Erythrophylloporus, a new bolete genus with a red-colored lamellate hymenophore, is erected within the family Boletaceae based on morphology and multigene phylogenetic analyses. Erythrophylloporus cinnabarinus is described as a new species and represents the type of the genus. Morphologically, E. cinnabarinus is distinguished by a combination of reddish orange to yellowish red basidiomes, reddish orange to yellowish red lamellae gradually changing grayish blue or grayish turquoise to grayish green when bruised, vivid yellow context slightly turning dark violet, blackish blue to dark blue when exposed to air, smooth and broadly ellipsoid, ellipsoid to nearly ovoid basidiospores and dark yellowish brown pigmented cystidia. Phylogenetic analyses based on the internal transcribed spacer region (ITS) and the combined sequences of the large subunit nuclear ribosomal RNA (nrLSU), translation elongation factor 1-alpha (tef1-α), the largest subunit of RNA polymerase II (rpb1) and the second largest subunit of RNA polymerase II (rpb2) confirm that Erythrophylloporus is an independent generic lineage in the Boletaceae. A detailed description, line drawings and comparisons with phenotypic similar genera are provided.
Two new species are reported: Phyllachora isachnicola on Isachne albens and P. sphaerocaryi on Sphaerocaryum malaccense. Descriptions, illustrations and ITS sequence data are provided.
Pinus sylvestris var. mongolica, a typical ectomycorrhizal-dependent arbor tree, is an important afforestation species for windbreak and sand fixation in northern China. To reveal the diversity of ectomycorrhizal fungi of Pinus sylvestris var. mongolica forests, three age groups of plantations (middle-aged, nearly mature, and mature) as well as nature forest were selected in the Hulunbuir Sandy Land. Based on field investigation, symbiotic ectomycorrhizal fungal populations were identified using molecular biology methods. The results indicated that there were 58 taxa of ectomycorrhizal fungi in 25 genera of 21 families. Among them, 48 taxa belonged to basidiomycetes, and the rest 10 taxa ascomycetes. The predominant genera were Amphinema, Cortinarius, and Suillus in the nature forest. Suillus was dominated genus in the plantation, and the relative abundance of other genera fluctuated with the stand age. Simpson, Shannon and Pielou evenness index of ectomycorrhizal fungi significantly differed between the nature forest and plantations of P. sylvestris (P<0.05). However, there were no significance among the P. sylvestris plantations (P>0.05). The population composition of ectomycorrhizal fungi between P. sylvestris natural forest and plantations is quite different in Hulunbuir Sandy Land. The ectomycorrhiza fungal community composition in the nearly mature plantation is closest to that in natural forest. This information provides theoretical basis for P. sylvestris plantation management in the sandy regions.
Vaccinium uliginosum is one of typical host plants of ericoid mycorrhizal (ERM) fungi, which also can be colonized with arbuscular mycorrhizal (AM) fungi and dark septate endophytic fungi (DSE) to form multiple symbionts. The purpose of the present study was to determine and evaluate the root symbiont development status of various cultivars of blueberry adult trees grown under different cultivation systems in flowering and fruiting stages, and the distribution of AM fungi in root zone soil in order to provide theoretical and technical bases for high quality cultivation management technology. Root and root zone soil samples from nine- to ten-year-old plants of Bluecrop, Duke and OʹNeal blueberry grown under plastic cover in the early spring and later autumn, open field and solar greenhouse in Jiawo blueberry production base of Qingdao were collected to observe and measure the quantity of root symbionts, number of AM fungal spores, and AM fungal species distribution. The results showed that ERM, AM, and DSE structures, and their multiple symbionts formed on roots of blueberry trees. Among them, AM gave the largest colonization percentages, followed by ERM, and DSE the lowest; while the colonization of multiple symbiont showed ERM+AM> ERM+DSE>ERM+AM+DSE. The multiple symbiont colonization quantity, AM fungal infection rate, arbuscule colonization percentage, and spore number of AM fungi showed greater in solar greenhouse as compared with those in plastic cover in the early spring, however, they showed decline in open field in later autumn. Different cultivars showed difference in quantity of root symbionts and AM fungal infection rate, manifesting progressive increase order of Bluecrop>Duke>OʹNeal, and ERM and DSE also gave the similar changing trend. Based on the morphological characters of AM fungal spores, totally eleven species in five genera of AM fungi were isolated and identified. More species were isolated from solar greenhouse from the root zone soil of Bluecrop plants. It is concluded that the root symbiont in the flowering and fruiting stage of the blueberry adult trees grown under solar greenhouse is well developed, and it is probably beneficial to the yield, quality and stress resistance of blueberry trees.
The genus Monascus has important economic and ecological value. In order to explore the systematic relationships of the genus Monascus, ITS and LSU genes of section Rubri were sequenced and analyzed and compared with the related genes of the related species and strains in GenBank. Three species in section Rubri and seven species in section Floridani were finally identified. The beta-ketoacyl synthase (pksKS) gene sequence of the Monascus pigment control gene cluster was also used to solve the problem that partial gene sequence analyses could not distinguish clearly the different morphological species M. ruber and M. purpureus. By comparing the results of TA cloning and direct sequencing, the single nucleotide polymorphism (SNP) of three M. sanguineus strains were further analyzed, and the genetic diversities of intra-species were firstly understood. Based on the results of sequence analyses of ITS, LSU and pksKS, in combination with morphological comparison between the related species, the genus Monascus is grouped into two major sections: Rubri and Floridani. Three species (M. ruber, M. purpureus and M. sanguineus) and three varieties (M. ruber var. albidulus, M. purpureus var. aurantiacus and M. purpureus var. rutilus) were identified within the section Rubri. Seven species (M. Floridanus, M. pallens, M. lunisporas, M. argentinensis, M. recifensis, M. flavipigmentosum and M. mellicola) were identified within the section Floridani. In addition, one single suspected nothospecies (M. ruber × sanguineus) was also found within the section Rubri. The results of the current study showed high level of species diversity in the genus of Monascus and the systematic relationship within the genus could be distinguished by polygenic and morphological analyses.
AM (arbuscular mycorrhiza) fungal community structure and diversity in the rhizosphere of Hedysarum laeve (Leguminosae) in desert steppe of Inner Mongolia were analysed by means of morphological identification, denaturing gradient gel electrophoresis (DGGE) fingerprinting technology and next generation sequencing (NGS) technology. Three classes, 4 orders, 10 families and 18 genera of AM fungi were revealed by NGS technique, and 2 classes, 2 orders, 2 families and 5 genera by DGGE, yet only 1 classes, 2 orders, 2 families and 3 genera were classified by morphological identification. There were significant difference between NGS, DGGE and morphological identification results, particularly in the genus level. Morphological method and DGGE fingerprint technology greatly underestimated AM fungal community composition and overestimated the relative abundance of dominant AM fungi, while molecular sequencing is a supplement and improvement over morphological classification.
Mitochondrium is an important organelle in eukaryotic cells for energy conversion and ATP formation. Many edible fungal mitochondrial genomes have been assembled and annotated, however, the mitogenome of Pleurotus pulmonarius is rarely reported. In this study, the mitogenome of the wild type P. pulmonarius strain X2 was assembled and annotated. Molecular markers were developed based on the mitogenomes of the strain X2 and commercial strain JX to classify a wide range of wild and commercial strains. The results showed that the mitogenome of P. pulmonarius is a closed ring structure with a size up to 75 709bp, containing small and large subunit genes of rRNA, 25 amino acids carrying tRNA genes and 14 common protein coding genes. The cox1 gene consists of nine introns mainly of IB type with conservative domain of LAGLIDADG_1 superfamily and GIY-YIG_C term superfamily. It was found that the differential fragment sequences are mainly located in the intron and intergenic regions, which are the main causes of mitochondrial polymorphism between strains X2 and JX. The phylogenetic tree was constructed with different fragment sequences, and the results indicated that the introns, rnl-trnX and trnD-atp6 sequences could be used to distinguish wild type strains from other commercial strains and could be used for the intraspecific identification of P. pulmonarius strains, however trnD-atp6 marker seemed more reliable in identification.
The immune response genes in Plutella xylostella against a high virulent strain of Beauveria bassiana was investigated. Illumina high-throughput sequencing technology was employed to sequence the transcriptome of uninoculated and B. bassiana-inoculated P. xylostella. Differentially expressed genes (DEGs) and their functions, classification and signal pathways were analyzed in 48h post inoculation using bioinformatic methods. Two thousand three hundred and thirty-four genes were significantly differentially expressed in the B. bassiana-inoculated insect as compared with the uninoculated control, including 1 080 up-regulated and 1 354 down-regulated genes. The DEGs were verified using qRT-PCR analysis. GO term enrichment analysis revealed that these DEGs could be assigned to 45 categories with 23 biological processes, 12 cellular components and 10 molecular functions. Four hundred and ninety-seven up-regulated genes and 811 down-regulated genes were enriched in 25 KEGG pathways. Some uncharacterized genes, such as histone, phosphopantothenoylcysteine decarboxylase, double stranded RNA binding protein, and myrosinase genes, were found to be dramatically upregulated during the infection process, suggesting that these genes might be related to the immune response in P. xylostella against the infection of B. bassiana.
Based on high-throughput sequencing technology, the fungal ecological niche of Metacordyceps neogunnii from five different regions, Yifeng (Jiangxi Province), Shiqian, Meitan, Fenggang and Shibing (Guizhou Province), was investigated. The results showed that the fungi from Metacordyceps neogunnii habitat soil (Cg1), surfaces of fruit body (Cg2) and endo-sclerotia (Cg3) belonged to four phyla [Ascomycota, Basidiomycota, Zygomycota and Rozellomycota] and some undetermined fungi, 17 classes, 43 orders, 77 families, 129 genera and 177 species. At genus level, the dominant genus in Cg1 was Mortierella, with relative abundance of 84.86%, followed by an uncertain genus (4.58%) and a genus of Chaetomiaceae (1.22%); the dominant genus in Cg2 was Metacordyceps neogunnii (83.66%), followed by Clonostachys rosea (9.48%), Sarocladium (2.07%) and Mortierella spp. (1.12%). The fungi in endo-sclerotia from Yifeng of Jiangxi (JX), and Shiqian (SX), Fenggang (FG), and Shibing of Guizhou (Cg3) possess the same dominant species, i.e. M. neogunnii, and the relative abundances were 99.74%, 99.50%, 99.73% and 98.82%, respectively. The relative abundances of the rest genera were less than 1%. The dominant genus colonized in a dried specimen of M. neogunnii from Meitan was an uncertain genus in Clavicipitaceae, and its relative abundance was up to 99.97%. Both the stroma and endo-sclerotia of M. neogunnii dried specimen from Shibing were colonized by the same dominant species, M. neogunnii, with the relative abundances of 99.96% and 99.38%, respectively. In conclusion, the endo-sclerotia of the specimens of M. neogunnii from four areas were multiply colonized by some other fungi. However, M. neogunnii did not exist in the habitat soil, and the dominant genera in the habitat soil were Mortierella and Trichoderma. The specimen similar to M. neogunnii from Meitan was actually not conspecific, but belonged the other uncertain genus of Clavicipitaceae, which was in need of further study on its taxonomic status.
The fermentation kinetics of chitin deacetylase (CDA) produced by Fusarium proliferatum was analyzed. The non-structured dynamic model of growth of F. proliferatum cell, synthesis of CDA and consumption of sugar were constructed by Logistic Equation. The model was simulated using 1stOpt software, and the nonlinear curve fitting graph and various model parameters were obtained using Origin 8.0 software. The results showed that the predicted values of the models were well fitted to the experimental data, and specific growth rate of F. proliferatum mycelium peaked at 15.52h (μm, x) 0.160h-1. The rate of substrate consumption peaked at 26.51h, and reached the peak value (μm, s) of 0.096 h-1. The specific synthesis rate of CDA peaked at (μm, p) 0.548U/(mL·h) in 19.40h. The model fitting and experimental data have a good adaptability, and the model made it possible to elucidate the kinetic characteristics of F. proliferatum during the enzyme production process. These models provide useful information for the industrialization scale production in the future.
Qualitative and quantitative analysis of ergosterol, and the study on fingerprint of liposoluble constituent in Ganoderma lingzhi spores were carried out by using the method of high performance liquid chromatography (HPLC). The method proved to be sensitive, accurate, reproducible and reliable, and the fingerprint of liposoluble constituent showed a flat baseline with abundant peaks, being possessed of high resolution. It was indicated that the developed method could be utilized for quality evaluation of ergosterol (at 282nm), and fingerprint analysis of liposoluble constituent (at 245nm). Wall-breaking treatment had an obvious influence on the extraction yield of ergosterol and other liposoluble constituent, therefore, the optimal wall-breaking duration should be kept in 20 to 30 minutes. Low storage temperature was better for extending the shelf life of the spores. The content of liposoluble constituent increased with the extension of duration of harvest period, along with mild decrease of ergosterol. The material origins had an important influence on the quality of spores. There was no correlation between the content of ergosterol and the fingerprint of liposoluble constituent in the spores, therefore the comprehensive analysis of both data was very important in evaluating the quality of the spore powder.
The effects of different extraction methods on the extraction of polysaccharide from Ophiocordyceps sinensis mycelium (PDOM) fermented in medium using the insect Dendrolimus punctatus (Lasiocampidae) as substrate were comparatively investigated. The results showed that the optimal extraction method was the application of microwave-assisted extraction technique, and the key factors influencing the extraction were the ratio of water to mycelium, microwave extraction period and microwave power. Box-Behnken test design combined with response surface methodology were applied to optimize the extraction process of PDOM. The polynomial model Y=6.87+0.058A+0.085B+0.075C+0.032AB+0.046AC+0.069BC-0.16A2- 0.37B2-0.11C2 is established to predict the extraction of PDOM. It was showed that the optimal process parameters were the ratio of water to mycelium (V/M, mL/g) 6.3:1, microwave power 520W, and microwave extraction period 326s. Under such a process the extraction rate of PDOM reached 6.76%.
Manganese peroxidase (MnP), one of the key enzymes secreted by white rot fungi, can degrade various xenobiotics efficiently. In the present study, the activity curve of MnP activity from Irpex lacteus was measured. The fermentation conditions of I. lacteus producing MnP were optimized using single factor and orthogonal experiments. Extracellular solutions of MnP were used to decolor five kinds of dye. The results showed that the MnP activity of I. lacteus was highest at day 5 of culture. The most optimal culture parameters for MnP production were soluble starch 20g/L, urea 1g/L, CaCl2 1mmol/L, FeCl3 1mmol/L, and pH 6.3. The MnP activity reached 29.24U/L after 5 days of culture, 1.25 times higher than that before optimization. The decolorization test showed that extracellular solutions of MnP from I. lacteus could decolor five types of dye. The decolorization effect against direct red and reactive red was particularly obvious, with decolorization rates of 82% and 81%, respectively, after 5 days of decoloration.
Three isolates with ability to decolorize methylene blue dye were screened from among 19 white-rot fungal isolates cultured in solid media demonstrating decolorant zone of 7.5, 6.8 and 5.5cm in diameter. They were identified as Trametes versicolor (ZT-197), Trametes pubescens (ZT-230) and Bjerkandera fumosa (ZT-307). ZT-230 showed the highest decolorizing ability, with decolorization rate of 100% for 100mL liquid media containing 50mg/L of methylene blue within six days, followed by ZT-197 with 98% within ten days and ZT-307 with 80% within ten days. Activities of extracellular enzymes (laccase, manganese peroxidase and lignin peroxidase) were also monitored. The results showed that ZT-197 and ZT-230 could secrete laccase and manganese peroxidase while ZT-307 lignin peroxidase only. Trametes pubescens ZT-230 has potential for dyeing effluent treatment application.
