Seven new species of Lactarius subg. Lactarius are described. These species are representative milk-caps in the subtropical-tropical or subalpine forests in southwestern China. Lactarius angustizonatus, L. pallidizonatus, L. pallido-ochraceus and L. sinozonarius belong to L. subsect. Zonarii, L. subbrevipes to L. subsect. Scrobiculati and L. purpureocastaneus to L. sect. Pallidini. Lactarius brachycystidiatus is tentatively placed in L. sect. Colorati due to the dry scaly pileus and thick hyphae in the pileipellis. Macro- and micro-scopical descriptions, morphological comparisons with their look-alikes and ITS sequences of these new species are provided.
Ellisembia bawanglingensis and Sporidesmium bannaense are two new species of hyphomycetes from southern China. Ellisembia bawanglingensis is distinguished by differentiated, solitary conidiophores and monoblastic conidiogenous cells producing 17-24-distoseptate, 115-145×15.5-20μm, obclavate conidia. Sporidesmium bannaense characterized by macronematous, mononematous conidiophores and monoblastic conidiogenous cells producing 11-25-euseptate, 215-430×12-16μm, obclavate conidia.
During 2004 to 2016 more than 1 000 specimens of wood-decaying fungi were collected in eight nature reserves or forest parks of Shanxi Province, northern China, and 160 species belonging to 77 genera were identified including 38 species new to the province and two species, Antrodiella onychoides and Phlebiopsis pilatii, new to China. Illustrated descriptions and ITS and nLSU sequences are provided for the new Chinese records.
A total of 152 strains of endophytic fungi were isolated from roots, branches and leaves of Blumea balsamifera. Based on morphological characters and ITS rDNA sequence analysis, 11 genera were identified, of which Fusarium, Aspergillus, Trichoderma and Curvularia were dominant. The leaves possessed the highest diversity and population. Luodian, Wangmo, Ceheng and Zhenfeng prefectures showed relatively higher in species diversity and population, especially Luodian. The highest species richness occurred in July, while the highest isolation frequency in December. The distribution of endophytic fungi seems comparatively similar in different localities and growing periods, whereas distribution similarity in branches, roots and leaves is quite different.
Seven hundred and seventy-four isolates of keratin-degrading fungi were isolated from soil samples collected in several hospitals by use of the hair-baiting technique. Fifty-eight species in 31 genera were identified through morphology, Barcode of Life Data Systems, UNITE Database, CBS Database and phylogenetic analysis. According to literatures and the International Society for Human and Animal Mycology rDNA-ITS Database, 512 isolates of 20 species in 13 genera (Alternaria, Aspergillus, Chaetomium, Cladosporium, Fusarium, Geomyces, Geotrichum, Microsporum, Penicillium, Purpureocillium, Talaromyces, Trichoderma and Trichophyton) were of potential pathogenicity to human.
The shiitake mushroom Lentinula edodes is the second popular edible and medicinal fungus in the world. Functional genomic research of this mushroom is developing gradually with the completion of its genome sequencing. Building a genetic transformation system is the foundation and hotspot of gene function research. In this study, we explored a stable genetic transformation system for random insertion mutation, which was mediated by Agrobacterium tumefaciens strains. A vector pYN6982 was constructed by employing hygromycin resistance gene hyg as selection marker, enhanced green fluorescent protein gene egfp as reporting gene. The Agrobacterium tumefaciens strains EHA105 and LBA4404 were both transformed, and then used to transform the mycelia of the sporulated monokaryon and protoplasted monokaryon together with their dikaryon parents simultaneously. The drug resistant transformants were obtained after selection. After five generations of subculture, PCR and sequencing data showed that insert fragment in pYN6982 has already integrated into the genome and could be inherited stably by partial transformants. Furthermore, green fluorescence signals can be observed under microscope. In this study, the feasible method for shiitake mushroom transformation was developed by using millet as a mycelium cultivation and infection medium, which will benefit future functional genetic studies of this fungus.
Simple sequence repeats (SSRs) in the Trametes versicolor genome were identified and characterized in terms of number, distribution and frequency using in silico approaches. SSR-containing genes were functionally annotated. The features of SSRs in T. versicolor were compared with those in other five species of Agaricomycetes, Agaricus bisporus, Bjerkandera adusta, Fomitopsis pinicola, Flammulina velutipes and Pycnoporus cinnabarinus. The results showed that 1 224 SSRs were detected in the T. versicolor genome with the relative abundance of 27 SSRs per Mb. Trinucleotides were the most frequent motifs. SSRs were found to be more frequent in UTR, intergenic regions and introns, though 299 SSRs occur in the CDS, ranking second after intergenic regions. Through blast analysis against the nr database, 485 SSR-containing genes were successfully annotated. Thus, 115 of them could be assigned to the Gene Ontology (GO) terms of molecular function, biological process and cell component, and 108 to KEGG pathways of metabolism, genetic information processing, cellular processes and environmental information processing. Fifty-eight pairs of primers were designed mainly for species identification, and other 23 pairs of primers were mainly used to study active substance metabolism and assist the breeding. Compared with other agaricomycete genomes, T. versicolor has lower number and reduced abundance of SSRs. SSR number was found to be unrelated to genomic size while some specific SSR motifs were related to GC content among the above six species of Agaricomycetes. The results of this study will benefit future population genetics and evolution investigation of T. versicolor.
In this study, seven out of ten endophytic fungal strains from Dongxiang wild rice were screened for producing β-glucosidase. The strain R57 has the highest conversion efficiency, and the biotransformation rate was 98.7% from geniposide to genipin. The strain R57 was identified as Eupenicillium javanicum R57 by morphological characteristics, microscopic observation and ITS rDNA, 18S rDNA, 28S rDNA sequence analysis. Using the combination of ammonium sulfate precipitation, dialysis, ultrafiltration, EzFast DEAE FF anion exchange chromatography and Sephadex G-150 gel column chromatography, β-glucosidase was purified. Molecular weight of β-glucosidase was determined to be 81kDa by SDS-PAGE. The β-glucosidase can hydrolyze p-nitrophenyl-β-D-galactopyranoside (PNPG) and geniposide, however, the efficiencies of hydrolyzing geniposide were higher than that of PNPG. The Km value of β-glucosidase to geniposide was 0.153mmol/L. β-glucosidase showed high pH stability and heat resistance, the highest reaction velocity was acheived at 60°C, pH 4.6. The hydrolysis activity of the β-glucosidase was activated by K+, Co2+, Zn2+, Mn2+, Al3+ and urea, while inhibited by Cu2+, Fe2+, Mo3+, Pb2+ and SDS.
In this study, four strains of endophytic fungi Alternaria sp. AS1113, Chaetomium sp. AS1221, Colletotrichum sp. AS1716 and Penicillium sp. AS1618 isolated from maize Zea mays, were back inoculated to the roots of maize seedlings in a pot experiment under well-watered and drought treatments in greenhouse. The results showed that inoculation of Chaetomium sp. AS1221 and Penicillium sp. AS1618 brought about the significant increase of the height of maize seedlings as compared with no-endophyte inoculation. Compared with no-endophyte inoculation, the aboveground biomass of maize seedlings was significantly increased as a result of inoculation of Chaetomium sp. AS1221, and the underground and total biomass of maize seedlings were significantly increased in case of inoculation of Chaetomium sp. AS1221 and Penicillium sp. AS1618 under the drought treatment. However, no significant difference of the aboveground, underground and total biomass of maize seedlings was observed under the well-watered treatment. The aboveground, underground and total biomass of maize seedlings inoculated with Chaetomium sp. AS1221 and Penicillium sp. AS1618 were significantly higher than those of seedlings inoculated with Alternaria sp. AS1113 under well-watered and drought treatments. Compared with no-endophyte inoculation, the leaf soluble sugar content was significantly lower in case of Alternaria sp. AS1113 inoculation, and the leaf peroxidase activity was significantly lower in case of Colletotrichum sp. AS1716 and Penicillium sp. AS1618 inoculation under well-watered treatment, but no significant difference of soluble sugar content and peroxidase activity of root and leaf was observed in case of inoculation of all four endophytic fungal strains under the drought treatment. This study suggests that Chaetomium sp. AS1221 and Penicillium sp. AS1618 have better effects on the growth of maize seedlings than Alternaria sp. AS1113 and Colletotrichum sp. AS1716 under drought stress.
The high-temperature tolerance is one of the most preferred indicators for evaluation of edible mushroom stress resistance. Five Pleurotus eryngii strains were used to investigate the effects of heat stress on the mycelial growth and their resistance to Trichoderma asperellum by detecting the colony growth rate, growth vigor, the hyphal morphological characters and the infection rate by T. asperellum. Results showed that the five P. eryngii strains responsed to heat stress differently. The mycelium growth recovered in 2-3 days after heat stress (37°C), and the growth vigor became stronger or unchanging. The cell length of distal hyphae, hyphal diameter, hyphal growing rate and mycelium branching frequency reduced significantly. The variation in mycelium growth rate was linearly and positively correlated with the decline in the ability of resistance to T. asperellum. We found that the infection rate of cultivation bags inoculated with the mycelium of T. asperellum attained 100%, while the infection rate was only 30% using spore suspension as inoculum under heat stress at 35°C for seven days. However, cultivation bags were uninfectious under treatment without heat stress. These results indicated that heat stress could reduce the resistance of P. eryngii to T. asperellum.
With the development of printing and dyeing industry in China, dyes have become a problematic class of pollutants to the environment. Exploration of a method which is cheap and effective to control dye-polluted water resources is extremely urgent. In the present study, the extracellular crude enzyme from Pleurotus ostreatus showed excellent activities to remove five synthetic dyes from water. The decolorization rates for crystal violet, acid fuchsin and Coomassie brilliant blue G-250 solutions (40mg/L) were 100%, 98.67%, and 92.5%, respectively. Three kinds of enzyme, viz. laccase, manganese peroxidase and lignin peroxidase were generated from the fermentation of P. ostreatus. Their maximum enzyme activities were measured on 12th, 7th and 8th day. The decolorization rate was the highest when the crude enzyme extract of the 12th day fermentation was employed. According to the relationship between the date of maximum enzyme activity and decolorization rate, we infer that laccase plays a main role in the decolorization process, while manganese peroxidase and lignin peroxidase play supporting role. The current method indicated that employment of crude enzyme was a simpler and cheaper way than that of pure enzyme for decolorizing synthetic dyes. The crude enzyme from P. ostreatus was found to be a potentially applicable decolorizer for effective treatment of effluent.
