Echinodontium Tinctorium Research Articles

Antiproliferative Fatty Acids Isolated from the Polypore Fungus Onnia tomentosa.

Onnia tomentosa is a widespread root rot pathogen frequently found in coniferous forests in North America. In this study, the potential medicinal properties of this wild polypore mushroom collected from north-central British Columbia, Canada, were investigated. The ethanol extract from O. tomentosa was found to exhibit strong antiproliferative activity. Liquid-liquid extraction and bioactivity-guided fractionation, together with HPLC-MS/MS and 1D/2D NMR analyses of the ethanol extract of O. tomentosa, led to the identification of eight known linoleic oxygenated fatty acids (1.1-1.4 and 2-5), together with linoleic (6) and oleic acids (7). The autoxidation of linoleic acid upon isolation from a natural source and compound 5 as an autoxidation product of linoleic acid are reported here for the first time. GC-FID analysis of O. tomentosa, Fomitopsis officinalis, Echinodontium tinctorium, and Albatrellus flettii revealed linoleic, oleic, palmitic, and stearic acids as the major fatty acids. This study further showed that fatty acids were the major antiproliferative constituents in the ethanol extract from O. tomentosa. Linoleic acid and oleic acid had IC50 values of 50.3 and 90.4 µM against human cervical cancer cells (HeLa), respectively. The results from this study have implications regarding the future exploration of O. tomentosa as a possible edible and/or medicinal mushroom. It is also recommended that necessary caution be taken when isolating unstable fatty acids from natural sources and in interpreting the results.

Isolation and characterization of an anti-proliferative polysaccharide from the North American fungus Echinodontium tinctorium

A novel polysaccharide EtGIPL1a was purified from fruiting bodies of Echinodontium tinctorium, a fungus unique to western North America. EtGIPL1a has an estimated weight average molecular weight of 275 kDa and is composed of glucose (54.3%), galactose (19.6%), mannose (11.1%), fucose (10.3%), glucuronic acid (4%), and rhamnose (0.6%). It has multiple glycosidic linkages, with 3-Glcp (28.9%), 6-Glcp (18.3%), 3,6-Glcp (13%), 4-GlcpA (9.2%), 6-Galp (3.9%), 2,6-Galp (2.6%), 3-Fucp (2.5%), 6-Manp (2.4%) being the most prominent, and unsubstituted glucose (15.3%), mannose (1.3%) and fucose (0.9%) as major terminal sugars. EtGIPL1a has a backbone containing mostly 3-substituted β-glucopyranose with 4-substituted glucopyranosyluronic acid. EtGIPL1a showed anti-proliferative activity against multiple cancer cell lines, with IC50 ranging from 50.6 to 1446 nM. Flow cytometry analyses confirmed that apoptosis induction is one mechanism for its anti-proliferative activity. EtGIPL1a should be further investigated for its potential anti-cancer activity in animal models, and for its possible utility in differentiation cancer therapy.

Deciphering the pharmacological potentials of Aganosma cymosa (Roxb.) G. Don using in vitro and computational methods

The methanolic leaf extract of A. cymosa yielded significant (60-75%) free radical scavenging activity in ABTS [2,2’-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid)] and DPPH (2,2’-diphenyl 1-picryl hydrazyl) assays, in the concentration range of 10-100 μg/mL. The extract showed antioxidant efficacy of 30-65% in reducing power assay and 55% inhibition in nitric oxide scavenging assay against ascorbic acid positive control. The plant extract exhibited modest inhibition against Escherichia coli, Pseudomonas aeruginosa and the fungal strain, Echinodontium tinctorium. The IC50 value of the extract was 100 μg/mL against MCF-7 breast cancer cells in MTT assay. The extract yielded 22-65% inhibition at 100-800 μg/mL concentration range in RBC membrane stabilization assay. Qualitative phytochemical screening revealed the presence of several classes of primary and secondary metabolites. Quantitative analysis revealed a high content of phenolic and flavonoid molecules. GC-MS profile of the plant extract revealed the presence of 44 bioactive compounds. Network analysis of key breast cancer proteins from differentially expressed genes aided in selecting the key proteins involved in breast cancer pathophysiology. ADMET-SAR analysis was done to assess the potential bioactivity of selected compounds. From the molecular docking results, coprostanol, 4-methylcatechol, pyrocatechol and indole were found to act as ligands of breast cancer target proteins. Furthermore, in molecular dynamics simulations (50 ns), the receptor-ligand complexes were stable for 50 nanoseconds. The results indicate that coprostanol is the principal component for binding to the selected breast cancer receptor proteins. Together, the results suggest that the phytochemicals explored herein exert the beneficial biological activities of A. cymosa.

Structural elucidation and immuno-stimulatory activity of a novel polysaccharide containing glucuronic acid from the fungus Echinodontium tinctorium

An immuno-stimulatory polysaccharide (EtISPFa) was purified from water extract of the fungus Echinodontium tinctorium. EtISPFa has an estimated weight average molecular weight (Mw) of 1354 kDa and is composed of glucose (66.2 %), glucuronic acid (10.1 %), mannose (6.7 %), galactose (6.4 %), xylose (5.6 %), rhamnose (3.1 %), fucose (1.8 %), and arabinose (0.2 %). It has multiple glycosidic linkages, with 3-Glcp (19.8 %), 4-GlcpA (10.8 %), 6-Glcp (10.7 %), and 3,6-Glcp (8.7 %) being the most prominent. NMR analysis showed that EtISPFa has a backbone containing mostly of 3-substituted β-glucopyranose with 4-substituted glucopyranosyluronic acid. Short side chains consisting of an average of two β-glycopyranose residues, connected through 1→6 linkages, are attached to the 6-position of about every 4th or 5th backbone glucose residue. EtISPFa is a novel glucuronic acid-containing β-glucan capable of significantly inducing the production of cytokines IL-17, IL-16, MIP-2, G-CSF,GM-CSF, LIF, MIP-1α, MIP-1β, and RANTES in vitro. EtISPFa should be further explored for its immuno-stimulatory activity in vivo.

Phytochemical composition, antioxidant and antimicrobial activities of Plecospermum spinosum Trecul.

Plecospermum spinosum Trecul. leaves collected from Eastern Ghats, India were assessed for antioxidant potential using the methanolic extract in vitro for free radical scavenging and phosphomolybdenum assays. The (2,2′-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid)) (ABTS+) assay exhibited potent free radical scavenging capacity at 100 μg/mL (86.48 ± 0.8 %). Antimicrobial activity of the methanolic leaf extract greatly inhibited the growth of Pseudomonas aeruginosa (9 ± 0.7 mm) and Echinodontium tinctorium (12 ± 0.9 mm) when compared to the other tested pathogens. The hemolysis (%) was calculated for anti-inflammatory activity and the results showed dose dependent effects. The lowest inhibition (53.08 ± 1.5 %) was obtained for 800 μg/mL extract when compared with control aspirin (90.41 ± 1.3 %) at the same concentration. Qualitative phytochemical tests confirmed the presence of triterpenoids, flavonoids, saponins, tannins, sterols, sugars and free aminoacids. Totally, 35 different phytochemical compounds were found using gas chromatography-mass spectrometry (GC–MS). Furthermore, the phenolics (70.34 ± 2.1 mg/g), flavonoids (164.28 ± 1.1 mg/g) and other metabolites were estimated quantitatively. These results exhibited considerable antioxidant, and antimicrobial activities due to the presence of bioactive phytochemicals. P. spinosum may be employed for the separation of novel bioactive metabolites with potential pharmaceutical activities.

Anti-Inflammatory Activity of the Wild Mushroom, Echinodontium tinctorium, in RAW264.7 Macrophage Cells and Mouse Microcirculation.

The aim of this study was to investigate the anti-inflammatory activity of a previously un-studied wild mushroom, Echinodontium tinctorium, collected from the forests of north-central British Columbia. The lipopolysaccharide (LPS)-induced RAW264.7 macrophage model was used to study the in vitro anti-inflammatory activity. The crude alkaline extract demonstrated potent anti-inflammatory activity, and was further purified using a “bio-activity-guided-purification” approach. The size-exclusion and ion-exchange chromatography yielded a water-soluble anti-inflammatory polysaccharide (AIPetinc). AIPetinc has an average molecular weight of 5 kDa, and is a heteroglucan composed of mainly glucose (88.6%) with a small amount of galactose (4.0%), mannose (4.4%), fucose (0.7%), and xylose (2.3%). In in vivo settings, AIPetinc restored the histamine-induced inflammatory event in mouse gluteus maximus muscle, thus confirming its anti-inflammatory activity in an animal model. This study constitutes the first report on the bioactivity of Echinodontium tinctorium, and highlights the potential medicinal benefits of fungi from the wild forests of northern British Columbia. Furthermore, it also reiterates the need to explore natural resources for alternative treatment to modern world diseases.

Phylogeny and taxonomy of Echinodontium and related genera

ABSTRACTThe phylogenetic relationship of eight species of Echinodontium, Laurilia, and Perplexostereum of Russulales were analyzed based on sequences of the nuc rDNA ITS1-5.8S-ITS2 (ITS [internal transcribed spacer]) and D1–D2 domains of nuc 28S rDNA (28S). Our results show that Echinodontium tinctorium, E. ryvardenii, and E. tsugicola represent Echinodontium sensu stricto. Based on morphological and phylogenetic evidence, the new genus Echinodontiellum is established to accommodate Echinodontium japonicum. Amylostereum, Echinodontium, Echinodontiellum, and Larssoniporia form the Echinodontiaceae clade. The Bondarzewiaceae clade includes Bondarzewia, Heterobasidion, Laurilia, and Lauriliella. The new genus Lauriliella is established for the species initially described as Stereum taxodii and Lauriliella taiwanensis new to science. The monotypic genus Perplexostereum forms a distinct clade. A key to the genera in the Echinodontiaceae and Bondarzewiaceae as well to Perplexostereum is provided.

Relationships among Amylostereum species associated with siricid woodwasps inferred from mitochondrial ribosomal DNA sequences

The genus Amylostereum currently includes four species, namely A. areolatum, A. chailletii, A. laevigatum and A. ferreum. Two of these species, A. areolatum and A. chailletii, are well known for their association with siricid woodwasps. Despite much interest in these fungus-woodwasp symbioses, the taxonomy and phylogeny of this genus received little attention in the past. The aim of this study was to investigate the phylogenetic relationship between the four species of Amylostereum. The placement of Amylostereum spp. among the Basidiomycetes was also investigated based on mt-SSU-rDNA sequence analyses. These data also clarify the taxonomic status of previously unidentified isolates. In this study, we have shown that A. areolatum is more distandy related to the three other species of Amylostereum, than they are to each other. Of the remaining three species, A. ferreum and A. laevigatum are more closely related to each other. One isolate that was collected from Sirex areolatus, and, therefore, expected to be A. chailletii, was more closely related to A. laevigatum and A. ferreum. As neither of the latter species have been implicated in associations with woodwasps, this finding warrants further investigation. Our data show that Amylostereum spp. group with neither Stereum nor Peniophora, as has been previously hypothesised, but rather with Echinodontium tinctorium. From this and other studies there was also an obvious relationship between Amylostereum/Echinodontium and Russula.

Characteristics of Black Bear Dens in Trees and Logs in Northeastern Oregon

Forty-one percent of 165 black bear (Ursus americanus) dens were in hollow trees or logs in northeastern Oregon from 1993 tol999. The remainder of the dens were in ground excavations (37%) and in caves or rock structures (22%). Hollow trees with the den entrance in the broken-off trunk were typically large-diameter grand fir (Abies grandis; x = 114 cm dbh) with Indian paint fungus (Echinodotium tinctorium) decaying the heartwood. Hollow trees with a base-entry were large-diameter grand fir and western larch (Larix occidentalis; (x = 108 cm dbh) with basal decay. Hollow logs in grand fir and western larch made up 11% of the 165 dens. Although a wide variety of habitat types were used, the majority of trees used for denning (88%) were in moist, late-seral stands of grand fir. Retaining large-diameter grand fir and western larch with evidence of decay will increase den site options for black bears in northeastern Oregon.

Influence of decay fungi on species composition and size class structure in mature Picea glauca × engelmannii and Abies lasiocarpa in sub-boreal forests of central British Columbia

Disturbance patterns in the sub-boreal spruce forests of central British Columbia have long been thought to result from frequent stand-initiating fires. However, recent evidence suggests that fires in the wetter areas of this region are infrequent (>500 years) and the uneven-aged stand structures have been shown to be self-maintaining in the absence of fire. The importance of decay fungi as agents of gap-formation and facilitators of uneven-aged stand structure was investigated. Three plots on each of two recently clearcut sites were established. Each stump in the plots was stem mapped and the species and diameter recorded. Decay at the stump top was recorded as white or brown rot, and the area occupied by decay was measured. Spruce stumps dominated the larger diameter classes but had less butt rot than sub-alpine fir stumps. Decay fungi contribute to stem breakage in living trees with heart rot, and as saprots of dead trees. Therefore, the results suggest that decay fungi play an important role in removing sub-alpine fir trees from the canopy of these wetter sub-boreal ecosystems, and in enabling spruce recruitment. The type of decay observed in sub-alpine fir suggests that breakage is predominantly due to white rot fungi causing heart rot, such as Echinodontium tinctorium and Stereum sanguinolentum. In spruce, brown rot fungi, which are predominantly saprot or wound-entry decay fungi, became more common in larger spruce, and may result from wounding as sub-alpine fir fall from the canopy. Spatial analysis indicated the stumps with stem decay were clumped. However, this is more likely due to strong clumping of the host, rather than spreading of butt rot pathogens through the roots.

Isolation of a fluorone pigment from the Indian paint fungus Echinodontium tinctorium and Pyrofomes albomarginatus

An orange pigment named echinotinctone was isolated from the wood-rotting polypores Echinodontium tinctorium and Pyrofomes albomarginatus. Its structure was elucidated as2, 6-dihydroxy-1, 8-dimethyl-3H-xanthen-3-one ( 1) on the basis of spectroscopic and chemical studies. 1 represents the first naturally occurring pigment with a simple fluorone chromophore. Echinotinctone ( 1) constitutes the first simple fluorone pigment from fungi. [Display omitted]

Roost Trees Used by Pileated Woodpeckers in Northeastern Oregon

We wanted to identify the types of trees and habitat used by pileated woodpeckers (Dryocopus pileatus) for roosting, and to develop management guidelines to provide this habitat. Thus, we located 123 roost trees used by 22 pileated woodpeckers and observed their roosting behavior in northeastern Oregon, 1989-90. In all 443 instances, adult pileated woodpeckers roosted inside a cavity at night. The majority (62%) of the roosts were in grand fir (Abies grandis), both live and dead, that were extensively decayed by Indian paint fungus (Echinodontium tinctorium). The decay had created a hollow chamber inside the tree that averaged 4.3 m long (SE=0.22) and 28 cm (0.29) wide where birds roosted

Somatic incompatibility in dikaryotic–monokaryotic and dikaryotic pairings of Echinodontium tinctorium

Somatic incompatibility in dikaryotic–monokaryotic (di–mon) and dikaryotic pairings of Echinodontium tinctorium was investigated in vitro on 4.5% malt agar. Antagonistic reactions of varying intensity occurred in all pairings between 12 allopatric dikaryons from Idaho and Arizona, between 14 sib-composed dikaryons from two Idaho sites, and in over 95% of pairings between sympatric dikaryons from separate trees at each location. Antagonistic reactions in dikaryotic pairings macroscopically appeared as dark reaction lines in the agar and aversion zones (barrage reactions) with hyphal massing on each side of the aversion zones. Self-crosses of dikaryons were somatically compatible, and hyphal anastomoses were common. Hyphal anastomoses were rare in the aversion zones between somatically incompatible dikaryons. Somatic incompatibility occurred in 51% of 89 di–mon pairings between 29 monokaryotic isolates and 7 sib-composed dikaryons from the same parent. Somatically incompatible di–mon pairings were characterized by hyphal massing in the contact zone and reaction lines on the reverse, but they lacked barrage reactions. Clamp connections formed in 54% of all di–mon crosses and at similar frequencies when no antagonistic reactions were present (55%) or when only hyphal massing occurred (53%), but they formed less frequently (33%) when dark reaction lines were present. The potential applications of these findings to epidemiological studies are discussed. Key words: Echinodontium, somatic incompatibility, epidemiology, Buller phenomenon, dikaryotic–monokaryotic, wood decay.

The Genetics of Sexual Incompatibility in the Indian Paint Fungus, Echinodontium Tinctorium

Basidiocarps of Echinodontium tinctorium were collected from three widely separated localities in Idaho and Arizona. Freezing basidiospore prints at -20 C for ten weeks stimulated germination. The role of low temperatures in breaking spore dormancy is discussed relative to environmental adaptation. Single-spore isolates of a single basidiocarp from each location were paired separately in all possible combinations. Four mating types were identified from each basidiocarp in Idaho. Four types of mac? roscopic interactions were observed in pairings between sympatric isolates. Basidiospores and cells of hyphae derived from basidiospores were monokaryotic (uninucleate), lacked clamp connections, and appeared haploid. Heterokaryotic hyphae derived from compatible matings were dikaryotic, exhibited clamp connections, and resembled generative contextual hyphae of basidiocarps. Complete intercompatibility between allopatric isolates demonstrated that the incompatibility factors were multiallelic. The fungus apparently does not fruit in culture, although mating results and cytological data indicate that it is heterothallic with a bifactorial (tetrapolar) mating system in which sexual incompatibility is controlled by multiple alleles at two loci on separate chromosomes.

The Genetics of Sexual Incompatibility in the Indian Paint Fungus, Echinodontium tinctorium

Basidiocarps of Echinodontium tinctorium were collected from three widely separated localities in Idaho and Arizona. Freezing basidiospore prints at -20 C for ten weeks stimulated germination. The role of low temperatures in breaking spore dormancy is discussed relative to environmental adaptation. Single-spore isolates of a single basidiocarp from each location were paired separately in all possible combinations. Four mating types were identified from each basidiocarp in Idaho. Four types of macroscopic interactions were observed in pairings between sympatric isolates. Basidiospores and cells of hyphae derived from basidiospores were monokaryotic (uninucleate), lacked clamp connections, and appeared haploid. Heterokaryotic hyphae derived from compatible matings were dikaryotic, exhibited clamp connections, and resembled generative contextual hyphae of basidiocarps. Complete intercompatibility between allopatric isolates demonstrated that the incompatibility factors were multiallelic. The fungus apparently does not fruit in culture, although mating results and cytological data indicate that it is heterothallic with a bifactorial (tetrapolar) mating system in which sexual incompatibility is controlled by multiple alleles at two loci on separate chromosomes.

Nitrogenase Activity Associated with Decayed Wood of Living Northern Idaho Conifers

Nitrogen fixation, as determined by the acetylene reduction technique, was demonstrated in decay columns of western larch and western white pine caused by Phellinus pini, western red cedar caused by Postia sericeomollis, and western hemlock and grand fir caused by Echinodontium tinctorium. Nitrogenase activity varied with tree species, fungal pathogen, wood decay stage, and seasonal moisturetemperature regimes within decaying stems. Nitrogen fixation potential in the productive forest eco? systems of northern Idaho was calculated between 0.06 kg/ha/yr and 4.91 kg/ha/yr and was primarily dependent on volume of decay in live standing trees on site.

TOXICITY OF SMOKE FROM SPOROPHORES OF THE INDIAN PAINT FUNGUS AND WOOD FROM WESTERN HEMLOCK TO YELLOWFEVER MOSQUITOES

AbstractTo mitigate the effects of mosquitoes, settlers in the Revelstoke area of British Columbia reportedly burned the sporophores of the Indian paint fungus, Echinodontium tinctorium (Ell. & Ev.) Ell. & Ev., a pathogen of western hemlock, Tsuga heterophylla (Raf.) Sarg., and true firs, Abies spp. Larval and adult yellowfever mosquitoes, Aedes aegypti (L.), were exposed to aqueous extracts of smoke (smoke-waters) from E. tinctorium sporophores, and from western hemlock sapwood and heartwood. Smoke-waters were of approximately equal toxicity to larvae. Fungus smoke-water, but not sapwood or heartwood smoke-waters, lost 50% of its potency in 5 months. Vapors from fungus smoke-water were significantly more toxic to adult mosquitoes than those from sapwood or heartwood. Thus smoke from E. tinctorium sporophores and T. heterophylla wood apparently contain different water-soluble combustion products toxic to A. aegypti.

Decay Fungi and Wounding in Advance Grand and White Fir Regeneration

Abstract A total of 464 living white and grand fir tree stems in 24 stands in Oregon and Washington were dissected to detect infections of hymenomycetes (decay fungi) and other microorganisms in woody tissue. Of 21,249 attempted isolations from dissected trunks, 43.2% yielded bacteria or yeasts, 38.6% were sterile, 11.0% were nonhymenomycetes, 3.1% were hymenomycetes, and 4.1% were contaminated or mixed cultures. Hymenomycetes most frequently isolated were Echinodontium tinctorium and Heterobasidion annosum, both of which caused the most discoloration and decay. Nearly 20% of all hymenomycetous isolations, particularly Hericium abietis and E. tinctorium, were from clear tissue not associated with discolored or decayed wood. More than 300 trunk wounds on 248 dissected trees were classified by size, location, age, and hymenomycetous associations. At least 45% of all trees with wounds had hymenomycetes present, of which E. tinctorium and Pholiota limonella were isolated most frequently. Discoloration caused by E. tinctorium was associated with wounds as recent as a year or as small as 56 cm2. All E. tinctorium infections that caused discoloration or decay were within 30 cm of a wound, whereas all infections within clear tissue were beyond 30 cm. These data support a hypothesis that E. tinctorium enters hosts via minute branchlet stubs, becomes dormant after the branchlet stubs occlude, and is activated by wounds in the vicinity of dormant infections. For. Sci. 33(2):347-355.

Decay capacities and interactions between isolates of an unidentified basidiomycete and Echinodontiumtinctorium Ell. & Ev. obtained from suppressed grand fir

Two isolates of an unidentified basidiomycete (UB) and two isolates of Echinodontiumtinctorium (ET), all from suppressed grand fir (Abiesgrandis (Doug.) Lindl.) in Oregon, were found to have an optimum growth temperature on malt agar medium of about 23 °C. In soil-block decay tests, conducted for 10 months at 23 °C. ET isolates caused average weight losses of 21.9 and 18.1%, while no decay was associated with UB. When UB was inoculated onto blocks precolonized with ET, it had a small negative effect upon the decay capacity of ET. However, when ET was inoculated onto blocks containing either living or dead (steam-sterilized) UB, no decay ensued. The UB isolates had apparently imparted a growth-inhibiting or fungitoxic material into the wood. As a result of this work, prior infection of twigs with UB is believed to inhibit or stop invasion of this infection court by ET.

Dinitrogen Fixation Associated with Sporophores of Fomitopsis Pinicola, Fomes Fomentarius, and Echinodontium Tinctorium

SUMMARYFixation of atmospheric dinitrogen by bacteria associated with contextual tissues of sporophores of Fomitopsis pinicola, Fomes fomentarius, and Echinodontium tinctorium is reported. Since ni...