Vanderbylia robiniophila (Huai’er) is one of the important medicinal fungi in China. Various compounds including polysaccharides, steroids and alkaloids have been isolated from this fungus. Huai’er is widely used in the adjuvant treatment of breast cancer, liver cancer, lung cancer, gastric cancer and other malignant tumors. It was reported that the mechanisms of the anti-neoplasia effect included inhibition of the growth and proliferation of cancer cells, inhibition of the invasion and migration of cancer cells, suppression of angiogenesis, induction of apoptosis of cancer cells and improving immunity, etc. Herein, the chemical constituents and anti-tumor effect of Huai’er are summarized as reference data for development of this medicinal fungus.
Orchidaceae plants are always interested by phytologists and ecologists due to their high species diversity and important social economy values. Orchid mycorrhizal fungi (OMF) play an important role in seed germination, nutrition absorption and population dynamics of orchids, and attract more and more people’s attention in recent years. Exploring the mechanisms of interaction between orchids and OMF is the hotspot in orchid mycorrhiza research field, and provides new insight into orchid conservation and population recovery. With the development and establishment of next generation sequencing and multi-omics databases, research on mechanisms of interaction between OMF and orchids has been entered a new era, and people know more about internal mechanisms of seed symbiotic germination, nutrient transportation, symbiotic signal transduction, host immunity and stress resistance by using the omics techniques such as genomics, transcriptomics, proteomics and metabolomics. In this paper, researches associated with the mechanisms of interaction between OMF and orchids by using omics technology in recent decade are summarized, and future advances in this field are prospected.
Diaporthe species are well-known as pathogens, endophytes or saprobes on a range of economically important crops, ornamentals and forest trees. Diaporthe species associated with symptomatic plant leaves from Yunnan Province in China were investigated. Two new species were described based on the analysis of DNA sequence data of the nuclear ribosomal internal transcribed spacer (ITS), β-tubulin (TUB), partial translation elongation factor-1α (TEF), calmodulin (CAL) and histone H3 (HIS) regions, in combination with their morphological features. Diaporthe conferta sp. nov. was described from symptomatic leaves of Elaeagnus conferta and D. chinensis sp. nov. was from those of Litchi chinensis.
Two new corticioid fungal species, Hyphoderma sinense and H. floccosum from Yunnan Province, southwestern China, are proposed based on a combination of morphological features and molecular evidence. Hyphoderma sinense is characterized by the resupinate basidiomata with smooth hymenial surface, presence of encrusted and moniliform cystidia, and cylindrical to allantoid basidiospores (8-11.5×3-5μm). Hyphoderma floccosum is characterized by the farinaceous hymenial surface, tubular cystidia and leptocystidia, and ellipsoid to allantoid basidiospores. Sequences of ITS+nLSU nrRNA gene regions of the studied samples were generated and phylogenetic analyses were performed with maximum likelihood, maximum parsimony and Bayesian inference methods. The phylogenetic analysis based on molecular data of ITS+nLSU sequences showed that two new species clustered into the genus Hyphoderma, in which H. sinense grouped with H. transiens, and H. floccosum grouped with a clade comprising H. pinicola, H. setigerum and H. subsetigerum. The new taxa are described, illustrated and compared with morphologically similar species. An identification key to 23 accepted species in China is provided.
Three species of the /pygmaea clade in the genus Psathyrella, P. panaeoloides, P. saponacea, and P. squarrosa, are reported in China. The new species, P. squarrosa, discovered in Guangdong Province, is characterized by its gregarious and small basidiomata, subcorniform and evanescent veil consisting of oblong-ellipsoid to subglobose cells and utriform pleurocystidia. The morphological characteristics and phylogenetic analyses (ITS, LSU, β-tub and tef-1α) show that P. squarrosa is unique and distinct from related species within Psathyrella. The detailed description, color photos, illustrations and a key to related species are presented.
Botryosphaeria spp. are plant endophytes, saprobes or parasites mostly associated with woody hosts. In this paper, two new species of Botryosphaeria causing stem canker of blueberries, B. dolichospermatii and B. fujianensis, are described and illustrated from southern China. Phylogenetic analysis based on combined ITS, LSU, tef1-α and TUB loci supports their separation from all other known species of Botryosphaeria. Koch’s postulate tests proved that both B. dolichospermatii and B. fujianensis were causal agents of stem canker of blueberries.
Diversity of fungi colonized in wheat seeds was investigated. Fifty-seven wheat seed samples from China and those imported from Australia, Canada, Kazakhstan, Russia were subjected to strain isolation, of which 21 samples were analyzed by high-throughput sequencing. In total 564 fungal strains were isolated, and 69 species in 31 genera were identified based on ITS sequence analyses and morphological observations. Species of Alternaria (with relative abundance of 51%), Pyrenophora (6%), Epicoccum (6%), Fusarium (5%), and Penicillium (5%) were the most abundant, and 17 genera were isolated from wheat seeds for the first time. In addition, several potentially important phytopathogens, e.g. Alternaria alternata, Bipolaris sorokiniana, Fusarium graminearum were also isolated. The diversity analysis of high-throughput amplicon sequences showed that only the samples from Russia were significantly different from those from China. The dominant genera revealed by high throughput sequencing were Alternaria, Cladosporium and Epicoccum, with relative abundances of 23.95%, 18.91% and 13.32% respectively. FUNGuilds analysis suggested that pathotrophs accounted for the highest portion of wheat seed fungi, and these potential phytopathogens may develop diseases under suitable conditions.
One thousand eight hundred and twenty-one isolates of endophytic fungi were isolated from 30 genera of Poaceae collected in Yunnan, Zhejiang and Inner Mongolia of China. They were identified by ITS rDNA sequence analysis and threshold for fungal delimitation. The sum total of 216 genera in 34 orders of 10 classes of 3 phyla were obtained, including 192 genera in 26 orders of 5 classes of Ascomycota, 21 genera in 6 orders of 3 classes of Basidiomycota and 3 genera in 2 orders of 2 classes of Mucoromycota. Sordariomycetes and Dothideomycetes were the dominant classes with the relative frequency of 54.8% and 30.9%, respectively. Pleosporales of Dothideomycetes and Hypocreales, Magnaporthales, Glomerellales and Xylariales of Sordariomycetes were the dominant orders with the relative frequency of 26.7%, 12.6%, 12.1%, 11.3% and 9.9%, respectively. The distribution of the dominant classes and orders were significantly different in regions with different latitude. Among the 216 genera identified, the relative frequency of seven known genera, Colletotrichum, Alternaria, Fusarium, Diaporthe, Penicillium, Arthrinium, Trichoderma, and a taxonomically uncertain genus, Pleosporales incertae sedis type 12, were over 2%. All the seven known genera were widely distributed. The community composition and diversity of endophytic fungi in Yunnan, Zhejiang and Inner Mongolia varied significantly, and only 58 genera were common when comparison was made between the two regions or among the three regions. The diversity of endophytic fungi in Yunnan was the highest, while that in Inner Mongolia was the lowest. The composition and diversity of endophytic fungi in roots and shoots varied significantly. Only 53 genera were common in roots and shoots, and the diversity of endophytic fungi in shoots was higher than that in roots. Further investigation in other regions, especially in tropical and subtropical regions, is necessary for enriching our understanding of the diversity of endophytic fungi in plants of Poaceae in China.
The high-throughput sequencing was used to determine the seasonal dynamics of community of arbuscular mycorrhizal fungi (AMF) in the root and rhizosphere soil of Vetiveria zizanioides in coal gangue dump with planting period of 17 years. Spearman, NMDS, RDA and Mantel test were used to analyze the correlation among AMF spore density, colonization rate, physicochemical properties of soil, richness and diversity index. The results showed that the spore density and colonization rate of AMF in summer were significantly higher than those in spring and autumn. In spring, summer and autumn, 127 OTUs, 258 OTUs and 96 OTUs were obtained from root system, rhizosphere soil and both of them, respectively, and 7 genera and 4 families of AMF were identified. Glomus was the dominant genus in the coal gangue dump. The AMF diversity and richness index of root first decreased and then increased with the seasonal change; the AMF diversity index of rhizosphere soil first decreased and then increased, while the richness index showed a decreasing trend with the seasonal change. Shannon, Chao1 and ACE indexes of rhizosphere soil in spring were significantly higher than those of root system, and the Chao1 index of rhizosphere soil was significantly higher than that of root in summer. NMDS analysis showed that the distribution of AMF community in summer was significantly different from that in other seasons. Spearman analysis showed that total potassium content in roots was significantly positively correlated with diversity and richness index of AMF. There was a significant negative correlation between available phosphorus and AMF richness index. The results of RDA analysis showed that AMF community composition was positively correlated with total phosphorus, total nitrogen, total potassium and available phosphorus in root system in spring and summer and in rhizosphere soil in summer and autumn, but it was negatively correlated with organic matter. Mantel test analysis showed that pH, organic matter, total nitrogen, total potassium, available phosphorus and sampling season significantly affected mycorrhizal colonization rate, and organic matter and total nitrogen were the dominant factors affecting spore density.
Diversity of community composition of endophytic fungi from Pinus massoniana in Guizhou Province was investigated. The endophytic fungi were isolated from the roots, stems and leaves of 30-year-old healthy P. massoniana by tissue isolation method in different seasons. Morphology identification combined with ITS sequence analysis resulted in acquisition of 1 099 isolates of endophytic fungi from 1 328 tissue blocks of P. massoniana. The total colonization rate was 93.98%, and the total isolation rate was 82.76%. Among them, 1 084 isolates belonged in 10 classes, 21 orders, 37 families, 55 genera and 69 species of Ascomycota, Basidiomycota and Mucormycota, Trichoderma (IR=10.84%, IF=13.10%), Pestalotiopsis (IR=9.55%, IF=11.57%), Fusarium (IR=8.58%, IF=10.38%), Alternaria (IR=7.07%, IF=8.56%) and Penicillium (IR=5.50%, IF=6.65%) were the dominant genera. Diversity and similarity analysis showed that the most abundance of endophytic fungi occurred in autumn (H′=3.02, D=0.94), but less in summer (H′=2.64, D=0.91). The abundance in winter and summer (Cswinter-summer=0.43) was most similar. The diversity of endophytic fungi in root (H′=2.96, D=0.93) was richer than that in stems (H′=2.69, D=0.43), while their similarity in stems and roots (Csstem-root=0.45) was the highest, that in leaves and stems (Csleaf-stem=0.33) was lower, and that in leaves and roots (Csleaf-root=0.25) lowest. Further exploration of the species composition, influencing factors and function development of endophytic fungi in P. massoniana needs to be carried out.
Thirty-seven wild isolates of Venturia inaequalis, causal agent of apple scab, were collected from abolished orchards without application of fungicides in Xinjiang for determining the baseline sensitivity of the pathogen to myclobutanil in order to monitor fungicide-resistance of the pathogen and disease control in the field. Baseline sensitivity of Venturia inaequalis to myclobutanil were tested by use of in vitro bioassay methods of determining conidial germ tube elongation and monoconidial hyphal growth rates of the fungus cultivated on fungicide-amended agar with various concentrations of myclobutanil. The EC50 of the baseline population was determined, ranging from 0.028mg/L to 1.017mg/L, with a mean value of 0.283mg/L. The tested isolates show certain sensitivities to myclobutanil, and no resistant isolates are found in the treated orchard. The in vitro tests imply that the baseline of 0.460mg/L could be used as a value to monitor V. inaequalis resistance to myclobutanil and disease control risk assessment for fungicide application in orchard.
The effects of different substances on the mycelial growth of Phallus impudicus were studied. P. impudicus-1 plate culture was used as tested materials, and the effects of five substances on the mycelial growth of P. impudicus-1 were analyzed. Brown sugar was the best carbon source, accelerating the mycelial growth rate to reach a maximum of 1.445mm/d, being superior to sucrose and glucose. Yeast extract was the best nitrogen source, stimulating the densification of mycelium and the mycelial growth rate could reached 1.413mm/d, being superior to beef peptone and trypton. Monopotassium phosphate, potassium chloride and calcium chloride could promote the mycelial growth of P. impudicus, while magnesium sulfate and sodium chloride was disadvantageous to the mycelium growth. When the concentration of potassium chloride was 0.5g/L, the mycelium growth rate reached the highest (1.034mm/d). VB1, VB2, VB6 and Vc could promote the mycelium growth. Under VB6 concentration of 2mg/L, the mycelial growth reached the highest rate of 1.673mm/d. Among the phytohormones tested, 6-BA had the strongest effect on mycelial growth, and NAA was the next best. The suitable medium formulation was the key factor to promote the growth and development of P. impudicus.
Botrytis disease is the most common and serious disease on Paris polyphylla var. chinensis in Hubei and Hunan provinces. In order to clarify the genetic diversity level of the pathogen Botrytis cinerea, 92 isolates from 10 regions of Hubei and Hunan provinces were analyzed by rDNA-ITS molecular identification and simple sequence repeats (SSR-PCR) molecular marker technique. rDNA-ITS molecular identification showed that these isolates were all B. cinerea and ITS phylogeny tree indicated significant genetic diversity. SSR-PCR diversity analysis showed that 109 alleles were detected by 9 primer pairs, and observed number of alleles and effective number of alleles average 12 and 3.6589 respectively. Shannon’s diversity index was 1.5048, and polymorphism information content 0.6196. For the investigated 10 regions, the values of effective number of alleles ranged from 1.622 to 3.971, with Shannon’s diversity index of 0.527-1.582, observed heterozygosity of 0.148-0.593, and expected heterozygosity of 0.321-0.708. The UPGMA phylogenetic tree based on Nei’s genetic distance showed that fungal isolates from 10 regions were divided into four groups. Gene flow value between populations was 0.19-4.67, with genetic differentiation value of 0.047-0.638, genetic identity value of 0.1531-0.9679, and genetic distance value of 0.0327-1.8766. In summary, the populations of B. cinerea on Paris polyphylla var. chinensis in Hubei and Hunan provinces were genetically diverse, and the results may benefit the control of the disease in these provinces.
Melampsora larici‐populina is a forest pathogenic fungus with wide distribution and serious damage potentials. Analyses of gene duplication modes and syntenic relationship are helpful for us to understand some biological points such as adaptive evolution of M. larici‐populina. There are few reports on gene duplications at the whole genome level in M. larici‐populina, and limited reports on the concerned synteny research. In this study, referenced genome of M. larici‐populina was used to analyze the gene duplication modes. The results indicated that gene number of transposed duplications was higher than that of segment, tandem and proximal duplications in M. larici‐populina. We inferred that whole genome duplication did not occur in M. larici‐populina, because of no large scale segmental duplications and no peaks found by detecting gene age distributions. The enrichment analysis showed that different duplicated genes were enriched in different functional categories, such as the tandem duplications were enriched in carbohydrate transport and metabolism, while transposed duplications were enriched in secondary metabolite biosynthesis, transport and catabolism. The levels of synteny were low among M. larici-populina, Cronartium quercuum f. sp. fusiforme and Puccinia striiformis f. sp. tritici, and one common synteny block was found among them by synteny analyses. There were six genes located in this conserved region, among which a putative small secreted protein was worthy of paying attention.
Anthracnose on Camellia oleifera is an important disease caused by Colletotrichum species, and always occurs in producing regions of C. oleifera in China. Our previous studies revealed that Colletotrichum fructicola was the major pathogen of anthracnose on C. oleifera, but the functions of the autophagy-related protein Atg8 in C. fructicola remains unknown. Based on homologous recombination principle and the PEG-mediated protoplast transformation, the CfATG8 gene-deletion mutants and the related complemented strains were obtained. It was found that the gene-deletion mutants showed reduction of growth rate, decrease in conidial production and appressorial formation, more sensitive to cell wall integrity stress, and no pathogenicity to C. oleifera. Our results reveal that CfAtg8 is functionally important to the vegetative growth, asexual reproduction, appressorial formation, external stress response and pathogenicity of C. fructicola.
Agaricus bisporus is an important species of edible mushrooms. To understand the influencing factors of codon bias and their relationship with gene expression, the codon usage bias of A. bisporus genome was analyzed. Based on the genome and transcriptome data, the codon usage of genomic genes, high expression genes (HEG) and low expression genes (LEG) in A. bisporus were analyzed. The results showed that the average GC content of coding genes was 49.08% in A. bisporus; the T3s value (35.59%) was the highest, and the average ENC (effective number of codons) value was high, while the expression potential of most A. bisporus genes was low. A total of fourteen optimal codons was identified, and all ended with the nucleotide C or T. The optimal codons adhered to the equilibrium of purine and pyrimidine. Comparatively, high expression genes have stronger codon bias and are affected by many factors, such as gene mutation, natural selection and so on. Gene expression was positively correlated with G/C content and codon adaptation index value. The highly expressed genes encode a variety of proteins and enzymes related to the growth and development of fungi. The results revealed the characteristics of the codon usage bias of A. bisporus genes, providing a reference for transgenic breeding and quality improvement of A. bisporus.
Antioxidant enzymes and heat shock proteins are important proteins for Agaricus bisporus to resist adversity stress. Under high temperature stress, mycelia can reduce the damage of itself through the differential expression of their genes. In this study, the mycelia of A. bisporus were treated with 40°C heat stress for 0-120min. It was found that with the extension of heat stress duration, the growth rate of mycelia decreased, aerial mycelia increased, and mycelia bifurcated significantly. Transcriptome analysis of the differential expression of antioxidant enzymes and heat shock protein genes indicated that the antioxidant enzyme genes gpx, ppo3 and cat3 and the heat shock protein genes hsp, hsp70-1 and hsp70-17 were significantly up-regulated under heat stress for 30-60 minutes, while the antioxidant enzyme genes ppo1, ppo2 and cat2 and the heat shock protein genes hsp16, hsp70-14, hsp70-3 and hsp70-16 were significantly up-regulated under heat stress for 90-120min. The result of tracking antioxidant enzyme activity showed that heat stress could activate catalase (CAT) and peroxidase (POD), and increase enzyme activity by 2-3 times; at the same time, heat stress could reduce superoxide dismutase (SOD) activity, but had little effect on polyphenol oxidase (PPO). The study also found that heat stress could make A. bisporus accumulate more O 2- which could damage the mycelia. Our research shows that A. bisporus mycelium can resist the damage caused by high temperature stress by activating the expression of different antioxidant enzymes and heat shock protein genes during heat stress. Under such circumstances, CAT and POD might play a major role in scavenging oxidative free radicals. Our preliminary research on high temperature tolerance genes of A. bisporus might provide reference for breeding varieties with high temperature tolerance.
Flammulina filiformis is a kind of edible fungus species that has been largely factory-produced domestically. However, its breeding work is still limited due to the low efficiency of phenotyping collection. In this study, the visible images of F. filiformis fruiting bodies were used as informative source, and a method for phenotyping collection and analysis was proposed by use of image recognition techniques and deep learning models to meet the needs of phenotypic information in breeding. A software for this method was consequently developed. Based on the visible light scanning images of F. filiformis fruiting bodies, this method is able to automatically calculate the phenotypic information of fruiting bodies in large quantities, including cap shape, area, position and color, and stipe length, width and color. The proposed method can greatly reduce observation errors, increase the reliability of phenotypic information, and extend to breeding of other edible fungi with similar appearance. The data obtained are instructive to quantitative analysis of phenotypic information of F. filiformis and useful to establish standardized phenotype data sets.
Sanghuangporus is currently recognized as one of the valuable medicinal macrofungi with antitumor activity. The metabolomics of S. vaninii cultivated on bran substrate was studied based on LC-MS. 355 metabolites were detected from the fruiting bodies of S. vaninii, of them 86 ones were differential metabolites, 51 ones were up-regulated and 35 were down-regulated. The main metabolites were amino acids and their derivatives, nucleotides and their derivatives, sugars and alcohols, lipids, flavonoids, alkaloids, organic acids and phenolic acids, etc. The analysis of KEGG metabolic pathway showed that different metabolites were enriched in 56 metabolic pathways, and seven of which were significantly different. They were ubiquinone and other terpenoid quinone biosynthesis, secondary metabolite biosynthesis, tryptophan metabolism, propionic acid metabolism, isoquinoline alkaloid biosynthesis, tyrosine metabolism and metabolic pathway. Based on UPLC-MS/MS technology, the metabonomics of the fruiting body of S. vaninii was determined. The result showed that the addition of bran could promote the synthesis and accumulation of the secondary metabolites. This study provides a reference for the comprehensive development and utilization of S. vaninii.
Triterpenoid was recognized as the major bioactive molecule of Ganoderma lingzhi. In this work, liquid superficial-static culture (LSSC) process was used to enhance the production of triterpenoid. After optimizing, the best condition to produce triterpenoid in LSSC was initial medium volume 2mL, inoculum size 7.0g/L, adding medium 2mL in 48h and fermentative duration 7d. Under the optimal conditions, the triterpenoid production scaled (65.91±0.84)mg/g, being 4.19-fold that in submerged culture. The results demonstrated that aerial mycelial formation was a key factor for triterpenoid biosynthesis of G. lingzhi. Compared with submerged culture, LSSC exhibited growing fast and high triterpenoid production and productivity needless to stir or shake, and had potential for industrial application.
The effects of growth age on the content of nutritional components (crude protein, crude fat, crude fiber and amino acid) and active components (polysaccharide and total triterpenes, flavonoid and phenol) and antioxidant activity (ABTS and FRAP) of one-, two- and three-year-old fruiting bodies of Sanghuangporus baumii (previously as Phellinus baumii) cultivated on oak segment were investigated. The result showed that the growth age had a significant effect on the nutritional components, content of active components and antioxidant activity of S. baumii fruiting bodies. The content of crude protein and fiber of one-year-old fruiting bodies is significantly higher than that of two- and three-year-old ones; the content of crude fat of three-year-old fruiting bodies is higher than that of one- and two-year-old ones; the content of total amino acid of two-year-old fruiting bodies is higher than that of one- and three-year-old ones. The content of leucine and isoleucine is significantly higher in one-year-old fruiting bodies than that in three-year-old ones, and the lowest in two-year-old ones. The content of polysaccharide of two-year-old fruiting bodies is the highest, that of three-year-old ones is secondary, and that of one-year-old ones is the lowest. The content of total flavonoid and phenol, and antioxidant activities (ABTS, FRAP) of one-year-old fruiting bodies are the highest, those of two-year-old ones secondary, and those of three-year-old ones the lowest. The content of total triterpenoid of one- and three-year-old fruiting bodies is higher than that of two-year-old ones. Correlation analysis showed that the crude protein content was negatively correlated with the crude fat content, and positively correlated with the total flavonoid and phenolic content. The crude fiber content was negatively correlated with the polysaccharide content. There were positive correlation between the total flavonoid and phenolic content, and antioxidant activities (ABTS and FRAP). The content of total amino acids and triterpenoids was no significant correlation with any other examined indicators. The results provided a reference for the quality control and evaluation of further development of Sanghuangporus product.
Hypoglycemic effect and active compounds in ethanolic extract of Sparassis latifolia fruiting bodies are investigated. Type II diabetes mellitus mouse model induced by combined administration of streptozotocin and high-sugar feed was adopted. Normal control, diabetic model control, three groups treated with different dosages of the fungal extract (300, 200 and 100mg/kg) and positive control were designed. Oral administration of the fungal extract was conducted daily for three weeks. Body weight, blood glucose, lipid levels and histopathological change of pancreas of the tested mice were investigated. Compared with the diabetic control group, the levels of blood glucose and cholesterol of mice treated with S. latifolia extract were significantly reduced. The protective effects were further supported by the histopathological observation of pancreas that the shape, amount, distribution of β cells were obvious ameliorated. The active compounds of S. latifolia extract were analyzed by UPLC-Triple-TOF/MS. Ten compounds were identified, and several of them might be involved in glucose and fat metabolism. The results may supply theoretical and technical support for the application and development of S. latifolia.
