Copper-spiked Research Articles

Anti-Hyperglycemic Effect of the Brown Slime Cap Mushroom Chroogomphus rutilus (Agaricomycetes) Crude Polysaccharide In Vitro and In Vivo.

The prevalence of diabetes is increasing worldwide, and it is very important to study new hypoglycemic active substances. In this study, we investigated the hypoglycemic effect of Chroogomphus rutilus crude polysaccharide (CRCP) in HepG2 cells and streptozotocin-induced diabetic mice. A glucose consumption experiment conducted in HepG2 cells demonstrated the in vitro hypoglycemic activity of CRCP. Furthermore, CRCP exhibited significant hypoglycemic effects and effectively ameliorated insulin resistance in insulin resistant HepG2 cells. In high-fat diet and streptozotocin-induced diabetic mice, after 4 weeks of CRCP administration, fasting blood glucose, fasting serum insulin, triglyceride, total cholesterol, low-density lipoprotein cholesterol, glutamate transaminase, alanine transaminase, and insulin resistance index significantly decreased, while high-density lipoprotein cholesterol and insulin sensitivity index (ISI) were markedly increased. Moreover, hematoxylin-eosin (HE) staining and immunofluorescence labeling of tissue sections indicated that CRCP attenuated the pathological damage of liver and pancreas in diabetic mice. These results indicate that CRCP is a potential hypoglycemic agent.

Structural characterization and osteogenic differentiation-promoting activity of polysaccharide purified from Chroogomphus rutilus

Chroogomphus rutilus (CR) possesses anti-inflammatory, antioxidant, and hypoglycemic properties. However, studies are yet to evaluate the anti-osteoporotic activity of the fungi and its polysaccharides. Therefore, this study is aimed at characterizing and evaluating the anti-osteoporotic effects of a novel polysaccharide from CR. The neutral polysaccharide CRP2 extracted and purified from the fruiting body of CR had a molecular weight of 20.41 kDa. Monosaccharide composition analysis revealed that CRP2 is composed of galactose, glucose, fucose, and mannose. The backbone of CRP2 primarily consisted of →6)-α-D-Galp-(1 → residues, with specific site substitutions speculated at partial positions, such as O-CH3 substitution at H-3 position, or a branch site located at C-2, including α-L-Fucp-(1 → 6)-β-D-Glcp-(1 → and α-D-Manp-(1→. CRP2 treatment increased trabecular bone density, restored a network-shaped structure, and upregulated the expression of osteoblast differentiation markers, including runt-related transcription factor 2, osterix, osteocalcin, and osteopontin in the femoral tissue of mice with osteoporosis (OP). Additionally, CRP2 treatment suppressed the expression of tumor necrosis factor-α and interleukin-1β in the femoral tissue of mice with OP. Mechanistically, CRP2 exerted anti-OP effect by inhibiting inflammation and promoting osteogenesis through the transforming growth factor β-1/Smad pathway. Conclusively, these findings augment our understanding of the potential role of CRP2 in OP treatment.

Chroogomphus rutilus Regulates Bone Metabolism to Prevent Periodontal Bone Loss during Orthodontic Tooth Movement in Osteoporotic Rats

Chroogomphus rutilus Regulates Bone Metabolism to Prevent Periodontal Bone Loss during Orthodontic Tooth Movement in Osteoporotic Rats

Insights into the Chemical Compositions and Health Promoting Effects of Wild Edible Mushroom Chroogomphus rutilus.

Chroogomphus rutilus is an edible mushroom that has been an important food source since ancient times. It is increasingly sought after for its unique flavor and medicinal value. It is one of the most important wild mushrooms for its medicinal and economic value. C. rutilus contains a variety of active ingredients such as vitamins, proteins, minerals, polysaccharides, and phenolics. C. rutilus and its active compounds have significant anti-oxidant, anti-tumor, immunomodulatory, anti-fatigue, hypoglycemic, gastroprotective, hypolipemic, and neuronal protective properties. This paper summarizes the fungal chemical compositions and health-promoting effects of C. rutilus by collecting the literature on the role of C. rutilus through its active ingredients from websites such as Google Scholar, Scopus, PubMed, and Web of Science. Current research on C. rutilus is limited to the cellular and animal levels, and further clinical trials are needed to conduct and provide theoretical support for further development.

Brown Slime Cap Mushroom (Chroogomphus rutilus, Agaricomycetes) Polysaccharide Resists Motion Sickness by Inhibiting the Activity of the Serotonin System in Mice.

Motion sickness (MS) is a disorder of the autonomic nervous system caused by abnormal exercise with symptoms such as nausea, vomiting and drowsiness. More than 90% of the human population has experienced different degrees of MS. At present, anticholinergics, antihistamines, and sympathomimetic drugs are used for treating MS, but these drugs generally have some adverse reactions and are not suitable for all people. Therefore, it is necessary to develop anti-MS drugs that have high efficiency and no adverse effects. Previous studies have found that Chroogomphus rutilus polysaccharide (CRP) is effective at preventing and treating MS in rats and mice. However, its mechanism of action is not clear. To clarify whether the CRP has anti-MS effects in mice, and to clarify its mechanism, we performed behavioral, biochemical, and morphological tests in a Kunming mouse model. Our results indicate that CRPs can significantly relieve the symptoms of MS, and their effect is equivalent to that of scopolamine, a commonly used anti-MS medicine. Our results indicate that CRPs may directly act on the gastrointestinal chromaffin cells to inhibit the synthesis and release of serotonin (5-hydroxytryptamine, or 5-HT) and thus reduce the signal from the gastrointestinal tract.

Ectomycorrhizal fungi enhance salt tolerance of Quercus mongolica by regulating ion balance

We explored the effects of salt stress on the growth of Quercus mongolica and the effects of ectomycorrhizal fungi (ECMF) on the ion balance of Q. mongolica. After inoculating four kinds of ECMFs (Gomphidius visci-dus, Suillus leteus, Suillus grevillea, Boletus edulis) on Q. mongolica seedlings, we treated the annual non-mycorrhizated and mycorrhizated seedlings with NaCl stress (0, 100, 200, 300 mmol·L-1) for 36 days, and then analyzed the mycorrhizal characteristics, growth, leaf injury symptoms, leaf electrolyte permeability, water content, and ion contents in roots, stems, and leaves. The results showed that the four ECMFs could establish a symbiotic system with Q. mongolica, and that root system of mycorrhizal seedlings was stronger than that of non-mycorrhizal seedlings. Under salt stress, the growth of Q. mongolica seedlings was inhibited, with the symptoms of scorched leaves. The damage to leaf plasma membrane and the degree of water loss were aggravated with the increases of salt stress. Under low salt stress (100 mmol·L-1), Q. mongolica preferentially accumulated Na+ in roots and stems. Under medium-high salt stress (200-300 mmol·L-1), roots became the primary organ for accumulating Na+. ECMF regulated ion balance in plant by increasing the Na+ level in roots and reducing the Na+ accumulation in stems and leaves, enhancing the absorption of K+ and Ca2+ to increase the K+/Na+ and Ca2+/Na+. The four ECMFs had different mitigation effects on salt poisoning of Q. mongolica. G. viscidus had the strongest effect, followed by S. leteus, while S. grevillei and B. edulis had relatively little effect.

Ectomycorrhizal Inoculation Enhances the Salt Tolerance of Quercus mongolica Seedlings.

Salt stress harms the growth and development of plants, and the degree of soil salinization in North China is becoming increasingly severe. Ectomycorrhiza (ECM) is a symbiotic system formed by fungi and plants that can improve the growth and salt tolerance of plants. No studies to date have examined the salt tolerance of Quercus mongolica, a typical ectomycorrhizal tree species of temperate forests in the northern hemisphere. Here, we inoculated Q. mongolica with two ectomycorrhizal fungi (Gomphidius viscidus; Suillus luteus) under NaCl stress to characterize the effects of ECM. The results showed that the symbiotic relationship of Q. mongolica with G. viscidus was more stable than that with S. luteus. The cross-sectional area of roots increased after inoculation with the two types of ectomycorrhizal fungi. Compared with the control group, plant height, soluble sugar content, and soluble protein content of leaves were 1.62, 2.41, and 2.04 times higher in the G. viscidus group, respectively. Chlorophyll (Chl) content, stomatal conductance (Gs), and intracellular CO2 concentration (Ci) were significantly higher in Q. mongolica inoculated with ectomycorrhizal fungi than in the control, but differences in the net photosynthetic rate (Pn), transpiration rate (Tr), and photosystem II maximum photochemical efficiency (Fv/Fm) were lower. The relative conductivity of Q. mongolica inoculated with the two ectomycorrhizal fungi was consistently lower than that of non-mycorrhizal seedlings, with the effect of G. viscidus more pronounced than that of S. luteus. The malondialdehyde (MDA) content showed a similar pattern. Peroxidase (POD) and catylase (CAT) levels in mycorrhizal seedlings were generally higher than those of non-mycorrhizal seedlings under normal conditions, and were significantly higher than those of non-mycorrhizal seedlings on the 36th and 48th day after salt treatment, respectively. Overall, the results indicated that the salt tolerance of Q. mongolica seedlings was improved by ectomycorrhizal inoculation.

Comparative study of structural properties and biological activities of polysaccharides extracted from Chroogomphus rutilus by four different approaches

Extraction processes significantly alter the structural and functional properties of polysaccharides. In this study, we extracted polysaccharides from Chroogomphis rutilus fruiting bodies (designated as CRP) using four methods, including hot water, ultrasound, microwave and sequential ultrasound-microwave, and designated these polysaccharides as CRP-H, CRP-M, CRP-U and CRP-UM, respectively. All CRPs were heteropolysaccharides with semblable monosaccharide types of glucose, mannose and galactose, mainly constituted of α-d-glucopyranosyl-(1 → 4). The extraction processes significantly affected the molecular weights, monosaccharide proportions, glycosidic bond ratios, branching degrees, triple-helix conformation and surface morphology of the CRPs. Among them, CRP-UM showed the highest yield and most potent antioxidative capacity in vitro and in HL-7702 cells, but the weakest activation of immunostimulatory response in RAW264.7 cells. In contrast, CRP-H exhibited the lowest yield but strongest immunostimulatory activity. Overall, microwave extraction could be utilized as a general and practical CRP extraction approach, based on its relatively high yield and bioactivities.

The first complete mitochondrial genome of edible and medicinal fungus Chroogomphus rutilus (Gomphidiaceae, Boletales) and insights into its phylogeny

In the present study, we assembled and annotated the complete mitochondrial genome of Chroogomphus rutilus. The complete mitochondrial genome of C. rutilus was composed of circular DNA molecules, with a size of 37,508 bp. The GC content of the C. rutilus mitogenome was 22.82%. A total of 18 protein-coding genes (PCGs), 2 ribosomal RNA (rRNA) genes, and 24 transfer RNA (tRNA) genes were detected in the C. rutilus mitogenome. Phylogenetic analysis based on combined mitochondrial gene dataset indicated that the C. rutilus exhibited a close relationship with species from the genus Rhizopogon. This study served as the first report on the complete mitochondrial genome from the family Gomphidiaceae, which will promote the understanding of phylogeny, evolution, and taxonomy of this important fungal species.

Effects of copper on biological treatment of NMF- and MDG-containing wastewater from TFT-LCD industry

This study aimed to evaluate the effects of copper on N-methylformamide (NMF)- and methyl diglycol (MDG)-containing wastewater treatment using batch experiments and a lab-scale anoxic–oxic (A/O) sequencing batch reactor (SBR). Batch experimental results indicated that aerobic degradation of NMF followed Monod-type kinetics. Copper inhibition on nitrification also followed Monod-type inhibition kinetics with copper-to-biomass ratio instead of copper concentration. Specific degradation rates of NMF and MDG under both aerobic and anoxic conditions decreased in the matrix of full-scale wastewater, and high copper dosage would further reduce the degradation rates. In the long-term presence of 0.5 mg/L copper, the A/O SBR could maintain stable and complete degradations of NMF and MDG, 95% of COD removal, and more than 50% of total nitrogen (TN) removal. High concentrations of copper spikes, including 40 mg/L and 110 mg/L, slowed down degradation rates for both NMF and MDG, but did not affect COD and TN removal efficiencies in the full 24 h-cycle operation. The long-term A/O SBR operation revealed that daily dosage of 0.5 mg/L copper was not detrimental to NMF/MDG degradations due to regularly wasting sludge, but 110 mg/L of copper spike obviously reduced NMF/MDG degradation rate although it could be recovered later by regularly wasting sludge and maintaining SRT at 20 days.

Antioxidant, Anti‐inflammatory and Cytotoxic Activities of Polyphenols Extracted from Chroogomphus rutilus

Chroogomphus rutilus is a rare fungal species that grows under pine trees and is now widely used as a functional food and pharmaceutical product. However, the chemical constituents and biological activities of Chroogomphus rutilus have been relatively limited. The present study aimed at determining the total polyphenols and flavonoids contents, biological activities and main phenolic compounds of Chroogomphus rutilus from different geographical origins at the stipe and pileus. The results suggested that Chroogomphus rutilus polyphenol extracts revealed a higher antioxidant, anti-inflammatory, and cytotoxic activities, and there were significant differences between samples from different locations and regions. Correlation analysis showed that the contents of total polyphenols and flavonoids were significantly correlated with antioxidant and anti-inflammatory activities. However, only the content of total flavonoids was significantly correlated with cytotoxicity, which means that the cytotoxicity of Chroogomphus rutilus polyphenol extracts may be regulated by flavonoids or other compounds. HPLC-DAD analysis revealed that the main phenolic compound was protocatechuic acid, followed by baicalin, p-hydroxyphenylacetic acid and p-hydroxybenzoic acid, but comparing with the pileus extracts, the stipe extracts can be considered as a higher concentration of phenolic compounds. Therefore, antioxidant, anti-inflammatory and cytotoxic activities of Chroogomphus rutilus polyphenol extracts could be due to the identified compounds. This study investigated a deep knowledge about the constituents and activities of Chroogomphus rutilus and provided the reference for its application in food and pharmaceutical.

The Genome Sequences of 90 Mushrooms

Macrofungus is defined as the fungus that grows an observable sporocarp. The sporocarps of many species are commonly called mushrooms and consumed by people all around the world as food and/or medicine. Most macrofungi belong to the divisions Basidiomycetes and Ascomycetes, which are estimated to contain more than 80,000 species in total. We report the draft genome assemblies of macrofungi (83 Basidiomycetes species and 7 Ascomycetes species) based on Illumina sequencing. The genome sizes of these species ranged from 27.4 Mb (Hygrophorus russula) to 202.2 MB (Chroogomphus rutilus). The numbers of protein-coding genes were predicted in the range of 9,511 (Hygrophorus russula) to 52,289 (Craterellus lutescens). This study provides the largest genomic dataset for macrofungi species. This resource will facilitate the artificial cultivation of edible mushrooms and the discovery of novel drug candidates.

Volatile components, total phenolic compounds, and antioxidant capacities of worm-infected Gomphidius rutilus

This study evaluated the effects of worm infection on the volatile components, total phenolic compounds, and antioxidant capacities of Gomphidius rutilus. G. rutilus without worms (GW), G. rutilus infected by a small amount of worms (GS; infected area <50%), and G. rutilus infected by a large amount of worms (GL; infected area>50%) were investigated. The volatile components of G. rutilus were analyzed by simultaneous distillation–extraction (SDE) and headspace solid-phase microextraction (HS-SPME) using gas chromatography–mass spectrometry (GC–MS). A total of 17 and 19 types of volatile compounds were detected, including ketones, alcohols, benzene, alkenes, aldehydes, esters, acids, and alkanes. Alcohols comprised the most abundant compound in GL, GS, and GW. The relative content of 1-octen-3-ol was the highest in all mushrooms. The concentration of eight-carbon (C8) compounds relative to the total volatile compounds varied widely, ranging from 40% (GW) to 64.34% (GS) and 84.42% (GS) and to 91.59% (GL), respectively, among the three samples. The antioxidant capability and the total phenolic contents of G. rutilus were evaluated in this study. The highest total phenolic content (TPC) of 192.23 mg GAE/g was found in GL, which differed significantly (P < 0.05) from the latter two samples, whereas the lowest value of 156.11 mg GAE/g was found in GW. ABTS radical cation scavenging activity, FRAP ferric reducing antioxidant capacity (FRAP) radical scavenging activity, and oxygen radical absorbance capacity (ORAC) were investigated to screen the antioxidant properties of extracts. The contents of total phenolic compounds and their antioxidant capacities in vitro showed significant correlations (P < 0.01). Among the three types of samples, the phenolic compounds of GL exhibited the highest antioxidant capacity, showing the values of 0.089 mM TE/g for ABTS, 0.949 mM Fe2+ E/g for FRAP, and 1.952 M TE/g for ORAC. However, regarding the total antioxidant capacity, GS exhibited the highest antioxidant capacity, showing the values of 0.002648 mM TE/g for ABTS, 0.004437 mM Fe2+ E/g for FRAP, and 0.256 μM TE/g for ORAC. In conclusion, HS-SPME was more suitable for the extraction of volatile aroma components from G. rutilus. GL had the most abundant aroma components. GL had the highest TPC and antioxidant capacity compared with those of GS and GW, whereas GS showed the opposite results. Interestingly, GS was found to have the highest total antioxidant capacity in vitro. Based on these measured indicators, worm infection had no negative effect on the quality of G. rutilus. Therefore, worm-infected G. rutilus can also be consumed by humans.

A Primary Study of the Antioxidant, Hypoglycemic, Hypolipidemic, and Antitumor Activities of Ethanol Extract of Brown Slimecap Mushroom, Chroogomphus rutilus (Agaricomycetes).

In vivo and in vitro treatments were carried out to investigate the effects of a 95% ethanol extract of Chroogomphus rutilus (CRE) on antioxidant, hypoglycemic, hypolipidemic, and antitumor properties. CRE showed potent radical scavenging activity against DPPH in vitro. It could increase antioxidant enzymatic activities (superoxide dismutase and glutathione peroxidase) and could reduce malondialdehyde content in vivo in mice in which aging was induced by D-galactose. CRE had hypoglycemic activity and could significantly inhibit α-glucosidase activity in vitro and decrease blood glucose concentration in vivo. CRE could decrease the serum total cholesterol, triglyceride, and low-density lipoprotein cholesterol levels and increase the high-density lipoprotein cholesterol level in diabetic mice. The MTT assay showed that CRE also had a certain inhibitory effect on the tumor cells. These results suggest that CRE may be beneficial for human health and could be useful for applications in medicine, the food industry, and agriculture.

Effects of Different Ectomycorrhizal Fungal Inoculates on the Growth of Pinus tabulaeformis Seedlings under Greenhouse Conditions

The tree species Pinus tabulaeformis Carr. (P. tabulaeformis) is commonly planted in China due to its economic and ecological value. In order to identify one or more ectomycorrhizal (ECM) fungal species for future P. tabulaeformis afforestation, we investigated the effects of five ECM fungal species: Laccaria laccata, Boletus edulis, Gomphidius viscidus, Suillus grevillei, and Suillus luteus on the growth of P. tabulaeformis seedlings under greenhouse conditions. The growth parameters of P. tabulaeformis seedlings were evaluated 90 days following fungal colonisation. The majority of seedlings were significantly affected by ECM inoculation. Mycorrhizal inoculated seedlings were taller, had more lateral roots, and a greater biomass compared with the non-mycorrhizal (CK) seedlings. With the exception of G. viscidus, inoculated seedlings exhibited higher phosphorus, potassium, and nitrogen content compared with the CK seedlings. In addition, ECM colonisation increased the enzymatic activity of catalase, acidic phosphatase, protease, and the urease content in the rhizosphere soil. Our study showed that Laccaria laccata, Suillus grevillei, and Suillus luteus may be useful for improving the growth and cultivation of P. tabulaeformis seedlings. Furthermore, we observed that S. luteus inoculation increased the gas exchange parameters of P. tabulaeformis seedlings under field conditions.

Penicillium chroogomphum, a new species in Penicillium section Ramosa isolated from fruiting bodies of Chroogomphus rutilus in China

During tissue culture from fruiting bodies of Chroogomphus rutilus, two Penicillium sp. strains were isolated that could not be assigned to any described species based on morphological and molecular phylogenetic analysis. Multigene phylogenetic analyses with the nuclear ribosomal internal transcribed spacer (ITS) region, β-tubulin (benA) and calmodulin (cmd) genes, as well as morphological analyses revealed that these isolates were in the Penicillium section Ramosa. These isolates were closely related to P. lenticrescens and P. soppii in terms of multigene phylogeny, but their colonies and micro-morphological characters differed from these closest related species. We consider that these isolates constitute a new species in the genus Penicillium section Ramosa and propose the name P. chroogomphum.

Chemical constituents of Chroogomphus rutilus (Schaeff.) O.K. Mill

This work describes the isolation of thirty-two compounds from Chroogomphus rutilus, including eight sterols (1–8), five phenols (9–13), three nucleosides (14–16), two monosaccharide derivatives (17–18) and fourteen coumarins (19–32). The chemical structures of the compounds were elucidated on the basis of spectral data (1H and 13C NMR) and by comparisons of spectroscopic data with reported values in the literatures. Twenty-eight compounds (1, 2, 4, 5, 8–15, 17–32) were first found in the family Gomphidiaceae. The chemotaxonomic significance of these compounds is discussed.

Isolation and Characterization of Extracellular Melanin Produced by Chroogomphus rutilus D447

Isolation and Characterization of Extracellular Melanin Produced by Chroogomphus rutilus D447

Copper Bioavailability to Beans (Phaseolus vulgaris) in Long-Term Cu-Contaminated Soils, Uncontaminated Soils, and Recently Cu-Spiked Soils

Copper solubility and its bioavailability to Phaseolus vulgaris in long-term copper-contaminated soils, uncontaminated soils, and copper-spiked soils were studied. The role of plant factors, total copper load in soils, and/or the aging effect on the uptake of copper was explored so as to assess health risks through contamination of the food chain associated with growing the crop on such soils. Contaminated soils and clean soils were collected from coffee-growing fields in Kilimanjaro and Arusha, Tanzania. Two bean seeds were sown per pot, replicated three times, and arranged in a randomized design in a glass house. Copper spiking significantly increased extractable copper, as expected. For all of the treatments except for the Mwanga and Arumeru control soils, the addition of CuSO4 did not significantly increase the humic-acid-bound copper, but it significantly increased the fulvic-acid-bound copper (p = 0.05). Moshi soils had significantly higher concentrations of copper in the bean shoots than was the case with other treatments (p = 0.05). For the respective soil types, there was no significant difference in the concentrations of copper in bean leaves between spiked and unspiked treatments (p = 0.05). Bean shoots did not accumulate copper beyond the normal concentrations.

Contents and Antioxidant Activities of Polysaccharides in 14 Wild Mushroom Species from the Forest of Northeastern China.

This study aimed to evaluate the polysaccharide contents and antioxidant activities of 14 important wild mushroom species in the Lesser Khingan Range Forest of northeastern China. The fungal species were identified by morphology and DNA matching, belonging to six families: Polyporaceae (four species), Tricholomataceae (three), Russulaceae (three), Lepiotaceae (two), Lycoperdaceae (one), and Paxillaceae (one). Polysaccharides were isolated and partially purified from the water extract of each m ushroom by ethanol precipitation, deproteinization, and dialysis. Antioxidant activities of the mushroom polysaccharides were found to vary with both the species and the assay methods, and the highest three were from Handkea utriformis (535.8 μmol trolox/g), Macrolepiota mastoidea (378.6), and Armillaria ostoyae (329.1) in radical scavenging; H. utriformis (5.94 mmol Fe/g), Lepista nuda (4.65), and A. ostoyae (4.42) in reducing power; and Armillariela cepistipes (484.6 μmol Fe2+/g), H. utriformis (274.8), and M. mastoidea (202.5) in Fe2+ chelating. Several of the polysaccharides showed notable anti-tyrosinase activities, and that of Chroogomphus rutilus was the most potent with an IC50 of 0.46 mg/ml. These wild mushrooms can be useful sources of polysaccharides as potential antioxidants and tyrosinase inhibitors. This is the first systematic study on the wild mushroom species, as well as their polysaccharide contents and biological activities, from this forest.