Tibet is one of the richest areas for biodiversity in China. Its complex topography and diverse habitat abound with fungal resources. The statistics of Tibetan fungi are based on published records and a total of 2 599 species in 551 genera of 185 families are included. Diversity and dominance analyses are made only for high-rank taxa. Up to the present, 12, 20, 70, 764 and 1 733 species are known in Chytridiomycota, Zygomycota, Glomeromycota, Ascomycota and Basidiomycota, respectively. For economic macrofungi, the number of edible, medicinal and poisonous ones are 468, 243 and 135 species, respectively, according to preliminary counting. The prospect for future work in further investigation of Tibetan fungal resources and biodiversity was proposed.
Collections of the mycoparasitic genus Hypomyces from different regions of China were surveyed. Three new species are introduced based on samples from Yunnan Province. Hypomyces amaniticola on fruitbodies of Amanita is characterized by perithecia semi-immersed to immersed in subiculum, oval to pyriform, orange yellow to brownish yellow; cylindrical asci containing 8 spores; and long ellipsoidal to ellipsoidal ascospores which are unicellular, non-apiculate and smooth. Hypomyces completiopsis on Boletus has pyriform to subglobose, orange yellow to blackish brown perithecia; cylindrical asci with 8 spores; and fusiform to lanceolate ascospores with a verrucose surface and a conspicuous apiculus at both ends. Hypomyces yunnanensis is distinguished by perithecia semi-immersed or nearly superficial, pyriform to subglobose, yellow-brown to warm brown; and ascospores subfusiform, apiculate unequally bicellular and with a verrucose surface. Morphological features of the new species are described and illustrated comprehensively and compared with their related fungi. In addition, Hypomyces microspermus and H. stephanomatis are reported for the first time in China.
Pilidiella guizhouensis sp. nov. on the leaf of Polygonum capitatum (Polygonaceae) was identified by combining morphological characters and phylogenetic analysis based on sequences of the ITS-5.8S rDNA. Its diagnosis characters were as follows: pycnidia were globose to subglobose; conidia were fusiform to long-ellipsoidal, sometimes truncate in the base, 27.9-34.2×5.4-7.6µm, length-width ratios were 4.7-5.8. The holotype and the ex-holotype were deposited in the Herbarium of Department of Plant Pathology, Agricultural College, Guizhou University (HGUP).
A new araneogenous fungus, Clonostachys aranearum sp. nov., was collected in Qianlingshan Park of Guiyang, Guizhou Province. The species is characterised by conidiophores dimorphic, A-conidiophores verticilium-like, cylindrical phialides divergent in whorls of 2-5, or single from lower levels, B-conidiophores penicillium-like, and conidia are often arranged in imbricate chains. Conidia are smooth-walled, hyaline, ellipsoid, 3.2-5.4×1.1-2.1μm, mostly curved. The type specimen GZAC QLS0625 has been deposited in GZAC, Guizhou University, Guiyang.
An entomopathogenic fungal specimen parasitized earwig was collected from Shennongjia of Hubei Province in China. The living culture GZUIFR-snj121022 was isolated from the specimen and the fungus was identified as a new species by morphological classification, phylogenic analysis and splits network analysis based on the ITS1-5.8S-ITS2 rDNA sequences. The species is named as Hirsutella shennongjiaensis. Morphological characters, illustration and phylogenetic analysis are provided in the paper.
The α-amanitin, β-amanitin and phalloidin were detected in 95 fungus samples from different genus excluding Amanita from Northeast China by using HPLC method. For the purpose of obtaining more accurate results, UPLC-MS/MS were used for further determination of 57 positive samples. It is particularly worth mentioning that Inocybe leiocephala is proved to be a fungus containing simultaneously α-amanitin, β-amanitin and phalloidin. The results of this study could provide a reference for preventing and warning poisoning, and screening and exploiting new toxin resources from poisonous mushrooms.
As important medicinal fungi, both Ophiocordyceps sinensis and Cordyceps militaris have gained significant scientific attention in recent decades. However, very little work on mycelial staining method, which is the basis of cytological research, has been reported. Fluorescent staining is a simple and fast staining technique. In the present study, four dyes were tested, singly and combinedly, for use on Ophiocordyceps sinensis and Cordyceps militaris as possible fluorescent stains of fungal nuclei and septa. The results showed that DAPI fluorescent stained better than PI for the nuclei, and the Calcoflour White is better than Congo Red in staining the septa. In the combined stainings, DAPI and Calcoflour White stained nuclei and septa, respectively, more effectively in O. sinensis than in C. militaris, whereas the combination of DAPI and Congo Red is more suitable for the latter by using the laser confocal microscope.
Transcription factors play an important role in the regulation of fungal environmental responses and physiological processes. The expression changes of transcription factor genes in light-induced brown mycelial film (BMF) formation were analyzed by the next-generation high-throughput sequencing technology. The results showed that there were 68 differential expressed transcription factor (TF) genes (48 up-regulated genes and 20 down-regulated genes) when comparing the BMF sample 313C formed under light inducement with white mycelial sample 313W formed in dark. Whereas, 80 differentially expressed TF genes were obtained (49 up-regulated genes and 31 down-regulated genes) when comparing the sample 313C (with BF) with initial white mycelial sample 118. These TFs belong to the families like WD40, MADS-box, MYB and GATA. The expression of transcription factor genes was different in light-induced BF formation, and there were stage-specific TF genes as well as those overlapping among the three samples. There were 14 TFs between the samples 313C and 313W, and 26 between the samples 313C and 118 being specifically transcibed by the fungus. There were 64 TF genes with overlapping expressional profiles, of which 39 TFs were up-regulated and 15 TFs down-regulated. The results of transcriptome analyses were examined by real-time quantitative PCR. The data of this study will facilitate future functional investigations of the TFs involved in Shiitake mushroom developmental controls.
Mitogen-activated protein kinases (MAPKs) have been considered as a family of serine/threonine protein kinases, which are involved in the transduction of a variety of extracellular signals, regulating growth and differentiation processes. To understand the stress-tolerance mechanism of Eurotium rubrum, members of mitogen-activated protein kinase (MAPK) superfamily were identified from E. rubrum genome using the Hidden Markov Models and BLAST method. The phylogenetic trees of the identified proteins of MAPK superfamily were constructed and the conserved motifs were predicted using MEME suite software. The E. rubrum contains four MAPK proteins, namely Hog1-type, MpkC-type, Slt2-type and Fus3/Kss1-type, three MAPK kinase (MAPKK) proteins, namely MKK1-type, Pbs2-type and Ste7-type, and three MAPK kinase kinase (MAPKKK) proteins, namely BCK1-type, Ste11-type and Ssk22-type. Prediction of the conserved motifs indicates that all of the MAPKs superfamily proteins contain protein kinase activity site “-D[L/I/V]K-” and ATP-binding site. The MAPK and MAPKK proteins contain “-TxY-” and “-SD[I/V]WS-” dual phosphorylation site respectively, and the former contains a conserved common docking motif (CD motif). In addition, all of the three MAPKKK proteins possess a conserved motif “-GTPYWMAPEV-” with unknown function. Our results could lay a foundation for elucidation of MAPK signal pathway-mediated cellular processes in E. rubrum.
Adding aromatic compounds in the culture medium could increase laccase activity and the utilization efficiency of lignin and cellulose in the suitable time and concentration. In order to increase the laccase activity in Hypsizygus marmoreus, the effect of eight aromatic compounds on the laccase activity and the relationship between the laccase activity and mycelial biomass were studied. It was found that the r value between laccase activity and mycelial biomass was 0.9956, indicating that they had positive correlation, but the laccase activity was relative low in the culture process. Most of these chemicals induced the laccase activity in different levels, reaching three times higher than the control by using 0.1mmol/L guaiacol, and these chemicals increased mycelial growth and biomass of H. marmoreus at the same time. However, some chemicals inhibited the laccase activity and mycelial biomass at different levels with the prolongation of culture period, probably resulting from the toxicity of these chemicals. It was also found that the transcription of three laccase genes were up-regulated at different levels under the affection of guaiacol in H. marmoreus. This study indicated that increasing laccase activity could stimulate the mycelial growth and biomass of H. marmoreus, and enhance the medium utilization efficiency.
The four extraction methods, hot water reflux, hot water-enzyme reflux, ultrasonic, and ultrasonic combined with enzyme hydrolysis, were compared for β-glucan extraction from Ganoderma lingzhi mycelia. The results showed that the ultrasonic combined with enzyme hydrolysis was the most efficient method. Single factor experiment reveals that the key parameters affecting β-glucan yield of G. lingzhi were ultrasonic power (A), ratio of water to material (B), pH (C) and enzyme concentration (D). Based on the results, Box-Benhnken design combined with response surface methodology were applied to optimize these four key parameters. The results showed that the parameters A, B and C were significant to extraction of β-glucan (P<0.01), however, correlation between parameter D and all the factors was not significant (P>0.05). A polynomial model was established to predict the β-glucan extraction yield, and analysis of response optimization suggested that the optimum conditions for β-glucan extraction were ultrasonic power 275.54W, ratio of water to material 25.62:1 (V/M, mL/g), pH 7.17, enzyme concentration 1.05%. The verified test showed that the extraction yield of β-glucan from G. lingzhi mycelia reached 5.50mg/g under the optimized conditions.
In recent years, a fungal disease known as grey leaf spot occurred severely in the suburbs of Hangzhou City, Zhejiang Province, China, and caused serious economic losses in local tomato production. There were no reports about pathogens causing the disease. The pathogen was identified as Stemphylium lycopersici based on combination of morphological characters, molecular data (sequence analysis of rDNA-ITS, gpd and EF-1α) and pathogenicity tests. This study provides scientific data for the disease decision making and effective control.