Isolation, structural characterization and immunomodulatory activity on RAW264.7 cells of a novel exopolysaccharide of Dictyophora rubrovalvata.

Fungal polysaccharides have been explored by many for both structural studies and biological activities, but few studies have been done on the extracellular polysaccharides of Dictyophora rubrovalvata, so a new exopolysaccharide was isolated from Dictyophora rubrovalvata and its structure and its immunological activity were investigated. The crude exopolysaccharide (EPS) was purified by DEAE52 cellulose and Sephadex G-200 to obtain a new acidic polysaccharide (DR-EPS). DR-EPS (2.66 × 103 kDa) was consisted mainly of mannose, glucose, galactose and glucuronic acid with a molar ratio of 1: 0.86: 0.20: 0.01. In addition, DR-EPS increased the phagocytic activity of RAW264.7 cells up to 2.67 times of the blank control group. DR-EPS improved intracellular nucleic acid and glycogen metabolism as observed by AO and PAS staining. DR-EPS(40 µg/mL) promoted NO production up to 30.66 µmol, enhanced acid phosphatase (ACP) and superoxide dismutase (SOD) activities, with activity maxima of 660 U/gprot and 96.27 U/mgprot, respectively, and DR-EPS (160 µg / mL) significantly increased the lysozyme content as 2.73 times of the control group. The good immunological activity of extracellular polysaccharides of Dictyophora rubrovalvata provides directions for the use of fermentation broths.

Two new species and one new combination of Ophiocordyceps (Hypocreales, Ophiocordycipitaceae) in Guizhou.

Ophiocordyceps is the largest genus in Ophiocordycipitaceae and has a broad distribution with high diversity in subtropical and tropical regions. In this study, two new species, pathogenic on lepidopteran larvae are introduced, based on morphological observation and molecular phylogeny. Ophiocordycepsfenggangensissp. nov. is characterised by having fibrous, stalked stroma with a sterile tip, immersed perithecia, cylindrical asci and filiform ascospores disarticulating into secondary spores. Ophiocordycepsliangiisp. nov. has the characteristics of fibrous, brown, stipitate, filiform stroma, superficial perithecia, cylindrical asci and cylindrical-filiform, non-disarticulating ascospores. A new combination Ophiocordycepsmusicaudata (syn. Cordycepsmusicaudata) is established employing molecular analysis and morphological characteristics. Ophiocordycepsmusicaudata is characterised by wiry, stipitate, solitary, paired to multiple stromata, yellowish, branched fertile part, brown stipe, immersed perithecia, cylindrical asci and cylindrical-filiform, non-disarticulating ascospores.

Reducing the mass and decreasing the bioavailability of heavy mental from organic wastes treated by black soldier fly larvae.

Black soldier fly larvae (BSFL), Hermetia illucens L., are widely used to reduce the mass of various wastes. However, the potential metal tolerance mechanisms during periods of waste bioconversion by BSFL remain largely unknown. To further reveal the mechanisms, BSFL were used to treat the agricultural organic wastes, including pig manure (PM), cow manure (COM), spent mushroom substrate (SMS), and wet distiller grains (WDG). After these individual and combined waste(s) were treated by BSFL, we investigated the waste reduction rates and evaluated the responses of BSFL gut microbes to heavy metals of agricultural organic wastes. Additionally, the colloidal particles of residual wastes were characterized by combing energy dispersive X-ray (EDX) spectroscopy, Size potential, Zeta potential, and excitation-emission matrix (EEM) spectroscopy. Results indicated that the waste reduction rates were up to 74% in COM+WDG and 69% in WDG, most of heavy metals (e.g., Zn and Co) from organic wastes were not accumulated in the bodies of mature larvae after treatment. Further, results obtained from the prediction of gene function on the basis of 16 S rRNA data revealed that the presence of multi-resistance genes in the gut of BSFL can help the larvae resist Zn and/or Co stress. In addition, the drug sensitivity test implied that BSFL5_L and BSFL6_L from BSFL gut bacterial strains have multi-resistance to Co and Zn. Additionally, EDX results revealed that the colloidal particles in five waste residues after BSFL treatment are mainly consisted of Fe, Ca and Si, which can capture heavy metals (e.g., Cu, Mn). Results from EEM spectroscopy and PARAFAC showed that tryptophan-like and humic-like accumulatively account for 56%- 68% of all components. Importantly, these two components could strongly bind the metal elements and form colloidal particles with high stability, and therefore reduce the heavy metal pollution of agricultural organic wastes. Our findings offered an environment-friendly method to treat agricultural organic wastes, which would be far-reaching influence to our environment.

Utilizing Black Soldier Fly Larvae to Improve Bioconversion and Reduce Pollution: A Sustainable Method for Efficient Treatment of Mixed Wastes of Wet Distiller Grains and Livestock Manure.

Widespread environmental contamination caused by huge amounts of wastes generated by human activities has become a critical global concern that requires urgent action. The black soldier fly (BSFL) has gradually been used to treat different wastes due to high efficiency and low cost. However, little information is available regarding the treatment of mixed wastes by BSFLs. The impact of BSFLs on conversion of cow manure (COM) and pig manure (PM) via the incorporation of wet distiller grains (WDG) was assessed. Results demonstrate that the waste reduction rate was increased by 20% by incorporating 45% WDG to COM and PM. The bioconversion rate of BSFLs in COM and PM also increased from 1.20 ± 0.02% and 0.92 ± 0.02% to 10.54 ± 0.06% and 10.05 ± 0.11%, respectively. Total nitrogen content and δ15N/14N ratios of WDG + COM and WDG + PM were found to be significantly lower than those of COM and PM alone (p < 0.01). The organic matter changes during manure degradation were further analyzed by combing ultraviolet-visible spectrum (UV-vis) with excitation-emission matrix (EEM) spectroscopy techniques and fluorescence area integration (FRI) method. The UV-vis spectra results indicate that the addition of WDG to manures resulted in the decreased aromaticity and molecular weight of the waste. EEM spectra demonstrated that the accumulative Pi,n values of regions III and V in COM, COM + WDG, PM, and PM + WDG were 58%, 49%, 52% and 63%, respectively. These results not only provide new insights into the potential of mixed wastes for BSFL treatment but also contribute to the basis for the formulation of effective management measurements that reduce and/or reuse these wastes.

A new species of Papiliomyces (Clavicipiteae, Hypocreales) from China.

Background: Papiliomyces (Hypocreales, Sordariomycetes) was introduced to accommodate two species: Papiliomycesliangshanensis and Papiliomycesshibinensis. Later, Papiliomycesliangshanensis was renamed Ophiocordycepsliangshanensis. However, the Papiliomycesliangshanensis molecular data (Nepalese) used to establish the Papiliomyces genus was different from Ophiocordycepsliangshanensis (China) strains. New information: This paper describes a new species, Papiliomyceslongiclavatus, found in Yangchang District, Guiyang City, Guizhou Province, China. It is proposed, based on morphology and multilocus phylogeny (ITS, SSU, LSU, TEF1, RPB1 and RPB2). The new species is phylogenetically most closely related to Papiliomycesliangshanensis (Nepalese collections). However, Papiliomycesliangshanensis (Nepalese collections) requires morphological details and additional detection. The new species differs from other Papiliomyces species in having robust stroma with completely immersed perithecia, multi-septate ascospores, cylindrical secondary ascospores, two types of phialides and two types of conidia:longer α-conidia and longer ß-conidia.

Regulation of Intracellular Reactive Oxygen Species Levels after the Development of Phallus rubrovolvatus Rot Disease Due to Trichoderma koningii Mycoparasitism.

Phallus rubrovolvatus is a unique mushroom used for medicinal and dietary purposes in China. In recent years, however, the rot disease of P. rubrovolvatus has seriously affected its yield and quality, becoming an economically important threat. In this study, samples of symptomatic tissues were collected, isolated, and identified from five major P. rubrovolvatus production regions in Guizhou Province, China. Based on combined analyses of phylogenies (ITS and EF1-α), morphological characteristics and Koch's postulates, Trichoderma koningiopsis and Trichoderma koningii were identified as the pathogenic fungal species. Among these, T. koningii exhibited stronger pathogenicity than the other strains; thus, T. koningii was used as the test strain in the follow-up experiments. Upon co-culturing T. koningii with P. rubrovolvatus, the hyphae of the two species were intertwined, and the color of the P. rubrovolvatus hyphae changed from white to red. Moreover, T. koningii hyphae were wrapped around P. rubrovolvatus hyphae, leading to their shortening and convolution and ultimately inhibiting their growth due to wrinkling; T. koningii penetrated the entire basidiocarp tissue of P. rubrovolvatus, causing serious damage to the host basidiocarp cells. Further analyses revealed that T. koningii infection resulted in the swelling of basidiocarps and significantly enhanced the activity of defense-related enzymes, such as malondialdehyde, manganese peroxidase, and polyphenol oxidase. These findings offer theoretical support for further research on the infection mechanisms of pathogenic fungi and the prevention of diseases caused by them.

A new monocyclic monoterpene derivative from volva of Phallus dongsun.

A new monoterpene derivative namely dongsunol A (1) and sixteen known compounds (2-17) were isolated from the volva of Phallus dongsun. All compounds were isolated from this fungus for the first time. Their structures and absolute configurations were determined by nuclear magnetic resonance (NMR), HRESIMS spectral data, and electronic circular dichroism (ECD). The new monoterpene derivative (1) exhibited antibacterial activity with a MIC of 200 µg/mL. Other compounds have inhibitory effects on Staphylococcus aureus and Pseudomonas aeruginosa, while have displayed moderate NO inhibitory activity and antineoplastic activity on SMMC-7721 and SW480 in vitro.

Three New Species of Lactifluus (Basidiomycota, Russulaceae) from Guizhou Province, Southwest China.

Lactifluus is a distinct genus of milkcaps, well known as ectomycorrhizal fungi. The characteristics of the genus Lactifluus include grayish-yellow, orange to orange-brown, or reddish-brown pileus, white latex from the damaged lamellae, discoloring to a brownish color, reticulate spore ornamentation, lampropalisade-type pileipellis, and the presence of lamprocystidia. Guizhou Province is rich in wild mushroom resources due to its special geographical location and natural environment. In this study, three novel Lactifluus species were identified through the screening of extensive fungal resources in Suiyang County, Guizhou Province, China, sampled from host species of mostly Castanopsis spp. and Pinus spp. Based on critical morphology coupled with nuclear sequences of genes encoding large subunit rRNA, internal transcribed spacer, and RNA polymerase II, these new species, Lactifluus taibaiensis, Lactifluus qinggangtangensis, and Lactifluus jianbaensis, were found to belong to Lactifluus section Lactifluus. A comparison with closely related species, Lactifluus taibaiensis was distinguished by its lighter-colored pileus, different colors of lamellae, and more subglobose basidiospores; Lactifluus jianbaensis was identified by the height of the spore ornamentation and its subglobose basidiospores; and Lactifluus qinggangtangensis was characterized by having smaller basidiospores, ridges, and pleurolamprocystid.

Lauric Acid Induces Apoptosis of Rice Sheath Blight Disease Caused by Rhizoctonia solani by Affecting Fungal Fatty Acid Metabolism and Destroying the Dynamic Equilibrium of Reactive Oxygen Species.

Rice sheath blight, caused by Rhizoctonia solani, is one of the major rice diseases. In order to better understand the inhibitory mechanism of lauric acid on the disease, RNA sequencing (RNA-Seq) was used to analyze the transcriptome changes in Rhizoctonia solani treated with lauric acid for 3 h, 6 h, 18 h, and 24 h, including 2306 genes; 1994 genes; 2778 genes; and 2872 genes. Based on gene ontology (GO) enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses, we found that protein processing in endoplasmic reticulum (KO04141), carbon metabolism (KO01200), and starch and sucrose metabolism were significantly enriched. Most oxidoreductase, dehydrogenase, reductase, and transferase genes are downregulated in this process. Lauric acid can affect ergosterol content, mitochondrial membrane potential collapse, hydrogen peroxide content, electrolyte leakage, reactive oxygen species balance, and can induce endoplasmic reticulum (ER) stress. Lauric acid also increased the expression levels of ER chaperone glucose regulatory protein Grp78 (BIP), protein disulfide isomerase (PDI), and Calpain (CNX), and decreased the expression levels of HSP40, HSP70, and HSP90 genes. Lauric acid affected the ergosterol content in the cell membrane of R. solani, which induces ER stress and increases the BiP level to induce the apoptosis of Rhizoctonia solani. These results indicated that lauric acid could be used to control rice sheath blight.

Ganodermaovisporum sp. nov. (Polyporales, Polyporaceae) from Southwest China.

Background: Ganoderma is a white-rot fungus with a cosmopolitan distribution and includes several economically important species. This genus has been extensively researched due to its beneficial medicinal properties and chemical constituents with potential nutritional and therapeutic values. Traditionally, species of Ganoderma were identified solely based on morphology; however, recent molecular studies revealed that many morphology-based species are conspecific. Furthermore, some type species are in poor condition, which hinders us from re-examining their taxonomic characteristics and obtaining their molecular data. Therefore, new species and fresh collections with multigene sequences are needed to fill the loopholes and to understand the biological classification system of Ganoderma. New information: In a survey of Ganoderma in Guizhou Province, southwest China, we found a new species growing on soil and, herein, it was identified by both morphology and phylogenetic evidence. Hence, we propose a new species, Ganodermaovisporum sp. nov. This species is characterised by an annual, stipitate, laccate basidiome, with a red-brown to brownish-black pileus surface and pale white pores, duplex context, clavate pileipellis terminal cells, trimitic hyphal system, ellipsoid basidiospores with dark brown eusporium bearing coarse echinulae and an obtuse turgid appendix. Phylogenetic analyses confirmed that the novel species sisters to G.sandunense with high bootstrap support. Furthermore, the RPB2 sequence of G.sandunense is supplied for the first time. Notably, we re-examined the type specimen of G.sandunense and provide a more precise description of the duplex context, pileipellis terminal cells and basidia. All species collected are described and illustrated with coloured photographs. Moreover, we present an updated phylogeny for Ganoderma, based on nLSU, ITS, RPB2 and TEF1-α DNA sequence data and species relationships and classification are discussed.

Structure analysis and antioxidant activity of polysaccharide-iron (III) from Cordyceps militaris mycelia.

Iron-enriched Cordyceps militaris was obtained by adding FeSO4 solution to the mycelia for biotransformation. The polysaccharide-iron (III) was extracted by water extraction and alcohol precipitation. High performance liquid chromatography showed that the crude polysaccharide-iron (III) had three components. The second component was purified by Sephadex G-150 and named as CPS-iron-II. The average molecular weight of CPS-iron-II was 44.136 kDa. The content of iron was 2.73%. The monosaccharide composition analysis indicated that the CPS-iron-II was composed of rhamnose, arabinose, galactose, glucose, mannose, galacturonic acid with percentage ratio of 0.94:3.12:27.01:36.62:30.20:2.12. The results of methylation analysis revealed that the CPS-iron-II was made of →2)-ß-D-Glcp-(1→, with →2, 4)-α-D-Glcp-(1→ highly branched. Congo-red test showed that CPS-iron-II can cause flocculation of Congo red solution. The anti-oxidative analysis showed that antioxidant activity of CPS-iron-II was almost equal to that of Vc. The manuscript provided a new way for the preparation of polysaccharide-iron(III) from Cordyceps militaris.

Ophiocordycepsaphrophoridarum sp. nov., a new entomopathogenic species from Guizhou, China.

BACKGROUND: Ophiocordyceps is the largest genus in the family Ophiocordicipitaceae, including many entomopathogenic species. In recent years, many species have been described in this genus, with a wide range of host insects. Entomopathogenic fungi include ecologically, economically and medicinally important species, but a large portion of their diversity remains to be discovered and described. NEW INFORMATION: In this study, a new species, Ophiocordycepsaphrophoridarum sp. nov, parasitising Aphrophoridae sp. (Hemiptera) is proposed from China, based on evidence from morphology and molecular phylogenetic analyses. This species is characterised by fibrous, pigmented stromata, cylindrical asci and filiform ascospores. Compared to its closest relative, O.tricentri, the new species has wider perithecia and longer asci. Molecular phylogenetic analyses of a multilocus dataset (consisting of SSU, ITS, LSU, TEF1, RPB1 and RPB2) confirm its placement in Ophiocordyceps. Ophiocordycepsaphrophoridarum is morphologically described and illustrated with colour photographs. Morphological comparisons with closely-related species are also presented in tabulated format.

Molecular evidence supports simultaneous association of the achlorophyllous orchid Chamaegastrodia inverta with ectomycorrhizal Ceratobasidiaceae and Russulaceae.

BACKGROUND: Achlorophyllous orchids are mycoheterotrophic plants, which lack photosynthetic ability and associate with fungi to acquire carbon from different environmental sources. In tropical latitudes, achlorophyllous forest orchids show a preference to establish mycorrhizal relationships with saprotrophic fungi. However, a few of them have been recently found to associate with ectomycorrhizal fungi and there is still much to be learned about the identity of fungi associated with tropical orchids. The present study focused on mycorrhizal diversity in the achlorophyllous orchid C. inverta, an endangered species, which is endemic to southern China. The aim of this work was to identify the main mycorrhizal partners of C. inverta in different plant life stages, by means of morphological and molecular methods. RESULTS: Microscopy showed that the roots of analysed C. inverta samples were extensively colonized by fungal hyphae forming pelotons in root cortical cells. Fungal ITS regions were amplified by polymerase chain reaction, from DNA extracted from fungal mycelia isolated from orchid root samples, as well as from total root DNA. Molecular sequencing and phylogenetic analyses showed that the investigated orchid primarily associated with ectomycorrhizal fungi belonging to a narrow clade within the family Ceratobasidiaceae, which was previously detected in a few fully mycoheterotrophic orchids and was also found to show ectomycorrhizal capability on trees and shrubs. Russulaceae fungal symbionts, showing high similarity with members of the ectomycorrhizal genus Russula, were also identified from the roots of C. inverta, at young seedling stage. Ascomycetous fungi including Chaetomium, Diaporthe, Leptodontidium, and Phomopsis genera, and zygomycetes in the genus Mortierella were obtained from orchid root isolated strains with unclear functional role. CONCLUSIONS: This study represents the first assessment of root fungal diversity in the rare, cryptic and narrowly distributed Chinese orchid C. inverta. Our results provide new insights on the spectrum of orchid-fungus symbiosis suggesting an unprecedented mixed association between the studied achlorophyllous forest orchid and ectomycorrhizal fungi belonging to Ceratobasidiaceae and Russulaceae. Ceratobasidioid fungi as dominant associates in the roots of C. inverta represent a new record of the rare association between the identified fungal group and fully mycoheterotrophic orchids in nature.

Two new entomopathogenic species of Ophiocordyceps in Thailand.

Ophiocordyceps is entomopathogenic and the largest studied genus in the family Ophiocordycipitaceae. Many species in this genus have been reported from Thailand. The first new species introduced in this paper, Ophiocordycepsglobiceps, differs from other species based on its smaller perithecia, shorter asci and secondary ascospores and additionally, in parasitising fly species. Phylogenetic analyses of combined LSU, SSU, ITS, TEF1α and RPB1 sequence data indicate that O.globiceps forms a distinct lineage within the genus Ophiocordyceps as a new species. The second new species, Ophiocordycepssporangifera, is distinguished from closely related species by infecting larvae of insects (Coleoptera, Elateridae) and by producing white to brown sporangia, longer secondary synnemata and shorter primary and secondary phialides. We introduce O.sporangifera based on its significant morphological differences from other similar species, even though phylogenetic distinction is not well-supported.

Two new endophytic Colletotrichum species from Nothapodytespittosporoides in China.

Two new endophytic species, Colletotrichumjishouense sp. nov. and. C.tongrenense sp. nov. were isolated from Nothapodytespittosporoides in Guizhou and Hunan provinces, China. Detailed descriptions and illustrations of these new taxa are provided and morphological comparisons with similar taxa are explored. Phylogenetic analysis with combined sequence data (ITS, GAPDH, ACT and TUB2) demonstrated that both species formed distinct clades in this genus. This is the first record of Colletotrichum species from N.pittosporoides in China.

Multigene phylogenetics of Polycephalomyces (Ophiocordycipitaceae, Hypocreales), with two new species from Thailand.

Polycephalomyces (Ophiocordycipitaceae) species are found in subtropical regions and are parasitic or hyperparasitic on insects. Two new species, P. aurantiacus and P. marginaliradians, parasitic on Ophiocordyceps barnesii and larva of Cossidae respectively, are introduced in this paper. Morphological comparison with extant species and DNA based phylogenies from analyses of a multigene dataset support the establishment of the new taxa. Polycephalomyces aurantiacus, exhibiting a hyperparasitic lifestyle on Ophiocordyceps barnesii, differs from other species in producing orange conidia in mass and have longer ß-phialides in culture. Polycephalomyces marginaliradians differs from other Ophiocordyceps species by producing single stromata with a stipe, smaller perithecia and branched α-phialides and catenate α-conidia and is parasitic on Cossidae. A combined nrSSU, nrLSU, ITS, tef-1a, rpb1 and rpb2 sequence data was analysed phylogenetically including Ophiocordyceps and Polycephalomyces taxa. The new species described herein are clearly distinct from other species in Polycephalomyces. We provide a key to the species of Polycephalomyces and discuss relevant interspecies relationships.

Endophytic Colletotrichum species from Dendrobium spp. in China and Northern Thailand.

Species of Colletotrichum are commonly found in many plant hosts as pathogens, endophytes and occasionally saprobes. Twenty-two Colletotrichum strains were isolated from three Dendrobium species - D.cariniferum, D.catenatum and D.harveyanum, as well as three unidentified species. The taxa were identified using morphological characterisation and phylogenetic analyses of ITS, GAPDH, ACT and ß-tubulin sequence data. This is the first time to identify endophytic fungi from Dendrobium orchids using the above method. The known species, Colletotrichumboninense, C.camelliae-japonicae, C.fructicola, C.jiangxiense and C.orchidophilum were identified as fungal endophytes of Dendrobium spp., along with the new species, C.cariniferi, C.chiangraiense, C.doitungense, C.parallelophorum and C.watphraense, which are introduced in this paper. One strain is recorded as an unidentified species. Corn meal agar is recommended as a good sporulation medium for Colletotrichum species. This is the first report of fungal endophytes associated with Dendrobiumcariniferum and D.harveyanum. Colletotrichumcamelliae-japonicae, C.jiangxiense, and C.orchidophilum are new host records for Thailand.

Beta-tubulin and Actin gene phylogeny supports Phaeoacremoniumovale as a new species from freshwater habitats in China.

A new species of Phaeoacremonium, P.ovale (Togniniaceae), was isolated during a diversity study of freshwater fungi from Yunnan Province in China. Morphological and cultural studies of the fungus were carried out and its sexual and asexual morphs (holomorph) are introduced herein. This species is characterised by peculiar long-necked, semi-immersed ascomata with oval to ellipsoid ascospores and ellipsoid to ovoid conidia. Phylogenetic analyses of a combined TUB and ACT gene dataset revealed that strains of P.ovale constitute a strongly supported independent lineage and are related to P.griseo-olivaceum and P.africanum. The number of nucleotide differences, across the genes analysed, also supports establishment of P.ovale as a new species.

Morphological and phylogenetic characterisation of novel Cytospora species associated with mangroves.

Mangroves are relatively unexplored habitats and have been shown to harbour a number of novel species of fungi. In this study, samples of microfungi were collected from symptomatic branches, stem and leaves of the mangrove species Xylocarpusgranatum, X.moluccensis and Lumnitzeraracemosa and examined morphologically. The phylogeny recovered supports our morphological data to introduce three new species, Cytosporalumnitzericola, C.thailandica and C.xylocarpi. In addition, a combined multi-gene DNA sequence dataset (ITS, LSU, ACT and RPB2) was analysed to investigate phylogenetic relationships of isolates and help in a more reliable species identification.

Anti-phytopathogenic sesquiterpenoid-xanthone adducts from potato endophytic fungus Bipolaris eleusines.

A bioguided separation on the cultures of the potato endophytic fungus Bipolaris eleusines led to the isolation of two anti-phytopathogenic (Alternaria solani) sesquiterpenoid-xanthone adducts, namely bipolins I and J (1 and 2). Their structures were established via extensive spectroscopic analysis. Compounds 1 and 2 exhibit potent inhibitory activity against A. solani with MIC values of 8 and 16 µg mL-1, respectively.