected by soil phosphorus availability.

AIMS: Intercropping can improve plant yields and soil phosphorus (P) use efficiency. This study compares inter- and intra-species intercropping, and determines whether P uptake and shoot biomass accumulation in intercrops are affected by soil P availability. METHODS: Four barley cultivars (Hordeum vulgare L.) and three legume species (Trifolium subterreneum, Ornithopus sativus and Medicago truncatula) were selected on the basis of their contrasting root exudation and morphological responses to P deficiency. Monocultures and barley-barley and barley-legume intercrops were grown for 6 weeks in a pot trial at very limiting, slightly limiting and excess available soil P. Above-ground biomass and shoot P were measured. RESULTS: Barley-legume intercrops had 10-70% greater P accumulation and 0-40% greater biomass than monocultures, with the greatest gains occurring at or below the sub-critical P requirement for barley. No benefit of barley-barley intercropping was observed. The plant combination had no significant effect on biomass and P uptake observed in intercropped treatments. CONCLUSIONS: Barley-legume intercropping shows promise for sustainable production systems, especially at low soil P. Gains in biomass and P uptake come from inter- rather than intra-species intercropping, indicating that plant diversity resulted in decreased competition between plants for P.

Response-based selection of barley cultivars and legume species for complementarity: Root morphology and exudation in relation to nutrient source.

Phosphorus (P) and nitrogen (N) use efficiency may be improved through increased biodiversity in agroecosystems. Phenotypic variation in plants' response to nutrient deficiency may influence positive complementarity in intercropping systems. A multicomponent screening approach was used to assess the influence of P supply and N source on the phenotypic plasticity of nutrient foraging traits in barley (H. vulgare L.) and legume species. Root morphology and exudation were determined in six plant nutrient treatments. A clear divergence in the response of barley and legumes to the nutrient treatments was observed. Root morphology varied most among legumes, whereas exudate citrate and phytase activity were most variable in barley. Changes in root morphology were minimized in plants provided with ammonium in comparison to nitrate but increased under P deficiency. Exudate phytase activity and pH varied with legume species, whereas citrate efflux, specific root length, and root diameter lengths were more variable among barley cultivars. Three legume species and four barley cultivars were identified as the most responsive to P deficiency and the most contrasting of the cultivars and species tested. Phenotypic response to nutrient availability may be a promising approach for the selection of plant combinations for minimal input cropping systems.

Organic Acids Regulation of Chemical-Microbial Phosphorus Transformations in Soils.

We have used an integrated approach to study the mobility of inorganic phosphorus (P) from soil solid phase as well as the microbial biomass P and respiration at increasing doses of citric and oxalic acid in two different soils with contrasting agronomic P status. Citric or oxalic acids significantly increased soil solution P concentrations for doses over 2 mmol kg-1. However, low organic acid doses (<2 mmol kg-1) were associated with a steep increase in microbial biomass P, which was not seen for higher doses. In both soils, treatment with the tribasic citric acid led to a greater increase in soil solution P than the dibasic oxalic acid, likely due to the rapid degrading of oxalic acids in soils. After equilibration of soils with citric or oxalic acids, the adsorbed-to-solution distribution coefficient (Kd) and desorption rate constants (k-1) decreased whereas an increase in the response time of solution P equilibration (Tc) was observed. The extent of this effect was shown to be both soil and organic acid specific. Our results illustrate the critical thresholds of organic acid concentration necessary to mobilize sorbed and precipitated P, bringing new insight on how the exudation of organic acids regulate chemical-microbial soil phosphorus transformations.

A Holistic Approach to Understanding the Desorption of Phosphorus in Soils.

The mobility and resupply of inorganic phosphorus (P) from the solid phase were studied in 32 soils from the UK. The combined use of diffusive gradients in thin films (DGT), diffusive equilibration in thin films (DET) and the "DGT-induced fluxes in sediments" model (DIFS) were adapted to explore the basic principles of solid-to-solution P desorption kinetics in previously unattainable detail. On average across soil types, the response time (Tc) was 3.6 h, the desorption rate constant (k-1) was 0.0046 h(-1), and the desorption rate was 4.71 nmol l(-1) s(-1). While the relative DGT-induced inorganic P flux responses in the first hour is mainly a function of soil water retention and % Corg, at longer times it is a function of the P resupply from the soil solid phase. Desorption rates and resupply from solid phase were fundamentally influenced by P status as reflected by their high correlation with P concentration in FeO strips, Olsen, NaOH-EDTA and water extracts. Soil pH and particle size distribution showed no significant correlation with the evaluated mobility and resupply parameters. The DGT and DET techniques, along with the DIFS model, were considered accurate and practical tools for studying parameters related to soil P desorption kinetics.

Performance of Seven Commercial Phytases in an in Vitro Simulation of Poultry Digestive Tract.

The aim of this study was to compare the biochemical properties of seven commercially available phytase products as well as their catalytic performance in an in vitro simulation of the digestive tract of poultry. Their enzymatic properties relevant with respect to phytate dephosphorylation in the digestive tract of birds were determined under identical assay conditions. All phytase products included in the study showed an acid pH optimum of activity and were capable of releasing the organically bound phosphate from phytate during the in vitro simulation. However, their overall biochemical properties and relative catalytic performances were remarkably different. The in vitro degradation system was considered as a simple and useful tool to evaluate the suitability of a phytase to be used as a feed supplement. Although relevant factors such as dietary P levels, intestinal phytase, and P absorption are not implemented in the system, this approach might help to reduce the number of feeding trials necessary in the search for a better suited phytase for animal feeding application.

Activity stabilization of Aspergillus niger and Escherichia coli phytases immobilized on allophanic synthetic compounds and montmorillonite nanoclays.

The aim of this work was to study the stabilization of the activity of two commercial microbial phytases (Aspergillus niger and Escherichia coli) after immobilization on nanoclays and to establish optimal conditions for their immobilization. Synthetic allophane, synthetic iron-coated allophanes and natural montmorillonite were chosen as solid supports for phytase immobilization. Phytase immobilization patterns at different pH values were strongly dependent on both enzyme and support characteristics. After immobilization, the residual activity of both phytases was higher under acidic conditions. Immobilization of phytases increased their thermal stability and improved resistance to proteolysis, particularly on iron-coated allophane (6% iron oxide), which showed activation energy (E(a)) and activation enthalpy (ΔH(#)) similar to free enzymes. Montmorillonite as well as allophanic synthetic compounds resulted in a good support for immobilization of E. coli phytase, but caused a severe reduction of A. niger phytase activity.

Identification of ß-propeller phytase-encoding genes in culturable Paenibacillus and Bacillus spp. from the rhizosphere of pasture plants on volcanic soils.

Phytate is one of the most abundant sources of organic phosphorus (P) in soils, but must be mineralized by phytase-producing bacteria to release P for plant uptake. Microbial inoculants based on Bacillus spp. have been developed commercially, but few studies have evaluated the ecology of these bacteria in the rhizosphere or the types of enzymes that they produce. Here, we studied the diversity of aerobic endospore-forming bacteria (EFB) with the ability to mineralize phytate in the rhizosphere of pasture plants grown in volcanic soils of southern Chile. PCR methods were used to detect candidate phytase-encoding genes and to identify EFB bacteria that carry these genes. This study revealed that the phytate-degrading EFB populations of pasture plants included species of Paenibacillus and Bacillus, which carried genes encoding ß-propeller phytase (BPP). Assays of enzymatic activity confirmed the ability of these rhizosphere isolates to degrade phytate. The phytase-encoding genes described here may prove valuable as molecular markers to evaluate the role of EFB in organic P mobilization in the rhizosphere.