Online Nano Solid Phase Extraction Fourier-Transform Ion Cyclotron Resonance Mass Spectrometry Workflow to Analyze Small Scale Gradients of Soil Solution Organic Matter in the Rhizosphere.

A new method combining online nano solid phase extraction coupled with Fourier-transform ion cyclotron resonance mass spectrometry (FT-ICR-MS) was developed to extract and analyze organic matter (OM) from microliter volumes of salt containing soil solution samples. This approach allows the reproducible analysis of only minute amounts of organic carbon (down to 10 ng C) without the need of further sample preparation. The new method was applied to unravel developing small-scale patterns of dissolved organic matter (DOM) in soil solutions of a soil column experiment in which Zea mays plants were grown for 3 weeks. Soil solution was sampled by micro suction cups from the undisturbed soil-root system once a week. Growth of the root system and, hence, position of individual roots relative to the suction cups was followed by X-ray computed tomography (X-ray CT). Our method makes it possible to resolve the chemical complexity of soil solution OM (up to 4300 molecular formulas from 2.5 µL sample). This allows to observe chemical gradients in the rhizosphere on a molecular level over time. The increasing influence of roots on soil solution OM is visible from higher molecular masses, an increasing degree of oxygenation and a higher fraction of formulas containing heteroatoms. The online nano solid phase extraction-FT-ICR-MS method provides novel insight into the processes affecting DOM in the rhizosphere, such as root exudation, microbial processes, and soil organic matter stabilization.

Experimental considerations in metal mobilization from soil by chelating ligands: The influence of soil-solution ratio and pre-equilibration - A case study on Fe acquisition by phytosiderophores.

The efficiency of chelating ligands in mobilizing metals from soils and sediments is generally examined under conditions remote from those under which they are exuded or applied in the field. This may lead to incorrect estimations of the mobilizing efficiency. The aim of this study was to establish the influence of the soil solution ratio (SSR) and pre-equilibration with electrolyte solution on metal mobilization and metal displacement. For this purpose a series of interaction experiments with a calcareous clay soil and a biogenic chelating agent, the phytosiderophore 2'-deoxymugineic acid (DMA) were carried out. For a fixed ligand concentration, the SSR had a strong influence on metal mobilization and displacement. Metal complexation was faster at higher SSR. Reactive pools of metals that were predominantly mobilized at SSR 6 (in this case Cu), became depleted at SSR 0.1, whereas metals that were marginally mobilized at SSR 6, were dominantly mobilized at SSR 0.1 (in this case Fe), because of large soil reactive pools. For a fixed "amount of ligand"-to-"amount of soil"-ratio, metal complexation scaled linearly with the SSR. The efficiency of ligands in mobilizing metals under field conditions can be predicted with batch experiments, as long as the ligand-to-soil-ratio is matched. In most previously reported studies this criterion was not met. Equivalent metal-complex concentrations under field conditions can be back-calculated using adsorption isotherms for the respective metal-complexes. Drying and dry storage created labile pools of Fe, Cu and Zn, which were rapidly mobilized upon addition of DMA solution to dry soil. Pre-equilibration decreased these labile pools, leading to smaller concentrations of these metals during initial mobilization, but did not reduce the lag time between ligand addition and onset of microbial degradation of the metal-complexes. Hence SSR and pre-equilibration should be carefully considered when testing the metal mobilizing efficiency of chelating ligands.

Retention of phytosiderophores by the soil solid phase - adsorption and desorption.

BACKGROUND AND AIMS: Graminaceous plants exude phytosiderophores (PS) for acquiring Fe. Adsorption of PS and its metal complexes to the soil solid phase reduces the FePS solution concentration and hence Fe uptake. In this study we aimed to quantify adsorption, and to determine to what extent adsorption depends on the complexed metal and on soil properties. Furthermore, we examined if adsorption is a reversible process. METHODS: Adsorption and desorption of PS and metal-PS complexes were examined in batch experiments in which the PS 2'-deoxymugineic acid (DMA) and its metal-complexes (FeDMA, CuDMA, NiDMA and ZnDMA) interacted with several calcareous soils. RESULTS: Adsorption of DMA ligand (0-1000 µM) and metal-DMA complexes (0-100 µM) was linear in the concentration range examined. Adsorption varied by a factor ≈2 depending on the complexed metal and by up to a factor 3.5 depending on the soil. Under field-like conditions (50 % water holding capacity), 50-84 % of the DMA was predicted to be retained to the soil solid phase. Alike adsorption, desorption of metal-DMA complexes is fast (approximate equilibrium within 1 hour). However, only a small fraction of the adsorbed FeDMA (28-35 %) could be desorbed. CONCLUSIONS: Despite this small fraction, the desorbed FeDMA still exceeded the amount in solution, indicating that desorption of FeDMA from soil reactive compounds can be an important process buffering the solution concentration.

Geochemical processes constraining iron uptake in Strategy II Fe acquisition.

Phytosiderophores (PS) are natural chelating agents, exuded by graminaceous plants (grasses) for the purpose of Fe acquisition (Strategy II). They can form soluble Fe complexes with soil-Fe that can be readily taken up. PS are exuded in a diurnal pulse release, and with the start of PS release a "window of iron uptake" opens. In the present study we examined how this window is constrained in time and concentration by biogeochemical processes. For this purpose, a series of interaction experiments was done with a calcareous clay soil and the phytosiderophore 2'-deoxymugineic acid (DMA), in which metal and DMA speciation were examined as a function of time and DMA concentration. Various kinetically and thermodynamically controlled processes affected the size of the window of Fe uptake. Adsorption lowered, but did not prevent Fe mobilization by DMA. Microbial activity depleted DMA from solution, but not on time scales jeopardizing Strategy II Fe acquisition. Complexation of competing metals played an important role in constraining the window of Fe uptake, particularly at environmentally relevant PS concentrations. Our study provides a conceptual model that takes into account the chemical kinetics involved with PS-mediated Fe acquisition. The model can help to explain how success or failure of PS-mediated Fe acquisition depends on environmental conditions.

Metal mobilization from soils by phytosiderophores - experiment and equilibrium modeling.

AIMS: To test if multi-surface models can provide a soil-specific prediction of metal mobilization by phytosiderophores (PS) based on the characteristics of individual soils. METHODS: Mechanistic multi-surface chemical equilibrium modeling was applied for obtaining soil-specific predictions of metal and PS speciation upon interaction of the PS 2'-deoxymugineic acid (DMA) with 6 soils differing in availability of Fe and other metals. Results from multi-surface modeling were compared with empirical data from soil interaction experiments. RESULTS: For soils in which equilibrium was reached during the interaction experiment, multi-surface models could well predict PS equilibrium speciation. However, in uncontaminated calcareous soils, equilibrium was not reached within a week, and experimental and modeled DMA speciation differed considerably. In soils with circum-neutral pH, on which Fe deficiency is likely to occur, no substantial Fe mobilization by DMA was predicted. However, in all but the contaminated soils, Fe mobilization by DMA was observed experimentally. Cu and Ni were the quantitatively most important metals competing with Fe for complexation and mobilization by DMA. CONCLUSION: Thermodynamics are unable to explain the role of PS as Fe carrier in calcareous soils, and the kinetic aspects of metal mobilization by PS need to be closer examined in order to understand the mechanisms underlying strategy II Fe acquisition.