Speciation and spatial distribution of Eu(III) in fungal mycelium.

Europium, as an easy-to-study analog of the trivalent actinides, is of particular importance for studying the behavior of lanthanides and actinides in the environment. Since different soil organisms can influence the migration behavior of these elements, a detailed knowledge of these interaction mechanisms is important. The aim of this study was to investigate the interaction of mycelia of selected wood-inhabiting (S. commune, P. ostreatus, L. tigrinus) and soil-inhabiting fungi (L. naucinus) with Eu(III). In addition to determining the Eu(III) complexes in the sorption solution, the formed Eu(III) fungal species were characterized using scanning transmission electron microscopy coupled with energy-dispersive X-ray spectroscopy, chemical microscopy in combination with the time-resolved laser-induced fluorescence spectroscopy. Our data show that S. commune exhibited significantly higher Eu(III) binding capacity in comparison to the other fungi. Depending on fungal strain, the metal was immobilized on the cell surface, in the cell membranes, and within the membranes of various organelles, or in the cytoplasm in some cases. During the bioassociation process two different Eu(III) fungal species were formed in all investigated fungal strain. The phosphate groups of organic ligands were identified as being important functional groups to bind Eu(III) and thus immobilize the metal in the fungal matrix. The information obtained contributes to a better understanding of the role of fungi in migration, removal or retention mechanisms of rare earth elements and trivalent actinides in the environment.

Uranium(VI) bioassociation by different fungi - a comparative study into molecular processes.

After the Chernobyl and Fukushima incidents it has become clear that fungi can take up and accumulate large quantities of radionuclides and heavy metals, but the underlying processes are not well understood yet. For this study, the molecular interactions of uranium(VI) with the white-rot fungi, Schizophyllum commune and Pleurotus ostreatus, and the soil-living fungus, Leucoagaricus naucinus, were investigated. First, the uranium concentration in the biomass was determined by time-dependent bioassociation experiments. To characterize the molecular interactions, uranium was localized in the biomass by transmission electron microscopy analysis. Second, the formed uranyl complexes in both biomass and supernatant were determined by fluorescence spectroscopy. Additionally, possible bioligands in the supernatant were identified. The results show that the discernible interactions between metals and fungi are similar, namely biosorption, accumulation, and subsequent crystallization. But at the same time, the underlying biochemical mechanisms are different and specific to the fungal species. In addition, Schizophyllum commune was found to be the only fungus that, under the chosen experimental conditions, released tryptophan and other indole derivatives in the presence of uranium(VI) as determined by nuclear magnetic resonance spectroscopy. These released substances most likely act as messenger molecules rather than serving the direct detoxification of uranium(VI).

Survival of the basidiomycete Schizophyllum commune in soil under hostile environmental conditions in the Chernobyl Exclusion Zone.

Radioactive contamination resulting from major nuclear accidents presents harsh environmental conditions. Inside the Chernobyl exclusion zone, even more than 30 years after the accident, the resulting contamination levels still does not allow land-use or human dwellings. To study the potential of basidiomycete fungi to survive the conditions, a field trial was set up 5 km south-south-west of the destroyed reactor unit. A model basidiomycete, the lignicolous fungus Schizophyllum commune, was inoculated and survival in the soil could be verified. Indeed, one year after inoculation, the fungus was still observed using DNA-dependent techniques. Growth led to spread at a high rate, with approximately 8 mm per day. This shows that also white-rot basidiomycetes can survive the harsh conditions in soil inside the Chernobyl exclusion zone. The unadapted fungal strain showed the ability to grow and thrive in the contaminated soil where both stress from radiation and heavy metals were present.

Effect of U(VI) aqueous speciation on the binding of uranium by the cell surface of Rhodotorula mucilaginosa, a natural yeast isolate from bentonites.

This study presents the effect of aqueous uranium speciation (U-hydroxides and U-hydroxo-carbonates) on the interaction of this radionuclide with the cells of the yeast Rhodotorula mucigilanosa BII-R8. This strain was isolated from Spanish bentonites considered as reference materials for the engineered barrier components of the future deep geological repository of radioactive waste. X-ray absorption and infrared spectroscopy showed that the aqueous uranium speciation has no effect on the uranium binding process by this yeast strain. The cells bind mobile uranium species (U-hydroxides and U-hydroxo-carbonates) from solution via a time-dependent process initiated by the adsorption of uranium species to carboxyl groups. This leads to the subsequent involvement of organic phosphate groups forming uranium complexes with a local coordination similar to that of the uranyl mineral phase meta-autunite. Scanning transmission electron microscopy with high angle annular dark field analysis showed uranium accumulations at the cell surface associated with phosphorus containing ligands. Moreover, the effect of uranium mobile species on the cell viability and metabolic activity was examined by means of flow cytometry techniques, revealing that the cell metabolism is more affected by higher concentrations of uranium than the cell viability. The results obtained in this work provide new insights on the interaction of uranium with bentonite natural yeast from genus Rhodotorula under deep geological repository relevant conditions.

Interaction of U(VI) with Schizophyllum commune studied by microscopic and spectroscopic methods.

Biosorption of actinides like uranium by fungal cells can play an important role in the mobilization or immobilization of these elements in nature. Sorption experiments of U(VI) with Schizophyllum commune at different initial uranium concentrations and varying metal speciation showed high uranium sorption capacities in the pH range of 4­7. A combination of high angle annular dark-field and scanning transmission electron microscopy analysis (HAADF-STEM) showed that living mycelium cells accumulate uranium at the cell wall and intracellular. For the first time the fluorescence properties of uranium accumulates were investigated by means of time-resolved laser-induced fluorescence spectroscopy (TRLFS) beside the determination of corresponding structural parameters using X-ray absorption fine structure spectroscopy (EXAFS). While the oxidation state of uranium remained unchanged during sorption, uranium speciation changed significantly. Extra and intracellular phosphate groups are mainly responsible for uranium binding. TRLFS spectra clearly show differences between the emission properties of dissolved species in the initial mineral medium and of uranium species on fungi. The latter were proved to be organic and inorganic uranyl phosphates formed depending on the uranyl initial concentration and in some cases on pH.

Chitin-based renewable materials from marine sponges for uranium adsorption.

Marine sponges of the order Verongida form three-dimensional networks of fibrous chitin, which can easily be extracted. In the hydrated state, these networks are flexible, mechanically stable and can be cut or pressed into any desired form. Here, we show for the first time that chitin-based networks of sponge origin are useful for effective uranium adsorption. They adsorb uranium from solution with a higher adsorption capacity than many other chitinous sorbents. Up to 288 mg/g could be achieved. Solid-state NMR, infrared, and Raman spectroscopy indicated that the uranyl is bound to the chitin by weak interactions. 90% of the uranyl could be desorbed using diluted hydrochloric acid. Uranium adsorption and desorption did not result in any destruction of the chitin-based material.

Spectroscopic investigations of U(VI) species sorbed by the green algae Chlorella vulgaris.

The green alga Chlorella vulgaris has the ability to bind high amounts of uranium(VI) in the pH range from 3 to 6. At pH 3 up to 40% of the uranium are bound by the algal cells. The uranium removal is almost complete at pH 5 and 6 under the given experimental conditions. Scanning electron microscopy and laser-induced fluorescence spectroscopy were used to characterize uranyl species formed in the selected pH range. The micrographs show a regular distribution of U(VI) on the cell surface. Fluorescence spectroscopic investigations of formed algal uranyl complexes indicate that the binding of U(VI) to carboxyl groups plays a dominating role at pH 3, whereas a minor impact of organic phosphate compounds on the U(VI) sorption cannot be excluded. In contrast, at pH 5 and 6 the phosphate groups are mainly responsible for the removal and binding of U(VI) by formation of organic and/or inorganic uranyl phosphates.