Aromatic compounds produced by Periconia atropurpurea, an endophytic fungus associated with Xylopia aromatica

6,8-Dimethoxy-3-(2'-oxo-propyl)-coumarin (1) and 2,4-dihydroxy-6-[(1'E,3'E)-penta-1',3'-dienyl]-benzaldehyde (2), in addition to the known compound periconicin B (3), were isolated from the ethyl acetate extract of Periconia atropurpurea, an endophytic fungus obtained from the leaves of Xylopia aromatica, a native plant of the Brazilian Cerrado. Their chemical structures were assigned based on analyses of MS, 1D and 2D-NMR spectroscopic experiments. Biological analyses were performed using two mammalian cell lines, human cervix carcinoma (HeLa) and Chinese hamster ovary (CHO). The results showed that compound 1 had no effect when compared to the control group, which was treated with the vehicle (DMSO). Compound 2 was able to induce a slight increase in cell proliferation of HeLa (37% of increase) and CHO (38% of increase) cell lines. Analysis of compound 3 showed that it has potent cytotoxic activity against both cell lines, with an IC50 of 8.0 microM. Biological analyses using the phytopathogenic fungi Cladosporium sphaerospermum and C. cladosporioides revealed that also 2 showed potent antifungal activity compared to nystatin.     

Novel polysaccharide produced by Azotobacter vinelandii isolated from plant rhizosphere

Summary A strain of Azotobacter vinelandii MTCC 2460 has been isolated from the rhizosphere of the rhizomes of lotus, which produces a novel exopolysaccharide (EPS) having a sugar composition of glucose : galactose : fucose: glucoronic acid (2.2 : 2.7 : 5.6 : 1.6). This composition has not been reported for any species belonging to this genus. The EPS gives high viscosity comparable to xanthan under specific conditions.     

Accumulation of secondary compounds in barley and wheat roots in response to inoculation with an arbuscular mycorrhizal fungus and co-inoculation with rhizosphere bacteria

 Colonization of Hordeum vulgare L. cv. Salome (barley)and Triticum aestivum L. cv. Caprimus (wheat) roots by the arbuscular mycorrhizal fungus Glomus intraradices Schenck & Smith leads to de novo synthesis of isoprenoid cyclohexenone derivatives with blumenin [9-O-(2′-O-β-glucuronosyl)-β-glucopyranoside of 6-(3-hydroxybutyl)-1,1,5-trimethyl-4-cyclohexen-3-one] as the major constituent and to transient accumulation of hydroxycinnamate amides (4-coumaroylagmatine and -putrescine). Accumulation of these compounds in mycorrhizal wheat roots started 2 weeks after sowing together with the onset of arbuscule formation and proceeded with mycorrhizal progression. Highest levels were reached in 3- to 4-week-old secondary roots (root branches of first and higher order) characterized by the formation of vesicles. In the final developmental stages, the fungus produced massive amounts of spores, enclosing the stele of older root parts (older than 5 weeks) characterized by cortical death. In these root parts, the secondary compounds were detected in trace amounts only, indicating that they were located in the cortical tissues. Some rhizosphere bacteria tested, i.e. Agrobacterium rhizogenes, Pseudomonas fluorescens, and Rhizobium leguminosarum, markedly stimulated both fungal root colonization and blumenin accumulation, thus, acting as mycorrhiza-helper bacteria (MHB). Application of blumenin itself strongly inhibited fungal colonization and arbuscule formation at early stages of mycorrhiza development. This was associated with a markedly reduced accumulation of the hydroxycinnamate amides 4-coumaroylputrescine and -agmatine. The results suggest that both the isoprenoid and the phenylpropanoid metabolism are closely linked to the developmental stage and the extent of fungal colonization. Their possible involvement in the regulation of mycorrhiza development is discussed. Accepted: 18 September 1998     

Production and characterization of antifungal compounds produced by Lactobacillus plantarum IMAU10014

Lactobacillus plantarum IMAU10014 was isolated from koumiss that produces a broad spectrum of antifungal compounds, all of which were active against plant pathogenic fungi in an agar plate assay. Two major antifungal compounds were extracted from the cell-free supernatant broth of L. plantarum IMAU10014. 3-phenyllactic acid and Benzeneacetic acid, 2-propenyl ester were carried out by HPLC, LC-MS, GC-MS, NMR analysis. It is the first report that lactic acid bacteria produce antifungal Benzeneacetic acid, 2-propenyl ester. Of these, the antifungal products also have a broad spectrum of antifungal activity, namely against Botrytis cinerea, Glomerella cingulate, Phytophthora drechsleri Tucker, Penicillium citrinum, Penicillium digitatum and Fusarium oxysporum, which was identified by the overlay and well-diffusion assay. F. oxysporum, P. citrinum and P. drechsleri Tucker were the most sensitive among molds.     

Inhibition of the phytopathogenic fungus Fusarium proliferatum by volatile compounds produced by Pseudomonas

The Fusarium head blight of grain cereals is a significant disease worldwide. In Argentina, high levels of contamination with Fusarium proliferatum have been found in crops. Many strains of the Pseudomonas genus antagonize the growth of fungi by different mechanisms, such as the production of antibiotics, siderophores, volatiles, and extracellular enzymes. In this work, we have designed a new system for studying the growth inhibition of F. proliferatum—namely by volatile compounds produced by Pseudomonas fluorescens MGR12. In both rich and minimal media, the bacterium released volatiles that negatively affected the mycelial growth of that phytopathogenic fungus. These bacterial compounds were analyzed by gas chromatography–mass spectrometry, but only a few could be identified by comparing their mass spectra with the libraries of the National Institutes of Standards and Technology MS search.     

Three oxygenated cyclohexenone derivatives produced by an endophytic fungus

Three cyclohexenone derivatives, (4S,5S,6S)-5,6-epoxy-4-hydroxy-3-methoxy-5-methyl-cyclohex-2-en-1-one (1), (4R,5R)-4,5-dihydroxy-3-methoxy-5-methyl-cyclohex-2-en-1-one (2), and (4R,5S,6R)-4,5,6-trihydroxy-3-methoxy-5-methyl-cyclohex-2-en-1-one (3), were isolated from unpolished rice fermented with an xylariaceous endophytic fungus (strain YUA-026). The structures of three compounds were established on the basis of spectroscopic analyses and chemical conversion. The minimum inhibitory concentrations of 1 and 3 were 100 microg/ml and 400 microg/ml against Staphylococcus aureus, 100 microg/ml and 200 microg/ml against Pseudomonas aeruginosa, and 200 microg/ml and >400 microg/ml against Candida albicans, respectively. In addition, 1 and 3 exhibited phytotoxic activity against lettuce.     

Nematicidal alkaloids and related compounds produced by the fungus Penicillium cf. simplicissimum

A new nematicidal alkaloid, peniprequinolone (1), together with the known alkaloids penigequinolones A and B (2a, 2b), 3-methoxy-4-hydroxy-4-(4'-methoxyphenyl)quinolinone (3), and 3-methoxy-4,6-dihydroxy-4-(4'-methoxyphenyl)quinolinone (4), were isolated from Penicillium cf. simplicissimum (Oudemans) Thom. Cyclopenin (5) and a compound (6a/6b) structurally related to cyclopenin also were isolated from the fungus, and their structures were established by spectroscopic analysis. The biological activities of 1, 2, 3, 4, and 5 were examined by a bioassay with root-lesion nematodes.     

Magenta pigment produced by fungus

A fungus producing magenta was isolated from cellulosic material by visual observation on Czapek's agar media and the product was conventionally analyzed. The fungal strain that produced magenta pigment was closely related to Phoma herbarum. The type of fibers added to Czapek's medium influenced which pigments were produced. Mycelia attached to the surface of nylon-6 and excreted magenta pigment into the fibers. The pigment structure was partially determined. This is the first report of the production of magenta pigment by a microorganism specifically in the presence of nylon-6 fibers, via an unknown mechanism. This phenomenon raises the question of why and how the fungus disperses the pigment inside the fiber and suggests that fabrics can be dyed using microorganisms.     

A previously unidentified leukotriene produced by human polymorphonuclear leukocytes

A previously unidentified leukotriene was isolated from incubations of human polymorphonuclear leukocytes with ionophore A23187. The compound eluted between LTB4 and 20-carboxy LTB4 on SP-HPLC and between the two 6-trans isomers of LTB4 on RP-HPLC. Ultraviolet spectroscopy revealed three absorption bands at 258 nm, 268 nm and 278 nm. 1802 was incorporated by the OH at both C5 and C12. Oxidative ozonolysis indicated the presence of 5S, 12S configuration. The structure of the newly identified leukotriene is 5S, 12S-dihydroxy-icosatetraenoic acid. Stereochemistry of the double bonds and biologic activity were not investigated.     

Bioactive metabolites produced by Chaetomium globosum,an endophytic fungus isolated from Ginkgo biloba

A novel cytotoxic chlorinated azaphilone derivative named chaetomugilin D (1), together with three known metabolites, chaetomugilin A (2), chaetoglobosins A (3) and C (4), has been isolated by a bioassay-guided fractionation from the EtOAc extract of the cultures of Chaetomium globosum, an endophytic fungus found in the leaves of Ginkgo biloba. Structure of 1 was established by analyses of spectroscopic methods, including 2D-NMR experiments (COSY, NOESY, HMQC, and HMBC). Compounds 1–4 displayed significant growth inhibitory activity against the brine shrimp (Artemia salina) and Mucor miehei.     

In vitro screening of taxol,an anticancer drug produced by the fungus,Colletotrichum capsici

The fungus Colletotrichum capsici was isolated from the diseased fruits of Chilli plant, Capsicum annuum. The isolated test fungus was identified by its morphological and molecular characteristic features. For the first time, the fungus was screened for the production of taxol on modified liquid medium. The presence of taxol was confirmed by the spectroscopic and chromatographic methods of analyses. The amount of taxol produced by this fungus was quantified by HPLC. The maximum amount of fungal taxol production was recorded as 687 μg/L. The production rate was 13 740‐fold higher than that, previously reported for the fungus Taxomyces andreanae. The extracted fungal taxol showed a strong cytotoxic activity in an in vitro culture of human cancer cells indicating that the increase in taxol concentration induces increased cell death. A PCR‐based screening for taxadiene synthase (ts), a unique gene in the formation of the taxane skeleton, confirmed the molecular blueprint for taxol biosynthesis. The results show that the fungus C. capsici is an excellent candidate for an alternate source of taxol supply and can serve as a potential species for genetic engineering to enhance the production of taxol to a higher level.     

Anthracobic acids A and B, two polyketides, produced by an endophytic fungus Anthracobia sp

Two new polyketides, anthracobic acids A and B (1 and 2, resp.), were isolated from a cultured endophytic fungus, Anthracobia sp. The NMR study revealed that both anthracobic acids contain a carboxylic acid conjugated tetraene attached to an octahydronaphthalene moiety. Both compounds showed antimicrobial activity.     

Hydrolytic enzymes and antifungal compounds produced by Tilletiopsis species, phyllosphere yeasts that are antagonists of powdery mildew fungi

Isolates of five species of the yeast-like fungus Tilletiopsis Derx (Tilletiopsis albescens Gokhale, Tilletiopsis fulvescens Gokhale, Tilletiopsis minor Nyland, Tilletiopsis pallescens Gokhale, and Tilletiopsis washingtonensis Nyland) were screened for exo- and endo--beta-1,3-glucanase and chitinase production in a liquid broth used to produce inoculum for biological control studies. There were significant differences among the species, and highest overall enzyme activity was present in T albescens and T. pallescens and lowest in T. washingtonensis. A time-course study of beta-1,3-glucanase and chitinase production in T pallescens ATCC 96155 in broth culture with 2.5% glucose as the carbon source showed that enzyme activity gradually increased over a 3- to 21-day period. Maximum enzyme activity was found between pH 4.0 and 5.0. SDS-PAGE of beta-1,3-glucanase isozymes revealed a range of molecular masses from 18 to 29 kDa. Five isozymes were present in both T albescens and T. pallescens and two in T washingtonensis. Antifungal compounds were also detected in ethyl acetate extracts of culture filtrates of T. pallescens ATCC 96155 after 6 days of incubation, while no activity was detected at 14 days. One active fraction was selected following fractionation and preparative chromatography and was bioassayed against Podosphaera (sect. Sphaerotheca) xanthii (Castagne) U. Braun & N. Shishkoff and a number of other fungi. A concentration of 130 microg/mL inhibited germ tube development in P. xanthii, and mildew spores appeared plasmolyzed. Other fungi were inhibited at higher concentrations. Collapse of hyphae and conidiophores was also observed on mildewed leaves treated with the active fraction. Proton NMR analysis indicated that the inhibitory compound was a fatty acid ester. In 3- to 6-day-old cultures of T pallescens ATCC 96155 demonstrating biological control activity, antifungal compound production may have a primary role in restricting growth of mildew fungi and other competitors when applied to leaves.