During a study of fungal diversity in the Puruogangri Glacier ecosystem on the Qinghai-Xizang Plateau, a strain was isolated from soil samples at the glacier forefields using the dilution plate method, and identified as Sinophaeosphaeria xizangensis gen. et sp. nov. Morphological analysis revealed that the fungus produced olive to olive-brown conidia on PDA medium. The conidia consist of two or more cells, arranged in curved or irregular chains. Some cells slightly swollen, forming distinctly prominent bulging regions or aggregating into dense, hammer-shaped structures. Phylogenetic analyses based on five genetic loci, including internal transcribed spacer (ITS), large subunit ribosomal DNA (LSU), small subunit ribosomal DNA (SSU), RNA polymeraseⅡsecond largest subunit (rpb2), and translation elongation factor 1-α (tef1-α), indicate that this fungus represents a previously undescribed lineage within the family Phaeosphaeriaceae. This genus is located at the base of a branch that includes Paraloratospora, Loratospora, Wingfieldomyces and Sulcispora, revealing its unique evolutionary position.
In present study, morphological observation and phylogenetic relationship analyses by using multiple loci DNA sequences, including the internal transcribed spacer (ITS) regions, the large subunit nuclear ribosomal RNA gene (nLSU), and the translation elongation factor 1-α gene (tef1), result in discovery of Sicyoideibasidia yunnanensis sp. nov. of Agaricales. The species is characterized by resupinate basidiomata with smooth, cream to yellowish hymenial surface, a monomitic hyphal system, generative hyphae bearing simple septa, and ellipsoid basidiospores. Phylogenetic analysis based on ITS+nLSU+tef1 data shows that the new species is closely related with S. bambusicola and S. punctata.
Dentiscutata hongfenghuana, a new species of Dentiscutata (Gigasporaceae, Diversisporales, Glomeromycotina), was discovered from rhizosphere soil of Capsicum annuum surrounding Hongfeng Lake in Qingzhen City, Guizhou Province. The species has colorless and transparent spores that are white to light greenish-yellow, with dimensions of 185-191 × 298-460 μm. The sporogenous cells are brownish-yellow, nearly spherical to flask-shaped, with a pale yellow to brownish-yellow germination shield 3-8-lobed. The spore wall consists of three layers: outer wall layers (OWL1-3), middle wall layers (MWL1-2), and inner wall layers (IWL1-3). The OW turns rust-colored to reddish-black in Melzer’s reagent, the MW shows no reaction, and the IW is stained light pink to deep purplish-red. The nuclear rDNA sequence [covering part of the small subunit (SSU), the entire internal transcribed spacer (ITS), and part of the large subunit (LSU)] supports its placement as a unique evolutionary branch within the genus Dentiscutata. Detailed morphological description, characteristic illustrations, and information of the type specimen are provided, and the distinguishing features of closely related species are discussed.
Based on morpho-anatomical and chemical identification in combination with phylogenetic reconstruction of rDNA-ITS sequences, a new species of Chlorangium in family Megasporaceae, Chlorangium longiconidicum sp. nov., is reported. Detailed morpho-anatomical descriptions and illustrations and comparative discussions with morphologically similar species within the genus are provided. The new species is characterized by peltate thallus with pruinose surface and strong stalk, conspicuous immersed pycnidia, long filiform conidia (14.0-22.0 µm), and the presence of three unknown chemical compounds. The genus Chlorangium is first reported from China.
Shitai County is located in south of Anhui Province, China. The rich vegetation in this county provides a favorable growth environment for various types of macrofungi. An in-depth investigation of macrofungal resources was conducted at 58 sampling sites in the county in 2024 by using the line transect method and random survey method. In total, 743 specimens were collected. Based on morphological and molecular methods, 285 species of macrofungi were identified, belonging to 2 phyla, 7 classes, 18 orders, 76 families, and 178 genera, of which 14 species of Ascomycota were referred to 3 classes, 4 orders, 8 families, and 13 genera, and 271 species of Basidiomycota to 4 classes, 14 orders, 68 families, and 165 genera. Five dominant families containing more than 10 species each are Polyporaceae, Hymenochaetaceae, Phanerochaetaceae, Russulaceae, and Omphalotaceae, which comprise 88 species accounting for 30.88% of all the known species. Seven dominant genera containing more than 5 species each are Russula, Trametes, Xylodon, Fuscoporia, Hymenochaete, Peniophora, and Marasmius, which comprise 50 species accounting for 17.54% of all the known species. There are 7 geographical distribution types at the genus level. The cosmopolitan genera and pantropic genera were found to be the most dominant, with 87 and 42 genera respectively. The resource evaluation showed that 19 species are edible, 26 medicinal, 19 simultaneously edible and medicinal, and 9 poisonous. The results provide basic data for the protection and sustainable utilization of macrofungal resources in Shitai County.
Walnut is an important economic tree in China, and its cultivation area and production rank first in the world. In recent years, pathogenic fungal diseases of walnuts in China have shown an increasing trend of damage, significantly impacting the yield and quality of product. To clarify the fungal species affecting walnuts in China, extensive collection of specimens was conducted in 2020-2024 in major walnut-producing areas in 9 provinces of the country. In total, 1 882 fungal isolates were obtained. Based on Koch’s postulate, 51 pathogenic species belonging to 9 families and 5 orders were identified. In addition, the fungal species previously reported in China to be pathogenic to walnuts were also compiled and revised. The results showed that a total of 135 pathogenic fungal species could cause walnut diseases, belonging to 2 phyla, 5 classes, 18 orders, and 31 families. The revised scientific names and Chinese names of the pathogens, Chinese disease names and distribution in China were listed based on the latest classification system and nomenclature regulations for the standardized use of researchers in China.
Wolfiporia hoelen is a traditional medicinal and edible fungus in China, with significant medicinal and economic values. Although large-scale cultivation of W. hoelen has been achieved domestically, some problems have to be settled urgently, including varietal admixture, unclear genetic backgrounds, and shortage of elite strains. Therefore, investigating the genetic diversity and establishing an efficient and accurate method for variety identification are of great importance. In this study, representative cultivated and wild strains of W. hoelen from China, as well as strains from Japan, were selected for genetic diversity analysis based on single nucleotide polymorphisms (SNPs) and a multi-nucleotide polymorphism (MNP) marker library was constructed. Population genetic structure analysis revealed that the tested strains could be divided into two major genetic groups, of which group A exhibited typical clonal genetic characteristics. The MNP marker library comprises 654 highly specific MNP markers, with genetic similarity (GS) among W. hoelen strains ranging from 9.6% to 100%, and the GS among strains within group A reach 99.8% to 100%. Genome resequencing of subcultured and protoplast-regenerated strains confirmed that the stability of MNP markers was not affected by short-term asexual reproduction. This study provides a reliable method for strain identification and offers support for parental selection in W. hoelen hybrid breeding.
Aspergillus cristatus is the dominant fungus in the fermentation process of Fuzhuan brick tea. Osmotic pressure can regulate the pure sexual and asexual reproduction of this fungus. MpdA plays a crucial regulatory role in spore formation and mannitol biosynthesis of fungi, which is also related to the response to osmotic pressure. To elucidate the role of the AcMpdA gene in the development and stress response of A. cristatus, and to investigate the connection between development and metabolites, the AcMpdA-deficient strain (∆AcMpdA) was constructed. By comparing the morphological differences and metabolite changes between the mutant and the wild-type strain, the following results were obtained. The mycelial branches of the mutant increased, ascospores were underdeveloped, and the number of conidia decreased. The mutant is more sensitive to congo red, osmotic stress, and heat stress. The mannitol test results showed that the mannitol content of the mutant strain was 61% less than that of the wild-type strain. Metabolomic result analysis revealed significant differences in 1 555 metabolites between the mutant and the wild-type strain. The differential metabolites included lipid and lipoid molecules, organic heterocyclic compounds, organic acids and their derivatives, among which lipid and lipoid substances accounted for a relatively high proportion. KEGG pathway enrichment analysis indicated that the differential metabolites were mainly enriched in metabolism, genetic information processing, environmental information processing, and cellular process pathways. The deletion of AcMpdA significantly affected the biosynthesis of unsaturated fatty acids and arachidonic acid metabolism. In conclusion, the AcMpdA gene of A. cristatus influences mycelial branching, ascospore and conidium formation, diminishes the ability to withstand environmental stress, and modulates the metabolic processes of lipids and lipoid molecules. The regulatory impact of this gene on lipid substances and their pathways has not been documented in the Aspergillus.
In nature, all orchids rely on symbiotic fungi to provide them with carbon sources and mineral nutrients for seed germination. The orchid mycorrhizae perform a unique endosymbiosis. However, there is still a lack of research on how the symbiotic fungi regulated the orchid mycorrhizal formation. Our previous study indicated that Epulorhiza repens ML01 was a typical orchid mycorrhizal fungus with a wide host range. In this study, the conditions for isolation and regeneration of ML01 were optimized, including culture duration, degrading enzyme combinations, enzymatic digestion time, and type of osmotic stabilizers. The protoplasts obtained were subjected to UV mutagenesis. Totally, 10 mutant strains with different growth characteristics or C/N utilization ability were obtained after screening. Symbiotic culture with seeds of the orchid Dendrobium officinale on oatmeal agar (OMA) medium showed that the mutant strain M3-12 displayed significantly weaker effect on seed germination and protocorm development than original strain ML01. This study provides reference for the practical application of orchid symbiotic fungi.
As a rare and traditional medicinal fungus in China, sanghuang is renowned for its multifaceted pharmacological properties, including antitumor, antioxidant, and hypoglycemic activities. Notably, its hypolipidemic effect has drawn significant interest, yet its specific mechanism of action remains unclear. This study employed an integrated approach combining in vitro assays and network pharmacology to investigate the lipid-lowering mechanism of Sanghuangporus quercicola. Pancreatic lipase inhibition assays demonstrated that the fruiting body extract of S. quercicola (40 mg/mL) exhibited potent inhibitory activity against pancreatic lipase, with an inhibition rate of 89.50%. Additionally, it showed sodium taurocholate and sodium glycocholate binding capacities of 34.59% and 49.12%, respectively, highlighting its hypolipidemic potential. Through ultra-high-performance liquid chromatography coupled with time-of-flight mass spectrometry (UHPLC-TOF/MS), 124 metabolites were identified, and 15 candidate bioactive compounds were selected based on drug-likeness rule of five. Subsequent target prediction using OMIM and GeneCards databases identified hyperlipidemia-associated targets, followed by protein-protein interaction (PPI) network construction and analysis, which pinpointed IL6, TNF, and AKT1 as core therapeutic targets. KEGG pathway enrichment analysis revealed significant associations with the PPAR signaling pathway, the lipid metabolism pathway, and the atherosclerosis pathway, suggesting that S. quercicola may alleviate hyperlipidemia by modulating inflammatory responses and lipid metabolism. Molecular docking analysis confirmed the binding capabilities of the identified compounds with core targets, with binding energies ranging from -3.8 to -9.3 kcal/mol. This study preliminarily elucidates the hypolipidemic mechanism of S. quercicola through a combination of in vitro experiments and network pharmacology, providing a theoretical foundation for the development of lipid-lowering drugs or functional foods.
Total amino acids are essential in Lentinula edodes for protein synthesis and physiological functions. In this study, protoplast fusion of eight parental strains, Le7, Le8, Le10, Le20, Le25, Le28, Le30 and XT08, resulted in the acquirement of 42 verified hybrids. Hybridization affinity averaged 80.8%. Among the hybrids, 25 (59.5%) exhibited amino acid heterosis, of which R23 was the highest (2.66 mg/g). Through combining ability analysis three elite monokaryons (Le7-8, Le10-46, Le28-3) and four superior hybrids (R23, R5, R32, R36) were identified. SSR analysis with nine primers revealed 100% polymorphism. UPGMA clustering divided the strains into two groups: most hybrids clustered with parents, while some showed significant genetic divergence. These results provide key materials and theoretical support for breeding high-amino-acid L. edodes strains.
Using three dominant Trichoderma species, T. atroviride, T. macrochlamydospora and T. subvermifimicola, contaminating substrates of Lentinula edodes as target species, a multiplex PCR molecular detection technique was optimized and established based on the specific primers designed according to their RNA polymerase Ⅱ subunit b (rpb2) gene sequences. This technique can simultaneously detect the three dominant Trichoderma species. Eight other Trichoderma species, two varieties of L. edodes, and common contaminative Penicillium and Mucor strains were used as reference strains for validation the specificity and sensitivity of the technique. Results showed that this technique had strong specificity, and three specific bands with the size of 358 bp, 805 bp and 614 bp were observed after agarose gel electrophoresis of the amplification products only when the genomic DNA of T. macrochlamydospora, T. subvermifimicola and T. atroviride were used as templates. Sensitivity verification found that the detection limit for the genomic DNA of T. macrochlamydospora and T. subvermificolola was 10 pg/μL, being consistent with sensitivity of single PCR detection. The detection limit of T. atroviride is 100 pg/μL, and the detection sensitivity is 10 times lower than that of single PCR. The detection technology was used to detect contaminated substrate samples of L. edodes from Beijing, Hebei and Henan Provinces, and the results met the actual production needs. This technology provides technical support for predicting the occurrence and scientifical prevention of substrate contamination from Trichoderma spp. of L. edodes.
The strain of Sanghuangporus mongolicus was isolated and purified from specimens collected on Hemiptelea davidii in the Horqin Sandy Lands of Inner Mongolia, China. Using this strain as experimental material, biological characteristics and fruiting conditions of the fungus were investigated. The results demonstrated that the optimal growth medium consisted of 200 g/L potato, 20 g/L elm branches, 20 g/L fructose, 2 g/L peptone, and 0.5 g/L potassium sulfate, and the fungus grew well at pH 6.0 and 35 ℃. Fruiting experiments revealed that the mycelial colonization phase required dark conditions at (30±3) ℃, while the fruiting stage necessitated (25±2) ℃ under moderate light exposure and ventilation. A substrate formulation containing 58% H. davidii sawdust/elm sawdust, 28% cottonseed hulls, and 10% wheat bran yielded fruiting bodies with superior agronomic traits and growth performance. These results provide reference for the commercial cultivation, development and utilization of S. mongolicus.
Growth conditions of Fomitiporia rhamnoidis are optimized, and bag-culture substrate for inducing and promoting the formation of fruiting bodies is further explored. The effects of carbon source, nitrogen source, temperature, pH and growth factors on the growth of F. rhamnoidis were studied by single factor test and orthogonal test. The results of single factor experiment showed that the optimum carbon source was soluble starch; the optimum nitrogen source was soybean powder, and the optimum growth factor was VB1. The fungus mycelia grew well at the optimum temperature of 28 °C and the optimum pH of 6.5. The results of orthogonal test showed that the mycelium growth rate was the fastest and the growth condition was the best when soluble starch was used as carbon source, wheat bran as nitrogen source, and VB7 as growth factor, and adjusting the pH to 6.0. Under the above optimal conditions, the mycelium bagfully colonized and yellowed, but the fruiting bodies were not successfully induced by using five different bag cultivation substrates. Domestication and cultivation of F. rhamnoidis need further study.
A strain of Irpex was isolated from Auricularia cornea-cultivated substrate using banboo shavings. Through morphological observation and ITS sequencing analysis, the strain was identified as Irpex laceratus. The orthogonal experiment results indicated that the main factor affecting the mycelial growth of I. laceratus was the nitrogen source, followed by the carbon source, pH and temperature. The optimal carbon to nitrogen ratio is 50:1-52:1. Addition of 14.5%-43.5% bamboo shavings significantly accelerated mycelial growth. The content of active substances varied with different formulations of cultivated substrates. Formula B (wood shavings 29%, bamboo shavings 29%, wheat bran 20%, cottonseed hulls 20%, lime 1%, gypsum 1%) exhibited the highest total triterpene content (66.10 mg/g); formula I (bamboo shavings 78%, wheat bran 20%, lime 1%, gypsum 1%) showed the highest total protein content (21.83%); formula CK (wood chips 58%, wheat bran 20%, cottonseed husks 20%, lime 1%, gypsum 1%) exhibited the highest total flavonoid content (0.99 mg/g); formula C (wood chips 14.5%, bamboo chips 43.5%, wheat bran 20%, cottonseed husks 20%, lime 1%, gypsum 1%) had the highest polysaccharide content of 2.80 g/100 g; formula F (bamboo chips 68%, wheat bran 20%, cottonseed husks 10%, lime 1%, gypsum 1%) demonstrated the highest total phenol content of 19.02 mg/g. The results provide valuable reference data for the cultivation and medicinal development of I. laceratus.
To augment feed resources and efficiently utilize straw, cellulose-degrading fungal strains were isolated from the rumen of Yajiang snow cattle in Xizang using a carboxymethyl cellulose selective medium. An investigation of their enzyme-producing properties and capabilities of degrading straw was carried out. The result was that six strains capable of degrading filter paper strips were discovered, of which one strain (XZ6) was classified as Pichia kudriavzevii. Single-factor optimization of enzyme production conditions showed that the optimal conditions for xylanase production by this strain were temperature of 33 °C, cultivation time of 18 h, shaker speed of 240 r/min, and inoculation volume of 1% (v/v). Under these conditions, xylanase activity peaked at 11.55 U/mL, which was 12 times higher than that before optimization. The β-glucanase activity peaked at 1.82 U/mL, a 7-fold increase over that before optimization; the β-mannanase activity reached a maximum of 2.07 U/mL, a 3-fold increase in the activity. Whole genome sequencing indicated that this strain included 408 glycoside hydrolase genes and 352 glycosyl transferase genes. The fermentation of highland barley straw and corn straw using the crude enzyme solution of strain XZ6 markedly diminished the levels of crude fiber, neutral detergent fiber, and acid detergent fiber in both types of straw. Strain XZ6 possesses the capacity to degrade straw fiber.
Twenty-four newly isolated strains of white-rot fungi were cultured on guaiacol- supplemented medium for screening laccase-producible strains, the capacity of laccase production of the 12 strains obtained was analysed in submerged fermentation and solid-state fermentation on different lignocellulosic substrates. The results indicated that there was no significant correlation between the laccase activity detected under submerged fermentation conditions and the ratio of colony diameters to photochromic laps among the 12 test strains (P>0.05). Similarly, no significant association was observed between the laccase activity detected during solid-state fermentation on different lignocellulosic substrates and the ratio of colony diameters to photochromic laps for the various test strains (P>0.05). In submerged fermentation, Cerrena unicolor LKY 798, Trametes hirsuta LKY 831, Funalia trogii LKY 846, and Neofavolus alveolaris LKY 850 exhibited comparatively high laccase activities keeping for a fairly long duration. N. alveolaris LKY 850 and Trametes sp. LKY 797 demonstrated the strongest laccase production capacities under solid-state fermentation condition using leaves of Quercus variabilis, Populus beijingensis, and needles of Pinus tabuliformis as substrates. It seems that leaves of Quercus variabilis and Populus beijingensis are more suitable than needles of Pinus tabuliformis for laccase production under solid-state fermentation condition. Additionally, Trametes sp. LKY 797, F. trogii LKY 846, and N. alveolaris LKY 850 showed relatively good laccase activities in fermentation using needles of Pinus tabuliformis as the substrate, with maximum laccase activities exceeding 3 700 U/mL.
To improve the utilization of Pseudostellaria heterophylla by-product resources and broaden the sources of feed, the bi-directional solid-state fermentation of medicinal P. heterophylla and Trametes versicolor was carried out. The substrate formula was optimized using mixture design with the mycelial growth rate as response value. The change in the content of bioactive compositions was assayed during the mycelial post-maturation stage. The content of nutritional and bioactive compositions and antioxidant activities of the fermented substrate were determined. The optimal substrate consisted of 59% corncob, 10% P. heterophylla roots, 29% wheat bran, 1% gypsum, and 1% lime. The content of polyphenols, flavonoids, polysaccharopeptide, and polysaccharides in the substrate increased with time during the mycelial post-maturation stage, while the triterpenoids decreased first and then increased. In scaled-up fermentation in plastic bags, the crude protein content of the fermented substrate was 8.20%, or increased by 31.41% as compared with that of the non-fermented substrate (6.24%). The crude ash content rose from 3.50% to 8.10% or increased by 131.43%. The content of polyphenols reached 6.87 mg/g, 8.70% higher than that of the non-fermented control (6.32 mg/g). The neutral detergent fiber content decreased considerably, while the acidic detergent fiber increased. The scavenging rate of ethanol extract of the fermented substrate against DPPH radicals was almost close to that of the non-fermented control, while that of the water extract was lower than that of the control. ABTS radical scavenging rate of both the water and ethanol extracts of the fermented substrate was lower than that of the non-fermented substrate. The present study provides a useful reference for the utilization of P. heterophylla by-product resources and the development of the fermented substrate as animal feed.
Dark septate endophytes (DSE) widely colonize the roots of plants in heavy metal- contaminated soils. The effects of DSE on the growth and medicinal components of Glycyrrhiza uralensis at different growth stages require further elucidation. In this study, pot soil containing different concentrations of cadmium (Cd) (0, 5, 10, and 15 mg/kg) was inoculated with Neocamarosporium betae (NB) and Cladosporium fusiforme (CF) to explore the effects of DSE inoculation on the growth and medicinal components of G. uralensis. The results showed that both CF and NB could effectively colonize the roots of G. uralensis. DSE inoculation increased the total root length, root volume, root surface area, and biomass of G. uralensis, as well as the content of available nitrogen in the soil. DSE colonization resulted in increase of soluble protein and glutathione levels in G. uralensis and decrease of malondialdehyde content. Variance partitioning analysis indicated that DSE explained 46.60%, 13.60%, and 7.80% of the variation of soil factors in the early, middle, and late stages respectively. Similarly, DSE accounted for 52.20% and 40.00% of the physiological variations of G. uralensis in the early and late stages, respectively. Soil factors contributed to 22.00%, 38.30%, and 58.40% of the variation of G. uralensis medicinal components in the three growth stages. Available potassium, phosphorus, and nitrogen in the soil significantly enhanced the biomass and root length of G. uralensis in the early stage. These data suggest that cadmium stress inhibited the growth of G. uralensis, while DSE inoculation could improve soil nutrient status and promote G. uralensis growth, and thereby facilitate the accumulation of glycyrrhizin and glycyrrhizic acid.
Remediation of aged soil contaminated by high molecular weight polycyclic aromatic hydrocarbons (HMW-PAHs) in a typical coal mining area was studied, using Fusarium sp. ZH-H2 as the remedial strain. A three-factor four-level orthogonal experiment was conducted to investigate the remediation efficiency of ZH-H2 under different environmental conditions. The study showed that the optimum environmental parameters for degradation of soil HMW-PAHs by ZH-H2 were 1 000-2 000 mg/kg of inoculation amount, 30-40 ℃ of temperature and 5%-15% of humidity. Under these environmental conditions, the total removal range of 10 HMW-PAHs was 22.72%-25.44%. Besides, building upon these optimized conditions, subsequent multi-stage inoculations revealed that both total HMW-PAHs and individual PAH removal rates peaked at the fourth inoculation cycle, achieving a total removal rate of 49.64% and individual removal rates ranging from 45.24% to 59.73%. Notably, benzo(a)pyrene (BaP) exhibited the highest degradation efficiency reaching 59.73%. This study innovatively proposes a ZH-H2-based multi-stage enhanced remediation strategy, establishing an “environmental optimization and inoculation enhancement” multi-stage remediation model for HMW-PAHs contaminated soils.
