Agaricus crocodilinus (Agaricaceae), an edible saprotrophic mushroom, accumulates high concentrations of cadmium (Cd) in unpolluted environments. This study investigates whether this species has evolved mechanisms to store Cd complexed with metallothioneins (MTs), proteins that bind heavy metal ions via cysteinyl (Cys)-thiolate bonds, how these MTs originated, and how similar mechanisms are present in other fungal species. Size exclusion chromatography revealed that a substantial fraction of Cd in A. crocodilinus sporocarps was sequestered in a 3.4 kDa complex containing Cys-rich peptides. Screening a sporocarp cDNA expression library in a Cd-sensitive Saccharomyces cerevisiae strain identified two MT transcripts, AcMT1 and AcMT2, encoding 49-amino acid (AA) AcMT1 with 10 Cys and 32-AA AcMT2 with 7 Cys. The presence of AcMT2 in the 3.4 kDa Cd-peptide complex isolated from sporocarp was confirmed by mass spectrometry. In mycelial isolates exposed to heavy metals, AcMT1 was more strongly upregulated, while AcMT2 was more expressed under normal conditions. Sequence comparisons revealed that AcMT2 is closer to the ancestral gene, whereas AcMT1 is a more recent duplicate. Combined bioinformatic and functional evidence supports AcMT2 as a constitutively expressed MT involved in Cd binding in the sporocarp, while AcMT1, though more inducible in mycelia and more protective in yeast, appears to serve a transient detoxification role. Moreover, the gene duplication and domain rearrangement mechanism underlying this MT diversification was also identified in other Agaricales and Boletales species.

• Klíčová slova
• Agaricaceae, Gene duplication, Gene internal duplication, Heavy metals,
• Publikační typ
• časopisecké články MeSH

With the advent rise is in urbanization and industrialization, heavy metals (HMs) such as lead (Pb) and cadmium (Cd) contamination have increased considerably. It is among the most recalcitrant pollutants majorly affecting the biotic and abiotic components of the ecosystem like human well-being, animals, soil health, crop productivity, and diversity of prokaryotes (bacteria) and eukaryotes (plants, fungi, and algae). At higher concentrations, these metals are toxic for their growth and pose a significant environmental threat, necessitating innovative and sustainable remediation strategies. Bacteria exhibit diverse mechanisms to cope with HM exposure, including biosorption, chelation, and efflux mechanism, while fungi contribute through mycorrhizal associations and hyphal networks. Algae, especially microalgae, demonstrate effective biosorption and bioaccumulation capacities. Plants, as phytoremediators, hyperaccumulate metals, providing a nature-based approach for soil reclamation. Integration of these biological agents in combination presents opportunities for enhanced remediation efficiency. This comprehensive review aims to provide insights into joint action of prokaryotic and eukaryotic interactions in the management of HM stress in the environment.

• Klíčová slova
• Cadmium (Cd), Eukaryotes, Heavy metal stress, Lead (Pb), Prokaryotes, Tolerance mechanism,
• MeSH
• Bacteria * metabolismus   účinky léků   genetika   MeSH
• biodegradace MeSH
• Eukaryota * metabolismus   účinky léků   MeSH
• houby metabolismus   účinky léků   MeSH
• kadmium * metabolismus   toxicita   MeSH
• látky znečišťující půdu metabolismus   MeSH
• olovo * metabolismus   toxicita   MeSH
• rostliny metabolismus   MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH
• Názvy látek
• kadmium * MeSH
• látky znečišťující půdu MeSH
• olovo * MeSH

As we have shown previously, the Cu and Ag concentrations in the sporocarps of Ag-hyperaccumulating Amanita strobiliformis are correlated, and both metals share the same uptake system and are sequestered by the same metallothioneins intracellularly. To further improve our knowledge of the Cu and Ag handling in A. strobiliformis cells, we searched its transcriptome for the P1B-1-ATPases, recognizing Cu+ and Ag+ for transport. We identified transcripts encoding 1097-amino acid (AA) AsCRD1 and 978-AA AsCCC2, which were further subjected to functional studies in metal sensitive Saccharomyces cerevisiae. The expression of AsCRD1 conferred highly increased Cu and Ag tolerance to metal sensitive yeasts in which the functional AsCRD1:GFP (green fluorescent protein) fusion localized exclusively to the tonoplast, indicating that the AsCRD1-mediated Cu and Ag tolerance was a result of vacuolar sequestration of the metals. Increased accumulation of AsCRD1 transcripts observed in A. strobiliformis mycelium upon the treatments with Cu and Ag (8.7- and 4.5-fold in the presence of 5 μM metal, respectively) supported the notion that AsCRD1 can be involved in protection of the A. strobiliformis cells against the toxicity of both metals. Neither Cu nor Ag affected the levels of AsCCC2 transcripts. Heterologous expression of AsCCC2 in mutant yeasts did not contribute to Cu tolerance, but complemented the mutant genotype of the S. cerevisiae ccc2Δ strain. Consistent with the role of the yeast Ccc2 in the trafficking of Cu from cytoplasm to nascent proteins via post-Golgi, the GFP fluorescence in AsCCC2-expressing ccc2Δ yeasts localized among Golgi-like punctate foci within the cells. The AsCRD1- and AsCCC2-associated phenotypes were lost in yeasts expressing mutant transporter variants in which a conserved phosphorylation/dephosphorylation site was altered. Altogether, the data support the roles of AsCRD1 and AsCCC2 as genuine P1B-1-ATPases, and indicate their important functions in the removal of toxic excess of Cu and Ag from the cytoplasm and charging the endomembrane system with Cu, respectively.

• Klíčová slova
• Amanita strobiliformis, P1-type ATPase, copper transporter, ectomycorrhizal fungi, metal homeostasis, silver transporter,
• Publikační typ
• časopisecké články MeSH