Status report of the first AMS laboratory in the Czech Republic at the Nuclear Physics Institute, Rez.

The first accelerator mass spectrometry (AMS) laboratory in the Czech Republic has been established and put into routine operation in February 2022. Here we briefly describe the facilities available, namely a 300 kV multi-isotope low-energy AMS system (MILEA) capable of determination 10Be, 14C, 26Al, 41Ca, 129I, isotopes of U, especially 236U, Pu and other actinoids, and accessories for 14C measurements, which include a gas interface system, a preparative gas chromatography system for compound-specific radiocarbon dating analysis, and an isotope-ratio mass spectrometer. The first results achieved for separation and measurement of the above radionuclides (except for 41Ca) are also reported, with the main focus on 14C measurements. A specimen breakdown of 729 graphitised samples analysed for 14C so far is presented, as well as a proof of measurement stability of the MILEA system obtained by analysis of radiocarbon standards and analytical blanks. For the other radionuclides, well proven or novel procedures for sample preparation and measurement are presented.

Is arsenic responsible for the toxicity of the hyperaccumulating mushroom Sarcosphaera coronaria?

The Violet Crown Cup, Sarcosphaera coronaria, is a rather inconspicuous mushroom, but with an interesting and unresolved mystery. In earlier days, the mushroom was considered edible, but several poisonings were reported in the early 20th century. The reason for the seemingly sporadic toxicity of S. coronaria is still unknown. One possible explanation is arsenic, since Crown Cups can take up high amounts of this element. We investigated the arsenic concentration and arsenic speciation in S. coronaria with inductively coupled plasma mass spectrometry (ICPMS) and HPLC coupled to ICPMS and found up to incredible 0.9% As (dry mass). Most of it was present as methylarsonic acid (MA), a less toxic form of this element. However, low concentrations of the highly toxic methylarsonous acid [MA (III)] were also detected. The amounts were too low to pose an acute risk for consumers, but the concentration of MA (III) significantly increased during simulated gastric digestion. We could not unambiguously identify arsenic as the toxic constituent of S. coronaria, but we demonstrated that the extremely toxic MA (III) can be formed under certain circumstances, which should be carefully investigated in future.

Zn overaccumulating Russula species clade together and use the same mechanism for the detoxification of excess Zn.

It has been firmly established that macrofungi can accumulate large amounts of heavy metals in their sporocarps. However, the mechanisms of the accumulation and storage are being uncovered only recently. We have previously documented that Russula bresadolae can accumulate over 1 g Zn kg-1 dry weight and that sequestration of a substantial proportion of overaccumulated Zn involves binding with peptides, RaZBPs, seen so far only in this species. In this work we examined Zn contents of 360 sporocarp collections from unpolluted environments covering 114 species of the genus Russula. Whilst the concentrations of Zn in most analysed species were in the range of 50-150 mg kg-1, the species of subgenera Brevipes and Compactae accumulate very low Zn (< 50 mg kg-1). We further identified five new Zn-overaccumulating species of subgenus Russula, which form with R. bresadolae a separate phylogenetic subclade in which the sporocarp Zn concentrations ranged from 326 to 845 mg kg-1. We demonstrate that R. pumila and R. ochroleuca express at least one ZBP gene and when expressed in metal-sensitive S. cerevisiae, all ZBPs protected the yeasts against Zn (and Cd) toxicity equally well. The respective ZBPs were confirmed in the native Zn-complexes of R. pumila and R. ochroleuca, which represented 80% of Zn extracted from their sporocarps. This study is the first extensive genus-wide report of metal accumulation in macrofungi, which further demonstrates that the Zn binding with cytosolic ZBP peptides is not a trait restricted only to R. bresadolae.

Speciation analysis of elements accumulated in Cystoderma carcharias from clean and smelter-polluted sites.

Collections of Cystoderma carcharias sporocarps were sampled from clean and smelter-polluted sites and analyzed for Ag, As, Cd, Cu, Pb, Se, and Zn contents. Concentrations of all elements were significantly higher in samples from the smelter-polluted area. Except for As and Pb, all elements were effectively accumulated in the sporocarps at both clean and polluted sites. With the highest concentration of 604 mg Cd kg-1, C. carcharias can be considered as Cd hyperaccumulator. As revealed by HPLC-ICPQQQMS analysis, the As species in sporocarps from clean and polluted areas involved besides the major arsenobetaine a variety of known and unknown arsenicals; the occurrence of dimethylarsinoylacetate and trimethylarsoniopropionate is reported for the first time for gilled fungi (Agaricales). Size-exclusion chromatography of C. carcharias extracts supported by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and RP-HPLC data indicated that detoxification of intracellular Cd and Cu may largely rely on metallothioneins (MT) or MT-like peptides, not phytochelatins.

Disentangling the factors of contrasting silver and copper accumulation in sporocarps of the ectomycorrhizal fungus Amanita strobiliformis from two sites.

Amanita strobiliformis (European Pine Cone Lepidella) is an ectomycorrhizal fungus of the Amanitaceae family known to hyperaccumulate Ag in the sporocarps. Two populations (ecotypes) of A. strobiliformis collected from two urban forest plantations in Prague, Czech Republic, were investigated. The concentrations of Ag, Cu, Cd, and Zn were determined in the mushrooms. The metal mobility and fractionation in the soils was investigated by single extractions and sequential extraction. The soil distribution of A. strobiliformis mycelium was assessed by quantitative polymerase chain reaction (qPCR). The metal uptake from the soil into the mushroom sporocarps was traced by Pb isotopic fingerprinting. The findings suggested that A. strobiliformis (i) accumulates primarily Ag from the topsoil layer (circa 12cm deep) and (ii) accumulates Ag associated with the "reducible soil fraction". The concentrations of all metals, particularly Ag and Cu, were significantly higher in the A. strobiliformis sporocarps from one of the investigated sites (Klícov). The elevated concentrations of Ag in the sporocarps from Klícov can possibly be attributed to the higher Ag content in the topsoil layer found at this site. However, the simultaneously elevated concentrations of Cu in A. strobiliformis from Klícov cannot be explained by the differences in the geochemical background and should be attributed to biological factors.

Arsenic hyperaccumulation and speciation in the edible ink stain bolete (Cyanoboletus pulverulentus).

The edible ink stain bolete (Cyanoboletus pulverulentus) was found to hyperaccumulate arsenic. We analyzed 39 individual collections determined as C. pulverulentus, mostly from the Czech Republic. According to our results, concentrations of arsenic in C. pulverulentus fruit-bodies may reach 1300mgkg-1 dry weight. In most collections, data for total and bioavailable arsenic in underlying soils were collected but no significant correlation between the soil arsenic content and arsenic concentrations in the associated fruit-bodies was found. Within the fruit-bodies, we found the majority of arsenic accumulated in the hymenium. Besides occasional traces of methylarsonic acid (MA), the arsenic speciation in all mushroom samples consisted solely of dimethylarsinic acid (DMA) and no inorganic arsenic was detected. Because of the carcinogenic potential of DMA, C. pulverulentus should not be recommended as an edible mushroom and its consumption should be restricted.

Mercury species in formerly contaminated soils and released soil gases.

Total mercury (T-Hg), elemental mercury (Hg0), methylmercury (MeHg+), phenylmercury (PhHg+), and gaseous elemental mercury (GEM) species were determined in soils formerly contaminated by different processes from two sites in the Czech Republic. Analytical methods involved atomic absorption spectrometry (AAS) using a single-purpose Advanced Mercury Analyser AMA-254 and radiochemical neutron activation analysis (RNAA) for T-Hg determination, a thermal desorption method was used for Hg0 determination, gas chromatography coupled with atomic fluorescence spectrometry (GC-AFS) was employed for assay of MeHg+ and PhHg+, while GEM measurement was carried out using a portable Zeeman-AAS device Lumex RA-915+. The first sampling site was in the surroundings of a former PhHgCl-based fungicide processing plant next to Príbram (central Bohemia). Although the use of Hg-based fungicides as seed mordant have been banned, and their production stopped at the end of 1980's, highly elevated Hg contents in soil are still observed in the vicinity of the former plant, reaching T-Hg values >13mgkg-1. The second sampling site was an abandoned mining area named Jedová hora Hill near Horovice (central Bohemia), where cinnabar (HgS) was occasionally mined as by-product of Fe ores hematite and siderite. Mining activities have been stopped here in 1857. Very high contents of T-Hg are still found at this site, up to 144mgkg-1. In most cases we found a statistically significant correlation between T-Hg and Hg0 values regardless of the pollution source. On the contrary, insignificant correlation was observed neither between T-Hg and GEM values, nor between GEM and Hg0. Concentrations of the investigated organomercury species were above a limit of detection (LOD) only in the most contaminated samples, where their levels were about two to three orders of magnitude lower compared to those of T-Hg.

Resurrection of Cortinarius coalescens: taxonomy, chemistry, and ecology.

Cortinarius coalescens Kärcher & Seibt is a rare European species of the subgenus Phlegmacium, section Phlegmacioides, neglected in recent molecular studies. New primers (CortF and CortR) designed for species in the section Phlegmacioides allowed to obtain ITS rDNA sequence data from the holotype collection of C. coalescens; according to the results, this epithet has priority over C. crassorum Rob. Henry ex Rob. Henry, C. pardinus Reumaux, and C. parargutus Bidaud, Moënne-Locc. & Reumaux. Morphological and ecological observations on recent collections of C. coalescens from the Czech Republic in comparison with the co-occurring C. largus are discussed. Nomenclatural and taxonomic comments on C. tomentosus Rob. Henry, C. balteatotomentosus Rob. Henry, and C. subtomentosus Reumaux are also provided. So far, C. coalescens is known with certainty from Germany, France, and the Czech Republic, where it grows in deciduous forests on acid to neutral soils. Arsenic and its compounds were determined in C. coalescens and related species of the section Phlegmacioides: C. largus, C. pseudodaulnoyae, and C. variecolor. Total arsenic concentrations were in the range 3.6-30.2 mg kg-1 (dry matter) and arsenobetaine was the major arsenic compound.