Novel Cyclic Peptide,Epichlicin, from the Endophytic Fungus,Epichloe typhina

The novel cyclic peptide, epichlicin, was isolated from Epichloe typhina, an endophytic fungus of the timothy plant (Phleum pretense L.). Its structure was determined by NMR studies and by mass spectrometry. Enantiomers of 3-amino tetradecanoic acid, a constituent amino acid of epichlicin, were synthesized as authentic standards. The stereochemistry of each amino acid was elucidated through a combination of the advanced Marfey method and chemical manipulation. Epichlicin showed inhibitory activity toward the spore germination of Cladosporium phlei, a pathogenic fungus of the timothy plant at an IC₅₀ value of 22 nM.     

Gamahonolides A,B, and Gamahorin,Novel Antifungal Compounds from Stromata of Epichloe typhina on Phleum pratense

Four novel antifungal compounds, gamahonolides A and B, gamahorin, and 5-hydroxyl-4-phenyl-2(5H)-furanone, were isolated from stromata of Epichloe typhina on Phleum pratense. Their structures were determined by spectroscopic methods. The absolute configuration of gamahonolide A was determined by its ORD spectrum and ¹H-NMR shift difference between the diastereomeric pair of its O-methylmandelates. The stereochemistry of gamahorin was determined by NOE difference spectra and its CD spectrum.     

Unguisin C,a GABA-containing cyclic peptide from the fungus Emericella unguis

Besides the known unguisins A and B, a new cyclic heptapeptide, unguisin C, containing a GABA-derived moiety in the ring, was isolated from the fungus Emericella unguis. The structure was determined by 1D and 2D NMR techniques. Marfey's method was used to determine the absolute stereochemistry. Precursor-directed biosynthesis of the unguisins was performed by supplementation of the culture medium with amino acids (L-Ala, L-Ser, L-Phe and L-Leu). A related cyclic heptapeptide, unguisin D, was detected by HPLC and characterized by sequence analysis using LC-QITMS.     

Unguisin C, a GABA-containing cyclic peptide from the fungus Emericella unguis

Besides the known unguisins A and B, a new cyclic heptapeptide, unguisin C, containing a GABA-derived moiety in the ring, was isolated from the fungus Emericella unguis. The structure was determined by 1D and 2D NMR techniques. Marfey's method was used to determine the absolute stereochemistry. Precursor-directed biosynthesis of the unguisins was performed by supplementation of the culture medium with amino acids ( -Ala, -Ser, -Phe and -Leu). A related cyclic heptapeptide, unguisin D, was detected by HPLC and characterized by sequence analysis using LC-QITMS.     

The distribution of Epichloe typhina in natural plant populations of the host plant Calamagrostis purpurea

The distribution of the systemic fungus Epichloe typhina was studied in natural populations of the host grass Calamagrostis purpurea. By searching for stroma in field and dissecting and staining plant parts in the lab I found that E. typhina was only present in nutrient rich and more wet habitats and totally absent from nutrient poor areas. A greenhouse experiment showed that under high nutrient conditions infected plants produced significantly more shoots than did healthy plants, while no difference was found when plants were grown in low nutrient conditions. Thus, the results suggest that the occurrence as well as the effect of E. typhina is strongly affected by environmental factors similar to the case of other systemic diseases, such as rusts and smuts.     

Benzophenone derivatives from the plant endophytic fungus,Pestalotiopsis sp.

Two new (1–2) and one known (3) benzophenone derivatives, along with five known ambuic acid analogues (4–8) were isolated from the solid cultures of a Pestalotiopsis sp. Compound 2 represented both enantiomeric and atropisomeric isomers, and the absolute configurations of enantiomers [(−)-2 and (+)-2] were determined by electronic circular dichroism (ECD) calculations. All the isolates were evaluated for their antimicrobial and cytotoxic activities. Chlorinated compounds 2 and 3 showed potent antimicrobial activities against four pathogenic bacteria, and compound 3 also displayed strong antifungal activity against Candida glabrata (ATCC 90030) with an MIC₅₀ value of 2.6 ± 0.1 μg/mL. Compound 1 exhibited moderate cytotoxicity against U2OS and MCF-7 with IC₅₀ values of 11.6 and 16.8 μM, respectively.     

Meroterpenes isolated from the endophytic fungus Guignardia mangiferae

Two new guignardones and one tricycloalternarene derivatives, named guignardone D, E (2–3) and tricycloalternarene F (4), and the known guignardone A (1) were isolated from Guignardia mangiferae, an endophytic fungus from the leaves of Viguiera arenaria (Asteraceae), after fermentation in Czapek medium. Structures were established on the basis of their spectroscopic data, including ¹H NMR, ¹³C NMR, HMQC, HMBC and HRESI-MS.     

Gentiopicrin-producing endophytic fungus isolated from Gentiana macrophylla

Gentiana macrophylla is a traditional Chinese medicinal plant. Its dominant active constituents are secoiridoids, mainly gentiopicrin. The objective of this study was to determine whether endophytic fungi isolated from this plant produce the bioactive ingredient gentiopicrin. Primary screening was done by Dragendorff's reaction and the strain re-selection was done with thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) to identify the fermentation products of the selected strains. In this study, 20 strains of endophytic fungi were isolated from G. macrophylla, and the extracts from five strains had a positive Dragendorff's reaction. Two strains (QJ16 and QJ18) had a component with the same R_f value in TLC as that of authentic gentiopicrin and one ingredient of the QJ18 extract had a retention time identical with that of authentic gentiopicrin in HPLC. Therefore, the fungus appears to produce the bioactive ingredient gentiopicrin, as does its host plant, and could be used for the production of gentiopicrin by fermentation.     

Pestalachlorides A-C, antifungal metabolites from the plant endophytic fungus Pestalotiopsis adusta

Pestalachlorides A-C (1-3), three new chlorinated benzophenone derivatives, have been isolated from cultures of an isolate of the plant endophytic fungus Pestalotiopsis adusta. The structures of these compounds were determined mainly by NMR spectroscopy, and the structures of 1 and 3 were further confirmed by X-ray crystallography. Pestalachloride A (1) was obtained as a mixture of two inseparable atropisomers (1a and 1b), whereas pestalachloride C (3) was found to be a racemic mixture. Compounds 1 and 2 displayed significant antifungal activities against three plant pathogens.     

源于植物内生无花果拟盘多毛孢真菌的新色原酮类次生代谢产物研究

从植物内生真菌无花果拟盘多毛孢菌株(Pestalotiopsis fici;AS3.9138=W106-1)的放大发酵粗提物中分离得到4个异戊二烯基取代的色原酮类新结构次生代谢产物pestaloficiolsM-P(1-4),并应用质谱和核磁共振技术确定了上述化合物的结构。生物活性测试结果表明化合物2能够抑制HIV-1病毒在C8166细胞中的复制;化合物3和4对宫颈癌细胞(HeLa)具有细胞毒活性;另外,化合物3对烟曲霉Aspergillus fumigatus也具有较强的抑制活性。     

Antifungal polyketide derivatives from the endophytic fungus Aplosporella javeedii

Six new polyketides aplojaveediins A–F (1–6) were isolated from the endophytic fungus Aplosporella javeedii associated with the host plant Orychophragmus violaceus (Brassicaceae). The structures of the new metabolites were elucidated by analysis of their NMR and MS data. Compound 1 exhibited antifungal activity against the hyphae form of Candida albicans strain ATCC 24433 in the agar plate diffusion assay and the microbroth dilution assay. The kinetic of killing of C. albicans cells for compound 1 was considerably faster than that of the positive control hygromycin B. Compounds 1 and 6 also exhibited moderate antibacterial activities against sensitive (ATCC 29213) and drug-resistant (ATCC 700699) strains of Staphylococcus aureus.     

Interaction between the endophytic fungus Epichloe bromicola and the grass bromus erectus: effects of endophyte infection, fungal concentration and environment on grass growth and flowering

Epichloe bromicola is an endophytic fungal species that systemically and perennially colonizes intercellular spaces of leaf blades, leaf sheaths and culms of Bromus grass species. E. bromicola causes choke disease in B. erectus, suppressing maturation of most, if not all, host inflorescences. In an investigation of the interaction between fungus and host, we used a quantitative polymerase chain reaction technique to estimate the amount of fungal DNA, and thereby fungal concentration, in host plants. Fungal concentration was directly correlated with vegetative vigour of the plant, as measured by longest leaf length, number of tillers and vegetative above-ground biomass, suggesting that, during vegetative growth, the endophytic fungus is most beneficial for the plant when present in high concentrations. In contrast, the reproduction of the plant, as measured by the number of functional inflorescences, was inversely correlated with fungal concentration: the majority of infected plants, and all that were associated with high concentrations of fungi, were diseased. Thus, the benefit of endophyte infection for the plant is coupled with the disadvantages of infertility. Fungal concentration was shown to be at least in part genetically determined because fungal concentration differed significantly in different plant-endophyte genotype combinations (symbiotum). In a field experiment with normal and CO2-enriched environments, elevated CO2 levels favoured fungal reproductive vigour over host reproductive vigour, suggesting that these plant endophytes would be at a selective advantage in a corresponding environmental-change scenario. We conclude that a dynamic and complex relationship between fungal endophyte infection, fungal concentration, genotype and environment affects growth and fecundity of B. erectus and should contribute to the evolution of these plant-fungal interactions.     

Metabolites from Microsphaeropsis olivacea, an endophytic fungus of Pilgerodendron uviferum

Seven compounds belonging to different structural skeletons were isolated from Microsphaeropsis olivacea grown in liquid and solid media. The enalin derivative 7-hydroxy-2,4-dimethyl-3(2H)-benzofuranone is reported for the first time, while additional spectroscopic information is provided for the acetates of botrallin and ulocladol. The activity of the isolated compounds was assessed towards the enzyme acetylcholinesterase (AChE) and their cytotoxicity against human lung fibroblasts. Graphislactone A and botrallin presented a moderate activity towards AChE, with IC50 of 8.1 and 6.1 microg/ml (27 and 19 microM, respectively). Under the same experimental conditions, the IC50 of the standard inhibitor galanthamine was 3 microg/ml. The cytotoxicity of both compounds was > 1000 and 330 microM, respectively. None of the compounds was promising as antibacterial or antifungic against phytopathogenic fungi and bacteria. Botrallin and graphislactone A were detected in the liquid potato-dextrose and yeast extract/malt extract/dextrose as well as on a solid substrate (rice). Butyrolactone I was obtained from the fungus growing on solid medium.     

Aspernolides F and G,new butyrolactones from the endophytic fungus Aspergillus terreus

Two new butyrolactones: aspernolides F (6) and G (7), together with three stigmasterol derivatives: (22E,24R)-stigmasta-5,7,22-trien-3-β-ol (1), stigmast-4-ene-3-one (2), and stigmasta-4,6,8(14), 22-tetraen-3-one (3), two meroterpenoids: terretonin A (4) and terretonin (5), and a butyrolactone derivative: butyrolactone VI (8) have been isolated from the endophytic fungus Aspergillus terreus isolated from the roots of Carthamus lanatus (Asteraceae). Their structures were determined by spectroscopic means (1D, 2D NMR, and HRESIMS), as well as optical rotation measurement and comparison with literature data. The isolated compounds were evaluated for their anti-microbial, anti-malarial, anti-leishmanial, and cytotoxic activities. Compound 1 displayed a potent activity against MRSA and C. neoformans with IC₅₀ values of 0.96 μg/mL and 4.38 μg/mL, respectively compared to ciprofloxacin (IC₅₀ 0.07 μg/mL) and amphotericin B (IC₅₀ 0.34 μg/mL), respectively. While, 6 showed good activity against C. neoformans (IC₅₀ 5.19 μg/mL) and mild activity against MRSA (IC₅₀ 6.39 μg/mL). Moreover, 1 and 2 exhibited very good anti-leishmanial activity towards L. donovani with IC₅₀ values of 4.61 and 6.31 μg/mL, respectively and IC₉₀ values of 6.02 and 16.71 μg/mL, respectively.     

Pestaloficiols A-E, bioactive cyclopropane derivatives from the plant endophytic fungus Pestalotiopsis fici

Pestaloficiols A-E (1-5), five new cyclopropane derivatives, have been isolated from cultures of the plant endophyte Pestalotiopsis fici. The structures of these compounds were determined by NMR spectroscopy, and the absolute configuration of 1 was assigned using the modified Mosher method. Compounds 1, 2, and 4 displayed inhibitory effects on HIV-1 replication in C8166 cells.     

Alkaloids from the mangrove endophytic fungus Diaporthe phaseolorum SKS019

Six new alkaloids including four new chromeno[3,2-c]pyridines, diaporphasines A-D (1–4), and two new isoindolinones, meyeroguillines C and D (6–7), as well as three known compounds meyeroguilline A (5), 5-deoxybostrycoidin (8), and fusaristatin A (9), were isolated from an endophytic fungus Diaporthe phaseolorum SKS019. Their structures were determined by analysis of 1D and 2D NMR and mass spectroscopic data. Compounds 1–9 are alkaloid components reported for the first time from the Diaporthe sp., and diaporphasines A-D (1–4) are the third examples of alkaloids possessing the unique chromeno[3,2-c]pyridine nucleus. All isolated compounds 1–9 were evaluated for their cytotoxic activity in vitro using MDA-MB-435, HepG2, MCF10A, HCT116, and NCI-H460 human cell lines. Compound 8 exhibited cytotoxicity against MDA-MB-435 and NCI-H460 human cancer cell lines with IC₅₀ values of 5.32 and 6.57 μM, respectively, and compound 9 showed growth-inhibitory activity against MDA-MB-435 human cancer cell line with IC₅₀ value of 8.15 μM.     

Furoic acid derivatives from the endophytic fungus Coniothyrium sp.

The endophytic fungus Coniothyrium sp. was isolated from leaves of Quercus robur. Fermentation of this fungus on solid rice medium yielded two new furoic acid derivatives ( 1 and 2 ) and two additional known compounds. The structures of the new compounds were determined by extensive analysis of 1D and 2D nuclear magnetic resonance spectra as well as high-resolution mass spectrometry data. Compound 1 , containing three aromatic chromophores attached by rotatable sigma bonds and a chirality center in benzylic position, was found to be a scalemic mixture with an excess of the (S) enantiomer, the absolute configuration of which was elucidated as by the solution time-dependent density functional theory-electronic circular dichroism approach. The ωB97X/TZVP PCM/MeCN and SOGGA11-X/TZVP SMD/MeCN methods were used for geometry reoptimization to reproduce the solution conformational ensemble. All isolated compounds were tested for their cytotoxicity but proved to be inactive.     

Dihydronaphthalenones from the endophytic fungus Botryosphaeria sp. BCC 8200

Five new dihydronaphthalenones (1, 2a/2b, 3a/3b), four of which were isolated as two inseparable mixtures of isomers, together with two known compounds, 7-hydroxy-2-(2-hydroxypropyl)-5-methylchromone (4) and siderin (5), were identified as secondary metabolites of the endophytic fungus Botryosphaeria sp. BCC 8200. The structures were elucidated by interpretation of NMR spectroscopic and mass spectrometry data. Compounds 2a/2b, 3a/3b, and 5 exhibited weak cytotoxic activities against cancer cell lines.     

Metabolites from the endophytic fungus Xylaria sp. PSU-D14

Glucoside derivatives, xylarosides A (1) and B (2), were isolated from the broth extract of the endophytic fungus Xylaria sp. PSU-D14 along with two known compounds, sordaricin (3) and 2,3-dihydro-5-hydroxy-2-methyl-4H-1-benzopyran-4-one (4). The structures were assigned by spectroscopic methods. Sordaricin (3), one of the known metabolites, exhibited moderate antifungal activity against Candida albicans ATCC90028 with a MIC value of 32 microg/ml.