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ObjectiveTo analyze the content of 6 kinds of amatoxins in 3 species of Lepiota collected from Chuxiong of Yunnan Province.MethodsThe collected mushrooms were baked and dried at 45 ℃， and species identification was carried out by morphological and molecular biology methods. The mushrooms were divided into three different growth periods （growth period， mature stage， aged stage） and different tissue parts （pileus， stipe， whole）， and the toxins were extracted by ultrapure water ultrasound. The determination was performed by ultra-performance liquid chromatography-triple quadrupole tandem mass spectrometry with electrospray （ESI-） in multiple-reaction monitoring （MRM） mode， and the content was quantitatively analyzed by external standard method.ResultsNo amatoxins were detected in Lepiota clypeolaria and Lepiota cristata， while α-amanitin and β-amanitin were detected in Lepiota brunneoincarnata， and the content of α-amanitin was higher than β-amanitin. The total content of α-amanitin was highest in the mature stage （1.67 mg/g）， followed by the grow stage （1.56 mg/g） and least in the aged stage （0.40 mg/g）. And the total content of β-amanitin was also highest in the mature stage （0.71 mg/g）， followed by the growth stage （0.60 mg/g） and lowest in the aged stage （0.14 mg/g）. The cap part was richer in α-amanitin and β-amanitin than the stipe part in the growth and mature stages， while the content of α-amanitin and β-amanitin in the aged stage was higher in the stipe than in the pileus.ConclusionIn Chuxiong Prefecture， Yunnan Province， the flesh-brown ring-stalk mushroom L. brunneoincarnata mainly contains Amatoxins， including α-AMA， β-AMA， Amanin， and Amaninamide， excluding phalloidin and virotoxin. The content of α-AMA and β-AMA varies in different growth periods and tissues， providing reference for the clinical diagnosis and treatment of poisoning by this type of mushroom.

Copyright of Medicina (Buenos Aires) is the property of Medicina (Buenos Aires) and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

During the last 20 years citizen science has largely contributed at promoting knowledge about mushroom diversity in Greece. In the frame of an ongoing collaboration among amateur and professional mycologists, hundreds of specimens have been subjected to DNA sequencing, followed by phylogenetic analyses and detailed morphoanatomical examination of selected collections. The study of recent findings revealed the existence of seven species which are new records for the Greek mycobiota, i.e. Amanita codinae, Cleistocybe carneogrisea, Hebeloma limbatum, Infundibulicybe alkaliviolascens, Lepiota farinolens, Leucoagaricus pilatianus and Pholiotina mediterranea. In particular, C. carneogrisea is a rare Mediterranean species, which has been proposed by the IUCN for inclusion in the Global Red List of Fungi. Moreover, these predominantly or exclusively southern European species are described and evaluated. All findings are discussed in respect to their most prominent features and closely related taxa, while ITS phylogenetic trees are presented for A. codinae and L. farinolens by including mainly species of the sections Lepidella and Ovisporae, respectively. [ABSTRACT FROM AUTHOR]

Abstract Background Amanitin-producing mushrooms, mainly distributed in the genera Amanita, Galerina and Lepiota, possess MSDIN gene family for the biosynthesis of many cyclopeptides catalysed by prolyl oligopeptidase (POP). Recently, transcriptome sequencing has proven to be an efficient way to mine MSDIN and POP genes in these lethal mushrooms. Thus far, only A. palloides and A. bisporigera from North America and A. exitialis and A. rimosa from Asia have been studied based on transcriptome analysis. However, the MSDIN and POP genes of many amanitin-producing mushrooms in China remain unstudied; hence, the transcriptomes of these speices deserve to be analysed. Results In this study, the MSDIN and POP genes from ten Amanita species, two Galerina species and Lepiota venenata were studied and the phylogenetic relationships of their MSDIN and POP genes were analysed. Through transcriptome sequencing and PCR cloning, 19 POP genes and 151 MSDIN genes predicted to encode 98 non-duplicated cyclopeptides, including α-amanitin, β-amanitin, phallacidin, phalloidin and 94 unknown peptides, were found in these species. Phylogenetic analysis showed that (1) MSDIN genes generally clustered depending on the taxonomy of the genus, while Amanita MSDIN genes clustered depending on the chemical substance; and (2) the POPA genes of Amanita, Galerina and Lepiota clustered and were separated into three different groups, but the POPB genes of the three distinct genera were clustered in a highly supported monophyletic group. Conclusions These results indicate that lethal Amanita species have the genetic capacity to produce numerous cyclopeptides, most of which are unknown, while lethal Galerina and Lepiota species seem to only have the genetic capacity to produce α-amanitin. Additionally, the POPB phylogeny of Amanita, Galerina and Lepiota conflicts with the taxonomic status of the three genera, suggesting that underlying horizontal gene transfer has occurred among these three genera.

Abstract Background Genomes of lethal Amanita and Galerina mushrooms have gradually become available in the past ten years; in contrast the other known amanitin-producing genus, Lepiota, is still vacant in this aspect. A fatal mushroom poisoning case in China has led to acquisition of fresh L. venenata fruiting bodies, based on which a draft genome was obtained through PacBio and Illumina sequencing platforms. Toxin-biosynthetic MSDIN family and Porlyl oligopeptidase B (POPB) genes were mined from the genome and used for phylogenetic and statistical studies to gain insights into the evolution of the biosynthetic pathway. Results The analysis of the genome data illustrated that only one MSDIN, named LvAMA1, exits in the genome, along with a POPB gene. No POPA homolog was identified by direct homology searching, however, one additional POP gene, named LvPOPC, was cloned and the gene structure determined. Similar to ApAMA1 in A. phalloides and GmAMA1 in G. marginata, LvAMA1 directly encodes α-amanitin. The two toxin genes were mapped to the draft genome, and the structures analyzed. Furthermore, phylogenetic and statistical analyses were conducted to study the evolution history of the POPB genes. Compared to our previous report, the phylogenetic trees unambiguously showed that a monophyletic POPB lineage clearly conflicted with the species phylogeny. In contrast, phylogeny of POPA genes resembled the species phylogeny. Topology and divergence tests showed that the POPB lineage was robust and these genes exhibited significantly shorter genetic distances than those of the house-keeping rbp2, a characteristic feature of genes with horizontal gene transfer (HGT) background. Consistently, same scenario applied to the only MSDIN, LvAMA1, in the genome. Conclusions To the best of our knowledge, this is the first reported genome of Lepiota. The analyses of the toxin genes indicate that the cyclic peptides are synthesized through a ribosomal mechanism. The toxin genes, LvAMA1 and LvPOPB, are not in the vicinity of each other. Phylogenetic and evolutionary studies suggest that HGT is the underlining cause for the occurrence of POPB and MSDIN in Amanita, Galerina and Lepiota, which are allocated in three distantly-related families.

Abstract The biosynthetic pathway for amanitins and related cyclic peptides in deadly Amanita (Amanitaceae) mushrooms represents the first known ribosomal cyclic peptide pathway in the Fungi. Amanitins are found outside of the genus in distantly related agarics Galerina (Strophariaceae) and Lepiota (Agaricaceae). A long-standing question in the field persists: why is this pathway present in these phylogenetically disjunct agarics? Two deadly mushrooms, A. pallidorosea and A. subjunquillea, were deep sequenced, and sequences of biosynthetic genes encoding MSDINs (cyclic peptide precursor) and prolyl oligopeptidases (POPA and POPB) were obtained. The two Amanita species yielded 29 and 18 MSDINs, respectively. In addition, two MSDIN sequences were cloned from L. brunneoincarnata basidiomes. The toxin MSDIN genes encoding amatoxins or phallotoxins from the three genera were compared, and a phylogenetic tree constructed. Prolyl oligopeptidase B (POPB), a key enzyme in the biosynthetic pathway, was used in phylogenetic reconstruction to infer the evolutionary history of the genes. Phylogenies of POPB and POPA based on both coding and amino acid sequences showed very different results: while POPA genes clearly reflected the phylogeny of the host species, POPB did not; strikingly, it formed a wellsupported monophyletic clade, despite that the species belong to different genera in disjunct families. POPA, a known house-keeping gene, was shown to be restricted in a branch containing only Amanita species and the phylogeny resembled that of those Amanita species. Phylogenetic analyses of MSDIN and POPB genes showed tight coordination and disjunct distribution. A POPB gene tree was compared with a corresponding species tree, and distances and substitution rates were compared. The result suggested POPB genes have significant smaller distances and rates than the house-keeping rpb2, discounting massive gene loss. Under this assumption, the incongruency between the gene tree and species tree was shown with strong support. Additionally, k-mer analyses consistently cluster Galerina and Amanita POPB genes, while Lepiota POPB is distinct. Our result suggests that horizontal gene transfer (HGT), at least between Amanita and Galerina, was involved in the acquisition of POPB genes, which may shed light on the evolution of the α-amanitin biosynthetic pathway.

A recent collection of Lepiota spiculata from the Dominican Republic is presented here. Macro- and micromorphological features of L. spiculata are described in detail, and its evolutionary (phylogenetic) position within Lepiota sect. Ovisporae, in the subincarnata/brunneoincarnata clade, is assessed on the basis of a combined nrLSU + nrITS + rpb2 + tef1 analysis. Additionally, high levels of deadly amatoxins were detected and quantified in L. spiculata for the first time by HPLC analysis; in particular, α-amanitin was found at concentrations up to approximately 4 mg/g dry weight, which render L. spiculata a potentially lethal mushroom, if ingested.

A recent collection of Lepiota spiculata from the Dominican Republic is presented here. Macro- and micromorphological features of L. spiculata are described in detail, and its evolutionary (phylogenetic) position within Lepiota sect. Ovisporae , in the subincarnata/brunneoincarnata clade, is assessed on the basis of a combined nrLSU + nrITS + rpb2 + tef1 analysis. Additionally, high levels of deadly amatoxins were detected and quantified in L. spiculata for the first time by HPLC analysis; in particular, α-amanitin was found at concentrations up to approximately 4 mg/g dry weight, which render L. spiculata a potentially lethal mushroom, if ingested. [ABSTRACT FROM AUTHOR]

The recent realistic estimate of fungal numbers which used various algorithms was between 2.2 and 3.8 million. There are nearly 100,000 accepted species of Fungi and fungus-like taxa, which is between 2.6 and 4.5% of the estimated species. Several forums such as Botanica Marina series, Fungal Diversity notes, Fungal Biodiversity Profiles, Fungal Systematics and Evolution—New and Interesting Fungi, Mycosphere notes and Fungal Planet have enhanced the introduction of new taxa and nearly 2000 species have been introduced in these publications in the last decade. The need to define a fungal species more accurately has been recognized, but there is much research needed before this can be better clarified. We address the evidence that is needed to estimate the numbers of fungi and address the various advances that have been made towards its understanding. Some genera are barely known, whereas some plant pathogens comprise numerous species complexes and numbers are steadily increasing. In this paper, we examine ten genera as case studies to establish trends in fungal description and introduce new species in each genus. The genera are the ascomycetes Colletotrichum and Pestalotiopsis (with many species or complexes), Atrocalyx, Dothiora, Lignosphaeria, Okeanomyces, Rhamphoriopsis, Thozetella, Thyrostroma (relatively poorly studied genera) and the basidiomycete genus Lepiota. We provide examples where knowledge is incomplete or lacking and suggest areas needing further research. These include (1) the need to establish what is a species, (2) the need to establish how host-specific fungi are, not in highly disturbed urban areas, but in pristine or relatively undisturbed forests, and (3) the need to establish if species in different continents, islands, countries or regions are different, or if the same fungi occur worldwide? Finally, we conclude whether we are anywhere near to flattening the curve in new species description. [ABSTRACT FROM AUTHOR]

Background: Amanitin-producing mushrooms, mainly distributed in the genera Amanita, Galerina and Lepiota, possess MSDIN gene family for the biosynthesis of many cyclopeptides catalysed by prolyl oligopeptidase (POP). Recently, transcriptome sequencing has proven to be an efficient way to mine MSDIN and POP genes in these lethal mushrooms. Thus far, only A. palloides and A. bisporigera from North America and A. exitialis and A. rimosa from Asia have been studied based on transcriptome analysis. However, the MSDIN and POP genes of many amanitin-producing mushrooms in China remain unstudied; hence, the transcriptomes of these speices deserve to be analysed. Results: In this study, the MSDIN and POP genes from ten Amanita species, two Galerina species and Lepiota venenata were studied and the phylogenetic relationships of their MSDIN and POP genes were analysed. Through transcriptome sequencing and PCR cloning, 19 POP genes and 151 MSDIN genes predicted to encode 98 non-duplicated cyclopeptides, including α-amanitin, β-amanitin, phallacidin, phalloidin and 94 unknown peptides, were found in these species. Phylogenetic analysis showed that (1) MSDIN genes generally clustered depending on the taxonomy of the genus, while Amanita MSDIN genes clustered depending on the chemical substance; and (2) the POPA genes of Amanita, Galerina and Lepiota clustered and were separated into three different groups, but the POPB genes of the three distinct genera were clustered in a highly supported monophyletic group. Conclusions: These results indicate that lethal Amanita species have the genetic capacity to produce numerous cyclopeptides, most of which are unknown, while lethal Galerina and Lepiota species seem to only have the genetic capacity to produce α-amanitin. Additionally, the POPB phylogeny of Amanita, Galerina and Lepiota conflicts with the taxonomic status of the three genera, suggesting that underlying horizontal gene transfer has occurred among these three genera. [ABSTRACT FROM AUTHOR]

Mushrooms containing amatoxins generate the highest number of fatal mycete poisonings on the planet. These toxins are produced not only by Amanita species, such as the well-known Amanita phalloides, but also by other genera, including Lepiota. In this work we report the treatment of a 51-year-old male patient weighing 79 kg who was referred to the hospital after 36 h of ingesting mushrooms. The mushrooms were identified as Lepiota brunneoincarnata, found for the first time in Argentina. The patient presented general malaise, nausea and repeated vomiting, abdominal pain, and diarrhea. Thanks to a quick anamnesis and early and accurate identification of the fungus, he was administered a nasogastric tube and serial activated charcoal. Additionally, N-acetylcysteine, phytomenadione and penicillin G EV were administered. The patient was discharged 11 days after admission. This case highlights the importance of obtaining a correct and sufficient anamnesis on fungi, enabling rapid analysis of them, and initiation of timely treatment for intoxication. Once again, the importance of having Toxicological Information and Advice Centers (CIAT) with experience and knowledge of micetisms is demonstrated.

Polyphenol oxidase (PPO) from the wild edible mushroom Lepiota procera is partially purified and biochemically characterized using three-phase partitioning (TPP), which is an easily applied and effective method. This method includes ammonium sulfate saturation at different concentrations, t-butanol addition (1:1; 1:5), and adjustment of pH. Optimum purification parameters with 20% ammonium sulfate saturation and 1:1 t-butanol conditions led to the highest activity at bottom phase with 8.4 fold purification. Sodium dodecyl sulfate- polyacrylamide gel electrophoresis (SDS-PAGE) analysis showed the molecular weight of the purified enzyme to be 35 kDa. The partially purified PPO presented a high level of activity with L-DOPA (Michaelis-Menten constant, Km – 0.12 mM), followed by caffeic acid (0.27 mM) and 4-methylcatechol (0.46 mM) and it is classified as a catecholase type of PPO. The enzyme had peak activity at a temperature of 40 °C and a pH value of 7.0. These results demonstrate a new enzyme source and easy purification method, useful for various industrial applications. [ABSTRACT FROM AUTHOR]

It is believed that Lepiota cristata is inedible and slightly poisonous, so it should always be avoided. This study analyzed the alpha, beta and gamma amanitin, phalloidin (PHN) and phallacidin (PCN) content of L. cristata collected from different regions of Turkey. Toxin levels have been assessed using a reversed-phase high-performance liquid chromatography (RP-HPLC) system with ultraviolet detectors. In this study, the L. cristata mushroom was not found to contain any amatoxins or phallotoxins. However, this finding does not mean that this mushroom is edible. New studies regarding the toxin content of L. cristata may be useful to understand the toxicity of this species. [ABSTRACT FROM AUTHOR]

Taxonomic and phylogenetic studies of Chlorophyllum were carried out on the basis of morphological differences and molecular phylogenetic analyses. Based on the phylogeny inferred from the internal transcribed spacer (ITS), the partial large subunit nuclear ribosomal DNA (nrLSU), the second largest subunit of RNA polymerase II (rpb2) and translation elongation factor 1-a (tef1) sequences, six well-supported clades and 17 phylogenetic species are recognised. Within this phylogenetic framework and considering the diagnostic morphological characters, two new species, C. africanum and C. palaeotropicum, are described. In addition, a new infrageneric classification of Chlorophyllum is proposed, in which the genus is divided into six sections. One new combination is also made. This study provides a robust basis for a more detailed investigation of diversity and biogeography of Chlorophyllum. [ABSTRACT FROM AUTHOR]

Species in the mushroom genus Lepiota can cause fatal mushroom poisonings due to their content of amatoxins such as α-amanitin. Previous studies of the toxin composition of poisonous Lepiota species relied on analytical methods of low sensitivity or resolution. Using liquid chromatography coupled to UV absorbance and mass spectrometry, we analyzed the spectrum of peptide toxins present in six Italian species of Lepiota, including multiple samples of three of them collected in different locations. Field taxonomic identifications were confirmed by sequencing of the internal transcribed spacer (ITS) regions. For comparison, we also analyzed specimens of Amanita phalloides from Italy and California, a specimen of A. virosa from Italy, and a laboratory-grown sample of Galerina marginata. α-Amanitin, β-amanitin, amanin, and amaninamide were detected in all samples of L. brunneoincarnata, and α-amanitin and γ-amanitin were detected in all samples of L. josserandii. Phallotoxins were not detected in either species. No amatoxins or phallotoxins were detected in L. clypeolaria, L. cristata, L. echinacea, or L. magnispora. The Italian and California isolates of A. phalloides had similar profiles of amatoxins and phallotoxins, although the California isolate contained more β-amanitin relative to α-amanitin. Amaninamide was detected only in A. virosa.

Lepiota lahorensis sp. nov. is described from Lahore, Pakistan. Its characterizing features are pinkish brown pileus with dark brown furfuraceous scales, oblong to ellipsoid spores, clavate cheilocystidia, and pileus covering made up of trichodermal elements without shorter elements at their base. The phylogenetic analysis based on nuclear ribosomal internal transcribed spacer sequence shows its unique position in sect. Ovisporae. [ABSTRACT FROM AUTHOR]

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Lepiota sect. Stenosporae (J.E. Lange) Kühner contains species with spurred or truncate basidiospores and pileus covering composed of adnate, ascending or erect long and slender elements. Seven species from southwestern, northeastern and southern China in this section have been identified, described and illustrated. Lepiota mandarina and L. subcastanea are described as new. Lepiota mandarina is characterized by densely orange red to brownish red squamules on pileus and stipe, subtriangular basidiospores with truncate bottom, narrowly clavate cheilocystidia and pileus covering a trichodermial palisade. Lepiota subcastanea has a pileus with yellow-brown to dark brown squamules, non-dextrinoid subtriangular basidiospores with a distinct sac-like spur, narrowly clavate or narrowly utriform cheilocystidia, and pileus covering a trichodermial palisade with short clavate elements. It is hypothesized that this section evolved from sect. Ovisporae subsect. Helveolinae. It also is speculated that subtriangular and spurred basidiospores have evolved from ellipsoidal basidiospores and that a trichodermial pileus covering composing long, erect elements and basal short, clavate elements might have evolved from a cutis or a trichoderm lacking such an under layer of short elements. [ABSTRACT FROM AUTHOR]

چهارگونه Lepiota anthomyces، L. serena، L. metulaespora وL. roseoalba از مناطق مختلف کرج ، تهران و اهواز جمع آوری و در این مقاله توصیف شده اند. هر چهار گونه فوق الذکر برای فلور ایران جدید می باشند، ترسیم هایی از خصوصیات میکروسکوپی و ماکروسکوپی از هر گونه نیز ارایه گردیده است. ضمناً گونه cristataL.که قبلاً از ایران گزارش شده در این مقاله مورد بحث و توصیف قرار می گیرد.

Amatoxin poisoning from the genus Lepiota may have a deadly outcome, although this is not seen as often as it is from the genus Amanita . In this report, we present a patient who was poisoned by a sublethal dose of Lepiota brunneoincarnata mushrooms. The patient was hospitalized 12 hours after eating the mushrooms. The patient’s transaminase levels increased dramatically starting on day 4. Aspartate transaminase peaked at 78 hours. Starting at 1265 IU/L, alanine transaminase peaked at 90 hours at 5124 IU/L. The patient was discharged on day 8 to outpatient care, and his transaminase levels returned to normal ranges in the subsequent days. A toxin analysis was carried out on the mushrooms that the patient claimed to have eaten. Using reversed-phase high-performance liquid chromatography analysis, an uptake of approximately 19.9 mg of amatoxin from nearly 30 g of mushrooms was calculated. This consisted of 10.59 mg of α-amanitin, 9.18 mg of β-amanitin, and 0.16 mg of γ-amanitin. In conclusion, we present a patient from Turkey who was poisoned by L. brunneoincarnata mushrooms. [ABSTRACT FROM AUTHOR]

We investigated the quantity and concentration of toxins in different parts and in different growth phases ofLepiota brunneoincarnatamushroom. The amatoxins and phallotoxin levels were measured using the reversed phase high-performance liquid chromatography (RP-HPLC) method. Alpha amanitin (2.38 ± 0.70 mg/g) was followed by beta amanitin (1.97 ± 0.52 mg/g) and gamma amanitin (0.04 ± 0.01 mg/g) in trace amounts; it did not contain any phallotoxin. The cap part is richer in amatoxins than the stipe part. While medium mushrooms were quite rich in amatoxins, less levels of toxin were measured in fully developed mushrooms. The study showed in detail the toxin concentrations ofL. brunneoincarnatawith regard to different developmental stages and different segments. [ABSTRACT FROM PUBLISHER]

Species in the mushroom genus Lepiota can cause fatal mushroom poisonings due to their content of amatoxins such as α-amanitin. Previous studies of the toxin composition of poisonous Lepiota species relied on analytical methods of low sensitivity or resolution. Using liquid chromatography coupled to UV absorbance and mass spectrometry, we analyzed the spectrum of peptide toxins present in six Italian species of Lepiota, including multiple samples of three of them collected in different locations. Field taxonomic identifications were confirmed by sequencing of the internal transcribed spacer (ITS) regions. For comparison, we also analyzed specimens of Amanita phalloides from Italy and California, a specimen of A. virosa from Italy, and a laboratory-grown sample of Galerina marginata. α-Amanitin, β-amanitin, amanin, and amaninamide were detected in all samples of L. brunneoincarnata, and α-amanitin and γ-amanitin were detected in all samples of L. josserandii. Phallotoxins were not detected in either species. No amatoxins or phallotoxins were detected in L. clypeolaria, L. cristata, L. echinacea, or L. magnispora. The Italian and California isolates of A. phalloides had similar profiles of amatoxins and phallotoxins, although the California isolate contained more β-amanitin relative to α-amanitin. Amaninamide was detected only in A. virosa. [ABSTRACT FROM AUTHOR]

Lepiota sanguineofracta sp. nov., a taxon with a hymeniform pileus covering, from Italy, is here described. A full description, colour pictures of basidiomata, line drawings of microscopic features, and ITS phylogenetic analysis are provided. It is morphologically and molecularly close (sister) to Lepiota coloratipes from which it differs mainly by olivaceus tinges on the pileus surface, basidiome surfaces and context strongly reddening on handling, a sweetish smell of withered rose and binucleate, not metachromatic spores. [ABSTRACT FROM AUTHOR]

In this paper, 2 lepiotaceous mushrooms [Lepiota indica sp. nov. and Lepiota attenuispora sp. nov. (Agaricaeae)] and 1 Limacella magna sp. nov. in the Amanitaceae are described. Lepiota indica has scurfy brownish carpophores, minute spores, and chains of golden brown-walled pileal elements. Lepiota attenuispora has a shaggy appearance, an appendiculate veil of the pileus margin, and a stipe sheathed with soft cottony scales. Limacella magna is a species characterized by prominent brown scales on the pileus and stipe, presence of a glutinous layer, and a trichodermal pileus cutis. [ABSTRACT FROM AUTHOR]

Lepiota acutesquamosa and L. cristata (Basidiomycota, Agaricales) collected from Himalayan moist temperate forests of Pakistan were characterized using internal transcribed spacers (ITS) of rNDA, a fungal molecular marker. The ITS-rDNA of both species was analyzed using polymerase chain reaction (PCR) and DNA sequencing. The target region when amplified using universal fungal primers (ITS1F and ITS4) generated 650-650bp fragments. Consensus sequences of both species were submitted for initial blast analysis which revealed and confirmed the identification of both species by comparing the sequences of these respective species already present in the GenBank. Sequence of Pakistani collection of L. acutesquamosa matched 99% with sequences of same species (FJ998400) and Pakistani L. cristata matched 97% with its sequences (EU081956, U85327, AJ237628). Further, in phylogenetic analysis both species distinctly clustered with their respective groups. Morphological characters like shape, size and color of basidiomata, basidiospore size, basidial lengths, shape and size of cheilocystidia of both collections were measured and compared. Both these species have been described first time from Pakistan on morph-anatomical and molecular basis. [ABSTRACT FROM AUTHOR]

The rare sequestrate fungus Amogaster viridiglebus is known from only one collection in California where it was discovered among Populus roots. Based on sporocarp coloration and spore morphology, this sequestrate taxon was putatively considered to be an ectomycorrhizal member of the Boletales. However, no molecular data were previously available to definitively determine the closest relatives of this fungus. Here we revisit the morphology of Amogaster viridiglebus and present a phylogenetic analysis based on ITS and 28S ribosomal DNA. Our phylogeny indicates that Amogaster viridiglebus is nested in the genus Lepiota, suggesting that this rare species has a saprobic trophic mode and does not form ectomycorrhizae with plants. A new combination, L. viridigleba, is made based on these phylogenetic results. [ABSTRACT FROM AUTHOR]

Abstract: An isolation procedure that comprised three ion-exchange chromatography steps on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and one gel-filtration step by fast protein liquid chromatography on Superdex 75 was utilized to isolate a laccase with a molecular mass of 65kDa from fresh fruiting bodies of the mycorrhizal fungus Lepiota ventriosospora. Laccase activity was adsorbed on both DEAE-cellulose and Q-Sepharose but unadsorbed on CM-cellulose. An overall 26.3-fold of purification was obtained. The enzyme demonstrated an optimum temperature at 60°C and an optimum pH 4.0. The purified laccase was quite stable at pH range of 3.6–4.4, but only 17.8% of total activity left after 1h incubating at 60°C. The ranking of its degradative activity toward aromatic substrates was catechol>hydroquinone>ABTS>2,6-dimethoxy-phenol. It demonstrated the highest inhibitory activity toward HIV-1 reverse transcriptase with an IC50 value of 0.60μM among fungal laccaes reported up to now. [Copyright &y& Elsevier]