Appraisal of Antioxidant Potential and Natural Bioactive Compound Determination through HPLC from Two Saprotrophic Amanita (Agaricomycetes) Species from Pakistan.

The present study was carried out to investigate and identify bioactive compounds along with antioxidant capacity, total flavonoids and total phenolic contents from two saprophytic Amanita species, i.e., mushrooms A. manicata (Berk. & Broome) Pegler and A. nana Singer. Antioxidant potential was assessed by DPPH (2,2-diphenyl-1-picrylhydrazyl) free radical scavenging and ABTS (2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid) assay, total phenolics, and flavonoids. Both mushrooms were found to possess antioxidants and wide range of phenolics and bioactive compounds. There was maximum percent inhibition (83.2 ± 0.120%) on DPPH by A. manicata. However, maximum percent inhibition using ABTS was found to be 79.5 ± 0.251% by A. nana. Similarly, A. nana possesses maximum amount of both flavonoids and phenolics i.e., 0.3473 ± 0.0088 mg/100 g of catechin and 0.097 ± 0.0011 mg/100 g of GAE equivalent, respectively. Both mushrooms exhibited a variety of natural compounds such as P-coumaric acid, M-coumaric acid, benzoic acid, ferulic acid etc. Ferulic acid, chlorogenic acid, and cinnamic acid were also detected in A. manicata. A. manicata exhibited best and greater antiradical potential than A. nana due the presence of excessive natural bioactive compounds. From the reported results, it was revealed that both inedible mushrooms could be a potential source of antioxidants and secondary metabolites and might be used for making novel drugs in future by pharmaceutical industries.

The research of liver failure in Banna miniature pigs caused by amanita exitialis / 中华劳动卫生职业病杂志

Objective: To explore the characteristics of Banna miniature pig liver failure induced by amanita exitialis. Methods: From September to October 2020, a reverse high performance liquid chromatography (RP-HPLC) method was used to determine the toxin content of amanita exitialis solution, and 2.0 mg/kg amanita exitialis solution (α-amanitins+β-amanitins) was administered orally to Banna miniature pigs. Toxic symptoms, blood biochemical indexes and histopathological changes of liver, heart and kidney were observed at each time point. Results: All Banna miniature pigs died within 76 h of exposure, and different degrees of digestive tract symptoms such as nausea, vomiting and diarrhea appeared between 6 and 36 h. The biochemical indexes of alanine aminotransferase, aspartate aminotransferase, total bilirubin, lactate dehydrogenase, myoglobin, creatine kinase isoenzyme, blood urea nitrogen and creatinine increased significantly at 52 h after exposure, and the differences were statistically significant compared with 0 h (P<0.05). The bleeding of liver and heart was obvious under macroscopic and microscopic observation, hepatocyte necrosis, renal tubule epithelial cell swelling. Conclusion: Large dose of amanita exitialis can cause acute liver failure of Banna miniature pigs, which is in line with the pathophysiological characteristics of acute liver failure, and lays a foundation for further research on the toxic mechanism and detoxification drugs of amanita exitialis induced liver failure.

Intracellular sequestration of cadmium and zinc in ectomycorrhizal fungus Amanita muscaria (Agaricales, Amanitaceae) and characterization of its metallothionein gene.

Amanita muscaria is an ectomycorrhizal mushroom that commonly grows at metal-polluted sites. Sporocarps from the lead smelter-polluted area near Príbram (Central Bohemia, Czech Republic) showed elevated concentrations of Cd and Zn. Size exclusion chromatography of the cell extracts of the sporocarps from both polluted and unpolluted sites indicated that substantial part of intracellular Cd and Zn was sequestered in 6-kDa complexes, presumably with metallothionein(s) (MT). When the cultured mycelial isolates were compared, those from Príbram were more Cd-tolerant and accumulated slightly less Cd and Zn than those from the unpolluted site. The analysis of the available A.muscaria sequence data returned a 67-amino acid (AA) MT encoded by the AmMT1 gene. Weak Cd and Zn responsiveness of AmMT1 in the mycelia suggested its metal homeostasis function in A.muscaria, rather than a major role in detoxification. The AmMT1 belongs to a ubiquitous peptide group in the Agaricomycetes consisting of 60-70-AA MTs containing seven cysteinyl domains and a conserved histidyl, features observed also in a newly predicted, atypical 45-AA RaMT1 of the Zn-accumulator Russula bresadolae in which the C-terminal cysteinyl domains VI and VII are missing. Heterologous expression in metal-sensitive yeast mutants indicated that AmMT1 and RaMT1 encode functional peptides that can protect cells against Cd, Zn, and Cu toxicity. The metal protection phenotype observed in yeasts with mutant variants of AmMT1 and RaMT1 further indicated that the conserved histidyl seems to play a structural, not metal binding role, and the cysteinyls of the C-terminal domains VI and VII are important for Cu binding. The data provide an important insight into the metal handling of site-associated ectomycorrhizal species disturbed by excess metals and the properties of MTs common in Agaricomycetes.

Natural Bridged Bicyclic Peptide Macrobiomolecules from Celosia argentea and Amanita phalloides.

Bridged peptide macrobicycles (BPMs) from natural resources belong to types of compounds that are not investigated fully in terms of their formation, pharmacological potential, and stereo- chemical properties. This division of biologically active congeners with multiple circular rings has merits over other varieties of peptide molecules. BPMs form one of the most hopeful grounds for the establishment of drugs because of their close resemblance and biocompatibility with proteins, and these bio-actives are debated as feasible, realistic tools in diverse biomedical applications. Despite huge potential, poor metabolic stability and cell permeability limit the therapeutic success of macrocyclic peptides. In this review, we have comprehensively explored major bicyclic peptides sourced from plants and mushrooms, including ßs-leucyl-tryptophano-histidine bridged and tryptophanocysteine bridged peptide macrobicycles. The unique structural features, structure-activity relationship, synthetic routes, bioproperties, and therapeutic potential of the natural BPMs are also discussed.

Diversity, Dispersal and Mode of Reproduction of Amanita exitialis in Southern China.

Amanita exitialis is a poisonous mushroom and has caused many deaths in southern China. In this study, we collected 118 fruiting bodies of A. exitialis from seven different sites in Guangdong Province in southern China and investigated their genetic relationships using 14 polymorphic molecular markers. These 14 markers grouped the 118 fruiting bodies into 20 multilocus genotypes. Among these 20 genotypes, eight were each found only once while the remaining 12 were each represented by two to 54 fruiting bodies. Interestingly, among the 12 shared genotypes, four were shared between/among local populations that were separated by as far as over 80 km, a result consistent with secondary homothallic reproduction and long-distance spore dispersal. Despite the observed gene flow, significant genetic differentiations were found among the local populations, primarily due to the over-representation of certain genotypes within individual local populations. STRUCTURE analyses revealed that the 118 fruiting bodies belonged to three genetic clusters, consistent with divergence within this species in this geographic region. Interestingly, we found an excess of heterozygous individuals at both the local and the total sample level, suggesting potential inbreeding depression and heterozygous advantage in these populations of A. exitialis. We discuss the implications of our results for understanding the life cycle, dispersal, and evolution of this poisonous mushroom.

An α-D-galactan and a ß-D-glucan from the mushroom Amanita muscaria: Structural characterization and antitumor activity against melanoma.

Polysaccharides α-D-galactan (GAL-Am) and ß-D-glucan (GLC-Am) were obtained from Amanita muscaria fruiting bodies. They were purified using different methodologies, such as Fehling precipitation (for both fractions), freeze-thawing process and ultrafiltration (for GLC-Am). Results showed that the GAL-Am has (1 â†’ 6)-linked Galp main chain branched at O-2 by terminal Galp units and has not been previously reported. Besides, GLC-Am has (1 â†’ 3)-linked Glcp in the main chain, substituted at O-6 by (1 â†’ 6)-linked ß-Glcp units. Both are water-soluble, with 9.0 × 103 g/moL and 1.3 × 105 g/moL, respectively. GAL-Am and GLC-Am presented a selective proliferation reduction against B16-F10 melanoma cell line, not affecting non tumoral BALB/3T3 fibroblast cell line. Furthermore, both fractions reduced clonogenic capacity of melanoma cell line over an extended period of time. These results were obtained without modulations in B16-F10 cell adhesion, reinforcing the biological activities towards cell proliferation impairment and eliciting these polysaccharides as promising compounds to further exploration of their antimelanoma properties.

Ibotenic Acid Biosynthesis in the Fly Agaric Is Initiated by Glutamate Hydroxylation.

The fly agaric, Amanita muscaria, is widely known for its content of the psychoactive metabolites ibotenic acid and muscimol. However, their biosynthetic pathway and the respective enzymes are entirely unknown. 50 years ago, the biosynthesis was hypothesized to start with 3-hydroxyglutamate. Here, we build on this hypothesis by the identification and recombinant production of a glutamate hydroxylase from A. muscaria. The hydroxylase gene is surrounded by six further biosynthetic genes, which we link to the production of ibotenic acid and muscimol using recent genomic and transcriptomic data. Our results pinpoint the genetic basis for ibotenic acid formation and thus provide new insights into a decades-old question concerning a centuries-old drug.

Metallic and metalloid elements in various developmental stages of Amanita muscaria (L.) Lam.

There is growing evidence that mushrooms (fruiting bodies) can be suitable for biogeochemical prospecting for minerals and as indicators of heavy metal and radioactive contaminants in the terrestrial environment. Apart from the nutritional aspect, knowledge of accumulation dynamics and distribution of elements in fruiting bodies, from emergence to senescence, is essential as is standardization when choosing mushroom species as potential bioindicators and for monitoring purposes. We studied the effect of fruitbody developmental stage on the contents of the elements (Li, K, V, Cr, Mn, Mg, Co, Ni, Cu, Zn, As, Rb, Sr, Ag, Al, Cd, Sb, Cs, Ba, Pb, Tl and U) in the individual parts of the Amanita muscaria fruiting body. Elements such as K, Mg, Mn, Ni, Co, Cu, Zn and Se remained similar throughout all developmental stages studied, however for K, differences occurred in the values of caps and stipes, as expressed by the cap to stipe concentration quotient (index QC/S). The other elements quantified, i.e., Li, V, Cr, As, Rb, Sr, Ag, Al, Cd, Sb, Cs, Ba, Pb, Tl and U are considered as nonessential or toxic (with the exception of V in A. muscaria). Their accumulation in the fruiting bodies and their distribution between cap and stipe did not show a uniform pattern. Pb, Sb, Tl, Ba, Sr, Li, Rb and Cs decreased with increasing maturity of the fruitbodies, implying that translocation, distribution and accumulation in stipes and caps was not a continuous process, while V, Cr, As, Ag, Cd, and U remained at the same concentration, similarly to the essential elements. Our results for A. muscaria confirm that elemental distribution in different parts of fruiting bodies is variable for each element and may change during maturation. Soil properties, species specificity and the pattern of fruitbody development may all contribute to the various types of elemental distribution and suggest that the results for one species in one location may have only limited potential for generalization.

A Convergent Total Synthesis of the Death Cap Toxin α-Amanitin.

The toxic bicyclic octapeptide α-amanitin is mostly found in different species of the mushroom genus Amanita, with the death cap (Amanita phalloides) as one of the most prominent members. Due to its high selective inhibition of RNA polymerase II, which is directly linked to its high toxicity, particularly to hepatocytes, α-amanitin received an increased attention as a toxin-component of antibody-drug conjugates (ADC) in cancer research. Furthermore, the isolation of α-amanitin from mushrooms as the sole source severely restricts compound supply as well as further investigations, as structure-activity relationship (SAR) studies. Based on a straightforward access to the non-proteinogenic amino acid dihydroxyisoleucine, we herein present a robust total synthesis of α-amanitin providing options for production at larger scale as well as future structural diversifications.

Transcriptome Analysis Provides Novel Insights into the Capacity of the Ectomycorrhizal Fungus Amanita pantherina To Weather K-Containing Feldspar and Apatite.

Ectomycorrhizal (ECM) fungi, symbiotically associated with woody plants, markedly improve the uptake of mineral nutrients such as potassium (K) and phosphorus (P) by their host trees. Although it is well known that ECM fungi can obtain K and P from soil minerals through biological weathering, the mechanisms regulating this process are still poorly understood at the molecular level. Here, we investigated the transcriptional regulation of the ECM fungus Amanita pantherina in weathering K-containing feldspar and apatite using transcriptome sequencing (RNA-seq) and validated these results for differentially expressed genes using real-time quantitative PCR. The results showed that A. pantherina was able to improve relevant metabolic processes, such as promoting the biosynthesis of unsaturated fatty acids and steroids in the weathering of K-containing feldspar and apatite. The expression of genes encoding ion transporters was markedly enhanced during exposure to solid K-containing feldspar and apatite, and transcripts of the high-affinity K transporter ApHAK1, belonging to the HAK family, were significantly upregulated. The results also demonstrated that there was no upregulation of organic acid biosynthesis, reflecting the weak weathering capacity of the A. pantherina isolate used in this study, especially its inability to utilize P in apatite. Our findings suggest that under natural conditions in forests, some ECM fungi with low weathering potential of their own may instead enhance the uptake of mineral nutrients using their high-affinity ion transporter systems.IMPORTANCE In this study, we revealed the molecular mechanism and possible strategies of A. pantherina with weak weathering potential in the uptake of insoluble mineral nutrients by using transcriptome sequencing (RNA-seq) technology and found that ApHAK1, a K transporter gene of this fungus, plays a very important role in the acquisition of K and P. Ectomycorrhizal (ECM) fungi play critical roles in the uptake of woody plant nutrients in forests that are usually characterized by nutrient limitation and in maintaining the stability of forest ecosystems. However, the regulatory mechanisms of ECM fungi in acquiring nutrients from minerals/rocks are poorly understood. This study investigated the transcriptional regulation of A. pantherina weathering K-containing feldspar and apatite and improves the understanding of fungal-plant interactions in promoting plant nutrition enabling increased productivity in sustainable forestry.

Bio-concentration potential and associations of heavy metals in Amanita muscaria (L.) Lam. from northern regions of Poland.

Fruiting bodies of Amanita muscaria and topsoil beneath from six background areas in northern regions of Poland were investigated for the concentration levels of Ag, Al, Ba, Ca, Cd, Co, Cu, Fe, Hg, K, Mg, Mn, Na, Rb, Sr, and Zn. In addition, the bioconcentration factors (BCF values) were studied for each of these metallic elements. Similar to studies from other basidiomycetes, A. muscaria showed species-specific affinities to some elements, resulting in their bioconcentration in mycelium and fruiting bodies. This mushroom growing in soils with different levels of the geogenic metallic elements (Ag, Al, Ba, Ca, Co, Cu, Fe, Hg, K, Mg, Mn, Na, Rb, Sr, and Zn) showed signs of homeostatic accumulation in fruiting bodies of several of these elements, while Cd appeared to be accumulated at a rate dependent of the concentration level in the soil substrate. This species is an efficient bio-concentrator of K, Mg, Cd, Cu, Hg, Rb, and Zn and hence also contributes to the natural cycling of these metallic elements in forest ecosystems.

Preferential accumulation of inorganic elements in Amanita muscaria from North-eastern Poland.

Concentrations of Al, Ba, Cd, Ca, Co, Cu, Fe, K, Mg, Mn, Hg, Rb, Ag, Na, Sr and Zn were analysed in fruiting bodies of A. muscaria. This mushroom is considered conditionally edible, since parboiling can detoxify its fruiting bodies from the hallucinogens and render it edible. The specific purpose of the research is the little-known phenomenon of the variability of mineral composition of mushrooms for generations harvested in the same forest areas - both in terms of their nutritional value and anthropogenic influences. Fungal materials were digested in nitric acid and analysed by a validated methods using inductively coupled plasma optical emission spectrometer (ICP-OES) and cold - vapor atomic absorption spectroscopy (CV-AAS). A. muscaria collected in the same area over the period of four years showed fluctuations in the concentrations of the essential elements Co, Cu, Fe, K, Mn, Na and Zn (p < 0.05; U Mann-Whitney test), while only Ca and Mg (p > 0.05) levels were similar. In addition, concentrations of Ag, Al, Ba, Cd, Rb and Sr in fruiting bodies fluctuated, while remained at a similar level for Hg. It is important to note that statistically significant variations in levels of several inorganic elements accumulated in A. muscaria would imply a more careful consideration would be required when assessing the nutritional value of mushroom species.

Study of three interesting Amanita species from Thailand: Morphology, multiple-gene phylogeny and toxin analysis.

Amanita ballerina and A. brunneitoxicaria spp. nov. are introduced from Thailand. Amanita fuligineoides is also reported for the first time from Thailand, increasing the known distribution of this taxon. Together, those findings support our view that many taxa are yet to be discovered in the region. While both morphological characters and a multiple-gene phylogeny clearly place A. brunneitoxicaria and A. fuligineoides in sect. Phalloideae (Fr.) Quél., the placement of A. ballerina is problematic. On the one hand, the morphology of A. ballerina shows clear affinities with stirps Limbatula of sect. Lepidella. On the other hand, in a multiple-gene phylogeny including taxa of all sections in subg. Lepidella, A. ballerina and two other species, including A. zangii, form a well-supported clade sister to the Phalloideae sensu Bas 1969, which include the lethal "death caps" and "destroying angels". Together, the A. ballerina-A. zangii clade and the Phalloideae sensu Bas 1969 also form a well-supported clade. We therefore screened for two of the most notorious toxins by HPLC-MS analysis of methanolic extracts from the basidiomata. Interestingly, neither α-amanitin nor phalloidin was found in A. ballerina, whereas Amanita fuligineoides was confirmed to contain both α-amanitin and phalloidin, and A. brunneitoxicaria contained only α-amanitin. Together with unique morphological characteristics, the position in the phylogeny indicates that A. ballerina is either an important link in the evolution of the deadly Amanita sect. Phalloideae species, or a member of a new section also including A. zangii.

Accumulation and distribution of metallic elements and metalloids in edible Amanita fulva mushrooms.

Baseline concentrations of Ag, Al, As, Ba, Ca, Cd, Co, Cu, Cr, Fe, K, Mg, Mn, Na, Ni, P, Pb, Rb, Sr, Tl, V, U and Zn were presented in Amanita fulva collected from unpolluted areas in Poland. There is no previous data published on the bio-element constituents of A. fulva. A very narrow range of values was determined by ICP-DRC-MS and ICP-AES for the trace elements Ag, Co, Cu, Cr, Ni, Rb, Sr, Tl and Zn in caps and of Ag, Co, Cu, Mn, Ni, Sr, U and Zn in stipes and also for the macro elements K, P, Na and Mg. The fruitbodies of A. fulva from the northern (Baltic Sea coastal forests) and southwestern (Lower Silesia forests) sites differed substantially in cadmium, lead and uranium, and those from the Lower Silesia region showed them in greater concentrations. This observation may imply that A. fulva under typical geochemical site conditions is able to regulate the accumulation of many of the elements mentioned in fruiting bodies.

Accumulation of Ag and Cu in Amanita strobiliformis and characterization of its Cu and Ag uptake transporter genes AsCTR2 and AsCTR3.

Macrofungi can accumulate in their sporocarps remarkably high concentrations of Cu and Ag. We have previously demonstrated that the non-essential Ag is in the ectomycorrhizal, Ag-hyperaccumulating Amanita strobiliformis sequestered by 3.4-kDa metallothioneins (MTs) produced as AsMT1a, 1b and 1c isoforms. Here, we describe two populations of wild-grown A. strobiliformis sporocarps, which showed certain correlation between the concentrations of accumulated Ag (284 ± 64 and 67 ± 15 mg kg(-1)) and Cu (76 ± 13 and 30 ± 12 mg kg(-1)), suggesting that an overlap may exist in the cell biology of Ag and Cu in this species. Metal speciation analysis revealed that the intracellular Cu in the sporocarps of both populations was, like Ag, associated with the 3.4-kDa MTs. A search of A. strobiliformis transcriptome for sequences encoding proteins of the Cu transporter (CTR) family identified four AsCTR cDNAs, which were, like AsMT1s, confirmed in both populations. The predicted AsCTR proteins showed homology to vacuolar (AsCTR1 and AsCTR4) and plasma membrane (AsCTR2 and AsCTR3) CTRs. Heterologous expression of AsCTR2, AsCTR3 and their translational fusions with green fluorescent protein (GFP) in Cu uptake-deficient S. cerevisiae indicated that both AsCTRs are functional Cu and Ag uptake transporters: recombinant genes complemented growth defects and increased Cu and Ag uptake rates in yeasts and the GFP-tagged protein localized to the cell periphery. Site directed mutagenesis revealed the importance of the conserved-among-CTRs M-X3-M motif for the AsCTR2- and AsCTR3-mediated transport of both Cu and Ag. These results provide the first evidence that fungal CTRs can recognize Ag for transport.

Characterization of three distinct metallothionein genes of the Ag-hyperaccumulating ectomycorrhizal fungus Amanita strobiliformis.

Mechanisms evolved in eukaryotes to handle heavy metals involve cytosolic, metal-binding metallothioneins (MTs). We have previously documented that the sequestration of silver (Ag) in the Ag-hyperaccumulating Amanita strobiliformis is dominated by 34-amino-acid (AA) AsMT1a, 1b, and 1c isoforms. Here we show that in addition to AsMT1a, 1b, and 1c isogenes, the fungus has two other MT genes: AsMT2 encoding a 34-AA AsMT2 similar to MTs known from other species, but unrelated to AsMT1s; AsMT3 coding for a 62-AA AsMT3 that shares substantial identity with as-yet-uncharacterized conserved peptides predicted in agaricomycetes. Transcription of AsMT1s and AsMT3 in the A. strobiliformis mycelium was specifically inducible by treatments with Ag or copper (Cu) and zinc (Zn) or cadmium (Cd), respectively; AsMT2 showed a moderate upregulation in the presence of Cd. Expression of AsMTs in the metal-sensitive Saccharomyces cerevisiae revealed that all AsMTs confer increased Cd tolerance (AsMT3 proved the most effective) and that, unlike AsMT1 and AsMT2, AsMT3 can protect the yeasts against Zn toxicity. The highest level of Cu tolerance was observed with yeasts expressing AsMT1a. Our data indicate that A. strobiliformis can specifically employ different MT genes for functions in the cellular handling of Ag and Cu (AsMT1s) and Zn (AsMT3).

Microanalysis characterization of bioactive protein-bound polysaccharides produced by Amanita ponderosa cultures.

Different compounds of edible mushrooms are responsible for their bioactivity. The ability to synthesize polysaccharides, namely protein-polysaccharide (PPS) complexes, is related to the antioxidant capacity of these compounds and present great interest in preventing a number of diseases, including cancer, cardiovascular and auto-immune diseases, and accelerated aging. Amanita ponderosa are wild edible mushrooms that grow in Mediterranean "montado" areas [Portuguese name given to cork oak (Quercus suber) and holm oak (Quercus ilex) forests]. The aim of this study was to evaluate the production of PPS complexes obtained from A. ponderosa cultures using a new microanalytical approach to quickly and easily monitor the production process. Microanalysis using Fourier-transform infrared using attenuated total reflection and Raman spectroscopy of PPS samples showed spectra compatible with identification of this type of compound in culture extracts. PPS separated by size-exclusion chromatography showed seven main complexes. Molecular weights of the main PPS complexes isolated from cultures ranged between 1.5 and 20 kDa and did not present toxicity against Artemia salina, demonstrating the potential of A. ponderosa as a source of biologically active compounds with nutraceutical value. Application of this microanalytical approach to monitoring the production of PPS compounds can be successfully applied in biotechnological processes.

Do differences in chemical composition of stem and cap of Amanita muscaria fruiting bodies correlate with topsoil type?

Fly agaric (Amanita muscaria) was investigated using a 1H NMR-based metabolomics approach. The caps and stems were studied separately, revealing different metabolic compositions. Additionally, multivariate data analyses of the fungal basidiomata and the type of soil were performed. Compared to the stems, A. muscaria caps exhibited higher concentrations of isoleucine, leucine, valine, alanine, aspartate, asparagine, threonine, lipids (mainly free fatty acids), choline, glycerophosphocholine (GPC), acetate, adenosine, uridine, 4-aminobutyrate, 6-hydroxynicotinate, quinolinate, UDP-carbohydrate and glycerol. Conversely, they exhibited lower concentrations of formate, fumarate, trehalose, α- and ß-glucose. Six metabolites, malate, succinate, gluconate, N-acetylated compounds (NAC), tyrosine and phenylalanine, were detected in whole A. muscaria fruiting bodies but did not show significant differences in their levels between caps and stems (P value>0.05 and/or OPLS-DA loading correlation coefficient <0.4). This methodology allowed for the differentiation between the fruiting bodies of A. muscaria from mineral and mineral-organic topsoil. Moreover, the metabolomic approach and multivariate tools enabled to ascribe the basidiomata of fly agaric to the type of topsoil. Obtained results revealed that stems metabolome is more dependent on the topsoil type than caps. The correlation between metabolites and topsoil contents together with its properties exhibited mutual dependences.

Mercury in the Grisette, Amanita vaginata Fr. and soil below the fruiting bodies.

This study examined the mercury concentration in the Grisette Amanita vaginata Fr. and soil below the fruiting bodies collected between 2000 and 2008 from the wild at seven distant sites across Poland. The Hg content in samples was determined by cold atomic absorption method (CV-AAS) at a wavelength of 253.7 nm. Mean Hg contents varied from 0.096 ± 0.052 to 0.48 ± 0.13 mg kg(-1) dry matter (dm) in caps (range, 0.043-0.73 mg kg(-1)), from 0.047 ± 0.02 to 0.23 ± 0.07 mg kg(-1) dm (range, 0.028-0.47 mg kg(-1)) in stipes, and in underlying soil were from 0.035 ± 0.018 to 0.096 ± 0.036 mg kg(-1) dm (range, 0.017 to 0.16 mg kg(-1)). The median Qc/s values ranged from 1.2 to 2.2 (mean 1.2 ± 0.4 to 2.1 ± 0.5) indicating that Hg content in stipes was generally lower than in caps. This mushroom species has some potential to bioconcentrate Hg in the fruiting bodies, as the values of the bioconcentration factor (BCF) varied for the sites between 1.2 ± 0.6 to 11 ± 5 for caps and 0.61 ± 0.26 to 7.4 ± 3.9 for stipes. Also available literature data on Hg in A. vaginata are reviewed and discussed.

Amanitin and phallotoxin concentration in Amanita phalloides var. alba mushroom.

Although rarely seen, Amanita phalloides var. alba, a variety of A. phalloides type mushrooms, causes mushroom poisoning resulting in death. Since it is frequently confused with some edible mushrooms due to its white colored cap and macroscopic appearance, it becomes important in toxicological terms. Knowledge of the toxin amount contained in this mushroom type is invaluable in the treatment of cases involving poisoning. In this study, we examined the toxin levels of various parts of the A. phalloides var. alba mushroom growing Duzce region of Turkey. Toxin analyses were carried out for A. phalloides var. alba, which were collected from the forests Duzce region of Turkey in 2011, as a whole and also separately in its spore, pileus, gills, stipe and volva parts. The alpha amanitin, beta amanitin, gamma amanitin, phalloidin and phallacidine analyses of the mushrooms were carried out using the RP-HPLC method. A genetic analysis of the mushroom showed that it had similar genetic characteristics as A. phalloides and was a variety of it. The lowest toxins quantity was detected in spores, volva and stipe among all parts of the mushroom. The maximum amount of amatoxins was measured in the gills. The pileus also contained a high amount of amatoxins. Generally, amatoxins and phallotoxin concentrations were lower as compared to A. phalloides, but interestingly all toxins other than gamma toxin were higher in the spores of A. phalloides var. alba. The amount of toxin in all of its parts had sufficient concentrations to cause death. With this study, the amatoxin and phallotoxin concentrations in A. phalloides var. alba mushroom and in its parts have been revealed in detail for the first time.