Atractosporocybe, Leucocybe and Rhizocybe: three new clitocyboid genera in the Tricholomatoid clade (Agaricales) with notes on Clitocybe and Lepista.

A molecular multigene analysis (ITS, 18S and 28S nrLSU ribosomal DNA, tef1, rpb2) was used to support the proposition of three new genera of clitocyboid fungi. Leucocybe is proposed to accommodate the clade formed by Clitocybe connata and C. candicans. Clitocybe inornata is invested as type species of Atractosporocybe, while the new genus, Rhizocybe, is proposed for the former species of section Vernae of Clitocybe, C. vermicularis, C. pruinosa and C. rhizoides. The three lineages are related to the families Lyophyllaceae and Entolomataceae and independent from the Clitocybeae clade. Morphologically Rhizocybe is characterized by the presence of conspicuous rhizomorphs, while Atractosporocybe presents long fusiform spores. Leucocybe includes two whitish species in the former section Candicantes of Clitocybe, but no relevant shared characteristic feature was detected. Other whitish clitocyboid species, such as C. phyllophila (= C. cerussata), C. dealbata, C. rivulosa, and Singerocybe hydrogramma, are shown to be genetically related to the core lineage of the Clitocybeae.

Bioclimatic Origin Shapes Phylogenetic Structure of Tirmania (Pezizaceae): New Species and New Record from North Africa.

The phylogenetic relationships among Tirmania were investigated using the internal transcribed spacer (ITS) and large subunit (LSU) regions of the nuclear-encoded ribosomal DNA (rDNA) and compared with morphological and bioclimatic data. The combined analyses of forty-one Tirmania samples from Algeria and Spain supported four lineages corresponding to four morphological species. Besides the two previously described taxa, Tirmania pinoyi and Tirmania nivea, here we describe and illustrate a new species, Tirmania sahariensis sp. nov., which differs from all other Tirmania by its distinct phylogenetic position and its specific combination of morphological features. We also present a first record of Tirmania honrubiae from North Africa (Algeria). Our findings suggest that restrictions imposed by the bioclimatic niche have played a key role in driving the speciation process of Tirmania along the Mediterranean and Middle East.

Comparison between Tuber gennadii and T. oligospermum lineages reveals the existence of the new species T. cistophilum (Tuberaceae, Pezizales).

New collections of T. gennadii, T. oligospermum and T. asa (Tuberaceae, Pezizales) from the Mediterranean basin were compared on the basis of morphology and DNA sequences. Phylogenetic inference based on the internal transcribed spacer region (ITS) and the 28S nuclear large ribosomal subunit (nrLSU) indicates that T. gennadii includes at least two lineages with characteristic morphological traits. T. gennadii s. str. has small subglobose ascomata with conspicuous glebal locules that form a continuous labyrinth when mature and has a prosenchymatic peridium. T. lacunosum has medium to large, deeply lobed ascomata with small to very small isolated locules when mature and has a pseudoparenchymatic peridium. Two major clades were identified within the T. oligospermum group. On the basis of morphological, ecological and biogeographical data, these are identified with the taxa T. oligospermum (= T. asa) and T. sphaerospermum. Tuber oligospermum forms small to medium globose or subglobose, whitish ascomata, with globose to broadly ellipsoidal ascospores and is found in alkaline, sandy soils. Tuber sphaerospermum forms medium to large, irregularly globose or deeply lobed, yellowish ascomata, with regularly globose ascospores and is found in acidic soils. Finally, a new truffle species with a prosenchymatic peridium and globose to ellipsoid reticulate spores, collected in acidic argillaceous soils of Spain under Cistus ladanifer is proposed here as the new species T. cistophilum.

Amanita Section Phalloideae Species in the Mediterranean Basin: Destroying Angels Reviewed.

In Europe, amatoxin-containing mushrooms are responsible for most of the deadly poisonings caused by macrofungi. The present work presents a multidisciplinary revision of the European species of Amanita sect. Phalloideae based on morphology, phylogeny, epidemiology, and biochemistry of amatoxins and phallotoxins. Five distinct species of this section have been identified in Europe to date: A. phalloides, A. virosa, A. verna, the recently introduced North American species A. amerivirosa, and A. vidua sp. nov., which is a new name proposed for the KOH-negative Mediterranean species previously described as A. verna or A. decipiens by various authors. Epitypes or neotypes are selected for species lacking suitable reference collections, namely A. verna and A. virosa. Three additional taxa, Amanita decipiens, A. porrinensis, and A. virosa var. levipes are here considered later heterotypic synonyms of A. verna, A. phalloides, and A. amerivirosa, respectively.

Cytotoxic action of Ganoderma lucidum on interleukin-3 dependent lymphoma DA-1 cells: involvement of apoptosis proteins.

Aqueous extracts and a semipurified fraction obtained by methanol extraction and column chromatography were isolated from Ganoderma lucidum [Ganoderma lucidum (Curtis) P. Karst.; Ganodermataceae Donk] and their effects on interleukin 3-dependent lymphoma cells (DA-1) were studied. Cell viability was reduced by the action of unboiled aqueous extract and by the methanol-extracted column-chromatography semipurified fraction, producing DNA fragmentation in DA-1 cells. Treatments with aqueous extracts showed increments of Bax after 13 h, increments of p53 and Mdm2 after 19 h and a reduction of these three proteins after 24 h. The methanol-extracted semipurified fraction also induced increments of p53 and Mdm2 factors at 19 h with a reduction after 24 h. The methanol-extracted column-chromatography semipurified fraction from Ganoderma lucidum produced minor changes in the level of Akt after treatments for 19 h in DA-1 cells with a slight reduction in the levels of NFkB-p65 factor. Both the unboiled aqueous extract and the methanol-extracted column-chromatography semipurified fraction produced cleavage of inactive caspase 3, as a clear indication of induction of apoptosis by compounds present in Ganoderma lucidum.

Tubariomyces, a new genus of Inocybaceae from the Mediterranean region.

The taxonomic position of Inocybe inexpectata is reconsidered on the basis of new molecular and morphological data from four published records. Up to three species can be recognized to which the name I. inexpectata has been applied. In addition to distinct morphological features present in this lineage it is proposed here as the new genus Tubariomyces. A complete taxonomical description of the genus and the species Tubariomyces inexpectatus and T. hygrophoroides sp. nov. also is provided.

Herbicidal effects of wood vinegar on nitrophilous plant communities.

In Europe, and many parts of the world, the number and variety of animal species on farmland is in marked decline. There is a need to search for alternatives that are safe for the environmental and are effective in controlling weeds. Wood vinegar from biomass pyrolysis may be an alternative for herb control. In this study, Wood vinegar (WV) pH, moisture content, and composition were analyzed, with subsequent assessment of the effects of WV on nitrophilous plant communities under natural conditions. The following three treatments were used: WV dissolved in water to form 25 vol% and 50 vol% dilutions and undiluted WV (100 vol%). The results showed a greater than 70% decrease in biomass at 7 days after WV application in all treatments. At the end of the sampling period (day 42), the plots treated with WV had four-times less biomass than the controls. No significant differences were observed among different treatments, thus indicating that a 25% dilution may suffice for use as an herbicide. However, this concentration also produced the highest variability in results. The area cleared by the affected species was colonized by perennial species. At the end of the sampling, 80% of the area of the treated plots was occupied by perennial species, whereas this percentage was 30% in control plots. Electron micrographs showed that the epidermis of the treated plants was severely affected within a few hours of the treatment, particularly of the stomatal cells. The most affected species were those with smooth leaves without protective structures and those with lighter stems and leaves. The good herbicidal performance of WV notwithstanding, regulations must be clarified for its use as an herbicide.

A new postural device for the treatment of positional obstructive sleep apnea. A pilot study.

BACKGROUND: Approximately 60% of obstructive sleep apnea (OSA) diagnoses are position-dependent, and avoidance of the supine position could represent an effective treatment. Nevertheless, the majority of the available anti-supine treatments result in discomfort and low adherence. This study evaluated the effectiveness of a new vibrating supine avoidance device in reducing time spent in the supine position and the apnea-hypopnea index (AHI) without affecting sleep structure. Furthermore, the tolerability and satisfaction were also scored. METHODS: Observational prospective study of patients suffering from positional OSA. They were treated with a vibrating device and followed up at the first and fourth weeks after starting the treatment, and further polysomnographic studies were conducted while patients' wore the device. The comparison of the results was carried out through non-parametric tests. Significance level was 5%. RESULTS: Twelve patients had complete data. The device reduced time spent in the supine position (from 51.5 ±â€¯14.8% to 25.2 ±â€¯21.0%, p = 0.005), median AHI (from 30.7 (23.2-38.2) at baseline to 21.5 (12.4-24.3) at the fourth week, p = 0.002). Also an improvement in the minimum SaO2 (from 82.2 ±â€¯7.5 to 87.2 ±â€¯3.6 at the 4th week) was also observed. No variations in sleep quality or quantity were identified. All patients evaluated the device positively. CONCLUSION: Our device was effective in reducing the time spent in the supine position and improving AHI, SaO2 variables and sleep architecture. The device was well tolerated by the patients.

Distribution of radionuclides in different parts of a mushroom: influence of the degree of maturity.

Mushrooms are known to be bioaccumulators of radionuclides, but little is known about their distribution within the fruiting bodies or the influence of the degree of maturity on uptake. We carried out a series of cultures of the species Pleurotus eryngii under controlled laboratory conditions to analyze these variables. The maximal uptake of 134Cs and 85Sr was found to occur in mature fruiting bodies, and with the growth of the mushroom the distribution of radionuclides within the fruiting bodies became inhomogeneous. In particular, there was an exponential increase in the percentage of the total activity of 134Cs, 85Sr, and 60Co in the cap+gills as the fruiting bodies matured, accompanied by a complementary decrease in the stem. Radiocaesium, potassium, calcium, (239+240)Pu, (234,238)U, (228,230,232)Th, and 226Ra were assayed in the cap, gills, and stem of fruiting bodies of Tricholoma equestre collected in a natural ecosystem and cultured P. eryngii. Potassium and radiocaesium were mainly located in the cap+gills, and 226Ra in the gills. There was a disequilibrium between (230,232)Th and 228Th in the different parts of the fungi, probably due to uptake of 228Ra and subsequent decay to 228Th. Finally, the distribution pattern of (239+240)Pu, (234,238)U, and (230,232)Th seemed to be species dependent.

Contrasted Genetic Diversity, Relevance of Climate and Host Plants, and Comments on the Taxonomic Problems of the Genus Picoa (Pyronemataceae, Pezizales).

The species concept within the genus Picoa Vittad. is here revisited in light of new molecular and ecological data obtained from samples collected throughout the Mediterranean basin. Two highly diverse widespread clades and four additional minor lineages were significantly supported by three genes dataset (ITS, 28s LSU and RPB2) inferences for 70 specimens. The two widespread clades occur in very different geographical and ecological areas associated with exclusive host plants in the genus Helianthemum. SEM study of spore surface morphology in these lineages revealed the existence of smooth ascospores in the majority of these clades. However the most frequent lineage in Europe and coastal North Africa displayed either smooth or verrucose spores. Hence this morphological criterion cannot be reliably used to discriminate between the different clades. In addition, SEM observations made on ascospores from several original collections of P. juniperi and P. lefebvrei supported the hypothesis that ornamentation depends on the degree of maturity in some of these lineages. Geographical and ecological, rather than morphological data are here suggested as the most useful characters to separate the different lineages in Picoa. Further studies focusing on these features are needed before the names P. juniperi and P. lefebvrei can be unambiguously linked with the genetic lineages observed.

Genetic variability of Pisolithus isolates associated with native hosts and exotic eucalyptus in the western Mediterranean region.

• Genetic diversity and host specificity of Pisolithus is reported here in exotic (Eucalyptus) and native hosts in the western Mediterranean region. • Polymorphism in the internal transcribed spacer (ITS) sequences of the nuclear rDNA of Pisolithus was analysed. Sequences for 17 isolates associated with native Mediterranean hosts and Eucalyptus were compared with those in the GenBank DNA database using distance and parsimony methods. • Bootstrap analysis showed clustering of all Pisolithus isolates associated with Mediterranean hosts. The ITS sequences suggest the occurrence of several ecological species adapted to exploit different soil types (basic, acid and clayey slate-derived soils), with specificity for particular indigenous hosts. Isolates from eucalypt plantations in Brazil, Kenya and the Mediterranean grouped together with eucalypt-associated Australian isolates. Transfer to native hosts did not occur; the host specificity range of these exotic strains might prevent out-competition and interbreeding with local species. • Pisolithus spp. in eucalypt plantations in the Mediterranean basin are of Australian origin; the co-introduction of the ectomycorrhizal fungi might explain the success of these exotic forest plantations.

Radiocaesium and natural gamma emitters in mushrooms collected in Spain.

Mushrooms can accumulate heavy metals in general, including radionuclides found in the nature. However, little attention has been paid to the radioactive content of mushrooms collected in Spain and the dose for the population due to their ingestion. To address this, we analysed the contents of 137Cs, 40K, 226Ra and 7Be present in different species of mushrooms, according to their genus and nutritional mechanism. We observed that mycorrhizal mushrooms accumulate 137Cs more than saprophytes, and vice versa for 7Be. 40K and 226Ra are accumulated to the same degree by the two groups of mushrooms. We estimated the dose due to ingestion of mushrooms in Spain to be 2 microSv/year, and the contribution of 40K and 226Ra to be generally greater than that of 137Cs. The contribution of 137Cs to the dose was calculated by taking into account the results of an experiment carried out under the controlled laboratory conditions, which showed that approximately 98% of 134Cs was associated with the readily digestible fraction of the mushrooms.

Molecular phylogeny of the mycorrhizal desert truffles (Terfezia and Tirmania), host specificity and edaphic tolerance.

Terfezia and Tirmania, so called desert truffles, are mycorrhizal fungi mostly endemic to arid and semi-arid areas of the Mediterranean Region, where they are associated with Helianthemum species. The aim of this work was to study the phylogenetic relationships in these pezizalean hypogeous fungi. The restriction fragment length polymorphism (RFLP) and DNA sequences of internal transcribed spacers (ITS) of the nuclear rDNA were studied for several morphological species, Terfezia arenaria, T. boudieri, T. claveryi, T. leptoderma, T. terfezioides (=Mattirolomyces terfezioides), Tirmania nivea and T. pinoyi. The sequences were analyzed with distance and parsimony methods. Phylogenetic analyses indicated a close genetic relationship between Tirmania and Terfezia. They may have arisen from a single evolutionary lineage of pezizalean fungi that developed the hypogeous habit as an adaptation to heat and drought in Mediterranean ecosystems. This analysis also supports the re-establishment of the genus Mattirolomyces. The genera Tirmania and Terfezia were monophyletic, and morphological species corresponded to phylogenetic species. The Tirmania clade comprises desert truffles with smooth spores and amyloid asci, which were found in deserts. The Terfezia clade grouped species found in semi-arid habitats having ornamented and spherical spores. These species are adapted to exploit different types of soil (either acid or basic soils) in association with specific hosts (either basophilous or acidophilous species). Although other factors might also play a role, host specialization and edaphic tolerances (fungus and/or host tolerances) might be the key in the species diversity of these genera.

Ganoderma lucidum induced apoptosis in NB4 human leukemia cells: involvement of Akt and Erk.

AIM OF THE STUDY: The final goal of this work was to study the toxic and apoptosis effects induced by fractions from Ganoderma lucidum [Ganoderma lucidum (Curtis) P. Karst.; Ganodermataceae Donk] on NB4 human leukemia cells. MATERIALS AND METHODS: Two aqueous extracts and a methanol-extracted column-chromatography semipurified fraction were obtained from Ganoderma lucidum fruiting body. Flow cytometry analyses were used to measure cell viability, cell cycle and DNA fragmentation and to quantify apoptosis. Western-blot analyses were used to quantify changes in apoptosis proteins and intracellular kinases. RESULTS: Aqueous extracts slightly reduce cell viability and induce DNA fragmentation in NB4 cells. Methanol-extracted semipurified fraction at dilutions down to 15% or 40% of the initial fraction concentration reduced significantly the viability of these leukemia cells (treated for 19h) with induction of DNA fragmentation and induction of apoptosis. Overmore, the dilution down to 15% of the initial E3 concentration induced a reduction of p53 levels, of the Bcl2/Bax relationship as well as reduced levels of both unphosphorylated and phosphorylated Akt (Protein kinase Akt, protein kinase B) and Erk (Erk1 and 2). CONCLUSIONS: Induction of apoptosis and alterations in signal transduction kinases (Akt and Erk) are produced by active fractions from Ganoderma lucidum on human leukemia cells. These data could be of important relevance from the viewpoint of antitumor actions of compounds from Ganoderma lucidum. Eventual therapy applications in leukemia cells might be developed.