The entomophthoroid fungi are important members in lower fungi, including more than 280 species, and mostly obligate arthropod parasites with the exception of few members occurring saprotrophically in soil and plant debris. Based on the taxonomic framework of entomophthoroid fungi by Humber, a taxonomic outline of entomophthoroid fungi was updated in combination with recently published taxonomic taxa. The taxonomic characteristics of one phylum, three classes, three orders, ten families and twenty-five genera were presented. The problems and prospects in the taxonomic study of entomophthoroid fungi were put forward.
Typical edible mushrooms of the Basidiomycota, such as Lentinula edodes, Ganoderma lingzhi, Flammulina filiformis, Pleurotus ostreatus, Auricularia heimuer, etc., exhibit a crucial form in their life cycle known as the dikaryon. The dikaryon can generate monokaryon through both sexual and asexual means, and the compatible monokaryons form dikaryon with the clamp connection through plasmogamy. Research on monokaryon of edible mushrooms has enriched the understanding of the biology of the monokaryon and the dikaryon. Existing as a haploid having two nuclei, the dikaryon possesses various biological traits distinct from diploids. Some studies have investigated the biological properties such as hierarchy, fitness, and stability of the dikaryon, offering insights for maintenance of strain characteristics, innovation of hybrid breeding, construction of genetic research paradigm. Analyzing biological characteristics of the dikaryon, such as reproductive ability, fertility, and totipotency, could provide insights into addressing the long-term stability issues in edible mushroom strains. The dominance-recessive relationship and complementary nature between nuclei in the dikaryon cells might hold significant guidance for innovative hybrid breeding. The presence of different expression levels of karyollele pairs in dikaryon cells offers a new perspective to explore scientific problems concerning edible mushroom. Research on the dikaryon could potentially form a more distinctive paradigm in edible mushroom studies.
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.
Anthracnose is one of the most important diseases of Camellia oleifera, caused by Colletotrichum fructicola. As an important epigenetic modification, histone acetylation plays an important role in the regulation of a variety of biological processes, which is currently unclear in C. fructicola. The research aims at exploring the biological function of histone acetyltransferase Rtt109 in C. fructicola in order to elucidate the molecular mechanism of CfRtt109 and provide theoretical basis for the prevention and control of oil-tea tree anthracnose. Based on homologous recombination principle and the PEG-mediated transformation method, the mutant strain of ΔCfrtt109 and the complemented strains were obtained. It was found that the vegetative growth and conidiation of mutant was significantly lower than that of wild type and the complemented strain. Compared with the wild type and the complemented strain, ΔCfrtt109 was more sensitive to 5.0 mmol/L DTT and 0.01% MMS, additives to PDA medium. The expression levels of HAC1, SCJ1 and PDI1 were significantly upregulated in ΔCfrtt109. RNA-seq combined with RT-qPCR analysis showed that loss of CfRTT109 would alter the expression of genes involved in DNA damage repair. Furthermore, the pathogenicity of the ΔCfrtt109 was significantly reduced on the wounded oil-tea leaf and apple. To sum up, our results revealed the critical roles of Rtt109 in growth, conidiation, tolerance to ER stress and DNA damage stress and pathogenicity of C. fructicola.
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.
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.
This study aims at investigating the effects of in vitro digestion and fecal fermentation on the chemical composition, structural characteristics and biological activities of Clitocybe squamulosa polysaccharides (D-CSFP) extracted by ultrasound-assisted deep eutectic solvents. The results showed that after saliva, stomach and intestine digestion, the content of D-CSFP increased from 0.44 mg/mL to 0.45, 0.55 and 0.57 mg/mL respectively and the molecular weight of D-CSFP decreased from 35 312 Da to 32 090, 25 519 and 25 473 Da respectively, but the structure of D-CSFP had no obvious change. After fecal fermentation in vitro, the OD600 of D-CSFP fermentation broth increased first and then decreased (0.63→ 1.24→1.19); pH decreased from 6.75 to 6.13 (P<0.05); the total carbohydrate content in D-CSFP fermentation broth decreased from 8.59 mg/mL to 5.05 mg/mL (P<0.05); the total short chain fatty acids (SCFA) content was significantly increased from 1.83 mmol/L to 11.15 mmol/L (P<0.05). 16S rRNA sequencing results showed that D-CSFP could effectively promote the relative abundance of beneficial bacteria (such as Bacteroides, Prevotella and Dialister), and inhibit the propagation of harmful bacteria (such as Klebsiella). In conclusion, D-CSFP can be used as a potential functional food to promote intestinal health.
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.
Using single-factor test and response surface design method, the biofermentation processes, including formulation of the liquid fermentation medium and incubation time of the wild strain of Auricularia sinodelicata were optimized. The results indicated that the optimal shake culture temperature for the strain was 28 ℃, the rotation speed was 230 r/min, and the pH value of the medium was 6.0. The optimal carbon and nitrogen sources, and inorganic salts, were formulated as 30 g/L of fructose, 2 g/L of yeast extract, and 2.5 g/L of potassium chloride, respectively. Using mycelial biomass as the response value, the optimal fermentation medium formulation based on the Box-Benhnken design was predicted to be 38.424 g/L of fructose as carbon source, 4 g/L of yeast extract as nitrogen source, and 2.736 g/L of potassium chloride. Under the same culture conditions, the measured dry weight of mycelial biomass in 7 d of fermentation was 2.163 6 g/100 mL or 166.18% of the theoretical value, indicating the feasibility of the study and the reliability of the conclusion. Maximum mycelial biomass was (2.530 0± 0.290 0) g/100 mL and the cellulase activity was 82.586 6 U/mL in the optimal incubation time of 8 d. This study aims at providing a reference for selecting high-yielding strains in artificial liquid cultivation of A. sinodelicata.
The mechanism of apoptosis and migration of renal cancer cell lines after stimulation by cordycepin administration was explored. Kidney cancer 786-O cell line and kidney cancer ACHN cell line were cultured in vitro, and MTT method, cell migration experiment, HE staining, immunofluorescence staining, and protein immunoimprinting were used to examine the inhibition rate of proliferation, cell migration, cell morphology and nuclear morphology of renal cancer cell lines under different concentration of cordycepin administration. The expression of apoptosis-related gene proteins was investigated, and the mechanism of apoptosis induced by cordycepin in renal cancer cells was explored. With the increase of concentration and dose, cordycepin can significantly promote apoptosis and inhibit migration of renal cancer cell lines. Morphological and HE staining observation showed that the number of cancer cells was significantly reduced, and the nucleus was significantly enlarged. Immunofluorescence staining showed that the expression of MMP-2, MMP-9 and BCL-2 proteins decreased significantly, the expression of Bax, Casepase-3 and Casepase-7 proteins increased significantly, and the apoptosis renal cancer cells could be induced through the AKT/mTOR signaling pathway.
SD rats were dosed with Russula subnigricans, and Horn method was used to calculate the median lethal dose (LD50). The effects of R. subnigricans on urea nitrogen (BUN), creatinine (CREA), potassium (K), sodium (Na), chlorine (Cl), white blood cell (WBC) count and classification, urinary protein, urine sugar and occult blood levels in SD rats were determined, and pathology of left kidney was observed. The LD50 and 95% confidence interval (95% CI) of female SD rats were 271 (169-441) mg/kg, and those of male SD rats were 233 (143-378) mg/kg. Compared with the control, the BUN, CREA, K, WBC counts, neutrophil ratio, the incidences and severity of urine protein, urine sugar and occult blood of SD rats dosed with R. subnigricans were increased, while Na and Cl were decreased. The renal tubular necrosis of SD rats in each treatment groups showed vacuolar degeneration, watery degeneration, and the brush margin disappearance of the epithelial cells, and exfoliated epithelial cells were visible in the lumen. With the increase of the dosage of R. subnigricans, the change of each index was more significant. The R. subnigricans could lead to kidney injury in rats in a dose-response relationship.
Stropharia rugosoannulata is a new cultivated edible and medicinal mushroom favourably received by consumers in recent years. Its total sugar and protein content, is closely related to its nutritional values. However, the existent content detection methods are complex and time-consuming, and it is greatly significant to develop simple and rapid prediction methods. In this study, a total of 420 infrared spectra from fruiting bodies of S. rugosoannulata cultivated on seven different substrates was collected by Fourier transform infrared spectroscopy FTIR. The spectral pretreatment method was established by comparing the original spectrum and the pretreated spectrum, while the total sugar and protein characteristic spectra were selected by competitive adaptative reweighted sampling (CARS). Random forest (RF), partial least square regression (PLS), and support vector machine (SVM) were used for modeling. The results proved that the best prediction model for total sugar content was PLS, in which Rc was 0.992 8 (the error was 0.007 2), RMSEC was 0.930 8, Rp was 0.981 4 (the error was 0.018 6), RMSEP was 1.166 2, and RPD was 7.341 1; the best prediction model for protein content was RF, with Rc at 0.994 7 (the error was 0.005 3), RMSEC at 0.380 3, Rp at 0.986 (the error was 0.014), RMSEP at 0.749 1, and RPD at 8.437 5. The results showed that infrared spectroscopy combined with stoichiometry could quickly and accurately predict the total sugar and protein content of fruiting bodies of S. rugosoannulata.
Based on morphological taxonomic examination and molecular phylogenetic analysis of partial calmodulin gene (CaM), DNA-dependent RNA polymerase II second largest subunit gene (Rpb2) and rDNA ITS1-5.8S-ITS2 (ITS) sequences, the moulds isolated from surface of the home-made dry-cured meat in Buyi and Miao Autonomous Prefecture and Liupanshui City in Guizhou Province are identified as eight Aspergillus species and one Penicillium species, namely, A. cumulatus, A. flavus, A. glaucus, A. jensenii, A. proliferans, A. pseudoglaucus, A. tennesseensis, A. venenatus and P. aurantioviolaceum; among which, A. cumulatus, A. venenatus and P. aurantioviolaceum are new to China.
