Phallus is one of the most important genera of macrofungi, and has important economic values and taxonomic status. China is one of the key regions of Phallus species diversity, where a quarter members of total Phallus species in the world are distributed. Phallus species reported previously in China were 32 among them, some taxa remain controversial at the taxonomic status. Based on previous research and voucher specimens, Phallus species distributed in China were reexamined, and the number of hitherto known species was affirmatively 25, including 17 with available ITS sequence and 15 with available LSU. Seven species were treated as uncertainty or unknown distribution. Our study outlined the species diversity and resources of Phallus in China, providing valuable references for Chinese researchers in taxonomic research, species identification and commercial cultivation.

Amanita cyclic peptide toxins are mainly found in mushrooms of genera Amanita, Galerina, and Lepiota, and probably pose serious health risks to humans and animals if they are ingested accidentally. Rapid and accurate identification of these toxins in mushrooms and biological samples is crucial for ensuring food safety and diagnosing and treating mushroom poisoning. This review discusses various reported detection methods, focusing on high-performance liquid chromatography coupled with mass spectrometry, which is the main analytical method for determining Amanita cyclic peptide toxins in complex matrices. Furthermore, current trends and future directions in the detection of Amanita cyclic peptide toxins are also presented.

In this paper three new species of Tremellodendropsis and one new species of Guepinia from southwestern China, namely T. fusispora, T. oblongispora, T. transseptata and G. montana, are described based on morphological characteristics, molecular phylogenetic evidence and ecological traits. These species are common elements of funga in southwestern China, but were mistakenly treated as species originally described from Europe. Our data indicated that they are different from similar species of Europe or other continents, and should be treated as separate species.

Ten fungal isolates from the healthy rhizomes of Polygonatum cyrtonema naturally distributed in Xianyushan Nature Reserve of Qimen County in Anhui Province were identified based on colony and mycelial characteristics in the absence of conidia and sporogenous structures and molecular phylogenetic analysis. Five species in Neosetophoma, namely new species, N. endophyticum, N. polygonatum and N. qimenensis, and known species, N. poaceicola and N. guiyangensis were identified; besides, Setophoma caverna was also identified. The isolates were deposited in the Research Center for Entomogenous Fungi (RCEF) at Anhui Agricultural University.

The Sanghuangporus species are well-known due to their medicinal values. Polysaccharides, flavonoids and triterpenes are the main active ingredients of Sanghuangporus, and the medicinal values of the three have received widespread attention. Phellinus orientoasiaticus is a wood-inhabiting fungus growing in the living trees of Rosaceae. It is phylogenetically related with Sanghuangporus and its medicinal value is rarely reported. To determine the medicinal value of P. orientoasiaticus, the natural basidiomata of Sanghuangporus baumii and S. lonicericola from Northeast China were selected as comparative studying materials for they are extensively investigated medicinal fungal species. The content of crude polysaccharides, total flavonoids, and total triterpenoids and their antioxidant activities were used as comparative research indicators. The determination results showed that the polysaccharide content for S. baumii was 4.64 mg/g, 2.94 mg/g for S. lonicericola, and 3.68 mg/g for P. orientoasiaticus. The total flavonoid content was 65.53 mg/g, 187.74 mg/g, and 46.19 mg/g respectively. The total triterpene content of S. baumii was 5.38 mg/g, of S. lonicericola 15.87 mg/g, and of P. orientoasiaticus 10.17 mg/g. The antioxidant activities included DPPH free radical scavenging capacity, ABTS free radical scavenging ability, superoxide anion scavenging ability, and hydroxyl radical scavenging ability. The polysaccharides of S. lonicericola and P. orientoasiaticus showed comparatively high antioxidant activity, and the antioxidant activity of total triterpenoids of S. lonicericola and P. orientoasiaticus was also comparatively strong. This study demonstrates that P. orientoasiaticus has a good application prospect for its comparatively high polysaccharide, total flavonoid and total triterpenes content and excellent antioxidant activity.

Ophiocordyceps, the most prominent genus in the Ophiocordycipitaceae, is renowned for its numerous entomopathogenic species with significant medicinal value. Despite the existing descriptions of numerous species, the complete diversity of this genus remains enigmatic. In this study, four Ophiocordyceps species previously unknown in China were collected, and a comprehensive approach, including multigene phylogenetic analyses (ITS, SSU, LSU, RPB1, RPB2, and TEF1α) along with morphological assessments was employed to determine their taxonomic status. Illustrated descriptions and taxonomic discussions for these newly discovered species are provided, contributing to our understanding of the diversity within the Ophiocordyceps genus.

Spider is a general term for all species of the order Arachnoidea in the phylum Arthropoda, and the fungi parasitized spider is called araneopathogenic fungi. Spider pathogenic fungi play an irreplaceable role in continuously controlling spider populations and maintaining ecological balance, and some species are rich in various biological activities and have important economic value. In this study, classical morphological and molecular systematics classification methods were combined to identify specimens and isolated strains from various forest ecosystems based on the latest entomopathogenic fungal classification system for revealing species diversity of spider pathogenic fungi in China. More than 80 specimens of spider pathogenic fungi from all over the country are available, and 2 families, 8 genera, and 21 species are identified, including 3 new species (published elsewhere), 7 unknown species and 1 new Chinese recorded species. Among them, Purpureocillium is dominant genus, with relative abundance of 26.25%; Secondly Gibellula, with relative abundance of 20.00%. Only one species (one specimen) is found for Akanthomyces. Some Isaria-like strains were referred to Akanthomyces sp. and Samsoniella spp. Chlorocillium griseum was reported for the first time in China. According to relevant literature on spider pathogenic fungi that has been published worldwide, a total of 18 genera and 119 species has been published, and all distributed in the Hypocreales of Ascomycota. Most species belong to Cordycipitaceae, with approximately 12 genera and 106 species. A few are distributed in the family Ophiocordycipitaceae, with 4 genera and 11 species; there is only one species in Bionectiaceae, and one species in Chlorocillium that classification status is situated between Ophiocordycipitaceae and the Clavicipitaceae. No species has been discovered in Clavicipitaceae yet.

Based on systematic resource investigation, species identification, and literature arrangement, the checklist of poisonous mushrooms in Beijing was complied. The poisonous mushrooms were classified according to the poisoning type. Scientific name, Chinese name, and references of each poisonous mushroom are provided. By 2023, 130 poisonous mushrooms have been found in Beijing, causing five known damage types. However, there are still some poisoning types difficult to confirm. The most toxic mushrooms to be concerned are Amanita subjunquillea and Lepiota brunneoincarnata, which can cause acute liver failure, as well as Amanita oberwinklerana, which can lead to acute renal failure.

A new species, Coltricia wuyiensis, is described from eastern China on the basis of morphological examination and molecular phylogeny. Phylogenetic analysis based on the internal transcribed spacer (ITS) regions and nuclear large subunit (nLSU) ribosomal RNA gene regions indicates that the new species is nested within the genus Coltricia in the order Hymenochaetales. Coltricia wuyiensis is characterized by the centrally stipitate basidiomata with even margin, orange brown to reddish brown pileal surface with concentric zones and clay-buff to fawn pore surface when fresh, loosely interwoven to subparallel tramal hyphae, sometimes septate basidioles, and broadly ellipsoid basidiospores measuring 4.3-5.5×3.7-4.4 µm. Phylogenetically it is closely related to C. minima, but the latter species has aseptate basidioles and bigger basidiospores (6-7×4-5 µm vs. 4.3-5.5×3.7-4.4 µm). An identification key to accepted species of Coltricia in China is given.

The inhibitory effects of different polar extracts of Sanghuangporus vaninii (water, ethanol, petroleum ether, ethyl acetate, n-butanol, and extraction residual water extracts) on the production of inflammatory cytokines IL-6 and TNF-α in RAW264.7 cells stimulated by LPS were evaluated. The inhibition of SARS-CoV-2 replication and entry by ethanol and water extract of S. vaninii was appraised. The expression level of nucleocapsid protein was detected by real-time fluorescent quantitative PCR and Western Blot using Vero as host cells. The results indicated that the ethanol, petroleum ether, ethyl acetate, n-butanol, and water extracts of S. vaninii showed good anti-inflammatory effects, and the inhibitory effect was significantly enhanced with the increase of concentration, and the levels of IL-6 and TNF-α produced by RAW264.7 cells stimulated by LPS were significantly reduced. The ethanol extract and water extract of S. vaninii significantly inhibited viral replication by inhibiting the expression level of nucleocapsid protein in SARS-CoV-2.

Seventy isolates of Diporthe fungi from healthy Camellia oleifera leaves were obtained by using tissue isolation method. Based on morphological characteristics and phylogenetic analysis of five fragment sequences (ITS, TEF, TUB, CAL, HIS), eight representative isolates were identified as six species, D. biguttulata, D. eres, D. hongkongensis, D. fusicola, D. qimenensis sp. nov. and D. reniformis sp. nov. β-Type conidia were not observed in the two new species. D. reniformis produces distinct reniform conidia.

Yunnan tropical areas of southwesten China with complex topography and diverse vegetations are endowed with abundant macrofungal resources. In this study, more than 2 100 specimens of wood-inhabiting macrofungi were collected. A total of 668 species, belonging to 245 genera, 67 families, 17 orders and 2 phyla, was identified based on morphological and molecular evidence, including 1 new family, 4 new genera and 64 new species previously published by the authors. Fourteen dominant families with more than 10 species for each family are Polyporaceae, Hymenochaetaceae, Xylariaceae, Steccherinaceae, Auriculariaceae, Hypoxylaceae, Incrustoporiaceae, Phanerochaetaceae, Meruliaceae, Mycenaceae, Fomitopsidaceae, Schizoporaceae, Stereaceae, and Dacrymycetaceae; they include 500 species, accounting for about 74.85% of the total species. There are 34 dominant genera with more that 5 species for each genus, including Hymenochaete, Xylaria, Trametes, Ganoderma, Fuscoporia, Skeletocutis and so on. They include 336 species, accounting for about 50.30% of the total species. Wood-inhabiting macrofungi of the studied areas are of typically tropical properties. There are 27 edible species, 75 medicinal species and 6 poisonous species.

Tulosesus is the second largest genus next to Coprinopsis in the number of known species of coprinoid fungi. Its main characteritics are that both the cap and the stipe present cystidiate. It is treated as an independent genus separating from Coprinellus recently, while its phylogenetic position and framework are still controversial. From 2021 to 2023, we have carried out a nationwide field research, and found more than 20 species of Tulosesus in China. In this paper, 10 species new to China are reported and illustrated with line drawings, viz. Tulosesus brevisetulosus, T. canistri, T. cinereopadillus, T. fuscocystidiatus, T. marculentus, T. pellucidus, T. plagioporus, T. sabulicola, T. sassii and T. velatopruinatus. Using ITS and LSU sequence datasets, the phylogenetic framework of this genus was inferred. The results show that Tulosesus is a monophyletic genus composed of three independent subgenus lineages. Combined with morphological characteristics, the characteristics of each lineage of this genus were preliminarily clarified.

Huishuhua (maitake) is a macrofungus with high edible and medicinal values in China. It was commonly recognized as Grifola frondosa. A total of 45 huishuhua samples from East Asia, Europe and North America was analyzed on the basis of morphological and multi-gene phylogenetic methods. The results showed that highly supported clades of huishuhua samples were corresponding to each continent, indicating that genetic differences existed to a certain extent among strains from different regions. All huishuhua strains isolated from China nested in the East Asian clade. The European and North American clade members are not yet found in China. The East Asian huishuhua was described in Japan as G. albicans, and this has been once regarded as a synonym of G. frondosa. Because most wild and cultivated huishuhua in China has dark-colored pileus, a new form G. albicans f. huishuhua is thereby proposed. Holotype specimen and extype strain of this form are designated (voucher specimen HBAU15778, voucher strain HBAU LM502).

Phyllosphere microorganisms play important roles in ecological functions, such as maintaining plant health, promoting plant growth and biogeochemical cycling. In recent years, the role and importance of phyllosphere microbes have received increasing attention. The diversity and community structure of epiphytic fungi in the phyllosphere of Rosa chinensis, a common greening plant in Beijing, were investigated. Four sites from the center to north of the city and near to far distance, Andingmen area (ADM), Hepingjie area (HPJ), Yayuncun area (YYC), and Beiqijia area (BQJ), were selected. The samples were taken in three seasons (April, July, and November) at different locations (the middle and sides of the road of the selected areas). A total of 3 425 amplicon sequence variants (ASVs) was obtained, and they were dominated by Dothideomycetes. The variance of Shannon index between sampling sites were ADM > BQJ > HPJ > YYC, indicating ADM is significantly higher than YYC. The diversity shows a decreasing and then increasing trend from the center to north of the city. The variance of Shannon index between seasons showed April > July > November, and April was significantly higher than July and November. There is no significant difference between the samples from the center and the sides of road. LEfSe analysis suggested the significant biomarkers of different regions and seasons. Significant biomarkers of Andingmen area were Neosetophoma, Neophaeococcomyces and Pseudocosmopora, while the significant biomarkers of Hepingjie area, Yayuncun area and Beiqijia area were Preussia, Orbilia and Pseudotomentella, respectively. Network analysis showed the dominant genera were Vacuiphoma, Erythrobasidium, Cladosporium, Mrakia, and Sphaerulina. The main functional groups of epiphytic fungi in the phyllosphere of Rosa chinensis were pathotroph and saprotroph. The prediction of fungal community function demonstrates that saprophytic functional groups were different in different seasons and sampling sites. The study revealed structural changes of phyllosphere fungi in urban plants and might be valuable to scientific management of urban greening.

Active substances of Ganoderma lingzhi are the material basis for the different functions of G. lingzhi. During the growth process of G. lingzhi, the content of active substances dynamically changes with the growth of basidiomata, and affects the quality and efficacy of G. lingzhi. Wood log cultivation is one of the two main cultivation modes of G. lingzhi, and the basidioma have a long growth cycle and accumulation time of active substances. Studying the changes of active substances during the growth stage has a positive significance for the efficient utilization of G. lingzhi materials. Currently, related reports have mainly focused on polysaccharides and triterpenoids, with little attention concerning other bioactive components of G. lingzhi, and the growth cycle covered was not complete enough. Therefore, there was limited understanding of the complete dynamic changes of each substance. This study analyzed the dynamic changes of active substances in the samples collected at scheduled time during nine stages of basidioma growth within 120 days from primordium formation stage to the end of sporulation. The summary is as follows: the content of total polysaccharides and twelve triterpenoid compounds in G. lingzhi was the highest in the primordium formation stage, rapidly decreased with growth, and became stable in the later stage of sporulation; the content of ergosterol was the highest during the primordium formation stage, rapidly decreased with growth, and slightly increased during the sporulation period; the content of uridine, uracil, and water-soluble protein/polypeptide continuously declined with the growth of basidiomata until the lowest, while adenosine changed undulately with the peak appearing in 60 days when G. lingzhi sporulation began. The total antioxidant capacity was weak during the primordium stage, and strong during the later stage of basidioma growth. This study elucidated the dynamic changes in the content and activity of the main active substances at the entire growth stage of G. lingzhi basidiomata, and provided a theoretical basis for guiding the production and efficient development and utilization of G. lingzhi.

Macrofungi play essential roles in ecosystems, but the seasonal dynamics of macrofunal communities in forests remain unclear. Macrofungal community componets of five forest types in different seasons in Ji’an National Nature Reserve (Jilin, China) were analyzed on the basis of collections made from 2019 to 2021. Environmental variables of selected plots in the five typical vegetation types were evaluated. In total, 1 325 specimens were identifed by using morphological and molecular biology methods, and 293 species belonging to 125 genera, 68 families, 18 orders, 6 classes, and 2 phyla were obtained. Russulaceae (37 species, 12.63% of the total), Amanitaceae (26 species, 8.87%), Tricholomataceae (20 species, 6.83%), and Boletaceae (19 species, 6.48%) were the most diverse family. Lactarius (19 species, 6.48% of the total), Amanita (16 species, 5.46%), Russula (16 species, 5.46%), and Cortinarius (14 species, 4.78%) were dominant. Diversity indices of macrofungi showed increasing trends from Pinus koraiensis forests to Quercus mongolica forests. The cumulative species richness was ranked as: Q. mongolica forest>broadleaf mixed forest>Q. mongolica and P. koraiensis mix forest (Q. mongolica as the dominant species) >P. koraiensis and Q. mongolica mix forest (P. koraiensis as the dominant species)>P. koraiensis forest. Agarics (204 species, 69.62% of the total) were the dominant group of macrofungi, and they were mainly found in Q. mongolica forest in August and influenced by soil temperature, air temperature, soil moisture, and the cover degree of Q. mongolica, especially significantly influenced by soil temperature (P<0.05). It is found that most macrofungi have a preference for forest types with a relatively high cover degree of Q. mongolica. Therefore, the deliberate protection of Q. mongolica forests is beneficial to maintaining macrofungal diversity.

Naematelia aurantialba, a rare jelly edible fungus of the Tremellales order, is endemic to China and is often used as traditional medicine and food due to its exceptional nutritional value and biological activity. It is a heterothallic basidiomycete with a tetrapolar mating system. However, the structures of mating loci have not been characterized. In this study, the genome of four different mating types of monokaryotic strains of N. aurantialba was sequenced by second generation sequencing technology, and mating type A and B loci were searched and compared with those of other macrofungi by bioinformatics method based on genomic data. The results showed that there were two A mating-type loci and two B mating-type loci in these monokaryons. All two A mating-type loci contain a pair of HD1 and HD2 genes arranged in a typical ‘head-to-head’ manner and the synteny of several genes’ upstream and downstream of A loci was higher as compared with that of other fungi, including conserved Oxidoreductase and PRL22 genes. However, the mip and β-fg genes are not closely linked to the HD gene and are not located on both sides of it. All two B mating-type loci contain a pheromone receptor and a pheromone precursor gene. Upon comparison of the B1 and B2 loci, it was discovered that the STE3 and phB genes at the B1 locus were situated in close proximity with one RVT_1 gene located upstream of phB. In contrast, at the B2 locus, three genes were inserted between the STE3 and phB genes, and the STE12 gene was absent from the B1 locus. Compared with B loci in other fungi, the synteny of B loci was poor, indicating that B loci were highly variable in different fungi. The gene for the pheromone receptor NaSTE3-3 is present in all four monokaryons, however, the gene for the pheromone precursor is not located in upstream or downstream, and the B mating type site is not present on the same contig. These results obtained would help us to understand the mating locus structures of N. aurantialba, being helpful to genetic breeding of this mushroom.

In order to explore the underlying mechanism of fruiting body formation and development of Pleurotus ostreatus, the small molecular metabolites in primordia and newly differentiated fruiting bodies were investigated through metabolomics approaches. All the 545 metabolites including phenolic acids, lipids, amino acids and their derivatives, nucleotides and their derivatives, organic acids, alkaloids, tannins, lignans and coumarins, and others were identified. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) revealed a significant difference between metabolites of primordia and newly differentiated fruiting bodies. A total of 253 significantly differential metabolites was screened and identified under the conditions of VIP (varible importance in the projection) ≥1 and fold change ≥2 or ≤0.5, involving in 76 pathways. It was inferred that the deep transformation and regulation of components were accomplished via tryptophan metabolism, phenylalanine, tyrosine and tryptophan biosynthesis, lysine degradation, d-arginine and d-ornithine metabolism, biotin metabolism, tropane, piperidine and pyridine alkaloid biosynthesis, tyrosine metabolism, purine metabolism, propanoate metabolism and isoquinoline alkaloid biosynthesis pathways. This study will provide a theoretical basis for exploring the developmental mechanism of Pleurotus ostreatus fruiting body.

Content of total polyphenols, triterpenoids and ergosterol of ethanol extracts from basidiomata of different sanghuang species, Sanghuangporus baumii, S. vaninii, S. sanghuang (wild) and Inonotus hispidus from Anhui, Zhejiang, Jilin, Shanxi and Shandong provinces, were compared. The results showed that there were apparent differences in the content of total polyphenols, triterpenoids and ergosterol in basidiomata from different sources. The highest content of total polyphenols was tested in S. vaninii from Zhejiang (1.99%), the highest content of triterpenoids was tested in I. hispidus from Shandong (1.32%), and the highest content of ergosterol was tested in S. vaninii from Jilin (0.19%). Malondialdehyde (MDA) was used as an index to compare the antioxidant activities of different species of sanghuang. The extracts of all tested species showed good inhibitory effects on the production of MDA in rat liver microsomal lipid peroxidase, of which the extract of S. vaninii from Zhejiang had the highest antioxidant activity (96.53%). Nineteen compounds from the aforementioned samples were identified by liquid chromatography-ion trap-time-of-flight mass spectrometry (LC-IT-TOF-MS). Among them, the metabolites of basidiomata of S. vaninii from Zhejiang, S. vaninii from Jilin and S. baumii from Anhui were similar, which were mainly composed of the hispidin derivatives. Sanghuangporus sanghuang (wild) basidiomata mainly contained hispidin derivatives, but the structure types of the derivatives were different from those of aforementioned samples. There was a large quantity of hispidin in basidiomata of I. hispidus from Shandong, and the content of hispidin derivatives was low. In conclusion, the apparent differences in structure types and content of active metabolites as well as their antioxidant activities were observed from different species or localities of sanghuang. This study provides scientific references for the development of antioxidant products of sanghuang species.

Flammulina filiformis is one of the edible fungi protected by the law of Protection List of New Varieties of Plants. The intellectual property protection of varieties is highly valued, and there is an urgent need for accurate, reliable, stable, and operable variety identification technologies to provide technical support for the protection of intellectual property rights of varieties. The study used the genome resequencing data of 42 F. filiformis strains to construct a database containing 428 polynucleotide polymorphism (MNP) molecular markers, and analyzed the genetic similarity between the test strains. The results showed that the genetic similarity of the 12 strains used by domestic cultivators was 100%, implying that domestic F. filiformis production varieties are highly homogeneous. The genetic similarity among the 12 wild strains ranged from 0.23% to 29.44% (except for F17 and F18, whose similarity was 100%), reflecting the rich genetic diversity of wild F. filiformis. The identification results of MNP molecular markers were also compared with the results of antagonistic experiments and ISSR molecular markers, reflecting the advantages of high sensitivity, good stability, and small workload of MNP molecular marker strain identification technology. Therefore, this identification technology can provide technical support for the intellectual property protection of F. filiformis varieties in China.

Species of Geosmithia (Ascomycota, Hypocreales) are commonly associated with bark beetles, and several species can infect tree hosts and cause serious diseases. In this study, a culture-dependent and pathogenic trial was conducted to classify a species of Geosmithia associated with Tomicus armandii infesting Pinus armandii in southwest China. In total, 387 Geosmithia isolates were obtained from 45 adults and 64 galleries of T. armandii in Yuxi and Fuyuan Cities, Yunnan Province, China. Ten representative isolates were further identified on the basis of morphological and physiological characteristics and the phylogenetic analyses were conducted based on ITS, TUB2, and TEF1-α sequences. Among these, 90 isolates were used for the establishment of the ITS tree, 66 of the TUB tree, and 77 of the TEF tree. As a result, a novel species, G. armandii was identified. This is the first species of Geosmithia reported to be associated with T. armandii. In vivo pathogenicity analysis of the two isolates selected indicated that in one month after inoculation, there was no significant difference between average lesion length caused by the inoculated isolates and the experimental control, showing that G. armandii is a weak pathogen of Pinus armandii.

In order to obtain the best preparation process of selenium polysaccharide from selenium-enriched Auricularia heimuer mycelium, A. heimuer strains Jiufeng, Heifeng and Xizang 6 were used as materials, and selenium tolerance and gradient concentration selenium enrichment screening, single factor test, response interview design analysis, and antioxidant determination were carried out for extraction and process-optimization of selenium polysaccharide from the mycelia. Antioxidant determination method was used to examine antioxidant activity of selenium polysaccharide. The results showed that ‘Jiufeng’ had the strongest selenium tolerance among the three A. heimuer strains. Selenium enriched fermentation experiment indicated that the total selenium content of A. heimuer mycelia was the highest when the selenium concentration was 60 μg/mL. The optimal conditions for selenium polysaccharide extraction were: ultrasonic time of 25.4 min, water bath time of 56 min, and material to liquid ratio of 1:49 (g:mL). Under this condition, the extraction rate of selenium polysaccharide was (17.49±0.10)%. Selenium-enriched A. heimuer mycelial polysaccharide has certain scavenging ability to DPPH and hydroxyl radicals, and shows strong reducing ability in the total reducing power test. The research results provide a preliminary theoretical basis for the selenium enriched culture of A. heimuer strains, and extraction of selenium polysaccharides from A. heimuer mycelia.

The blue fluorescent protein expression plasmid PJW-EXP-intron-opbfp and yellow fluorescent protein expression plasmid PJW-EXP-intron-opyfp were constructed and transformed into protoplasts of Ganoderma lingzhi. Transformants were selected by PCR and subsequently microscopically screened for fluorescent signals. The pJW-EXP-intron-opbfp and pJW-EXP- intron-opyfp transformants respectively exhibited blue and yellow fluorescence in the mycelia. However, no fluorescence was detected in wild-type (WT) strains. These results showed that blue and yellow fluorescent proteins were successfully expressed in G. lingzhi. Moreover, the activity of the cellulase Ⅱ gene promoter (Pcbh2) was evaluated in G. lingzhi using yfp as a reporter gene. The yfp-Pcbh2 transformants were obtained by genetic transformation of the plasmid yfp-Pcbh2 into protoplasts of G. lignzhi. Under induction of inducer avicel, the mycelia of the yfp-Pcbh2 transformants emitted yellow fluorescence signals, while no fluorescence was detected in the mycelia without the inducer. These results showed that Pcbh2 was successfully activated by avicel in G. lingzhi.

Pyrvinium pamoate (PP) is an antiparasitic drug, which has been found to exert synergistic effect with azoles against Aspergillus fumigatus. The putative protein encoded by the af-fadO gene in A. fumigatus is a FAD binding oxidoreductase, which might be involved in the respiration chain in the mitochondria. af-fadO knockout strain was constructed to investigate the role of the gene in antifungal susceptibility, osmotic and oxidative stress adaption of A. fumigatus, and the synergistic effect of PP and azoles. af-fadO knockout strain was constructed via homologous gene replacement method. The effect of af-fadO disruption on the in vitro antifungal susceptibility, oxidative and osmotic stress adaption as well as the effect on the combinational interactions between PP and azoles (posaconazole, itraconazole, voriconazole) in the mutant Δaf-fadO and wild-type (WT) (ku80, pyrG⁺) strains was observed. The effect of the gene on synergism of PP and azoles was investigated via broth microdilution checkerboard technique and disk diffusion method. In vitro antifungal susceptibility assay revealed that fractional inhibitory concentration index (FICI) value of PP combined with posaconazole against Δaf-fadO was 0.563, indicating no interaction, whereas the FICI against WT was 0.5, indicating synergistic effect. The results of disk diffusion method showed that the inhibitory zone diameter of PP combined with posaconazole against Δaf-fadO was significantly smaller as compared with that against WT. In addition, Δaf-fadO was more sensitive to the oxidants H₂O₂ and menaphthoquinone, osmotic agents NaCl and d-sorbitol as compared with WT. The disruption of af-fadO gene in A. fumigatus results in mutant Δaf-fadO showing more sensitivity to oxidative and osmotic stress, reversing the synergistic effect of PP and posaconazole. The present study enhances the knowledge of PP and azole antifungal mechanism, and is valuable for future clinical application.

Polycephalomyces ramosus is a rare species of Cordyceps-like fungi. Many species in Polycephalomyces have medicinal or health protection values. The synnemata of P. ramosus are domestically cultivated for the first time and the biological characteristics and active components of the fungus are studied. The mycelia have been identified by ITS sequence. The carbon source, nitrogen source, cultivating temperature and pH for mycelial growth were optimized by single factor experiment and orthogonal experiment. Rice, wheat, millet and coix seed were used as cultivated media. The content of polysaccharide was detected by phenol-sulfuric acid method and the content of adenosine and cordycepin was detected by high performance liquid chromatography (HPLC). The optimum medium for solid culture was sucrose 20 g/L, peptone 15 g/L, agar 20 g/L, KH₂PO₄ 1g/L, MgSO₄·7H₂O 1 g/L, and Vitamin B₁ 10 mg/L; the optimum pH was 6.5 and growth temperature 23 ℃. The optimal liquid culture medium was sucrose 30 g/L, peptone 15 g/L, KH₂PO₄ 1 g/L, MgSO₄·7H₂O 1 g/L, and Vitamin B₁ 10 mg/L; the mycelia grew well under pH 6.5 and temperature 25 ℃. The yield of synnemata on wheat as culture medium was the highest, with average biological efficiency of 9.824%. The polysaccharide content of synnemata on rice media was the highest (4.693%). The content of adenosine in the synnemata produced on millet media was the highest (0.130%), while the content of cordycepin was the lowest (0.018%). The study provides a reference for utilization and artificial cultivation of P. ramosus.

Fungi, integral components of the gut microbiota, exert direct or indirect influence on host health. Extensive research has focused on the anti-aging effects of gut bacteria, however, investigations into the impact of intestinal fungi on anti-aging are limited. In this study, a combination of broad-spectrum antibiotics was employed to deplete bacteria from the feces of young mice, which were subsequently transplanted into the intestines of naturally aged mice. Physiological parameters, including learning and memory abilities, body weight, organ indices, and histopathological changes were assessed. Real-time quantitative PCR and enzyme-linked immunosorbent assay were utilized to evaluate the expression levels of aging-related genes, proteins, and inflammatory factors in various tissues of aged mice. Results demonstrated that fecal microbiota transplantation (FMT) following bacterial depletion significantly improved spatial learning and memory abilities in aged mice without adversely affecting body weight and other organ indices. Further analysis revealed that FMT ameliorated tissue conditions in the brain, liver, spleen, lung, kidney, and small intestine in aged mice. FMT upregulated genes associated with intestinal barrier integrity while downregulated the expression of aging-related genes p53 and p21 in the brain, liver, and kidney of aged mice, with no significant impact observed on the lung, spleen, and small intestine in aged mice. Additionally, FMT significantly increased antioxidant enzyme levels in the serum and multiple tissues of aged mice and reduced the expression of aging biomarkers and pro-inflammatory factors. In conclusion, the gut fungal microbiota from young donor can effectively delay the aging process in serum, brain, liver, and kidney. This study provides a scientific foundation for further exploration of the potential mechanisms underlying the anti-aging effects of gut fungi.

Lentinula edodes is edible and medicinal mushroom with the second largest yield and output value in the world. It is necessary to explore the regulatory mechanism of qualified characteristics and the important functional genes related to characteristics in L. edodes. In this study, a known functional gene is edited by using CRISPR/Cas9 technology, and CRISPR/Cas9 is confirmed to work in L. edodes. By predicting and screening a U6 snRNA promoter in L. edodes, sgRNA was designed using a pyrG encoding gene of orotidine-5ʹ-monophosphate decarboxylase as the target gene for CRISPR/Cas9. According to the codon preference of L. edodes genome, the cas9 gene that is the core component of CRISPR/Cas9 system is optimized. A binary expression vector with both sgRNA and cas9 gene was constructed. The vector was transformed into the monokaryon by Agrobacterium-mediated genetic transformation of L. edodes mycelium to make the mutation of pyrG gene. The L. edodes monokaryon mutant with base deletion in pyrG gene was obtained successfully and the mutant was determined by functional screening as an uracil trophic deficiency strain. In this study, a binary expression vector was successfully constructed, which could simultaneously express the two important components of CRISPR/Cas9, and the purpose of using CRISPR/Cas9 to edit functional genes was realized.

Coumestrol and psoralen as important derivatives of coumarins were mainly found in plants and play important roles in anticancer and anti-inflammation but were less reported in fungi. Our study successfully isolated and characterized Inonotus hispidus L-1, a strain with high polysaccharides and flavonoids. Metabolomic analysis indicated that the content of coumestrol and psoralen in basidiomata of I. hispidus L-1 was significantly higher than that in mycelia and primordia. Combining transcriptomics and metabolomics approaches, the synthetic pathways and regulatory genes involved in the production of coumestrol and psoralen were elucidated. The synthesis of coumarin and psoralen in I. hispidus L-1 has a common precursor, L-phenylalanine. L-phenylalanine synthesis was co-regulated by the glycolysis/gluconeogenesis pathway and the phenylalanine pathway, and its levels were significantly elevated in the primordium and basidioma stages. More interestingly, the downstream metabolites in the P-coumarate acid synthesis pathway, by using L-phenylalanine as a precursor, were significantly increased. These downstream metabolites include 2,7,4ʹ-trihydroxyisoflavanone and coumestrol in the isoflavonoid biosynthesis pathway, as well as 7-hydroxycoumarine and psoralen in the coumarin biosynthesis pathway. The synthesis of coumestrol and psoralen was mainly regulated by the phenylalanine ammonia-lyase (APL), APL1, and glutathione S-transferase omega-like 2 (ECM4) genes. The results highlight the potential of I. hispidus L-1 as a valuable source of coumarins and psoralens for the development and application of nutraceuticals.

As an agricultural byproduct, the husks of lotus seeds can be used to cultivate edible fungi. This study aims at utilizing lotus seed husks to cultivate Ganoderma lingzhi in order to reduce the costs and increase the efficiency of cultivation. The 10%, 20%, 40%, 60% and 80% lotus seed husks were used to replace sawdust and corncob in the basic formula (45% sawdust, 45% corncob, 5% wheat bran, 4% corn flour, and 1% gypsum) of conventional cultivating substrate. The agronomic traits, nutrient content and production benefits were investigated to select the high-efficient formula. Results showed that certain proportion of lotus seed husks in the substrate could promote the mycelium growth rate of G. lingzhi. The growth rate reached 4.44 mm/d under the replacement of 60% lotus seed husks. Ganoderma lingzhi cultivated in the lotus-seed-husk substrate displayed shorter stipe, thinner cap and lower moisture content of basidiomata, while the cap size, stipe diameter and fresh weight of basidiomata showed no significant difference. The determination of nutrients revealed that the polysaccharide and protein content in basidiomata significantly increased with the increase of lotus seed husk proportion in the substrate. The polysaccharide content in basidiomata cultivated in the substrate with 80% lotus seed husks (10.57 mg/g) was 1.66 times higher than that in the conventional substrate, and the protein content (15.14%) was also significantly higher than that in basidiomata cultivated in the conventional substrate (14.12%). However, the triterpenoid acids displayed a trend of “low proportion promotion, high proportion inhibition” in the substrate with lotus seed husk replacement. Besides, lotus-seed-husk substrate significantly improved total amino acid content in G. lingzhi basidiomata, particularly in the substrate with 10% lotus seed husks (32.44%). In total, 13 amino acids were detected. Production benefit analysis showed that G. lingzhi cultivated in lotus seed husk substrate significantly improve the cultivation profit, and the profit was the highest (1.22 yuan per bag) when cultivation was carried out in the substrate with 80% lotus seed husks.

The genus Diplodia (Botryosphaeriales, Botryosphaeriaceae) comprises many important pathogens or latent pathogens associated with stem canker, shoot blights and die-back of plants across a wide geographic distribution. In this study, a total of 13 Diplodia isolates was obtained from diseased branches of loquat (Eriobotrya japonica) trees in Yunnan Province. Based on phylogenetic analyses of ITS, tef1 and tub2 loci, combined with morphological characteristics, two species were identified, D. mutila new to China and D. pipa sp. nov. The pathogenicity tests conducted on potted loquat trees demonstrated that both species can infect loquat trees, with D. mutila exhibiting a higher level of virulence as compared with D. pipa. To our knowledge, this is the first report of Diplodia from loquat trees in China.

Sanguinoderma is an important group of Ganodermataceae, many physiological functions have been found in the species of the genus, which are better than those of many well-known edible and medicinal fungi. Domestication of five wild Sanguinoderma species was attempted, and the cultivation characteristics and antioxidant activities of the species were studied, including the optimal temperature of mycelial growth, basidioma development process, polysaccharide and triterpene content and biological efficiency. The results showed that the optimum temperature for mycelium growth of the five species ranged from 25 to 30 ℃, and 25-38 days were needed for the fruiting body growth from primordium formation to harvest of basidiomata. S. rugosum growth is fastest and sporulation is early, while Sanguinoderma sp. growth is slowest and no sporulation was observed during cultivation. The morphological characters of cultivated basidiomata were quite different among different strains. The pilea were fan-shaped, reniform to nearly circular, with surface of greyish brown, ash-black, black and beige red, and length ranged from 4.96 cm to 12.56 cm, while width ranged from 3.84 cm to 10.76 cm, and height ranged from 0 cm to 10.1 cm. The first harvest biological efficiencies were 6.59%-18.34% for different strains, and the crude polysaccharide content and the total triterpene content were 1.89%-2.95% and 0.78%-1.68%, respectively. The antioxidant activities of polysaccharides and total triterpenes were compared with those of six Ganoderma species. It was found that Sanguinoderma spp. have better antioxidant activity. Sanguinoderma sp. and S. microporum have radical scavenging rate of over 80% to DPPH, and of 28% to total oxidant, under concentration of 3 mg/mL crude polysaccharide and 1 mg/mL total triterpene, respectively. In conclusion, Sanguinoderma spp. grow fast, contain rich polysaccharide and triterpene in basidiomata, and have good antioxidant activities for potential utilization.

The response to phosphorus fertilizer level and planting pattern of soil fungal community in wheat field was analyzed. Three phosphorus (P) application rations, 90 kg/hm² (P1), 135 kg/hm² (P2) and 180 kg/hm² (P3), and two planting modes mono-cropping (WS) and intercropping (WI), were set. Illumina high-throughput sequencing was used to investigate the effects of different phosphorus fertilization levels and intercropping on soil nitrogen and phosphorus content and fungal community composition and diversity in the field of winter wheat. The results showed that different phosphorus application levels had significant effects on soil phosphorus content in intercropping and mono-cropping wheat fields. Intercropping was beneficial to increase available phosphorus content of the soil with low phosphorus level. In total, 1 157 759 effective tags and 847 OTUs were obtained by high-throughput sequencing. All the treated OTUs were classified into 8 phyla, 26 classes, 54 orders, 82 families and 108 genera. There were 196 OTUs in 6 treatments. There were 147 and 102 unique OTUs in intercropping and monoculture modes, respectively. There were 113, 70 and 66 OTUs unique to P1, P2 and P3 treatments, respectively. Phosphorus application and intercropping had little effect on fungal alpha diversity in wheat rhizosphere soil. The results of PCoA analysis showed that the level of phosphorus application and intercropping and their interaction had certain effects on fungal community structure. The results of redundancy analysis showed that soil available phosphorus had a greater impact on the fungal community structure, and Ascomycota, Basidiomycota, Mortierellomycota and Glomeromycota were positively correlated with available phosphorus, and Chytridiomycota was negatively correlative.

An endophytic fungus strain GZY-Z-02 was isolated from the leaves of Paraboea sinensis and identified as Purpureocillium lilacinum by molecular biological method. The fungus was fermented on rice medium for 30 days at room temperature, and the crude extracts were obtained by extraction with ester acetate. By means of silica gel column chromatography, Sephadex LH-20 and HPLC twelve metabolites were obtained. The twelve metabolites were identified through NMR, MS and CD spectra, which all belong to alkaloids, including a new compound lilacinumopin (1), five diketopiperazines (2-6), two indole analogs (7-8), three nucleoside derivatives (9-11) and Vitamin B₂ (12). An antimicrobial activity assay proved that the compounds 1, 7, and 8 displayed selectively moderate antibacterial activity to Staphylococcus aureus.

The Shaluli Mountains are located in the central part of the Hengduan Mountains, southwest China. This is an area with wide ranges of altitudes and vertical distributions of vegetations. Macrofungal resources in this area are rich due to its diverse climate, varied topography, and diverse vegetation types. A three-year survey of macrofungal species diversity in the Shaluli Mountains was conducted to explore fundamental data on macrofungal resources in the region. The survey area includes eight nature reserves, and eight counties. In total, 3 633 specimens were collected. The specimens were identified by DNA barcode sequencing combined with morphological examination and phylogenesis of some taxa was analysed. In the study, 503 species were identified, and they belong to 19 orders, 73 families and 162 genera. Among them, 395 species were newly recorded in the Shaluli Mountains and 18 species were newly recorded in China. There were 14 families containing more than 10 species, including Russulaceae, Cortinariaceae, Amanitaceae, Pluteaceae, Tricholomataceae, Inocybaceae, Agaricaceae, etc. There were 18 genera containing more than 5 species, including Cortinarius, Russula, Lactarius, Amanita, Tricholoma, etc. Cosmopolitan, north temperate, and pan-tropical genera account for 56.7%, 37.6%, and 4.9% of the total number of known genera, respectively. The resource evaluation showed that 60 species were edible, 54 simultaneously edible and medicinal, 20 medicinal, and 79 poisonous, showing increase of 50, 38, 9 and 62 species, respectively as compared with the previous record. Our study provides reference material for the conservation, utilization and management of macrofungal resources in the region.

Schisandra phenanthera is an important medicinal plant. Numerous species of Pestalotiopsis having wide-ranging distribution are associated with plant hosts as pathogens, endophytes, or saprobes. Ten isolates of Pestalotiopsis were obtained from samples of S. phenanthera collected from Ningshan County, Ankang City, Shaanxi Province through tissue separation method. Taxonomic identification were conducted based on morphological characteristics and phylogenetic analyses of multi-gene sequence data from three loci (ITS, tef1, and tub2). These isolates were identified as P. abietis, P. jiangxiensis, P. lushanensis, P. orzae and P. schisandrae sp. nov. The species of Pestalotiopsis are first reported in Schisandra phenanthera.