A hundred and thirty-three collections of cup fungi from the Qilian Mountains in Gansu Province, China were examined. Forty-five species were identified, which belong to the families Ascobolaceae, Ascodesmidaceae, Cudoniaceae, Gelatinodiscaceae, Helotiaceae, Helvellaceae, Lachnaceae, Mollisiaceae, Pyronemataceae, Rhytismataceae and Thelebolaceae. Of which, Cheilymenia qilianshanica, Coprotus qilianshanicus, Neobulgaria qilianshanica and Trichophaea sinobullata are described and illustrated as new species, and Hymenoscyphus chloophilus, H. sulphuratus and Sowerbyella brevispora are newly reported in China. The distinctive characteristics of the new species and their allies were compared in detail.
Investigations on dematiaceous hyphomycetes on plant debris in Jiangxi Province have yielded three new species of Ellisembia, viz. E. jiangxiensis sp. nov., E. jinggangshanensis sp. nov., and E. lushanensis sp. nov. They are described and illustrated from specimens collected on dead branches of unidentified plants. In total, 69 species have been reported in Ellisembia. The genus is worldwide in distribution, commonly found as a saprobe on plant debris or submerged wood, but not reported as plant pathogens.
Plantations of Pinus sylvestris var. mongolia, one of the most important conifer species for environmental restoration in the desertified northern China, have become increasingly declined. Root-associated fungi (RAF) are closely related to the growth of P. sylvestris var. mongolia, and they are probably potentially connected with the decline. Field investigation on P. sylvestris var. mongolia plantations of three stand age, middle-age (27a), nearly mature (33a), mature (44a), was conducted to reveal the community structure and functional group of RAF in the desertified lands of Yulin in Shaanxi Province. Root tip samples were identified by high-throughput sequencing and FUNGuild platform. 855 OTUs were obtained, and there was no significant difference in the RAF’s diversity indices in different stand ages (P>0.05). The RAF were assigned to six phyla, 18 classes, 48 orders, 87 families, and 197 genera. Ascomycota and Basidiomycota accounted for 97.31% in relative abundance. The dominant genera were Penicillium and Archaeorhyzomyces. With the stand aging, the proportion of pathotrophic and saprophytic fungi increased first and then decreased, meanwhile symbiotic fungi were in a converse trend. The community structure and functional group of the RAF associated with P. sylvestris var. mongolia are diversiform, and ectomycorrhizal fungi are dominant functional group. The relative abundance of saprophytic fungi is higher than that of symbiotic fungi in the near-mature plantation, implying the potential decline of P. sylvestris var. mongolia.
Alternaria alternata is an important phytopathogen producing a variety of mycotoxins, such as alternariol (AOH). Essential oils are the important volatile plant extracts that could suppress the pathogenic infection. The active components of essential oils include citral and other compounds. The results of this study showed that citral could effectively inhibit the growth of A. alternata and AOH production. Citral fumigation could lead to the fracture of hyphae and affecting their elongation, but had no obvious effect on the conidial structure. Citral could disturb the generation of reactive oxygen species in A. alternata, resulting in significant reduction of AOH production. Citral could be a potential alternative of traditional fumigant to control the disease and mycotoxin contamination induced by A. alternata. The study provides a new way for the development and application of citral.
Comparative transcriptomic analysis of Sanghuangporus sanghuang mycelia and fruiting body was performed for studying related growth and development mechanism of the fungus. The mycelia and fruiting body of the strain S23 were sequenced by using Illumina sequencing platform. The mapping results showed that 82.89% reads from mycelia were aligned to the initial reference transcriptome dataset, while 83% from fruiting body. Analysis of differentially expressed genes showed that there were 2 898 significantly up-regulated genes and 1 965 significantly down-regulated genes expressed in fruiting body as compared with mycelium. Functional annotation with Blast nr database indicated that up-regulated genes in S. sanghuang fruiting body were involved in oxidoreductase activity and fungal hydrophobin, while down-regulated genes were involved in binding and transport of carbohydrate, amino acids etc. Gene ontology functional enrichment analysis showed that these differentially expressed genes involved in transmembrane transport were significantly enriched in fruiting bodies. KEGG metabolic pathway analysis showed that there were significant differences in gene enrichment in metabolic pathways, such as steroid biosynthesis, arginine biosynthesis and MAPK (mitogen-activated protein kinase) signaling pathway.
Colletotrichum fructicola is a major pathogen causing anthracnose on Camellia oleifera. In previous study, we characterized that the bZIP transcription factor CfHac1 controlled the development and pathogenicity of C. fructicola. To identify the role of CfHac1 in the endoplasmic reticulum (ER) stress in C. fructicola, the growth of a ΔCfhac1 mutant strain of C. fructicola was determined on the PDA medium containing dithiothreitol, an ER stress factor. The results revealed that the ΔCfhac1 strain was sensitive to dithiothreitol. To uncover the potential mechanisms of CfHac1 contributions to pathogenicity and response to ER stress, the transcriptome profiles of the wild type strain and mutant ΔCfhac1 of C. fructicola were characterized with high-throughput sequencing technology. The data indicated that, relative to the wild type, there were 2 680 differentially expressed genes (DEGs) in mutant ΔCfhac1, of which 1 181 were up-regulated and 1 499 were down-regulated. The Gene Ontology classification showed that the DEGs were mainly related to catalytic activity, binding, metabolic process, cellular process, cellular component organization or biogenesis, regulation of biological process and response to stimulus pathway. The KEGG pathway analysis also revealed that the up-regulated expressed genes were mainly related to ribosome, ribosome biogenesis in eukaryotes, RNA transport and cyanoamino acid metabolism pathways; the down-regulated expressed genes were mainly related to protein processing in ER, N-glycan biosynthesis, steroid biosynthesis, and protein export pathways. Genes involved in ER stress response and virulence were down-regulated at transcriptional level in the ΔCfhac1 strain. This study provides useful data concerning the effect of CfHac1 in endoplasmic reticulum stress response at the genome level and a basis for illuminating pathogenic mechanism of C. fructicola.
The transformation system of Macrocybe gigantea was successfully established for the first time by using Agrobacterium tumefaciens EHA105 bearing binary plasmid plasmid4. The results of hygromycin resistance screening, PCR identification and fluorescence microscopy analysis indicated that Hyg was integrated into M. gigantea genome, and enhanced green fluorescent protein gene (eGFP) was expressed in M. gigantea. The development of Agrobacterium-mediated genetic transformation system can facilitate further gene function research in M. gigantea.
As an obligate biotrophic fungus, Puccinia striiformis (Pst) secretes bunches of effectors into host cells, where they interfere with host defense response and promote pathogen virulence. Identification and functional analyses of Pst effectors are of great significance in revealing the fungal pathogenesis. In this study, as a result of analysing haustorial transcriptome of Pst CYR31, an haustoria specific induced secreted protein Hasp68 was identified as a candidate effector, which was capable of suppressing cell death caused by the mouse pro-apoptotic protein Bax in Nicotina benthamiana cells. The effector Hasp68 is of 105_aa in length, containing a 20_aa signal peptide at its N-terminus, with no transmembrane region or any predicted conserved motifs. BlastX analyses revealed that Hasp68 specifically existed in Pst with no homologues in other plant pathogenic fungi. Sequence alignment among the available Pst genomes and re-sequencing data of 16 Pst races showed low sequence polymorphism, indicating that Hasp68 was relatively conserved during the evolution of Pst. Delivery of Hasp68 into wheat cells via the Pseudomonas fluorescences EtHAn strain type three secretion system (TTSS) suppressed the callose deposition, the typical character of PAMP-triggered immunity (PTI), caused by the nonpathogenic bacteria. Meanwhile transient expression of Hasp68 in wheat cells could also inhibit the reactive oxygen species (ROS) accumulation and hypersensitive response (HR) induced by the avirulent Pst isolate. It was suggested that the virulence function of Hasp68 was possessed of suppressing host immunity including PTI and effector triggered immunity (ETI). Yeast two hybrid (Y2H) screening revealed that Hasp68 interacted with the host cathepsin B TaCTSB. Moreover, BiFC (bimolecular fluorescent complimentary) analysis further verified the interaction in vivo of co-expression of Hasp68 and TaCTSB in Nicotiana benthamiana. It is presumable that Pst effector Hasp68 targets wheat TaCTSB to modulate plant immunity.
Ultra performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-QTOF-MS) combined with multivariate statistical methods was used to analyze the differences of intracellular metabolites of Grifola frondosa mycelium cells in submerged fermentation of 7 days by the addition of compound solution of Gastrodia elata and Fagopyrum tataricum. The results showed that the principal component (PCA) model revealed a significant differences between metabolites of mycelium cells cultured in compound solution and solution of experiment control (P<0.05). Through orthogonal partial least squares discriminant analysis (OPLS-DA), 44 differential metabolites were screened and identified under the conditions of VIP (varible importance in the projection)>1 and P<0.05, including 6 sugars, 13 amino acids, 5 vitamins, 7 nucleotides, 10 organic acids and 3 fatty acids. The content of 7 substances such as L-rhamnose monohydrate, D-galactose, D-mannitol, fructose-6-phosphate was significantly reduced, the content of 37 substances such as D-xylitol, isoleucine, lysine, pantothenate, dihydroxyacetone, D-glucuronic acid, succinate increased significantly as compared with the experimental control. Pathway analysis of differential metabolites indicated that there were 14 metabolic pathways with significant impact and the syuthetic pathway of Grifola frondosa extracellular polysaccharide was constructed. The results proved the synergistic effect of the addition of Gastrodia elata and Fagopyrum tataricum compound solution on the production of extracellular polysaccharide of Grifola frondosa thereby improving the nutritional quality. Further study on the effect of exogenous additives on the fermentation process of Grifola frondosa is needed.
The volatile components in pileus and stipe of Oudemansiella raphanipes were determined by head space solid phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS). Respectively 101 and 102 compounds were identified from the pileus and stipe, 44 of which were the same components, accounting for 43% of the total components. They were mainly anesane, ester and acid compounds, of which the relative content of tetradecane, octamethylcyclotetrasiloxane, hexanoic acid, l-(+)-ascorbic acid 2,6-dihexadecanoate and linalool is the highest. Nutritional analysis showed that the O. raphanipes fruiting body contained 1.2% coarse fiber, total sugar 28.7%, fat 0.3g/100g, protein 3.58g/100g, ash 1.6%, essential amino acid 0.871g/100g, and 31.30% of total amino acids (2.783g/100g). Per kg fruiting body contains 29.2mg Na, 4.37×10 3mg K, 115mg Mg, 12.6mg Zn, 6.19mg Ca, 46.5mg Fe, 6.93mg B1, 17.9mg B2, and 33.6mg B6. This is the first analysis of the nutritional and volatile components in O. raphanipes, aiming at providing theoretical support for further study and the improvement of the compound spectrum library.
Laccase is a green and efficient oxidase for biodegradation of polyphenol, which has a great potential in the degradation of bisphenol A. For reducing the production cost of laccase, the solid-state fermentation conditions for Trametes sp. were optimized by using bran and grapefruit peel as the main substrates, and the degradation ability of the laccase product to bisphenol A was further investigated. As a result, by using the optimized medium containing components formulated in the proportion of bran and grapefruit peel ratio of 6:4 (W/W), solid-liquid ratio of 1:2.5 (W/V), copper ion 2% (W/W), sucrose 3% (W/W), potassium nitrate 2% (W/W), and rice husk 20% (W/W), the enzyme activity of the laccase product reached 38.4U/gds in 11 days of fermentation. Under initial concentration of 10μg/mL, bisphenol A was almost completely degraded by laccase at 55°C within incubation duration of 140min.
Flavonoids are a kind of important secondary metabolites, which widely exist in various plants and fungi, and have antibacterial, anticancer, antioxidant and other biological activities. In the present study, in order to obtain the optimal technology of extracting total flavonoids from the fruiting bodies of Auricularia cornea, response surface methodology was used to investigate the effects of ultrasonic power, extraction time, ethanol concentration and powder-liquid ratio on the extraction of total flavonoids. Response surface analysis indicated that the influence of various factors on the response value ranged in seguence of extraction time＞ ultrasonic power＞ethanol concentration＞ratio of powder-liquid, and the influence of ultrasonic power, extraction time and ethanol concentration showed the most significant. The optimum conditions for extracting total flavonoids from the fruiting bodies of A. cornea were ultrasonic power 168W, extraction time 50min, ethanol concentration 60%, and ratio of powder-liquid 1:7, and the maximum predicted extraction quantity of total flavonoids was 2.11%. The results of verification test showed that the average content of total flavonoids was 2.08%, being consistent with the theoretical value.
A high-performance liquid phase was established to determine the content of liposoluble components in Ganoderma lingzhi spore powder. The determination of 12 liposoluble components was carried out by optimizing the column, elution conditions and ELSD parameters. The method proved to be simple, accurate, stable, and appropriate to the simultaneous quantitative detection of triglycerides, fatty acids and sterols in G. lingzhi spore powder. The analysis of multiple samples showed that the liposoluble components were 301.49-397.37mg/g in the broken spore powder, and 626.00-713.07mg/g in the spore oil product, being higher than the content of triterpenes. 1,2-Olein-3-palmitin, triolein and 1,2-Olein-3-linolein were the main liposoluble components, and the fatty acids were mainly unsaturated fatty acids including linoleic acid and oleic acid. The content of ergosterol was the highest in sterols. The research clarified the liposoluble components in G. lingzhi spore powder, and provided reference for further investigation of active ingredients and comprehensive evaluation of quality control of G. lingzhi spore powder.
Four new Chinese records, Dialonectria ullevolea, Pseudocosmospora eutypellae, Thelonectria nodosa and T. rubi, are reported based on specimens collected from Shandong, Henan, Sichuan and Yunnan provinces. Their descriptions and illustrations are provided. Morphological and molecular comparisons are made between the Chinese materials and the original descriptions.
