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Nitrogen and carbon stable isotope values for fungi from the Upton site. Ectomycorrhizal fungi are represented by shaded circles; saprotrophic fungi are represented by open squares; samples for Calostoma cinnabarinum are represented by shaded triangles.
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Calostoma cinnabarinum Corda belongs to the suborder Sclerodermatineae (Boletales), which includes many well-known ectomycorrhizal basidiomycetes, but the genus Calostoina has been described as saprotrophic. This study combines isotopic, molecular, and morphological techniques to determine the mode of nutrition of C. cinnabarinuln. delta C-13 and d...
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This study was conducted to investigate the effect of thinning on ectomycorrhizal fungal communities in a forest. Ectomycorrhizal root tips were collected from forest soils in thinning and non-thinning sites and identified using morphological characteristics and molecular analysis of ITS rDNA sequences. As a result, species richness of ectomycorrhi...
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... These morphologically unique basidiomycetes fruits grow with minimal nutrients on sandy soil with decomposing deciduous tropical forest leaves ( Figure 1B). The main species of Dipterocarp trees are Shorea and Hopea [29]. The young mushrooms are commonly eaten raw by the local people as a Thai dessert in coconut milk ( Figure 1C, left panel) and as "Koi-Hed-Ta-Lo", a spicy condiment, mixed with chili powder, roasted rice, and fish sauce ( Figure 1C, right panel). ...
... The structure of basidiospores at a microscopic level demonstrated the significance of functional morphology in relation to their ability to reproduce. The star-shaped formation served as a protective barrier for the basidiospore, shielding it from adverse conditions, aiding in its attachment to the substrate, and enabling its survival in a less than ideal environment [29]. Deloya-Olvera et al. (2023) [35] found that C. insigne differed from C. lutescens, C. ravenelii, and C. cinnabarinum in macroand micromorphology. ...
Calostoma insigne puffball mushrooms are only found in forests with rich biodiversity in very few countries including Thailand, and their biofunctions remain largely unexplored. This study used the agar disk diffusion assay, the anti-glucosidase assay, and the 3, 4, 5-dimethylthiazol-2-yl-2-5-diphenyltetrazolium bromide (MTT) assay to evaluate the bioactive potential of these endangered puffball mushrooms. Internal transcribed spacer (ITS) gene analysis identified C. insigne, a puffball mushroom with green, globose, and spiny spores. Fourier-transform infrared spectroscopy (FTIR) analysis confirmed the polysaccharide structure while scanning electron microscopy (SEM) revealed a fiber-like network. The ethanolic gelatinous fruiting body extract exhibited 1,1-diphenyl-2-picrylhydrazyl (DPPH)-scavenging capacity (57.96%), a ferric ion-reducing antioxidant power (FRAP) value of 1.73 mg FeSO4/g, and α-glucosidase inhibition (73.18%). C. insigne cytotoxicity was effective towards HT-29 colon cancer cells using the MTT assay (IC50 of 770.6 µg/mL at 72 h) and also showed antiproliferative capacity (IC50 of 297.1 µg/mL). This puffball mushroom stimulated apoptotic genes and proteins (caspase-3, Bax, and p21) via an intrinsic apoptotic pathway in HT-29 cells. In the laboratory, the medium formula consisting of 20% potato, 2% sucrose, and 0.2% peptone was optimal to increase fungal mycelial biomass (2.74 g DW/100 mL), with propagation at pH 5.0 and 30 °C. Puffball mushrooms are consumed as local foods and also confer several potential health benefits, making them worthy of conservation for sustainable utilization.
... fuscum and C. cinnabarinum) are used as edible and medicinal mushrooms by different ethnic groups in Thailand, Malaysia and Mexico (Abdullah & Rusea 2009, Alatorre 1996, Bandoni et al. 1998, Bautista-Nava & Moreno-Fuentes 2009. The ectomycorrhizal life strategy has been described only for three species of the genus in association with Fagaceae, Myrtaceae, and Nothofagaceae (Wilson et al. 2007(Wilson et al. , 2012. The genus Calostoma belongs to the family Calostomataceae, suborder Sclerodermatinae (Binder & Bresinsky 2002), and order Boletales (Gilbert 1931), according to their ribosomal, mitochondrial and nuclear DNA sequences (Hughey et al. 2000. ...
... The type species of the genus C. cinnabarinum Desvaux (1809) is ectomycorrhizal and forms a pigment called calostomal (Gruber & Steglich 2007). This species has been reported in several countries in the Americas (Wilson et al. 2007, Baseia et al. 2007, Calonge et al.2005, Arzu et al. 2012) and in China (Wilson et al. 2012). The specific epithet cinnabarinum comes from the Ancient Greek word "kinnábari" (κιννάβαρι), which refers to its "cinnabar-red" color of the spherical head of the species. ...
During a study of fungal species with biocultural importance in the Sierra Mixe in the state of Oaxaca, Mexico, specimens belonging to the genus Calostoma were collected in 2015. Morphological characteristics, electron micrographs and phylogenetic analyses of the collected specimens were used to identify the fungi at the species level. Herein we describe Calostoma naaxtutus and Calostoma tooteic as new species and compared them morphologically and phylogenetically with their most similar species, C. cinnabarinum. This constitutes one of the few records of any mushroom species that the Mexican Ayuuk jä'äy ethnic group uses.
... Some species of the genus were thought to be ectomycorrhizal symbionts; Quercus L. is believed to be the host of C. cinnabarinum, while Castanopsis Spach and Dipterocarpus C.F. Gaertn. are regarded as the hosts of C. sarasinii Lim (1969: 109) and C. berkeleyi Massee (1888: 39), respectively (Wilson & Hibbett 2006;Wilson et al. 2007;Fui et al. 2018). Although Calostoma is a small genus, it is distributed worldwide, and has attracted much attention (Minakata 1903;Kobayasi 1962;Liu 1979;Castro-Mendoza et al. 1983;Baseia et al. 2007;Wilson et al. 2007;Trierveiler-Pereira et al. 2013;Liu et al. 2018). ...
... are regarded as the hosts of C. sarasinii Lim (1969: 109) and C. berkeleyi Massee (1888: 39), respectively (Wilson & Hibbett 2006;Wilson et al. 2007;Fui et al. 2018). Although Calostoma is a small genus, it is distributed worldwide, and has attracted much attention (Minakata 1903;Kobayasi 1962;Liu 1979;Castro-Mendoza et al. 1983;Baseia et al. 2007;Wilson et al. 2007;Trierveiler-Pereira et al. 2013;Liu et al. 2018). For example, a pigment named calostomal was extracted from the fresh fruitbodies of C. cinnabarinum and is responsible for the red-orange colour of the basidiomata (Gruber & Steglich 2007;Zhou & Liu 2010). ...
Several collections of Calostoma (Calostomataceae, Boletales) from the south of China were investigated using morphology, and molecular phylogenetic analyses of DNA sequences from the nuc 28S rDNA D1-D2 domains (28S), and the nuc rDNA internal transcribed spacer (ITS). Two species of the genus are recognized, one is the previously described taxon, C. areolatum, the other, C. sinocinnabarinum, is described as new: Detailed descriptions, color photos of fresh basidiomata and line-drawings of microstructures of the two taxa are presented.
... Accurately identifying hosts can be tedious if done through inoculation studies, or misleading if done in the field. While the most straight-forward way to identify a host is by molecular means (Muir and Schl€ otterer 1999;Sato et al. 2007;Wilson et al. 2007), this step is not done routinely. This also concerns the correct and detailed annotation of sequences deposited in GenBank, which often lacks isolation source data, including geographic location and host (Vilgalys 2003;Bidartondo et al. 2008;Tedersoo et al. 2011). ...
Historical biogeography helps us keep track of organisms over time and space. However, microbial ecology and evolutionary studies are fraught with challenges due to their unknown life histories, a poor fossil record, and problematic taxonomy. Mycorrhizal fungi are key players for most terrestrial ecosystems and interact with a vast number of plants, including many woody trees that dominate temperate, tropical, and boreal ecosystems. They are also highly polyphyletic and disproportionally diverse globally and across the fungal tree of life, making them biogeographically intriguing. In this review, we focus on describing phylogenetic approaches that have been or could be applied in mycorrhizal biogeographic studies. We start by summarizing molecular-based studies and methods for species delimitation, pointing out the need for robust phylogenetic hypotheses in a phylogenomic or multi-locus framework. We describe methods for the reconstruction of ancestral states or areas and their use in biogeography, also synthesizing some of the progress with respect to generalized patterns in mycorrhizal biogeography. Next, we go through aspects related to time-calibrating fungal phylogenies and their time-of-origin, as well as downstream analyzes involving the estimation of diversification rates. Diversification rate, trait evolution, and phylo-community analyses are also put in the context of relevant evolutionary ecology hypotheses. Finally, we dedicate a section to methodological biases and caveats, which are often associated with global mycological fieldwork and sampling, and some phylogenetic methods.
... Because the carbon and nitrogen sources of saprotrophic and ectomycorrhizal fungi often differ (Lindahl et al. 2007;Hobbie et al. 2014a, b), another tool uses natural isotopic tracers to distinguish between these two functional groups. Stable isotope measurements use small differences in ratios of 13 C: 12 C and 15 N: 14 N (expressed as δ 13 C and δ 15 N values) among different sources as markers for those sources (Hobbie et al. 2001, Wilson et al. 2007. They also can be used as markers of differences between saprotrophic and ectomycorrhizal fungi in how they process nitrogen and carbon within fungi once assimilated (Hobbie et al. 2005(Hobbie et al. , 2012. ...
We assessed the nutritional strategy of true morels (genus Morchella) collected in 2003 and 2004 in Oregon and Alaska, 1 or 2 y after forest fires. We hypothesized that the patterns of stable isotopes (δ13C and δ15N) in the sporocarps would match those of saprotrophic fungi and that radiocarbon (∆14C) analyses would indicate that Morchella was assimilating old carbon not current-year photosynthate. We compared radiocarbon and stable isotopes in Morchella with values from concurrently collected foliage, the ectomycorrhizal Geopyxis carbonaria (Alb. & Schwein.) Sacc., the saprotrophic Plicaria endocarpoides (Berk.) Rifai, and with literature to determine isotopic values for ectomycorrhizal or saprotrophic fungi. Geopyxis, Plicaria and Morchella, respectively, were 3‰, 5‰ and 6‰ higher in 13C than foliage and 5‰, 7‰ and 7‰ higher in 15N. High 15N enrichment in Morchella indicated that recent litter was not the primary source for Morchella nitrogen, and similar 13C and 15N enrichments to Plicaria suggest that Morchella assimilates its carbon and nitrogen from the same source pool as this saprotrophic fungus. From radiocarbon analyses Morchella averaged 11 ± 6 y old (n = 19), Plicaria averaged 17 ± 5 y old (n = 3), foliage averaged 1 ± 2 y old (n = 8) and Geopyxis (n = 1) resembled foliage in ∆14C. We conclude that morels fruiting in postfire environments in our study assimilated old carbon and were saprotrophic.
... In general, stable carbon and nitrogen isotope ratios (expressed as δ 13 C and δ 15 N values) in ECM are usually, but not always, lower in δ 13 C and higher in δ 15 N than in saprotrophic fungi (Gebauer and Taylor 1999;Hobbie 2005;Hobbie et al. 1999Hobbie et al. , 2001Hobbie et al. , 2006Kohzu et al. 1999;Henn and Chapela 2001;Taylor et al. 2003;Mayor et al. 2009;Seitzman et al. 2011;Hou et al. 2012). Therefore, patterns in stable carbon and nitrogen isotope values can be used to assign fungal trophic status in taxa where status is uncertain (Hobbie et al. 2001;Wilson et al. 2007;Mayor et al. 2009;Hou et al. 2012). ...
Phlebopus portentosus is one of the most popular wild edible mushrooms in Thailand and can produce sporocarps in the culture without a host plant. However, it is still unclear whether Phlebopus portentosus is a saprotrophic, parasitic, or ectomycorrhizal (ECM) fungus. In this study, Phlebopus portentosus sporocarps were collected from northern Thailand and identified based on morphological and molecular characteristics. We combined mycorrhizal synthesis and stable isotopic analysis to investigate the trophic status of this fungus. In a greenhouse experiment, ECM-like structures were observed in Pinus kesiya at 1 year after inoculation with fungal mycelium, and the association of Phlebopus portentosus and other plant species showed superficial growth over the root surface. Fungus-colonized root tips were described morphologically and colonization confirmed by molecular methods. In stable isotope measurements, the δ(13)C and δ(15)N of natural samples of Phlebopus portentosus differed from saprotrophic fungi. Based on the isotopic patterns of Phlebopus portentosus and its ability to form ECM-like structures in greenhouse experiments, we conclude that Phlebopus portentosus could be an ECM fungus.
... Hydnomerulius pinastri (formerly Leucogyrophana), a single species that is not placed in any of the suborders, is sister to the remaining Boletineae members (Jarosch and Besl 2001). Current research is focused on the ecology of Sclerodermatineae ( Wilson et al. 2007Wilson et al. , 2012) and the taxonomic structure of Boletineae, especially the Boletaceae. Monographic work has led to a better definition of Boletus and its being restricted to the B. edulis group ( Dentinger et al. 2010;Nuhn et al. 2013), the revision of gilled boletes in Phylloporus ( Neves et al. 2012), and Xerocomus, which has been split into several new genera (S ˇ utara 2008). ...
Agaricomycetes includes ca. 21,000 described species of mushroom-forming fungi that function as decayers, pathogens, and mutualists in both terrestrial and aquatic habitats. The morphological diversity of Agaricomycete fruiting bodies is unparalleled in any other group of fungi, ranging from simple corticioid forms to complex, developmentally integrated forms (e.g., stinkhorns). In recent years, understanding of the phylogenetic relationships and biodiversity of Agaricomycetes has advanced dramatically, through a combination of polymerase chain reaction-based multilocus phylogenetics, phylogenomics, and molecular environmental surveys. Agaricomycetes is strongly supported as a clade and includes several groups formerly regarded as Heterobasidiomycetes, namely the Auriculariales, Sebacinales, and certain Cantharellales (Tulasnellaceae and Ceratobasidiaceae). The Agaricomycetes can be divided into 20 mutually exclusive clades that have been treated as orders. This chapter presents an overview of the phylogenetic diversity of Agaricomycetes, emphasizing recent molecular phylogenetic studies.
... Literature – Baseia et al. (2006: 114, 2007: 278), Cortez (2009: 03), Rick (1961b: 456, as Mitremyces zanchianus Rick). Comments – This species is considered putative ectomycorrhizal since its close relative C. cinnabarinum was proven to be mycorrhizal with oaks (Wilson et al. 2007). However, no direct evidence exists for the ectomycorrhizal status of C. zanchianum. ...
Sulzbacher MA, Grebenc T, Jacques RJS, Antoniolli ZI 2013 – Ectomycorrhizal fungi from southern Brazil – a literature-based review, their origin and potential hosts. Mycosphere 4(1), 61– 95, Doi 10.5943 /mycosphere/4/1/5 A first list of ectomycorrhizal and putative ectomycorrhizal fungi from southern Brazil (the states of Rio Grande do Sul, Santa Catarina and Paraná), their potential hosts and origin is presented. The list is based on literature and authors observations. Ectomycorrhizal status and putative origin of listed species was assessed based on worldwide published data and, for some genera, deduced from taxonomic position of otherwise locally distributed species. A total of 144 species (including 18 doubtfull species) in 49 genera were recorded for this region, all accompanied with a brief distribution, habitat and substrate data. At least 30 collections were published only to the genus level and require further taxonomic review.
... However, our cloning of PCR amplicons from ectomycorrhizae at Sicamous Creek indicates that A. purpurea is an ECM fungus; additionally, the finding that it was much more abundant (relative to other ECM taxa) on root tips than as hyphae suggests that it is an especially successful colonizer of fine roots. The ECM fungus Coltricia perennis was recently included in the Hymenochaetoid clade along with A. purpurea (Dentinger and McLaughlin, 2006), and the nutritional mode of Calostoma cinnabarium, a member of the Boletales previously thought to be saprotrophic, was recently confirmed to be ECM based on its 13 C signature (Wilson et al., 2007). The ␦ 13 C and ␦ 15 N of A. purpurea sporocarps from our study was within the range of other ECM fungi from the site, but was peripheral enough in that range that a discriminant analysis did not classify it as such. ...
Shifts in ectomycorrhizal (ECM) fungal community composition occur after clearcut logging, resulting in the loss of some dominant forest fungi. Because decaying wood is a remnant of the original forest and an important habitat for ECM fungal species, we examined ECM spruce roots and hyphae in 1-ha coarse woody debris (CWD) retention and removal plots (N = 3) at a high elevation spruce forest 12 and 13 years after clearcut harvesting to test for a medium-term effect of CWD retention. Root tips from ten Picea engelmannii (Parry ex. Engelm.) saplings per plot were grouped morphologically, and the ECM fungal symbiont identified by Sanger sequencing. Sand-filled hyphae-trapping mesh bags were buried amongst the roots of saplings for one year. PCR product from all bags per plot was pooled and submitted for GS-FLX Titanium sequencing. Forty-six of 89 operational taxonomic units (OTUs) from root tips and 50 OTUs from hyphae were identified as ECM taxa. The most abundant taxa amongst root tip OTUs were Thelephora terrestris, Alloclavaria purpurea, Amphinema byssoides, and Tylospora asterophora. The most abundant OTUs from mesh bags were ectomycorrhizal taxa, and the most abundant of these were A. byssoides, T. terrestris, Wilcoxina mikolae, and T. asterophora. The retention of CWD had no detectable effect on taxon richness, evenness or diversity of ECM fungi on root tips or in mesh bags; however, there was a detectable shift in species composition. The relative abundance of A. byssoides root tips was significantly higher at removal plots while A. purpurea was a significant indicator species for CWD removal. Our results suggest that the retention of CWD at the time of harvest has affected ECM habitat at this site, and has resulted in altered ECM species composition, even though the logs are still hard and intact.
... Most Sclerodermatineae are considered to be ectomycorrhizal (Binder & Hibbett, 2006;Wilson et al., 2007). For example, Pisolithus and Scleroderma are used in reforestation projects because they can form ectomycorrhizas with multiple species of host trees (Molina & Trappe, 1982b;Danielson, 1984;Sanon et al., 2009). ...
... However, the methods used to identify ectomycorrhizal hosts vary widely among studies. Astraeus, Calostoma, Pisolithus and Scleroderma have been conclusively shown to form ectomycorrhizas with angiosperms and gymnosperms through synthesis studies (Molina, 1981;Molina & Trappe, 1982a;Danielson, 1984) and molecular analyses (Tedersoo et al., 2007;Wilson et al., 2007). Unfortunately, there are many cases where the determination of ectomycorrhizal host is based on the observed proximity of fungus and putative hosts. ...
... Fungal and plant DNA were isolated from ectomycorrhizal roots. PCR and cycle sequencing of fungal DNA from ectomycorrhizal root tips followed protocols described in Wilson et al. (2007), while analyses of plant DNA used primers rbcL-F1 and rbcL-R1 and protocols described in Sato et al. (2007) to amplify the ribulose biphosphate carboxylaze chloroplast gene (rbcL). Fungal ITS and plant rbcL sequences were used as queries in BLAST searches of GenBank. ...
This study uses phylogenetic analysis of the Sclerodermatineae to reconstruct the evolution of ectomycorrhizal host associations in the group using divergence dating, ancestral range and ancestral state reconstructions. Supermatrix and supertree analysis were used to create the most inclusive phylogeny for the Sclerodermatineae. Divergence dates were estimated in BEAST. Lagrange was used to reconstruct ancestral ranges. BayesTraits was used to reconstruct ectomycorrhizal host associations using extant host associations with data derived from literature sources. The supermatrix data set was combined with internal transcribed spacer (ITS) data sets for Astraeus, Calostoma, and Pisolithus to produce a 168 operational taxonomic unit (OTU) supertree. The ensuing analysis estimated that basal Sclerodermatineae originated in the late Cretaceous while major genera diversified near the mid Cenozoic. Asia and North America are the most probable ancestral areas for all Sclerodermatineae, and angiosperms, primarily rosids, are the most probable ancestral hosts. Evolution in the Sclerodermatineae follows the biogeographic history of disjunct plant communities associated with early Cenozoic mesophytic forests and a boreotropical history. Broad geographic distributions are observed in the most promiscuous Sclerodermatineae (those with broad host ranges), while those with relatively limited distribution have fewer documented ectomycorrhizal associations. This suggests that ectomycorrhizal generalists have greater dispersal capabilities than specialists.
