A new Terriera species, T. intraepidermalis on Photinia prunifolia is described in English and Chinese with an illustration and discussion. The type specimen is deposited in the Reference Collection of Forest Fungi of Anhui Agricultural University, China (AAUF).
A new species of Coccomyces, C. jiangxiensis on fallen leaves of Rhododendron latoucheae, is reported from the Sanqingshan National Park, Jiangxi Province. Description, illustration, and comments are provided for this taxon. The type specimen is deposited in the Reference Collection of Forest Fungi of Anhui Agricultural University, Hefei (AAUF).
A new brown spot lesion on bagged apples was discovered in Penglai, Qixia and Yiyuan of Shandong Province in 2011 and 2012. Diseased apple fruits were sampled and two strains of Acremonium were isolated from the brown lesions. One of them produced conidia arranged in capitiform and the other in chains. Both strains could infect ripe apple fruits through wounds, resulting in brown sunken lesions, but could not infect unwounded fruits. ITS, β-tubulin, nucLSU and nucSSU sequences of the two strains did not show any variation. The two strains were identified as Acremonium sclerotigenum according to ITS and nucLSU sequences, and morphological character. A. sclerotigenum is an opportunistic pathogen. The pathogen would infect apple fruits before harvest when conditions were favorable. The new disease was named Acremonium brown spot.
The main property difference of soybean Sclerotinia sclerotiorum isolates from Heilongjiang Province is studied. Sclerotinia sclerotiorum was isolated and purified on PDA medium, and the RAPD and rDNA ITS methods were used to analyze the isolates. A total of 50 isolates of Sclerotinia sclerotiorum were obtained from diseased soybean. Using RAPD marker the genetic similarity coefficient ranged from 0.54 to 0.98, and the average value was 0.76, indicating that there were genetic difference among the isolates. Analyzing the sequence of the 5.8S region and nrDNA ITS of 32 Sclerotinia sclerotiorum isolates showed that there was large variation at the ITS1 region of 1-40bp, mainly in the form of base transversion and transition. There was no mutation at the ITS2 region of most of the isolates. There was significant genetic variation among Sclerotinia sclerotiorum isolates in Heilongjiang Province at the DNA level and ITS spacer region.
The samples of tobacco roots, stems and leaves were collected in Yunnan “World Tobacco Cultivar Garden” for investigation of endophytic fungi. From these plant tissues, 199 isolates of endophytic fungi were generated and 25 species of 17 genera were identified by rDNA-ITS sequence analysis. The majority of the fungi belong to Pleosporales, and Phoma, Alternaria and Fusarium were dominant with the relative frequency of 25.1%, 24.6% and 11.6%, and the dominance (Y) values of 0.251, 0.172 and 0.104, respectively. The community composition of the fungi in different tissues varied significantly. The species of Fusarium and Phoma were frequently isolated from roots with the dominance (Y) values of 0.235 and 0.123, respectively. Species frequently isolated from stems were those of Phoma and Alternaria with the dominance (Y) values of 0.186 and 0.155, respectively. Species frequently isolated from leaves were those of Alternaria with the dominance (Y) value of 0.286. Phoma could be isolated from all tissues of tobacco, however, Alternaria were restricted within stems and leaves, while Fusarium within roots and stems, indicating colonization selectivity of the fungi to the plant tissues.
Symbiotic compatibility of Polyporus umbellatus and Armillaria mellea is the key factor for the successfully artificial cultivation of Polyporus umbellatus sclerotia. In order to obtain the combination with high compatibility, five isolates of Polyporus umbellatus and four isolates of Armillaria mellea were screened. The growth rate, antagonism line, and mycelium morphology of the combinations were compared separately after co-cultivation on a plate or in a culture bottle. The result showed that the fastest growth of mycelia reaching 3.36mm/d was the combination GU-06&AM-08. The antagonistic line could be observed in each co-cultivation of P. umbellatus isolate GU-06 or GU-07-2 and A. mellea isolates. After co-culture for 28 days, more branches and hyphal strands emerged in mycelia of P. umbellatus. Rhizomorph of the A. mellea was found obviously in the sclerotia of P. umbellatus, with more branches, hollow, and slender epidermal cells. Seven combinations with infectivity were GU-04&AM-06, GU-06&AM-06, GU-06&AM-08, GU-06&AM-13, GU-07-2&AM-06, GU-07-2&AM-08 and GU-15&AM-06. The most serious infection happened in the GU-06&AM-06 and GU-06&AM-13, and no infection was found in the other combinations. In conclusion, the four group of highly compatible symbiotic combinations, GU-06&AM-08, GU-06&AM-06, GU-07-2&AM-08, and GU-06& AM-13, could be regarded as most suitable for artificial production of P. umbellatus sclerotia.
“jiuyao” is an important saccharification and fermentation starter used in the production of Shaoxing ricewine. Samples of starter (jiuyao) used for the fermentation of different brands of Shaoxing ricewine, including Guyuelongshan, Kuaijishan, Tapai, Shenyonghe, Nüerhong and Jianhu, were collected during the winter of 2013 and then subjected for quantitative yeast isolation. The result showed that the colony counts of yeasts in the starter samples of Kuaijishan and Shenyonghe were significantly higher than those in the other samples (P<0.01), while the Tapai samples showed the lowest yeast counts among the samples compared (P<0.05). Three yeast species, Saccharomycopsis fibuligera, Saccharomyces cerevisiae and Cryptococcus sp., were identified from the yeast strains isolated using the 26S rDNA D1/D2 domain sequence analysis. Saccharomycopsis fibuligera was found to be an dominant species in all the samples examined, suggesting that this species may be crucial for the quality of starter.
Pycnial and aecial formation of Puccinia recondita on Clematis intricata are observed by means of light and electron microscopies. The pycnium of P. recondita is flask-shaped, producing a large number of monokaryotic pycniosporophores and periphyses inside. Numerous pycniospores are generated successively from the top of pycniosporophores in chain. After a mature pycniospore released from a pycniosporophores, an annular scar was observed on the top surface of a pycniosporophore. A pycniospore is oval in shape with no special ornaments on the surface. An aecium of P. recondita is cylinder-like in shape, producing numerous long, and dikaryotic aeciosporophores that generate successively a number of aeciospores. An aeciosporophore divided to form an aeciospore initial, and a septum separated the initial into an immature aeciospores and an intercalary cell, with the further development of aeciospores, the intercalary cell disintegrated and the spores separated from each other. The surface of a mature aeciospore is covered with numerous verrucas.
For the purposes of exploring the invasion patterns of Fusarium oxysporum strains in Rehmannia glutinosa, plasmid pCT74 carrying GFP gene was successfully transferred into the pathogen by using protoplast transformation method mediated by PEG-CaCl2. The infection histological process in R. glutinosa was observed by using transformants labeled with GFP. Yellowing and wilt was observed in the infected plants due to root and leaf infection. Microscopic examination showed that stem vascular bundles of the seedling were mainly infected which led to the disagglomeration of spongy tissues and the black rot of the underground rootstock. However, leaf infection showed a significantly higher severity as compared with root infection at all inoculum level tested. Spore concentration above a level of 2-5×103spores/mL did not further enhance disease severity. F. oxysporum DNA were extracted from leave, stems and roots of the root-infected plants. The pathogen invasion was detected in roots in 60h after inoculation, in stem tissue in 84h, and in leaves in 96h. The labeled strain may serve as a better material for understanding rhizospheric biological processes in continuous cropping system of R. glutinosa.
Glomerella leaf spot, an apple disease caused by Glomerella cingulata, is newly found in China. In order to understand the sexual reproduction, biological characteristics and pathogenicity of the fungal ascospores, diseased leaves were sampled from Dangshan of Anhui and Muping of Shandong, and mono-conidium cultures were obtained. The mono-conidium cultures produced asci on PDA and each had 8 ascospores. Twelve asci were separated from cultures, of which 10 asci crowded 4 spores with “+” mating type and 4 spores with “-” mating types, and the remainder 2 asci produced only spores with “-” mating type. After 72 hours’ single ascospore cultivation, colonies of “+” type strains were white and the mycelia were flourishing. The colonies of “-” type strains were gray with white edge and a large number of orange conidia on center. Colony diameter of “-” strains was slightly smaller and the mycelia were thinner than those of “+” strain. Heterothallism of “+” and “-” strains produced abundant perithecia at the junction of the two colonies. The colonies of “+” strains produced sparse fertile perithecia, while, single “-” strain only produced sterile perithecia. There were no significant difference in growth rate between “+” and “-” strains and in sensitivity to temperature, nutrition, illumination and pH. Sexual reproduction of “+” strain did not result in variation of sequences of rDNA-ITS and β-tubulin genes.
Two primers among 300 RAPD primers were obtained to specialize CYR33 of Puccinia striiformis f. sp. tritici using SCAR-PCR technique. Two specific DNA fragments of CYR33 were recovered from the agarose gel, cloned, and sequenced (GenBank Deposit Number: AB914691 and AB914692). Based on the sequences, two specific primers (S261F33/S261R33, and S300F33/S300R33) were designed and they could amplify 247bp and 763bp specific bands from genomic DNA of CYR33 and total DNA of wheat leaves infected by CYR33, respectively. The identification results using the two primer pairs were the same as those using differential host. Thus, the two primer pairs could be used for CYR33 identification and monitoring rapidly.
DNA polymorphism of arbuscular mycorrhizal fungi (AMF) in root system and those in rhizosphere soil of Prunus mume was analyzed through DNA amplification by nested PCR based on AFLP marker. The results show that the purified DNA can be extracted from 28 soil samples, accounting for 93.3% of totally 30 samples, collected from rhizosphere of 18 cultivars, averaging 6.5 loci for each sample. The average genetic identity was 0.3559±0.1382, Shannon information index was 0.5299±0.1676. The number of loci in rhizosphere soil was much more than that of loci in root sample, and most of loci in root sample could be found in soil sample. It was proved that the AMF in root might be developed from soil AMF. The clustered groups of AMF DNA by AFLP marker both from roots and from soils were not connected with cultivar groups or cultivars of P. mume, and indicated that there was no specific symbiotic relationship between AMF and P. mume.
The goal of this study was to detect the direct evidence of somatic recombination of Puccinia striiformis through development and characterization experimental isolates using virulence tests and molecular markers under controlled greenhouse conditions. A total of nine pairwise combinations involving seven isolates of P. striiformis f. sp. tritici (wheat stripe rust) and two isolates of P. striiformis f. sp. hordei (barley stripe rust) were selected for this study. “Parental” and selected variant “progeny” isolates were tested for virulence patterns on the seedlings of wheat and barley differential cultivars. Eighty-four progeny isolates with virulence patterns were different from the parental isolates. The results preliminarily indicated that somatic recombination might occur in rust fungi. Simple sequence repeat (SSR) markers were used to identify somatic recombinant isolates. A total of 28 progeny isolates from the five pairwise combinations were tested by 6 of 11 SSR markers. Three isolates had SSR patterns being different from their “parental” isolates and the combined bands from both parental isolates, indicating that the three isolates were recombinants. The three isolates were also determined as recombinants in the virulence tests. The results demonstrate that somatic recombination is a mechanism by which new variants can be generated in P. striiformis and lead to virulence variation.
This work was designed to obtain the mutant strain of Volvariella volvacea with low-temperature tolerance and analyze the differentially expressed genes between the mutant and parental strain. By UV mutagenesis, a mutant strain Vtlt-1 was obtained with low-temperature tolerance. Using cDNA microarray, differently expressed genes between Vtlt-1 and parental strain V23 were selected and analysed by GO (Gene ontology) and KEGG (Kyoto encyclopedia of genes and genomes). There were 1 600 significant differently expressed genes in all, among which 704 genes were up-regulated and 896 genes down-regulated. These differently expressed genes were classified according to the GO database. The genes concerned in biological process were enriched in involvement of metal ion binding, oxidation-reducing process, carbohydrate metabolic process and nucleic acid binding, while those concerned in cellular component were related to nucleus. The genes with molecular function were enriched in oxidoreductase activity, helicase activity, ATP-dependent helicase activity and DNA-directed RNA polymerase activity. To determine the function and relationship of these differentially expressed genes, molecular pathway analysis based on the KEGG database were performed. These genes were enriched in amino acid and nitrogen metabolism, fatty acid and alkaloid biosynthesis, ribosome biogenesis in eukaryotes, cytochrome P450, sulfur and amino acid metabolism and nucleotide repair. These results provided a molecular basis for future analysis of the mechanism of low-temperature tolerance in V. volvacea.
In order to reveal the protein expression changes that occur at the different developmental stages of the button mushroom Agaricus bisporus fruiting body, the technique of iTRAQ-MS/MS (i.e. isobaric tags for relative and absolute quantification coupled two-dimensional liquid chromatography tandem mass spectrometry) was used to analyze the fruiting body proteomes of the primordium, button, harvest and opening stages of A. bisporus As2796 strain. As a result, 1 059 proteins, included a total of 5 869 unique peptides, were identified. Among them, 1 007 proteins were detected with quantitative information. As compared with the proteins of primordium stage, the samples of button, harvest and opening stages produced 242, 200, and 240 different proteins, respectively, accounting for 24.0%, 19.9% and 23.8% of the proteins identified, respectively. Meanwhile, the differentially expressed proteins between different stages, as well as all of the identified proteins, were analyzed via bioinformatics. The genes of 8 proteins either up- or down-expressed continuously at different developmental stages were analyzed by real-time PCR (qPCR), and 3 of them, i.e., mismatched base pair and cruciform DNA recognition protein, Cytochrome C1 and a hypothetical protein, were confirmed to have similar expression trends with the corresponding proteins. The data of this study provided a basis for future functional research of the genes involved in fruiting body development in A. bisporus.
Taxol, a well-known anti-cancer medicine, is popularly studied due to its unique anti-cancer mechanism, excellent anti-cancer effects, and its tight market. Although taxol has great economic benefits, its price is quite high because of its restricted production. Producing taxol through endophytic fungus fermentation method can solve this difficult situation. In this paper, we reported the effects of the addition of precursors and elicitors to taxol-producing endophytic fungus Aspergillus fumigatus TMS-26 broth, and the optimization of inoculum density, loading volume, initial pH value and fermentation duration for taxol production. Results showed that the maximum taxol yield could reach 446.28µg/L, with 46.61% increase as compared with that by traditional method. Maximum yield was achieved when concentrations of phenylalanine, sodium benzoate, sodium acetate, glycine, CuSO4, H2O2 and 3,5-dinitrosalicylic acid were 20mg/L, 30mg/L, 8g/L, 15mg/L, 0.05mg/L, 6mmol/L and 15mg/L respectively. Optimal fermentation condition for the strains was obtained under pH7.5, inoculation quantity of 5%, liquid volume of 120mL/250mL and fermentation duration of 10 days.
MREEs was prepared by enriching the low molecule weight bioactive components extracted from the fruiting bodies of Hericium erinaceus using 95% ethanol in HPD-100 macroporous resin with the metronidazole as positive control. The MREEs were leached by petroleum ether (60-90°C) and chloroform for several times and three extractions were isolated which were encoded as A1, A2 and B and were also detected by GC-MS. The anti-Helicobacter pylori activities of the MREEs, A1, A2 and B were performed through antimicrobial susceptibility test, and the minimum inhibitory concentration (MIC) of five Helicobacter pylori strains was then determined. The results indicated that the MREEs had obvious inhibition effect on the growth of Helicobacter pylori and the inhibition rate of the three extracts (A1, A2, and B) was higher than 40.0%, especially for A1 of which the inhibition rate was as high as 62.9%. The main components of A1, A2, B analyzed by GC-MS were DBP (A1, 4.53% and A2, 6.93%) and palmitic acid (2.87%). In addition, The MIC of A1 against Helicobacter pylori W2504, Helicobacter pylori 9 and Helicobacter pylori ATCC 43504 was 0.25mg/mL, and MIC of A2 against Helicobacter pylori 78 was 0.25mg/mL. B was the most potent fraction against Helicobacter pylori W2504 and Helicobacter pylori 9, whose MIC was only 0.125mg/mL. In conclusion, extractions with petroleum ether and chloroform from Hericium erinaceus presented strongly inhibitory activity against Helicobacter pylori.
The metabolite 11’-deoxyverticillin A (C42), which belongs to epipolythiodioxopiperazines (ETPs), has various bioactivities including viral replication inhibition. Autophagy, a phenomenon that participates in the degrading long-half-life proteins, exists extensively in eukaryotic cells. Autophagy regulates many physiological and pathological processes, and has been demonstrated to play a critical role in the replication of hepatitis B virus (HBV). However, it remains almost unknown whether C42 has a regulatory role in HBV replication via altering autophagy. In this study, we found that, in contrast to primary HepG2 cells, HepG2.215 cells have an increase in the formation of autophagosomes concurring with upregulation of Akt signaling. C42 down-regulated the levels of LC3-II and p62, both of them are autophagic genes, accompanying with a decrease in Akt signaling. The presentation of chloroquine (CQ), a compound often used in autophagic flux detection, was found to block C42-induced LC3-II degradation, suggesting that C42 activates autophagy in the cells. The inhibition of Akt activity or depletion of LC3 and p62 reduced the expression of HBV-X, which is believed to be essential for HBV replication. Notably, CQ, which inhibits the fusion between autophagosome and lysosome, was demonstrated to increase the expression of HBV-X protein. Thus, we consider that C42 inhibits viral replication by decreasing the expression of autophagic genes and altering Akt signal pathway.
In order to evaluate the application potential of Ceriporia lacerata in dealing with the azo dye, methyl orange dye with stable performance was used as material in batch experiments for investigating the effect of initial dye concentration, fungal biomass, temperature and pH on the decolorization of methyl orange dye by C. lacerata. Decolorization mechanism was probed by means of mycelium reverse inoculation, dye solution absorption spectrum and mycelium microexamination. The toxicity test for original dye and decolorizing products using plant seed germination method was conducted. Results showed that C. lacerata effectively decolorized methyl orange dye, and the optimal temperature and pH were 35°C and 6 respectively. The decolorization of dye was caused by both mycelium absorption and enzyme degradation. During the course of decolorization methyl orange dye had little effect on the mycelium. Toxicity test suggested that the products in 48h of decolorization by C. lacerata had more toxicity than original dye. C. lacerata was a useful potentially fungus in the treatment of azo dye.
One newly recorded genus, Dendrosporium, and three newly recorded species, D. lobatum, D. candelabroides and Ochroconis cordanae of aquatic hyphomycetes from China are reported in this paper. The descriptions given below are based on Chinese materials. The examined cultures are deposited in the Herbarium of the Key Laboratory for Conservation and Utilization of Bio-resources of Yunnan University.
Three new Chinese fungal records, Ilyonectria estremocensis, I. pseudodestructans and I. robusta, isolated from Arisaema erubescens and Aconitum kongboense in Tibet are reported in this paper. The morphological characteristics and phylogenetic analysis are provided. The examined specimens are desposited in Biotechnological Center, Institute of Medicinal Plant Development (IMPLD), Chinese Academy of Medical Science (CAMS).
A new record genus Thuemenella for North Asia is reported based on collections from Jilin, China. Thuemenella cubispora is characterized by pulvinate to hemispherical, lemon-yellow, Hypocrea-like stromata and four one-septate ascospores that disarticulate into eight smooth, angular to cuboid, brown part-ascospores in ascus. Morphological features of the fungus are described and illustrated. This extends the distribution of T. cubispora to North Asia.
Due to obscure knowledge to ascomycete phylogenetics, genetic relationship, and its complexity of anamorph and teleomorph, mycologists still hold different opinions on the ascomycete classification. The anagenesis of mating type (MAT) gene is conservative, and the protein coded regulates the sexual reproductive process of ascomycetes. Sclerotinia sclerotiorum is a typical filamentous homothallic fungus which belongs to Ascomycota, Discomycetes. The mating type genes MAT1-1 and MAT1-2, which regulate fungal sexual reproduction, are closed linked to each other at the same chromosome and the fungi do not maintain complexity of anamorph and teleomorph. Therefore, based on the cloned S. sclerotiorum mating type gene MAT1-1, this study generated a phylogenic analysis by PAUP* with other 81 mating type genes containing Alpha-box domain from fungi. Through phylogenic analysis on the level of nucleotide and amino acid and comparative study under Ainsworth’s classification system and the latest classification system of Deep Hyphae, it is found that the evolutionary phylogenic tree constructed illustrates basically the same evolutionary relationship in traditional classification, and the function of mating type gene MAT1-1 is relatively conservative in the evolutionary process. The result is helpful to the study of cloning, phylogenetic classification and anagenesis of other mating type genes in ascomycetes. Moreover, it has great significance on the phylogeny, disease prediction and prevention of S. sclerotiorum.
