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Amanita is one of the most salient mushroom genera due to its cultural, economic, and medical importance. Recently, many new Amanita species have been described worldwide, increasing the genus richness. However, several clades have cryptic diversity, and many species complexes have not yet been resolved. This is the case of the rubescent species in the Validae section, which have been widely cited under the name Amanita rubescens s.l. We used a four-locus matrix (nuc rDNA internal transcribed spacer [ITS] and 28S regions and genes for RNA polymerase II subunit 2 [ rpb2 ], translation elongation factor 1-α [ tef1-α ], and β-tubulin [ tub2 ]) to solve the phylogenetic relationships within the Amanita section Validae . To analyze the diversity and distribution patterns of species, we used an extensive ITS sequence sampling including environmental DNA databases. The phylogenetic analyses demonstrated that the Validae section is divided into three monophyletic and highly supported major clades: Mappae, Validae , and Rubescentes . At least 11 species-level clades within the Rubescentes clade were highly supported: A. cruentilemurum nom. prov. A. brunneolocularis, A. rubescens s.s. (European clade), A. rubescens s.s. (Asiatic clade), A. orsonii s.s. A . ' orsonii ,' A. aureosubucula nom. prov., A. novinupta, A. flavorubens , and two undescribed North American species. We proved that A. rubescens s.s. has two segregated populations (European and Asiatic) and it is not naturally distributed in America. Furthermore, we found that America has more cryptic species within the Rubescentes clade than Eurasia.

Amanita exitialis, a deadly mushroom found in eastern Asia, causes the highest death rates among all poisonous mushrooms in China. The aim of the present study was to develop an efficient, accurate, and user-friendly PCR-based method for identifying A. exitialis that could facilitate the prevention, diagnosis, and treatment of associated food poisoning. A. exitialis-specific primers and probes were designed based on the internal transcribed spacer region variations of 27 mushroom species. Specificity was confirmed using conventional and real-time PCR for 23 non-target mushroom species, including morphologically similar and closely related species. Compared to conventional PCR, real-time PCR was more sensitive (detectable DNA concentration: 1.36 × 10 -2 ng/μL vs. 1.36 × 10 -3 ) and efficient (analysis time: 1 h vs. 40 min). Furthermore, the real-time PCR results could be immediately visualized using amplification curve analysis. The results present two robust PCR-based methods for A. exitialis identification that can facilitate food safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Food poisoning caused by toxic mushrooms, such as species in the Amanita genus, occurs frequently around the world. To properly treat these patients, it is important to rapidly and accurately identify the causal species. Matrix-assisted laser desorption/ionization time-of-flight (MALDI-ToF) mass spectrometry is a rapid technique that has been used in medical laboratories for the past three decades to identify bacteria, yeasts, and filamentous fungi.Matrix-assisted laser desorption ionization time of flight mass spectrometry (MALDI-Tof MS) is a rapid method used for the past three decades to identify microorganisms. In this study, we created and internally validated a MALDI-Tof MS reference database comprising 15 Amanita species frequently encountered in France, and we challenged this database with 38 Amanita specimens from four French locations, using a free online application for MALDI-ToF spectra identifications.Assessment of the database showed that mass spectra can be obtained by analyzing any portion of a carpophore and that all portions enabled identification of the carpophore at the species level. Most carpophores were correctly identified using our database, with the exception of specimens from the Vaginatae section. Decay tests also demonstrated that decayed portions (like those found in the kitchen garbage can) of Amanita phalloides mushrooms could be properly identified using MALDI-ToF MS.Our findings provide important insight for toxicology laboratories that often rely on DNA sequencing to identify meal leftovers implicated in food poisoning. In future developments, this technique could also be used to detect counterfeit mushrooms by including other genera in the reference database., Lay Summary: MALDI-ToF MS is a powerful identification tool for microorganisms. We demonstrate that the technique can be applied to Amanita specimens. This will prevent food intoxications as a rapid and definite identification can be obtained, and it can also be used for food remnants., (© The Author(s) 2021. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

Four epitypes and three new species of Amanita (Amanitaceae, Agaricales, Agaricomycetes, Basidiomycota) are described from Guineo-Congolian rainforests of Cameroon. Amanita echinulata, A. fulvopulverulenta, A. robusta , and A. bingensis are epitypified based on collections that are the first since the species were described nearly a century ago. Morphological features of the epitypes are described and enumerated. Amanita minima, Amanita luteolamellata , and A. goossensfontanae are described as new and added to the known macromycota of tropical Africa. Habit, habitat, and known distribution are provided for each species. Sequence data for the internal transcribed spacer (ITS) locus are provided for types and other collections of all taxa, and a molecular phylogenetic analysis across the genus Amanita corroborates morphology-based infrageneric placement for each.

Amanita cf. lavendula collections in eastern North America, Mexico, and Costa Rica were found to consist of four cryptic taxa, one of which exhibited consistently unreadable nuclear rDNA ITS1-5.8S-ITS2 (fungal barcode) sequences after ITS1 base 130. This taxon is designated here as Amanita cf. lavendula taxon 1. ITS sequences from dikaryotic basidiomata were cloned, but sequences recovered from cloning did not segregate into distinct haplotypes. Rather, there was a mix of haplotypes that varied among themselves predominantly at 28 ITS positions. Analysis of each of these 28 variable bases showed predominantly two alternate bases at each position. Based on these findings and additional sequence data from the nuclear rDNA 28S, RNA polymerase II subunit 2 (RPB2) and mitochondrial rDNA small subunit (SSU) and 23S genes, we speculate that taxon 1 represents an initial hybridization event between two divergent taxa followed by failure of the ribosomal repeat to homogenize. Homogenization failure may be a result of repeated hybridization between divergent internal transcribed spacer (ITS) types with inadequate time for concerted evolution of the ribosomal repeat or, alternately, a complete failure of the ribosomal homogenization process. To our knowledge, this finding represents the first report of a geographically widespread taxon (Canada, eastern USA, Costa Rica) with apparent homogenization failure across all collections. Findings such as these have implications for fungal barcoding efforts and the application of fungal barcodes in identifying environmental sequences.

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.

Amanita is a diverse and cosmopolitan genus of ectomycorrhizal fungi. We describe Amanita nouhrae sp. nov., a new hypogeous ('truffle-like') species associated with Nothofagus antarctica in northern Patagonia. This constitutes the first report of a sequestrate Amanita from the Americas. Thick-walled basidiospores ornamented on the interior spore wall ('crassospores') were observed consistently in A. nouhrae and its sister epigeous taxon Amanita morenoi, a rarely collected but apparently common species from northern Patagonia that has sometimes been misidentified as the Australian taxon Amanita umbrinella. Nuclear 18S and 28S ribosomal DNA and mitochondrial 16S and 26S DNA placed these two species in a southern temperate clade within subgenus Amanita, together with other South American and Australian species. Based on a dated genus-level phylogeny, we estimate that the southern temperate clade may have originated near the Eocene/Oligocene boundary (ca. 35 Ma ± 10 Ma). This date suggests a broadly distributed ancestor in the Southern Hemisphere, which probably diversified as a result of continental drift, as well as the initiation of the Antarctic glaciation. By comparison, we show that this clade follows an exceptional biogeographic pattern within a genus otherwise seemingly dominated by Northern Hemisphere dispersal., (Copyright © 2017 British Mycological Society. Published by Elsevier Ltd. All rights reserved.)

Some Amanita specimens collected from Malaysia are critically investigated by morphological examination and molecular analysis of two gene fragments, the nuc rDNA partial 28S (28S) gene and the internal transcriber spacer (ITS1-5.8S-ITS2 = ITS) regions. Six phylogenetic species of Amanita section Caesareae are recognized among the studied collections. One of them is described as new, A. malayensis. Four of the phylogenetic species correspond with existing morphology-based taxa: A. aporema, A. javanica, A. princeps, and A. similis. The remaining species is not described because of the paucity of material. Detailed descriptions and the distribution of these southeastern Asian species are provided, along with a key to the species of section Caesareae from Malaysia.

Amanita subpallidorosea is a recently discovered lethal Amanita sect. Phalloideae species found in China that is clustered with A. virosa in the same clade based on molecular phylogenetic analysis. However, the cyclopeptide toxin contents of these lethal mushrooms remain poorly studied. In this study, the cyclopeptide toxins in A. subpallidorosea were reported for the first time and the cyclopeptide compositions of A. subpallidorosea and A. virosa species were systematically analyzed. Thirteen cyclopeptides and two unknown compounds were identified or observed from these two lethal mushrooms by high-performance liquid chromatography coupled with high-resolution mass spectrometry. Of the known cyclopeptides, the virotoxins alaviroidin, viroisin, and viroidin, which were previously thought to be restricted to A. virosa, were identified in A. subpallidorosea. The cyclopeptide compositions showed that there are diversities in the kinds and levels of amatoxins, phallotoxins, and virotoxins between A. subpallidorosea and A. virosa species, and that the amount of total toxins in the tested A. subpallidorosea is significantly higher than that in the tested A. virosa. Furthermore, consistency of the cyclopeptide toxins with the molecular phylogenetic relationships was demonstrated., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Amanita is a worldwide-distributed fungal genus, with approximately 600 known species. Most species within the genus are ectomycorrhizal (ECM), with some saprotrophic representatives. In this study, we constructed the first comprehensive phylogeny including ECM species from Colombia collected in native Quercus humboldtii forests and in introduced Pinus patula plantations. We included 8 species (A. brunneolocularis, A. colombiana, A. flavoconia, A. fuligineodisca, A. muscaria, A. rubescens, A. sororcula, and A. xylinivolva) out of 16 species reported for the country, two new reports: A. citrina and A. virosa, and a new variety A. brunneolocularis var. pallida. Morphological taxonomic keys together with a phylogenetic approach using three nuclear gene regions: partial nuc rDNA 28S nuc rDNA internal transcribed spacers ITS1 and ITS2 and partial translation elongation factor 1-α gene (TEF1), were used to classify the specimens. Several highly supported clades were obtained from the phylogenetic hypotheses obtained by Bayesian inference and maximum likelihood approaches, allowing us to position the Colombian collections in a coherent infrageneric level and to contribute to the knowledge of local Amanita diversity.

Lethal amanitas (Amanita sect. Phalloideae) are responsible for 90% of all fatal mushroom poisonings. Since 2000, more than ten new lethal Amanita species have been discovered and some of them had caused severe mushroom poisonings in China. However, the contents and distribution of cyclopeptides in these lethal mushrooms remain poorly known. In this study, the diversity of major cyclopeptide toxins in seven Amanita species from Eastern Asia and three species from Europe and North America were systematically analyzed, and a new approach to inferring phylogenetic relationships using cyclopeptide profile was evaluated for the first time. The results showed that there were diversities of the cyclopeptides among lethal Amanita species, and cyclopeptides from Amanita rimosa and Amanita fuligineoides were reported for the first time. The amounts of amatoxins in East Asian Amanita species were significantly higher than those in European and North American species. The analysis of distribution of amatoxins and phallotoxins in various Amanita species demonstrated that the content of phallotoxins was higher than that of amatoxins in Amanita phalloides and Amanita virosa. In contrast, the content of phallotoxins was significantly lower than that of amatoxins in all East Asian lethal Amanita species tested. However, the distribution of amatoxins and phallotoxins in different tissues showed the same tendency. Eight cyclopeptides and three unknown compounds were identified using cyclopeptide standards and high-resolution MS. Based on the cyclopeptide profiles, phylogenetic relationships of lethal amanitas were inferred through a dendrogram generated by UPGMA method. The results showed high similarity to the phylogeny established previously based on the multi-locus DNA sequences., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Lethal amanitas (Amanita sect. Phalloideae) cause many casualties worldwide. Recent molecular phylogenetic studies revealed diverse lethal Amanita spp. in China. Here a 5-gene phylogeny (nuc rDNA region encompassing the internal transcribed spacers 1 and 2 with the 5.8S rDNA, the D1-D3 domains of nuc 28S rDNA, and partial RNA polymerase II second largest subunit, translation elongation factor 1-α and β-tubulin genes) is used to investigate the phylogenetic lineages and species delimitation in this section. Thirteen species are recognized, including four new species, namely A. griseorosea, A. molliuscula, A. parviexitialis, and A. subfuliginea They are documented with morphological, multigene phylogenetic, and ecological evidence, line drawings, and photographs and compared with similar species. A key to the Chinese lethal Amanita species is provided., (© 2016 by The Mycological Society of America.)

Cases of mushroom poisoning in Thailand have increased annually. During 2008 to 2014, the cases reported to the National Institute of Health included 57 deaths; at least 15 died after ingestion of amanitas, the most common lethal wild mushrooms inhabited. Hence, the aims of this study were to identify mushroom samples from nine clinically reported cases during the 7-year study period based on nuclear ITS sequence data and diagnose lethal peptide toxins using a reversed phase LC-MS method. Nucleotide similarity was identified using BLAST search of the NCBI database and the Barcode of Life Database (BOLD). Clade characterization was performed by maximum likelihood and Bayesian phylogenetic approaches. Based on BLAST and BOLD reference databases our results yielded high nucleotide similarities of poisonous mushroom samples to A. exitialis and A. fuliginea. Detailed phylogenetic analyses showed that all mushroom samples fall into their current classification. Detection of the peptide toxins revealed the presence of amatoxins and phallotoxins in A. exitialis and A. fuliginea. In addition, toxic α-amanitin was identified in a new provisional species, Amanita sp.1, with the highest toxin quantity. Molecular identification confirmed that the mushrooms ingested by the patients were members of the lethal amanitas in the sections Amanita and Phalloideae. In Thailand, the presence of A. exitialis was reported here for the first time and all three poisonous mushroom species provided new and informative data for clinical studies.

Some of the effects of past climate dynamics on plant and animal diversity make-up have been relatively well studied, but to less extent in fungi. Pleistocene refugia are thought to harbour high biological diversity (i.e. phylogenetic lineages and genetic diversity), mainly as a product of increased reproductive isolation and allele conservation. In addition, high extinction rates and genetic erosion are expected in previously glaciated regions. Some of the consequences of past climate dynamics might involve changes in range and population size that can result in divergence and incipient or cryptic speciation. Many of these dynamic processes and patterns can be inferred through phylogenetic and coalescent methods. In this study, we first delimit species within a group of closely related edible ectomycorrhizal Amanita from North America (the American Caesar's mushrooms species complex) using multilocus coalescent-based approaches; and then address questions related to effects of Pleistocene climate change on the diversity and genetics of the group. Our study includes extensive geographical sampling throughout the distribution range, and DNA sequences from three nuclear protein-coding genes. Results reveal cryptic diversity and high speciation rates in refugia. Population sizes and expansions seem to be larger at midrange latitudes (Mexican highlands and SE USA). Range shifts are proportional to population size expansions, which were overall more common during the Pleistocene. This study documents responses to past climate change in fungi and also highlights the applicability of the multispecies coalescent in comparative phylogeographical analyses and diversity assessments that include ancestral species., (© 2015 John Wiley & Sons Ltd.)

Background: Lethal amanitas (Amanita section Phalloideae) are a group of wild, fatal mushrooms causing many poisoning cases worldwide. However, the diversity and evolutionary history of these lethal mushrooms remain poorly known due to the limited sampling and insufficient gene fragments employed for phylogenetic analyses. In this study, five gene loci (nrLSU, ITS, rpb2, ef1-α and β-tubulin) with a widely geographic sampling from East and South Asia, Europe, North and Central America, South Africa and Australia were analysed with maximum-likelihood, maximum-parsimony and Bayesian inference methods. Biochemical analyses were also conducted with intention to detect amatoxins and phalloidin in 14 representative samples., Result: Lethal amanitas were robustly supported to be a monophyletic group after excluding five species that were provisionally defined as lethal amanitas based on morphological studies. In lethal amanitas, 28 phylogenetic species were recognised by integrating molecular phylogenetic analyses with morphological studies, and 14 of them represented putatively new species. The biochemical analyses indicated a single origin of cyclic peptide toxins (amatoxins and phalloidin) within Amanita and suggested that this kind of toxins seemed to be a synapomorphy of lethal amanitas. Molecular dating through BEAST and biogeographic analyses with LAGRANGE and RASP indicated that lethal amanitas most likely originated in the Palaeotropics with the present crown group dated around 64.92 Mya in the early Paleocene, and the East Asia-eastern North America or Eurasia-North America-Central America disjunct distribution patterns were primarily established during the middle Oligocene to Miocene., Conclusion: The cryptic diversity found in this study indicates that the species diversity of lethal amanitas is strongly underestimated under the current taxonomy. The intercontinental sister species or sister groups relationships among East Asia and eastern North America or Eurasia-North America-Central America within lethal amanitas are best explained by the diversification model of Palaeotropical origin, dispersal via the Bering Land Bridge, followed by regional vicariance speciation resulting from climate change during the middle Oligocene to the present. These findings indicate the importance of both dispersal and vicariance in shaping the intercontinental distributions of these ectomycorrhizal fungi.

Amanita caesareoides is a sister species of Amanita caesarea, also known as Caesar's mushroom and one of the most desirable edible mycorrhizal mushrooms. However, cultivation of Caesar's mushrooms has not yet been successful due to the difficulties involved in establishing pure cultures. In this study, we established pure cultures of four Asian Caesar's mushroom species, i.e., A. caesareoides, Amanita javanica, Amanita esculenta, and Amanita similis, which were identified by sequence analysis of their rDNA internal transcribed spacer (ITS) region. Five selected isolates in A. caesareoides, A. javanica, and A. esculenta were tested for ectomycorrhizal syntheses with axenic Pinus densiflora seedlings in vitro. Ectomycorrhizal tips of each fungal isolate tested were observed on pine lateral roots within 5 months of inoculation. Seventeen pine seedlings that formed ectomycorrhizas in vitro with these three Amanita species were acclimatized under non-sterile conditions. Seven months following acclimatization, ectomycorrhizal colonization by A. caesareoides was observed on newly grown root tips, which was confirmed by polymerase chain reaction restriction fragment length polymorphism analysis of the fungal rDNA ITS region. Two other Amanita species also survived during ectomycorrhizal acclimatization. These results suggest that the cultivation of A. caesareoides and its relatives can be attempted through mycorrhizal synthesis using P. densiflora as a host. This is the first report of in vitro mycorrhization of Asian Caesar's mushrooms and their acclimatization under non-sterile conditions.

In Facultad de Ciencias Exactas y Naturales,Universidad de Buenos Aires there is a service called Servicio de Identificación de Hongos Tóxicos, directed by researchers of the Program of Medicinal Plants and Fungi Involved in Biological Degradation (PROPLAME-PRHIDEB, CONICET) that assist hospitals and other health establishments, identifying the different samples of fungi and providing information about their toxicity, so that patients can receive the correct treatment. The objective of the present study was to analyze all the cases received from 1985 to 2012. This analysis permitted the confection of a table identifying the most common toxic species. The information gathered revealed that 47% of the patients were under 18 years of age and had eaten basidiomes; the remaining 53% were adults who insisted that they were able to distinguish edible from toxic mushrooms. Chlorophyllum molybdites turned out to be the main cause of fungal intoxication in Buenos Aires, which is commonly confused with Macrolepiota procera, an edible mushroom. In the second place Amanita phalloides was registered, an agaric known to cause severe symptoms after a long period of latency (6-10 hours), and which can lead to hepatic failure even requiring a transplant to prevent severe internal injuries or even death, is not early and correctly treated.

The occurrence of the exotic ectomycorrhizal fungus Amanita muscaria in a mixed Nothofagus-Eucalyptus native forest was investigated to determine if A. muscaria has switched hosts to form a successful association with a native tree species in a natural environment. A mycorrhizal morphotype consistently found beneath A. muscaria sporocarps was examined, and a range of morphological and anatomical characteristics in common with those described for ectomycorrhizae formed by A. muscaria on a broad range of hosts were observed. A full description is provided. The likely plant associate was determined to be Nothofagus cunninghamii based upon anatomy of the roots. Analysis of ITS-1 and ITS-2 regions of nuclear ribosomal DNA sequences confirmed the identities of both fungal and plant associates. These findings represent conclusive evidence of the invasion of a non-indigenous ectomycorrhizal fungus into native forest and highlight the ecological implications of this discovery.

Objective: To investigate the anti-tumor activity of two wild Amanita strains, and to determine the inhibition ratio of their effective chemical components on tumor cells in vitro., Methods: Two wild Amanita strains collected in Sichuan province were identified through morphology and molecular methods. The chemical components from the fruit body of the two strains were extracted with petroleum ether, chloroform, methanol and water. The anti-tumor activity of the extracts were tested on the cancer cells of MGC and Lovo strains. The polysaccharides were isolated with hot water and tested for its anti-tumor activity in vitro., Results: The two wild Amanitas were identified as Amanita subjunquillea and Amanita pantherina. The water-soluble extracts from the Amanitas had anti-tumor effect, with polysaccharide as one of the main effective components. The IC50 value of the polysaccharide isolated from M1 was 3.790 mg/mL for the MGC cells., Conclusion: Polysaccharides isolated from the two wild Amanitas have considerable anti-tumor activity against MGC cells in vitro.

Some species in the genus Amanita have a great variety of toxic secondary metabolites. They are characterized macroscopically by having a white spore print and free gills, and microscopically by the presence of a divergent hymenophoral trama. Some species of Amanita present in Colombia were chemically characterized by analyzing their toxin composition using HPLC. Samples were collected in oak (Quercus humboldtii) and pine (Pinus radiata) forests. Twelve species were recovered, Amanita fuligineodisca, Amanita xylinivolva, Amanita flavoconia, Amanita rubescens, Amanita bisporigera, Amanita muscaria, Amanita humboldtii, Amanita sororcula, Amanita brunneolocularis, Amanita colombiana, Amanita citrina, Amanita porphyria as well as two unreported species. Results showed that most of the analyzed species have α -amanitin in concentrations ranging from 50 ppm to 6000 ppm. Concentrations of α-amanitin in the pileus were significantly greater than in the stipe. Phalloidin and phallacidin were only present in A. bisporigera. Chromatographic profiles are proposed as an additional taxonomic tool since specific peaks with similar retention times were conserved at the species level., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The systematic position of secotioid (Torrendia) and gasteroid (Amarrendia) forms within the agaricoid Amanita lineage (Agaricales, Basidiomycota) was studied with molecular (nLSU, ITS) data. Secotioid and gasteroid forms occur in four independent clades nested within agaricoid forms. One clade corresponds to the secotioid T. pulchella from southern Europe and northern Africa. The others correspond to Torrendia and Amarrendia species from Australia. Mediterranean climatic conditions are postulated as a force driving the convergent evolution of these secotioid and at least one of the gasteroid forms in geographically distant areas. Species formerly placed in Torrendia and Amarrendia are transferred to Amanita. A new species of Torrendia from Australia was discovered during the revision of the collections originally identified as T. arenaria and is described here as Amanita pseudoinculta.

Amanita phalloides, Lepiota cristata, Lepiota brunneoincarnata and Inocybe asterospora are among the most important species responsible of mushroom poisoning in northern Italy. A real time PCR method for the identification of samples containing DNA from each of these species was developed. To test specificity all protocols were applied on DNA extracted from various mushroom species; sensitivity was assessed performing serial dilutions on all samples; versatility of the protocols was evaluated performing tests on DNA extracted from different matrices. The protocols showed high sensitivity (32 ng dried mushroom), high specificity and sensitive detection of DNA extracted from difficult samples, including pasta with mushroom, cooked mushrooms and gastric aspirates.

Picking mushrooms, especially in summer and autumn, is still very popular in Poland. Despite raising awareness of poisonous mushrooms in the Polish society, year after year hospitals treat many patients diagnosed with poisoning with the most common toxic species of mushroom found in our country. Furthermore, growing interest in hallucinogenic mushrooms among young people has become a serious medical problem of our time. Websites make it incredibly easy for people to obtain information on the morphology and appearance of mushrooms with psychoactive properties, which leads inexperienced pickers to misidentification, resulting frequently in a fatal outcome. The article explores the subject of poisoning with the most common mushrooms with neurotropic effects, these are: Amanita muscaria, Amanita pantherina, Inocybe rubescens, Clitocybe dealbata, Clitocybe rivulosa and Psilocybe semilanceata. Toxins found in these species show symptoms that affect the central nervous system, parasympathetic system as well as the gastro-intestinal system. The effects of poisoning in the mushroom species mentioned above are mild in general, liver and kidney damage occur rarely, but the symptoms depend on both the dosage of the consumed toxins and individual susceptibility. In most cases the treatment is of symptomatic nature. There is no specific treatment. Medical procedures mainly involve induced gastrolavage--stomach pumping (providing that the patient is conscious), prescription of active carbon as well as replacement of lost body fluids and electrolytes. If the muscarinic symptoms prevail it is generally advised to dose atropine. Patients showing the signs of hyperactivity receive tranquilizers or narcoleptics to eliminate psychotic symptoms.

Amanita lippiae is described as a new species from a semi-arid (caatinga) region of northeastern Brazil. It possesses clampless basidia and elongate to nearly cylindrical inamyloid basidiospores, bears irregular remains of universal veil on the stipe bulb and is a gemmatoid species assignable to Amanita sect. Amanita.

We described and illustrated Amanita viscidolutea sp. nov. from specimens collected in the state of Rio Grande do Norte, northeastern Brazil. The main characteristics of the new species are its yellow pileus with white margin, the viscidity of the pileal surface, an exannulate stipe and inamyloid basidiospores. We also present an artificial dichotomous key to Central and South American species of Amanita (subgenus Amanita) section Amanita.

In order to demonstrate the ability of Fourier transform infrared (FTIR) spectroscopy for the differentiation of macro-fungi at variety level, FTIR spectroscopy was used to obtain vibrational spectra of Amanita hemibapha (Berk. & Broome) Sacc. subsp. hemibapha and Amanita hemibapha var. ochracea Zhu L. Yang. The results show that their spectra are very similar to each other, with a small difference in the relative intensity of absorbance. For the purpose of enhancing the spectral resolution and amplifying the differences, the first-derivative spectra and second-derivative spectra were selected for evaluating the correlation coefficients respectively. The results show that the second-derivative spectra of two fruit bodies of Amanita hemibapha (Berk. & Broome) Sacc. subsp. hemibapha and one fruit body of Amanita hemibapha var. ochracea Zhu L. Yang have obvious differences, the correlation coefficients are only 0.245 and 0.356 respectively, the second-derivative spectra of two fruit bodies of Amanita hemibapha (Berk. &Broome) Sacc. subsp. hemibapha are very similar, and the correlation coefficient is 0.865. The authors' results show that Fourier transform infrared spectroscopy in combination with correlation analysis method can be used to identify Amanita hemibapha (Berk. & Broome) Sacc. subsp. hemibapha and Amanita hemibapha var. ochracea Zhu L. Yang rapidly and accurately.

A growing number of molecular studies show that many fungi have phylogeographic structures and that their distinct lineages are usually limited to different continents. As a conservative test of the extent to which wind-dispersed mycorrhizal fungi may exhibit phylogeographic structure, we chose to study Amanita muscaria, a host-generalist, widespread, wind-dispersed fungus. In this paper, we document the existence of several distinct phylogenetic species within A. muscaria, based on multilocus DNA sequence data. According to our findings, A. muscaria has strong intercontinental genetic disjunctions, and, more surprisingly, has strong intracontinental phylogeographic structure, particularly within North America, often corresponding to certain habitats and/or biogeographic provinces. Our results indicate that the view of A. muscaria as a common, widespread, easily identifiable, ecologically plastic fungus with a wide niche does not correctly represent the ecological and biological realities. On the contrary, the strong associations between phylogenetic species and different habitats support the developing picture of ecoregional endemisms and relatively narrow to very narrow niches for some lineages.

Two ectomycorrhizal macrofungal Amanita species of the section Lepidella, A. strobiliformis and A. solitaria, were found to hyperaccumulate silver (Ag). All samples were collected from non-argentiferous areas with background Ag content in soils (0.07-1.01 mgkg(-1) Ag). The Ag contents of both Amanita species were mostly in the range of 200-700 mgkg(-1)D.W. with the highest Ag content of 1253 mgkg(-1) in one sample of A. strobiliformis. Silver concentrations in macrofungal fruit bodies were commonly 800-2500 times higher than in underlying soils. A. strobiliformis and A. solitaria are the first eukaryotic organisms known to hyperaccumulate Ag.

The molecular phylogeny and biogeography of two widely distributed Amanita species, A. muscaria and A. pantherina, were studied based on specimens from diverse localities. Analyses of both a partial sequence of the ITS region of nuclear DNA and a partial sequence of the beta-tubulin gene were able to resolve specimens of each species. Analyses revealed a greater divergence of the beta-tubulin region than the ITS region. Based on molecular phylogeny of the combination of the ITS and beta-tubulin regions, A. muscaria could be separated into at least three groups (Eurasian, Eurasian subalpine, and North American), and A. pantherina could be separated into at least two groups (North American and Eurasian). We hypothesize that the speciation of A. muscaria occurred in Eurasia with subsequent migration to North America via land bridges. However, it is impossible to determine whether A. pantherina moved from Eurasia to North America or vice versa. For both A. muscaria and A. pantherina, the intracontinental relationships of both Eurasia and North America were closer than the relationships between eastern Asia and eastern North America.

We determined the size of genets of late-stage ectomycorrhizal fungi in field sites in coastal Northern California. Basidiocarps were collected, mapped and subjected to genetic fingerprinting using amplified fragment length polymorphisms (AFLPs). The minimum size estimates for the largest genets of Amanita francheti, Lactarius xanthogalactus and Russula cremoricolor were 1.5, 9.3 and 1.1 m2, respectively. The molecular markers also showed that R. cremoricolor is dimorphic, with red- and white-capped morphotypes of this species forming a continuous population. Our results suggest that spore propagation plays a much more important role in the life history of the Russulaceae in undisturbed forest settings than previously recognized. Fungi appearing late in the succession sequence and systems without obvious disturbance therefore do not necessarily colonize primarily by mycelium.

The toxic peptides in Amanita virosa and Amanita verna from various region were analyzed by a reversed-phase high-performance liquid chromatography. It is showed that there were great differences in the concentrations of toxic peptides between species and strains of A. virosa and A. verna.

Amanita pantherina and Amanita muscaria are commonly occurring mushrooms in Polish forests. They contain ibotenic acid and muscimol: the substances reacting with neurotransmitter receptors in central nervous system. The ingestion of these mushrooms produces a distinctive syndrome consisting of alternating phases of drowsiness and agitation with hallucinations, and sometimes with convulsions. The diagnosis of Amanita pantherina or Amanita muscaria poisoning is established by means of mycologic investigation of gastric lavage. The treatment is only symptomatic, and the prognosis is usually good.

Mushroom poisonings caused by amatoxins are mostly lethal. Information about mycetisms caused by white species of Amanita is scarce. The present paper describes a case of mushroom poisoning caused by A. virosa. A prolongated latency period (6-10 hours), followed by cholera-like, improvement and visceral complication phases confirmed the amatoxin poisoning. The consumption of about 3 pounds of the toadstool by seven persons caused the death of five. Two patients survive the ingestion.

To our knowledge Amanita proxima poisoning has never been reported. Amanita proxima is a mushroom seldomy encountered, similar to a common and edible species: Amanita ovoïdae. During October 1992, we had the opportunity to care for five cases of intoxications with Amanita proxima. In all cases early digestive disorders, cytolytic hepatitis and acute renal failure were noted. Outcome was favourable for all patients within three weeks with total recovery of both renal and hepatic functions with symptomatic treatment. Temporary dialysis was required in four patients.