A total of 27 endophytic fungal strains were isolated from Huperzia serrata,which were richly distributed in the stems and leaves while less distributed in roots. The 27 strains were identified by Internal Transcribed Spacer( ITS) r DNA molecular method and one of the strains belongs to Basidiomycota phylum,and other 26 stains belong to 26 species,9 general,6 families,5 orders,3 classes of Ascomycota Phylum. The dominant strains were Colletotrichum genus,belonging to Glomerellaceae family,Glomerellales order,Sordariomycetes class,Ascomycota Phylum,with the percentage of 48. 15%. The inhibitory activities of the crude extracts of 27 endophytic fungal strains against acetylcholinesterase( ACh E) and nitric oxide( NO) production were evaluated by Ellman's method and Griess method,respectively. Crude extracts of four fungi exhibited inhibitory activities against ACh E with an IC50 value of 42. 5-62. 4 mg·L~(-1),and some fungi's crude extracts were found to inhibit nitric oxide( NO) production in lipopolysaccharide( LPS)-activated RAW264. 7 macrophage cells with an IC50 value of 2. 2-51. 3 mg·L~(-1),which indicated that these fungi had potential anti-inflammatory activities.The chemical composition of the Et OAc extract of endophytic fungus HS21 was also analyzed by LCMS-IT-TOF. Seventeen compounds including six polyketides,four diphenyl ether derivatives and seven meroterpenoids were putatively identified.
Acetylcholinesterase ; Animals ; Anti-Inflammatory Agents ; isolation & purification ; pharmacology ; Ascomycota ; chemistry ; classification ; isolation & purification ; Cholinesterase Inhibitors ; isolation & purification ; metabolism ; Endophytes ; classification ; isolation & purification ; Huperzia ; microbiology ; Mice ; RAW 264.7 Cells

Peach brown rot, caused by Monilinia fructicola, is one of the most serious peach diseases. A strain belonging to the Actinomycetales, named Streptomyces blastmyceticus JZB130180, was found to have a strong inhibitory effect on M. fructicola in confrontation culture. Following the inoculation of peaches in vitro, it was revealed that the fermentation broth of S. blastmyceticus JZB130180 had a significant inhibitory effect on disease development by M. fructicola. The fermentation broth of S. blastmyceticus JZB130180 had an EC₅₀ (concentration for 50% of maximal effect) of 38.3 µg/mL against M. fructicola, as determined in an indoor toxicity test. Analysis of the physicochemical properties of the fermentation broth revealed that it was tolerant of acid and alkaline conditions, temperature, and ultraviolet radiation. In addition, chitinase, cellulase, and protease were also found to be secreted by the strain. The results of this study suggest that S. blastmyceticus JZB130180 may be used for the biocontrol of peach brown rot.
Ascomycota/pathogenicity* ; Bacterial Proteins/metabolism* ; Cell Wall/metabolism* ; Cellulase/metabolism* ; Chitinases/metabolism* ; Fermentation ; Fruit/microbiology* ; Pest Control, Biological/methods* ; Phylogeny ; Plant Diseases/prevention & control* ; Prunus persica/microbiology* ; Siderophores/metabolism* ; Streptomyces/physiology*

An extracellular lipase from Aureobasidium pullulans was obtained and purified with a specific activity of 17.7 U/mg of protein using ultrafiltration and a DEAE-Sepharose Fast Flow column. Characterization of the lipase indicated that it is a novel finding from the species A. pullulans. The molecular weight of the lipase was 39.5 kDa, determined by sodium dodecyl sulfonate-polyacrylamide gel electrophoresis (SDS-PAGE). The enzyme exhibited its optimum activity at 40 °C and pH of 7. It also showed a remarkable stability in some organic solutions (30%, v/v) including n-propanol, isopropanol, dimethyl sulfoxide (DMSO), and hexane. The catalytic activity of the lipase was enhanced by Ca²⁺ and was slightly inhibited by Mn²⁺ and Zn²⁺ at a concentration of 10 mmol/L. The lipase was activated by the anionic surfactant SDS and the non-ionic surfactants Tween 20, Tween 80, and Triton X-100, but it was drastically inhibited by the cationic surfactant cetyl trimethyl ammonium bromide (CTAB). Furthermore, the lipase was able to hydrolyze a wide variety of edible oils, such as peanut oil, corn oil, sunflower seed oil, sesame oil, and olive oil. Our study indicated that the lipase we obtained is a potential biocatalyst for industrial use.
Ascomycota/enzymology* ; Calcium ; Catalysis ; Corn Oil/metabolism* ; Detergents/chemistry* ; Enzyme Stability ; Fungal Proteins/chemistry* ; Glucans/chemistry* ; Hexanes/chemistry* ; Hydrogen-Ion Concentration ; Hydrolysis ; Industrial Microbiology ; Lipase/chemistry* ; Manganese/chemistry* ; Olive Oil/metabolism* ; Peanut Oil/metabolism* ; Sesame Oil/metabolism* ; Substrate Specificity ; Sunflower Oil/metabolism* ; Surface-Active Agents ; Temperature ; Zinc/chemistry*

Phytochemical investigation of the culture of Epicoccum nigrum,an endolichenic fungus inhabiting Leptogium masiaticum,led to the isolation of 11 compounds. Based on NMR spectroscopy and HRESIMS data,their structures were determined as one alkaloid fusaricide( 1),and seven benzofuran derivatives including epicoccone( 2),4,6-dihydroxy-5-methoxy-7-methyl-1,3-dihydro isobenzofuran( 3),5-methyl-epicoccone B( 4),3,6,7-trihydroxy-5-methoxy-4-methylisobenzo furan-1( 3 H)-one( 5),3-methoxyepicoccone B( 6),2,3,4-trihydroxy-6-( hydroxymethyl)-5-methylbenzyl-alcohol( 7),and isoochracinic acid( 8),together with three epicoccolide analogs epicocconigrones A( 9),epicoccolide B( 10),and epicocconigrones B( 11). Compounds 1,9 and 10 showed potent microorganism inhibitory effects. These results indicated the potential perspective of this endophytic fungus as an eco-friendly biocide.
Ascomycota/chemistry* ; Endophytes/chemistry* ; Magnetic Resonance Spectroscopy ; Microbial Sensitivity Tests ; Secondary Metabolism

To develop a genetic transformation method of Aureobasidium pullulans and T-DNA insertion for high-efficient screening of polymalic acid (PMA) producing strain. Agrobacterium tumefaciens-AGL1, containing the selection genes encoding hygromycin B phosphotase or phosphinothricin acetyltranferase, was used to transform Aureobasidium pullulans CCTCC M2012223 and transformants were confirmed by colony PCR method. Transferred DNA (T-DNA) insertional mutants were cultured in microwell plate, and screened for high-titer PMA producing strain according to the pH response model. DNA walking was used to detect the insertion sites in the mutant. Results show that the selection markers could stably generated in the transformants, and 80 to 120 transformants could be found per 10(7) single cells. A high-titer PMA mutant H27 was obtained, giving a good PMA production caused by the disruption of phosphoglycerate mutase, that increased by 24.5% compared with the control. Agrobacterium tumefaciens-mediated transformation and high-efficient screening method were successfully developed, which will be helpful for genetic transformation of Aureobasidium pullulans and its functional genes discovery.
Agrobacterium tumefaciens ; Ascomycota ; genetics ; metabolism ; DNA, Bacterial ; Malates ; metabolism ; Polymerase Chain Reaction ; Polymers ; metabolism ; Transformation, Genetic

To investigate the secondary metabolites of endophytic fungi Pericinia sp. F-31. Column chromatography on silica gel, Sephadex LH-20 and semi-preparative HPLC were used to separate and purify the compounds. Two compounds were isolated from the fermentation broth of Periconia sp. Their structures were identified as 5-(1-hydroxyhexyl) -6-methyl-2H-pyran-2-one (1) and 2-(3-hydroxy-4-methylphenyl) -propanoic acid (2). Compound 1 was a new lactone compound, compound 2 was new natural product, and the NMR data of compound 2 was reported for the first time.
Annona ; microbiology ; Ascomycota ; chemistry ; genetics ; isolation & purification ; metabolism ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; metabolism ; Endophytes ; chemistry ; genetics ; isolation & purification ; metabolism ; Lactones ; chemistry ; isolation & purification ; metabolism ; Mass Spectrometry ; Molecular Structure

To investigate the mechanism of agarwood formation in Aquilaria sinensis induced by Lasiodiplodia theobromae, the fermentation liquor of L. theobromae was analyzed qualitatively and quantitatively by gas chromatography-mass spectrometry (GC-MS). JAs were detected in the fermentation liquor. The effect of the fermentation liquor on the abundance of sesquiterpenes in the callus of A. sinensis was analyzed by solid phase microextraction-gas chromatography-mass spectrometry (SPME-GC-MS). And the fermentation liquor stimulated alpha-guaiene, alpha-humulene and delta-guaiene biosynthesis in calli. It was inferred that L. theobromae produced JAs, which resulted in a significant increase of sesquiterpenes in A. sinensis.
Ascomycota ; physiology ; Fermentation ; Sesquiterpenes ; metabolism ; Thymelaeaceae ; metabolism ; microbiology

Podostroma cornu-damae is a rare fungus that houses a fatal toxin in its fruit body. In this case report, two patients collected and boiled the wild fungus in water, which they drank for one month. One patient died, presenting with desquamation of the palms and soles, pancytopenia, severe sepsis and multiple organ failure. The other patient recovered after one month of conservative care after admission. We found a piece of Podostroma cornu-damae in the remaining clusters of mushrooms. Mushroom poisoning by Podostroma cornu-damae has never been previously reported in Korea.
Agaricales/metabolism ; Anti-Bacterial Agents/therapeutic use ; Ascomycota/*metabolism ; Fatal Outcome ; Female ; Fever ; Hospitalization ; Humans ; Male ; Middle Aged ; Mushroom Poisoning/*diagnosis ; Pancytopenia/chemically induced ; Republic of Korea

A total of 52 endophytic fungi were isolated from roots and stems of Tibetan medicinal plant Phlomis younghusbandii Mukerjee. These fungal isolates were molecularly identified based on ITS sequnces and 28S sequences distributed to 12 genera, including Phoma, Chaetosphaeronema, Fusarium and Leptosphaeria, etc. Among them, the dominant genus was Phoma. Extracts of all strains were evaluated for anti-HIV-1 integrase activity by using soluable integrase expressed in E. coli BL21 (DE3). The results showed that seven samples from five fungal endophytes PHY-24, PHY-38, PHY-40, PHY-51, PHY-53, which belonged to genus Chaetosphaeronema, inhibited strand transfer reaction catalyzed by HIV-1 integrase with IC50 values, of 6.60, 5.20, 2.86, 7.86, 4.47, 4.56 and 3.23 microg x mL(-1) respectively. In conclusion, the endophytic fungi of Phlomis younghusbandii Mukerjee are valuable for further screening anti-HIV-1 integrase agents.
Ascomycota ; enzymology ; isolation & purification ; Chaetomium ; enzymology ; isolation & purification ; Endophytes ; enzymology ; isolation & purification ; Escherichia coli ; enzymology ; HIV Integrase ; genetics ; metabolism ; HIV Integrase Inhibitors ; pharmacology ; Phlomis ; microbiology ; Phylogeny ; Plant Roots ; microbiology ; Plant Stems ; microbiology ; Plants, Medicinal ; microbiology ; Plasmids ; Recombinant Proteins ; genetics ; metabolism

AIM: To isolate new and/or bioactive constituents from EtOAc extract of liquid culture of endophyte Guignardia sp. from the leaves of Undaria pinnatifida (Harv.) Sur. METHODS: Isolation and purification were performed through silica gel column chromatograph, Sephadex LH-20 and reversed-phase ODS column and the structures of the compounds obtained were identified through a combination of spectral and chemical methods (IR, MS, (1)H and (13)C NMR). In vitro bioactive assays including antifungal activity against three human pathogenic fungi Microsporum canis, Tricophyton rubrum and Epidermophyton floccosom and cytotoxic activity against the human nasopharyngeal epidermoid tumor KB cell line were evaluated. RESULTS: Seven compounds have been obtained from the liquid culture of the title endophyte: ergosterol peroxide (6, 22-diene-5, 8-epidioxyergosta-3-ol) (1), ergosterol (2), cyclo-(Tyr-Leu) (3), cyclo-(Phe-Phe) (4), cyclo-(Val-Leu) (5), cyclo-(Phe-Pro) (6) and cyclo-(Leu-Ile) (7). Compounds 1-3 and 6 inhibited the growth of M. canis with MICs of 10.0, 20.0, 50.0 and 5.0 μg·mL(-1), respectively and compounds 1, 2 and 6 against T. rubrum with MICs of 15.0, 20.0 and 10.0 μg·mL(-1), respectively and 1 and 6 against E. floccosom with MICs of 20.0 and 50.0 μg·mL(-1), respectively. In addition, compounds 1, 3 and 6 exhibited cytotoxic activity against KB cell line with IC(50) of 20.0, 10.0, 10.0 μg·mL(-1), respectively. CONCLUSION Compounds 1-7 were obtained from Guignardia sp. of U. pinnatifida for the first time, and compounds 1 and 6 had potent cytotoxic and antifungal activity.
Antifungal Agents ; chemistry ; isolation & purification ; pharmacology ; Antineoplastic Agents ; chemistry ; isolation & purification ; pharmacology ; Ascomycota ; chemistry ; metabolism ; Biological Products ; chemistry ; pharmacology ; Carcinoma, Squamous Cell ; drug therapy ; Cell Line, Tumor ; Endophytes ; Epidermophyton ; drug effects ; Humans ; Inhibitory Concentration 50 ; Microbial Sensitivity Tests ; Microsporum ; drug effects ; Nasopharyngeal Neoplasms ; drug therapy ; Phytotherapy ; Plant Leaves ; microbiology ; Undaria ; microbiology