Nuclear conditions of basidiospores and hyphal cells in the edible mushroom Oudemansiella aparlosarca.

Oudemansiella aparlosarca is an edible mushroom possessing medicinal and health benefits. Although there are studies on the cultivation of O. aparlosarca, only a few studies have focused on its genetics and life cycle. Therefore, the main objective of this study was to identify the nuclear conditions of basidiospores and homokaryotic and heterokaryotic hyphal cells and to determine the influence of different nuclear conditions on basidiospore diameter in O. aparlosarca. Two parental strains: strain-55 and strain-81 were used. Staining of basidiospores and hyphal cells in the apical region was performed. We observed the following nuclear conditions: non-nucleate, mononucleate, binucleate, and multinucleate. In both parental strains, binucleate spores were predominant, while the number of non-nucleate spores was the lowest. The diameter of non-nucleate spores was the smallest, being 11.52 µm and 12.15 µm in parental strain-81 and strain-55, respectively, while multinucleate spores had the largest diameter, being 14.78 µm in both parental strains. Both homokaryotic and heterokaryotic strains were identified in isolated single spores from parental strains. Binucleate cells were majorly present in heterokaryotic hyphal cells, and multinucleate cells were predominant in homokaryotic hyphal cells. We conclude that O. aparlosarca contains homokaryotic and heterokaryotic basidiospores, which indicates an amphithallic life cycle. The observed binucleate spores might be the result of post-meiotic mitosis.

A precise relationship among Buller's drop, ballistospore, and gill morphologies enables maximum packing of spores within gilled mushrooms.

Basidiomycete fungi eject basidiospores using a surface tension catapult. A fluid drop forms at the base of each spore and, after reaching a critical size, coalesces with the spore and launches it from the gill surface. It has long been hypothesized that basidiomycete fungi pack the maximum number of spores into a minimal investment of biomass. Building on a nascent understanding of the physics underpinning the surface tension catapult, we modeled a spore's trajectory away from a basidium and demonstrated that to achieve maximum packing the size of the fluid drop, the size of the spore, and the distance between gills must be finely coordinated. To compare the model with data, we measured spore and gill morphologies from wild mushrooms and compared measurements with the model. The empirical data suggest that in order to pack the maximum number of spores into the least amount of biomass, the size of Buller's drop should be smaller but comparable to the spore size. Previously published data of Buller's drop and spore sizes support our hypothesis and also suggest a linear scaling between spore radius and Buller's drop radius. Morphological features of the surface tension catapult appear tightly regulated to enable maximum packing of spores. If mushrooms are maximally packed and Buller's drop radii scale linearly with spore radii, we predict that intergill distance should be proportional to spore radius to the power 3/2.

Ultrastructure and development of acanthocytes, specialized cells in Stropharia rugosoannulata, revealed by scanning electron microscopy (SEM) and cryo-SEM.

Acanthocytes are special cells with a distinct spiky shape produced exclusively by the fungi of Stropharia and can be used to defend against nematodes. In the present study, the ultrastructure and development of acanthocytes were revealed by scanning electron microscopy (SEM) and cryo-SEM in S. rugosoannulata, a popular cultivated mushroom both in China and Europe. The acanthocytes were abundant on the surface of rhizomorph, casing soils, and vegetative mycelia of homokaryotic and heterokaryotic strains in S. rugosoannulata. The development of the acanthocyte was investigated with cryo-SEM, which has distinct advantage for observation of the ultrastructure of live, hydrated structures. Three distinct stages, including formation of lateral branch that was covered with patches, spiky structure formation, and maturation of acanthocytes, were identified and described. The irregular patches deposited on the surface of lateral branches and the holes in the spiky branches of the acanthocytes were reported for the first time. The environmental nitrogen level showed impact on acanthocyte production, but it seemed not to be the indispensable factor. Acid medium could delay the initiation of the acanthocyte formation but did not affect the overall morphology and structure, indicating that the central deposit of acanthocytes should be acid soluble. Acanthocytes of S. rugosoannulata have similar hydrophobicity to mycelia. The observation of ultrastructure and development process of acanthocytes provides insights into the ecological function and evolution of this special structure.

Actualización del conocimiento del género Cyttaria Berk. (Cyttariales, Ascomycota) en Chile / Update of the knowledge of the genus Cyttaria Berk (Cyttariales, Ascomycota) in Chile

El género Cyttaria pertenece a la familia Cyttariaceae y sus especies son parásitos obligados de árboles del género Nothofagus. Se distribuye naturalmente en el hemisferio sur, encontrándose siete especies de Cyttaria presentes en nuestro país: C. berteroi, C. darwinii, C. espinosae, C. hariotii, C. hookeri, C. johowii y C. exigua, que comúnmente se conocen como "Digüeñes" y que tienen una gran importancia desde el punto de vista alimenticio, ya que han sido consumidos desde la prehistoria por pueblos originarios. En Chile se han realizado estudios sobre la taxonomía, ecología y propiedades medicinales de algunas especies del género Cyttaria y en esta revisión, se describen los caracteres macro y microscópicos e información relevante de las siete especies de este género presentes en nuestro país, de acuerdo a la literatura disponible y observaciones personales de la autora. Además, se presenta un resumen sobre los resultados de un estudio reciente de las propiedades bioactivas de las especies más consumidas en nuestro país. (AU)

The genus Cyttariabelongs to the family Cyttariaceae; its species are obligate parasites of trees of the genus Nothofagus. It is naturally distributed in the Southern Hemisphere, with seven species of Cyttariapresent in Chile: C. berteroi, C. darwinii, C. espinosae, C. hariotii, C. hookeri, C. johowiiand C. exigua, which are commonly known as "Digüeñes" and are of great nutritional importance, since they have been consumed from prehistory by native people. Studies have been carried out in Chile on the taxonomy, ecology and medicinal properties of some species of the genus Cyttaria. In this review, the macro and microscopic characteristics and relevant information of the seven species of this genus present in our country are described, according to the available literature and personal observations of the author. A summary of the results of a recent study of the bioactive properties of the most consumed species in our country is also presented. (AU)

Medicinal Properties of Clitocybe brunneocaperata (Agaricomycetes) from India.

The present research documented the first evidence of Clitocybe brunneocaperata from Tripura, Northeast India. The main purposes of this study were identification of the mushroom and qualitative toxicity screening along with antibacterial and antioxidant activities of its methanolic extracts. The species has been recognized on the basis of the morphometric and molecular studies. Toxicity test revealed its inedibility nature. Methanolic extracts of C. brunneocaperata showed potent antibacterial, free radical scavenging effect (65.42%), chelating effects on ferrous ion (65.93%), total phenolic content (0.028 mg GAE/g), and flavonoid content (0.013 mg CE/g). The lowest EC50 values of the free radical scavenging effect (5.94 mg/ml) and chelating effects on ferrous ion (0.072 mg/ml) were observed at 16 and 1.5 mg/ml concentrations, respectively, which implied stronger capability of free radical scavenging. Therefore, further research is also required to isolate and characterize the principal components of C. brunneocaperata.

Multiple evolutionary origins of sequestrate species in the agaricoid genus Chlorophyllum.

Chlorophyllum accommodates lamellate agaricoid species as well as sequestrate angiocarpic taxa, which do not form a monophyletic lineage within the genus. To clarify phylogenetic affinities and delimit species boundaries among sequestrate representatives of the genus, we analyzed historical and contemporary material from a broad geographic range, encompassing North America, southern Africa, eastern Asia, the Greek and Iberian peninsulas, and the Mediterranean islands of Cyprus and Lesvos. Six sequestrate lineages of Chlorophyllum were identified, which appear to have evolved in at least three independent gasteromycetation events. Multigene analysis of the nuc rDNA internal transcribed spacer region ITS1-5.8S-ITS2 (ITS) and portions of the nuc 28S rDNA (28S), RNA polymerase II second largest subunit (rpb2), and translation elongation factor 1-α (tef1) genes revealed the presence of a previously undescribed species, introduced here as Chlorophyllum levantinum, sp. nov. Its sister species C. lusitanicum, previously known only from Spain, is shown to have a wider distribution throughout the Mediterranean basin. A South African collection of Secotium gueinzii, a poorly known taxon described in 1840 and seldom appearing in literature since, was also shown to nest within Chlorophyllum. An epitype for this rare species is designated, Secotium becomes a priority synonym of Chlorophyllum, and nomenclatural implications are discussed. The phylogenetic placement of C. arizonicum is confirmed after successful sequencing of the century-old holotype and an undescribed sister lineage of this species detected. Emended descriptions of sections Chlorophyllum, Endoptychorum, and Sphaerospororum are provided to reflect current results, along with updated descriptions and extensive imagery for all known sequestrate taxa of Chlorophyllum.

Clitopilus, Clitocella, and Clitopilopsis in China.

Within the family Entolomataceae, many reports of Entoloma and Rhodocybe have been published, but few reports on Clitopilus and allied genera are available. In this study, Clitopilus, Clitocella, and Clitopilopsis from China were studied by morphological and phylogenetic methods. Portions of nuc 28S rDNA (28S), RNA polymerase II second largest subunit (rpb2), translation elongation factor 1-alpha (tef1), and ATPase subunit 6 (atp6), were employed to elucidate the relationships of Clitopilus and allies. Results indicate that Clitopilus should be divided into three sections, namely, sect. Clitopilus, sect. Scyphoides, and sect. Crispi, the last of which is newly described. In total, 17 species of Clitopilus, Clitocella, and Clitopilopsis are found in China; 14 species belong to Clitopilus, of which 5 are new: C. yunnanensis, C. brunniceps, C. rugosiceps, C. sinoapalus, and C. umbilicatus. Two species of Clitocella, including one new species, Clitocella orientalis, are described. One new species of Clitopilopsis, namely, Clitopilopsis albida, is proposed.

New species and reports of Cuphophyllus from northern North America compared with related Eurasian species.

This study describes four gray or brown species of Cuphophyllus (Hygrophoraceae, Agaricales), two of them new species, restricted to arctic-alpine and northern boreal zones of North America, and relates them morphologically and phylogenetically using multigene and nuc rDNA internal transcribed spacer ITS1-5.8S-ITS (ITS barcode) analyses to their similar, known counterparts. Cuphophyllus cinerellus, epitypified here, is shown to be a pan-palearctic species with sequence-confirmed collections from Fennoscandia and easternmost Asia. Occupying a similar habitat in the Nearctic is its sister species, the morphologically similar but novel C. esteriae, so far known only from eastern North America, including Greenland. Sister to the C. cinerellus-C. esteriae lineage, and known only from boreal raised Sphagnum bogs in Newfoundland, is a new medium-sized light cinereous brown species, C. lamarum. It has a yellow stipe but is phylogenetically distant from the yellow-stiped European C. flavipes and its North American sister species, Hygrophorus pseudopallidus. As cryptic speciation was discovered within C. flavipes, we lecto- and epitypify the name and transfer H. pseudopallidus to Cuphophyllus based on ITS analysis of the holotype. We also transfer the small European Hygrocybe comosa to Cuphophyllus based on morphology. Cuphophyllus hygrocyboides is reported from North America with the first sequence-confirmed collections from arctic-alpine British Columbia and Greenland. In addition, sequencing the holotype of C. subviolaceus identifies it as the sister species to the putative C. lacmus. Both species seem to have an intercontinental distribution. In total, we add new sequences to GenBank from 37 Cuphophyllus collections, including the holotypes of C. hygrocyboides and C. subviolaceus, the two new epitypes, and the two novel species.

Adventurous cuisine in Laos: Hebeloma parvisporum, a new species in Hebeloma section Porphyrospora.

Hebeloma parvisporum is described as new and placed within H. sect. Porphyrospora. This mushroom is sold as an edible in markets of Laos under the local name "wai khom." Hebeloma sect. Porphyrospora is discussed and expanded to include the species formerly included in the genus Anamika and recently transferred to Hebeloma. Hebeloma sect. Porphyrospora currently comprises 16 species, 14 of which are known only from the western Pacific and Indian subcontinent. All species in this section share the character of having red-brown spores when fresh, atypical for other sections of Hebeloma, which causes the lamellae to be red-brown. However, this red-brown color fades when the material is dried. The close links, morphologically and molecularly, between H. parvisporum and other members of H. sect. Porphyrospora, particularly H. victoriense, are shown.

Cortinarius section Thaumasti in South American Nothofagaceae forests.

We studied the South American species of Cortinarius section Thaumasti based on morphological and molecular data. Members of this group can easily be identified in the field because the basidiomata are small and Phlegmacium-like with a bulbous stipe and the universal veil in most species forms a distinct volva at the base of the stipe. The phylogenetic delimitation of the clade was mostly in concordance with the earlier, morphology-based grouping of the South American taxa except that C. chrysophaeus was resolved outside of the clade. Altogether nine species were recognized in the section. Four species, C. chlorophanus, C. coleopus, C. cosmoxanthus, and C. vaginatus, were previously described by other authors, whereas three species, C. chlorosplendidus, C. olivaceovaginatus, and C. subcosmoxanthus, are described here as new. We were able to identify two remaining taxa, but we do not have sufficient morphological data to allow for a formal description. All of the species in C. section Thaumasti form ectomycorrhizal associations with Nothofagaceae. They have been documented from South America and New Zealand. The Patagonian species are considered endemic to the region. A key to the described species is provided.

Rare Fungal Keratitis Caused by Coprinellus Radians.

A case of fungal keratitis due to Coprinellus radians is reported. To our knowledge, fungal keratitis caused by this species was rare. Fungal hyphae were detected in corneal scrapings, and isolates were identified by morphology and by sequencing the internal transcribed spacer region of ribosomal DNA. The patient was treated with systemic and local antifungal therapy for 5 days, and lamellar keratoplasty was performed after no obvious improvement in symptoms. The in vitro antifungal susceptibilities of the case strain were tested for six antifungal agents. The results showed that 5-fluorouracil was resistant, fluconazole was moderately sensitive, and the other drugs assayed (amphotericin B, posaconazole, itraconazole and voriconazole) were highly effective against this fungus.

Nodulose-spored Inocybe from the Rocky Mountain alpine zone molecularly linked to European and type specimens.

Inocybe (Inocybaceae) is one of the most diverse ectomycorrhizal genera in arctic and alpine habitats where the primary hosts are Salix, Betula, and Dryas. Subgenus Inocybe is common in these habitats and typically characterized by the presence of thick-walled pleurocystidia. Here, we focus on species that have angular or nodulose spores. Historically, over 30 taxa from this group have been reported from arctic and alpine habitats. Many names have been synonymized, whereas molecular analysis has revealed new species. Nuc rDNA internal transcribed spacer ITS1-5.8S-ITS2 (ITS) sequence data of 26 type specimens in this group now allow for further taxonomic clarification and comparison across continents of disjunct populations. Here, we compare ITS sequence data and the D1-D2 portion of nuc 28S rDNA (28S) from Rocky Mountain specimens with those of types and European reference material. We report 10 species from the Rocky Mountain alpine zone, all of which are conspecific with known European boreal, montane, or alpine species, and four are described as new; all have intercontinental distributions. Nodulose-spored Inocybe taxa that occur in the Rocky Mountain alpine zone include I. alpinomarginata, sp. nov., I. arctica, I. giacomi, I. leonina, I. murina, sp. nov., I. occulta, I. paragiacomi, sp. nov., I. phaeocystidiosa, I. purpureobadia, and I. subgiacomi, sp. nov. Remarkably, these species occur at elevations up to 4000 m and at latitudes as low as 36°N, hundreds of miles from the Arctic, the European alpine, and original type localities. Distributions are explained in part by host distributions and historical glaciation patterns. A key and full descriptions for Rocky mountain species are provided to promote species recognition.

Hydrolysis of oligosaccharides by a fungal α-galactosidase from fruiting bodies of a wild mushroom Leucopaxillus tricolor.

A novel acidic α-galactosidase (EC 3.2.1.22) designated as Leucopaxillus tricolor α-galactosidase (LTG) has been purified to homogeneity from the fruiting bodies of L. tricolor to 855-fold with a specific activity of 956 U mg-1 by the application of chromatography and gel filtration. The molecular mass of LTG was estimated to be 60 kDa as determined by both SDS-PAGE and by gel filtration. The purified enzyme was identified by LC-MS/MS and four inner amino acid sequences were obtained. When 4-nitrophenyl α-D-glucopyranoside (pNPGal) was used as substrate, the optimal pH and optimal temperature of LTG were pH 5.0 and 50 °C, respectively. The enzyme activity was strongly inhibited by Hg2+ , Fe3 , Cu2+ , Cd2+ , and Mn2+ ions. The chemical modification agent N-bromosuccinimide (NBS) completely inhibited the enzyme activity of LTG, indicating the paramount importance of tryptophan residue(s) to its enzymatic activity. Besides, LTG displayed wide substrate diversity with activity toward a variety of substrates such as stachyose, raffinose, melibiose, locust bean gum, and guar gum. Given the good ability of degrading the non-digestible and flatulence-causing oligosaccharides, this fungus may become a useful source of α-galactosidase for multiple applications.

Hymenogaster macmurphyi and Splanchnomyces behrii are sequestrate species of Xerocomellus from the western United States.

Hymenogaster is an ectomycorrhizal genus of brown-spored sequestrate fungi that is related to the mushroom-forming genera Hebeloma and Alnicola (Agaricales). However, because of difficulties in morphological taxonomy of sequestrate fungi, Hymenogaster has become a polyphyletic repository for a variety of unrelated brown-spored sequestrate species. During studies of ectomycorrhizal ecology and sequestrate fungal evolution in the western USA, we encountered specimens of a morphologically unique species. It was originally described as Hymenogaster macmurphyi, but our morphological and molecular analyses indicate that it is not closely related to Hymenogaster. Phylogenetic analyses of multiple gene regions indicate that H. macmurphyi is actually a member of the Boletineae (Boletales, Basidiomycota) and is nested within the epigeous genus Xerocomellus, distantly related to any of the other known genera of sequestrate Boletales. While examining additional herbarium collections, we came upon isotype material of Splanchnomyces behrii, which represents a closely related species. Here we document the morphology and phylogenetic affinities of these unusual sequestrate Boletineae and transfer both species to Xerocomellus as X. macmurphyi and X. behrii. During our study, we also noted that the sequestrate taxon Rhopalogaster transversarius is nested within the epigeous genus Suillus.

Morphological and molecular evidence for two new species in Lepiota from China.

Two new species in Lepiota sect. Ovisporae are described from tropical China. Lepiota angusticystidiata has a pileus with brownish yellow squamules covering a trichodermial palisade, ellipsoidal basidiospores, and narrowly clavate cheilocystidia; L. brunneosquamulosa has a tomentose, squamulose pileus with brownish yellow to yellowish brown concentric zones covering a trichodermial palisade, rarely with short elements, ellipsoidal or subcylindrical basidiospores with straight adaxial side, and no cheilocystidia. Phylogenetic relationships among species of sect. Ovisporae were inferred based on DNA sequences of the internal transcribed spacer region (ITS1-5.8S-ITS2 = ITS), the 5' end of the 28S gene (D1-D2-D3 variable domains), and partial sequences of the intergenic spacer (IGS1) of the nuc rDNA and the mitochondrial small ribosomal RNA gene (mtSSU), supporting the delineation of these new species.

The Inocybe geophylla group in North America: a revision of the lilac species surrounding I. lilacina.

The Inocybe geophylla group is circumscribed based on phylogenetic analysis of DNA sequences largely sampled from North America and Europe. Twenty-nine phylogenetic species are uncovered after analysis of combined nuc 28S rDNA (28S) and RNA polymerase II second largest subunit (rpb2) DNA sequence data. Species in the I. geophylla group share the presence of a cortina, silky-fibrillose pileus and stipe, pruinose stipe apex, spermatic odor, thick-walled hymenial cystidia, and smooth amygdaliform or elliptical basidiospores. Within the group, as many as five phylogenetic species attributable to I. lilacina and allies form a strongly supported clade based on analysis of nuc ITS1-5.8S-ITS2 rDNA (ITS [internal transcribed spacer]), 28S, and rpb2 data. However, all lilac-colored species do not form a monophyletic group. Sufficient morphological and ecological data are present to document four of the I. lilacina subgroup species, two of which are described from North America as new: I. ionocephala and I. sublilacina. Inocybe lilacina is recircumscribed based on sequencing the holotype and is distributed in the eastern United States under pines and/or hardwoods. Inocybe pallidicremea is a widespread and common conifer associate in mostly northern parts of North America, to which the name I. lilacina was previously applied. Descriptions, photographs, line drawings, and a taxonomic key to lilac species in the I. lilacina subgroup from North America are provided. Well-documented collections, especially notes on gross morphology and ecology, are needed to continue to assess and describe the high taxonomic variation in the I. lilacina subgroup and its allies worldwide.

Morphological variants of Moniliophthora roreri on artificial media and the biotroph/necrotroph shift.

Moniliophthora roreri (Mr) causes frosty pod rot of Theobroma cacao in a hemibiotrophic association. The Mr biotroph-like phase has not been studied in culture. Mr spores (isolates Co12, Co52, and B3) were germinated on high (V8) and low (BPMM) nutrients with different media hardness (0.5% to 3% agarose). Germination was high on V8 media. Hardness affected germination on BPMM. Most colonies on V8 were slow-growing, failing to sporulate. Colony morphology depended on the isolate. On BPMM, exaggerated mycelia formed of limited length with enlarged cells. On agarose, rapidly expanding sporulating necrotrophic colonies formed rarely. Co12 and B3 spores were germinated on V8 and BPMM with low melting point (LMP) agarose. Slow-growing colonies of B3 on BPMM were unstable on LMP agarose, often forming slow-growing/rapidly expanding hybrids. Slow-growing colonies are hypothesized to represent the biotrophic phase. One nucleus was common in Mr cells, other than spores. Binucleate cells were occasionally observed in aged cells of slow-growing mycelia. Co52 cells often had more than two nuclei per cell after germination. Mr mycelia cells typically carry a single nucleus, being considered haploid. Biotroph- and necrotroph-like mycelia displayed differential gene expression but results were inconsistent with published in vivo results and require further study.

Anatomical characterization of PDF-tri neurons and peptidergic neurons associated with eclosion behavior in Drosophila.

The peptidergic Pigment-dispersing factor (PDF)-Tri neurons are a group of non-clock neurons that appear transiently around the time of adult ecdysis (=eclosion) in the fruit fly Drosophila melanogaster. This specific developmental pattern points to a function of these neurons in eclosion or other processes that are active around pupal-adult transition. As a first step to understand the role of these neurons, we here characterize the anatomy of the PDF-Tri neurons. In addition, we describe a further set of peptidergic neurons that have been associated with eclosion behavior, eclosion hormone (EH), and crustacean cardioactive peptide (CCAP) neurons, to single cell level in the pharate adult brain. PDF-Tri neurons as well as CCAP neurons co-express a classical transmitter indicated by the occurrence of small clear vesicles in addition to dense-core vesicles containing the peptides. In the tritocerebrum, gnathal ganglion and the superior protocerebrum PDF-Tri neurites contain peptidergic varicosities and both pre- and postsynaptic sites, suggesting that the PDF-Tri neurons represent modulatory rather than pure interneurons that connect the subesophageal zone with the superior protocerebrum. The extensive overlap of PDF-Tri arborizations with neurites of CCAP- and EH-expressing neurons in distinct brain regions provides anatomical evidence for a possible function of the PDF-Tri neurons in eclosion behavior.

Morphological and molecular characterization of coprinoid fungi newly recorded for the mycobiota of Iran.

Twenty-five specimens of coprinoid fungi were collected during an ongoing survey of agaric fungi in Kermanshah Province, western Iran. The specimens were identified based on morphological characteristics and molecular analysis of internal transcribed spacer sequences. Five species of Coprinellus viz C. disseminates, C. flocculosus, C. micaceus, C. radians and C. xanthothrix, three species of Coprinopsis viz C. atramentaria, C. insignis and C. semitalis and two species of Coprinus viz C. pinetorum and C. sterquilinus were identified. Among the species identified in this research, three unreported species from Iran namely C. insignis, C. semitalis and C. pinetorum are reported. Detailed morphological descriptions and illustrations of this three newly-recorded species were provided here and their evolutionary relationships were presented by the constructing of a phylogenetic tree.

The genus Parasola: phylogeny and the description of three new species.

Parasola represents an enigmatic lineage of veil-less, coprinoid fungi in Psathyrellaceae (Agaricales). The species-level taxonomy of the genus has been in a flux recently, resulting in the elimination of some long-established names and the description of new taxa. Here, we reconstruct the phylogeny of Parasola using two nuc rDNA loci, the internal transcribed spacer region (ITS1-5.8S-ITS2) and 28S and identify several putatively undescribed species, of which three are formally described here (Parasola crataegi, P. ochracea, and P. plicatilis-similis) based on molecular and morphological data. Morphological descriptions for the new species and an identification key to accepted Parasola species are given. We revise and discuss our current understanding of the phylogeny of Parasola.