Unveiling fungal diversity in uranium and glycerol-2-phosphate-amended bentonite microcosms: Implications for radionuclide immobilization within the Deep Geological Repository system

Uranium (U) represents the preeminent hazardous radionuclide within the context of nuclear waste repositories. Indigenous microorganisms in bentonite can influence radionuclide speciation and migration in Deep Geological Repositories (DGRs) for nuclear waste storage. While bacterial communities in bentonite samples have been extensively studied, the impact of fungi has been somewhat overlooked. Here, we investigate the geomicrobiological processes in bentonite microcosms amended with uranyl nitrate and glycerol-2-phosphate (G2P) for six-month incubation. ITS sequencing revealed that the fungal community was mainly composed of Ascomycota (96.6 %). The presence of U in microcosms enriched specific fungal taxa, such as Penicillium and Fusarium, potentially associated with uranium immobilization mechanisms. Conversely, the amendment of U into G2P-suplemented samples exhibited minimal impact, resulting in a fungal community akin to the control group. Several fungal strains were isolated from bentonite microcosms to explore their potential in the U biomineralization, including Fusarium oxysporum, Aspergillus sp., Penicillium spp., among others. High Annular Angle Dark-Field Scanning Transmission Electron Microscopy (HAADF) analyses showed the capacity of F. oxysporum B1 to form U-phosphate mineral phases, likely mediated by phosphatase activity. Therefore, our study emphasizes the need to take into account indigenous bentonite fungi in the overall assessment of the impact of microbial processes in the immobilization of U within DGRs environments.