RESUMEN
Eragrostis plana (Nees) is an allelopathic plant with invasive potential in South American pastures. To isolate and identify phytotoxic compounds from leaves and roots of E. plana, a bioassay-directed isolation of the bioactive constituents was performed. This is the first report on a new diterpene carbon skeleton, neocassane, and of three new neocassane diterpenes, neocassa-1,12(13),15-triene-3,14-dione, 1; 19-norneocassa-1,12(13),15-triene-3,14-dione, 2; and 14-hydroxyneocassa-1,12(17),15-triene-3-one, 3, identified from the roots. Compounds 1, 2, and 3 inhibited the growth of duckweed by 50% at concentrations of 109 ± 28, 200 ± 37, and 59 ± 15 µM, respectively. Compound 2 was fungicidal to Colletotrichum fragariae, Colletotrichum acutatum, and Colletotrichum gloeosporioides. The compounds identified here could explain the allelopathy of E. plana. The description of the newly discovered compounds, besides contributing to the chemical characterization of the species, may be the first step in the study of the potential of these compounds as bioherbicides.
Asunto(s)
Diterpenos/aislamiento & purificación , Eragrostis/química , Herbicidas , Agrostis/efectos de los fármacos , Agentes de Control Biológico , Colletotrichum/efectos de los fármacos , Diterpenos/química , Diterpenos/farmacología , Fungicidas Industriales/farmacología , Herbicidas/química , Herbicidas/aislamiento & purificación , Herbicidas/farmacología , Lactuca/efectos de los fármacos , Extractos Vegetales/química , Extractos Vegetales/farmacología , Hojas de la Planta/química , Raíces de Plantas/químicaRESUMEN
BACKGROUND Endophytic fungi, present mainly in the Ascomycota and Basidiomycota phyla, are associated with different plants and represent important producers of bioactive natural products. Brazil has a rich biodiversity of plant species, including those reported as being endemic. Among the endemic Brazilian plant species, Vellozia gigantea (Velloziaceae) is threatened by extinction and is a promising target to recover endophytic fungi. OBJECTIVE The present study focused on bioprospecting of bioactive compounds of the endophytic fungi associated with V. gigantea, an endemic, ancient, and endangered plant species that occurs only in the rupestrian grasslands of Brazil. METHODS The capability of 285 fungal isolates to produce antimicrobial and antimalarial activities was examined. Fungi were grown at solid-state fermentation to recover their crude extracts in dichloromethane. Bioactive extracts were analysed by chromatographic fractionation and NMR and displayed compounds with antimicrobial, antimycobacterial, and antimalarial activities. FINDINGS Five fungi produced antimicrobial and antimalarial compounds. Extracts of Diaporthe miriciae showed antifungal, antibacterial, and antimalarial activities; Trichoderma effusum displayed selective antibacterial activity against methicillin-resistant Staphylococcus aureus and Mycobacterium intracellulare; and three Penicillium species showed antibacterial activity. D. miriciae extract contained highly functionalised secondary metabolites, yielding the compound epoxycytochalasin H with high antimalarial activity against the chloroquine-resistant strain of Plasmodium falciparum, with an IC50 approximately 3.5-fold lower than that with chloroquine. MAIN CONCLUSION Our results indicate that V. gigantea may represent a microhabitat repository hotspot of potential fungi producers of bioactive compounds and suggest that endophytic fungal communities might be an important biological component contributing to the fitness of the plants living in the rupestrian grassland.
Asunto(s)
Plasmodium/efectos de los fármacos , Pruebas de Sensibilidad Microbiana , Magnoliopsida/clasificación , Magnoliopsida/microbiología , Hongos Mitospóricos/efectos de los fármacos , Bacterias Aerobias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Antibacterianos/aislamiento & purificación , Antibacterianos/farmacología , Antifúngicos/aislamiento & purificación , Antifúngicos/farmacología , Antimaláricos/aislamiento & purificación , Antimaláricos/farmacología , Clima Tropical , Bioensayo , Candida/efectos de los fármacos , Endófitos/químicaRESUMEN
BACKGROUND: Endophytic fungi, present mainly in the Ascomycota and Basidiomycota phyla, are associated with different plants and represent important producers of bioactive natural products. Brazil has a rich biodiversity of plant species, including those reported as being endemic. Among the endemic Brazilian plant species, Vellozia gigantea (Velloziaceae) is threatened by extinction and is a promising target to recover endophytic fungi. OBJECTIVE: The present study focused on bioprospecting of bioactive compounds of the endophytic fungi associated with V. gigantea, an endemic, ancient, and endangered plant species that occurs only in the rupestrian grasslands of Brazil. METHODS: The capability of 285 fungal isolates to produce antimicrobial and antimalarial activities was examined. Fungi were grown at solid-state fermentation to recover their crude extracts in dichloromethane. Bioactive extracts were analysed by chromatographic fractionation and NMR and displayed compounds with antimicrobial, antimycobacterial, and antimalarial activities. FINDINGS: Five fungi produced antimicrobial and antimalarial compounds. Extracts of Diaporthe miriciae showed antifungal, antibacterial, and antimalarial activities; Trichoderma effusum displayed selective antibacterial activity against methicillin-resistant Staphylococcus aureus and Mycobacterium intracellulare; and three Penicillium species showed antibacterial activity. D. miriciae extract contained highly functionalised secondary metabolites, yielding the compound epoxycytochalasin H with high antimalarial activity against the chloroquine-resistant strain of Plasmodium falciparum, with an IC50 approximately 3.5-fold lower than that with chloroquine. MAIN CONCLUSION: Our results indicate that V. gigantea may represent a microhabitat repository hotspot of potential fungi producers of bioactive compounds and suggest that endophytic fungal communities might be an important biological component contributing to the fitness of the plants living in the rupestrian grassland.
Asunto(s)
Antibacterianos/farmacología , Antifúngicos/farmacología , Antimaláricos/farmacología , Endófitos/química , Magnoliopsida/microbiología , Antibacterianos/aislamiento & purificación , Antifúngicos/aislamiento & purificación , Antimaláricos/aislamiento & purificación , Bioensayo , Candida/efectos de los fármacos , Bacterias Gramnegativas/efectos de los fármacos , Bacterias Grampositivas/efectos de los fármacos , Magnoliopsida/clasificación , Pruebas de Sensibilidad Microbiana , Hongos Mitospóricos/efectos de los fármacos , Plasmodium/efectos de los fármacos , Clima TropicalRESUMEN
The endophytic fungal community associated with the ethnomedicinal plant Echinacea purpurea was investigated as well as its potential for providing antifungal compounds against plant pathogenic fungi. A total of 233 endophytic fungal isolates were obtained and classified into 42 different taxa of 16 genera, of which Alternaria alternata, Colletotrichum dematium, and Stagonosporopsis sp. 2 are the most frequent colonizers. The extracts of 29 endophytic fungi displayed activities against important phytopathogenic fungi. Eight antifungal extracts were selected for chemical analysis. Forty fatty acids were identified by gas chromatography-flame-ionization detection (GC-FID) analysis. The compounds (-)-5-methylmellein and (-)-(3R)-8-hydroxy-6-methoxy-3,5-dimethyl-3,4-dihydroisocoumarin were isolated from Biscogniauxia mediterraneaEPU38CA crude extract. (-)-5-Methylmellein showed weak activity against Phomopsis obscurans, P. viticola, and Fusarium oxysporum, and caused growth stimulation of C. fragariae, C. acutatum, C. gloeosporioides, and Botrytis cinerea. (-)-(3R)-8-Hydroxy-6-methoxy-3,5-dimethyl-3,4-dihydroisocoumarin appeared slightly more active in the microtiter environment than 5-methylmellein. Our results indicate that E. purpurea lives symbiotically with different endophytic fungi, which are able to produce bioactive fatty acids and aromatic compounds active against important phytopathogenic fungi. The detection of the different fatty acids and aromatic compounds produced by the endophytic community associated with wild E. purpurea suggests that it may have intrinsic mutualistic resistance against phytopathogen attacks in its natural environment.
Asunto(s)
Antifúngicos/análisis , Antifúngicos/farmacología , Asteraceae/microbiología , Bioprospección , Endófitos/química , Hongos Mitospóricos/efectos de los fármacos , Filogenia , Antifúngicos/química , Relación Dosis-Respuesta a Droga , Endófitos/aislamiento & purificación , Ácidos Grasos/análisis , Ácidos Grasos/química , Ácidos Grasos/farmacología , Medicina Tradicional , Pruebas de Sensibilidad Microbiana , Hongos Mitospóricos/clasificación , Conformación Molecular , Relación Estructura-ActividadRESUMEN
This study assessed the diversity of cultivable rock-associated fungi from Atacama Desert. A total of 81 fungal isolates obtained were identified as 29 Ascomycota taxa by sequencing different regions of DNA. Cladosporium halotolerans, Penicillium chrysogenum and Penicillium cf. citrinum were the most frequent species, which occur at least in four different altitudes. The diversity and similarity indices ranged in the fungal communities across the latitudinal gradient. The Fisher-α index displayed the higher values for the fungal communities obtained from the siltstone and fine matrix of pyroclastic rocks with finer grain size, which are more degraded. A total of 23 fungal extracts displayed activity against the different targets screened. The extract of P. chrysogenum afforded the compounds α-linolenic acid and ergosterol endoperoxide, which were active against Cryptococcus neoformans and methicillin-resistance Staphylococcus aureus respectively. Our study represents the first report of a new habitat of fungi associated with rocks of the Atacama Desert and indicated the presence of interesting fungal community, including species related with saprobes, parasite/pathogen and mycotoxigenic taxa. The geological characteristics of the rocks, associated with the presence of rich resident/resilient fungal communities suggests that the rocks may provide a favourable microenvironment fungal colonization, survival and dispersal in extreme conditions.
Asunto(s)
Ascomicetos/metabolismo , Cladosporium/metabolismo , Cryptococcus neoformans/efectos de los fármacos , Sedimentos Geológicos/microbiología , Staphylococcus aureus Resistente a Meticilina/efectos de los fármacos , Penicillium/metabolismo , Ascomicetos/clasificación , Ascomicetos/genética , Ascomicetos/aislamiento & purificación , Chile , Cladosporium/clasificación , Cladosporium/genética , Cladosporium/aislamiento & purificación , Clima Desértico , Ecología , Ecosistema , Datos de Secuencia Molecular , Penicillium/clasificación , Penicillium/genética , Penicillium/aislamiento & purificaciónRESUMEN
Endophytic fungi represent ubiquitous microbial organisms able to live in the tissues of different plants around the world and represent a prolific source of bioactive metabolites. In the present study, the endophytic fungus Aspergillus calidoustus was isolated from the medicinal plant Acanthospermum australe (Asteraceae), and identified using molecular, physiological and morphological methods. A methylene chloride crude extract of A. calidoustus has been produced and subjected to antifungal bioassay-directed fractionation which resulted in the isolation of the two bioactive compounds: ophiobolin K and 6-epi-ophiobolin K. These pure compounds displayed antifungal activity against fungal plant pathogens, protozoal activity against Trypanosoma cruzi, and cytotoxic activity against human tumoral cell lines. The results show that A. calidoustus was able to produce the antifungal and cytotoxic metabolites ophiobolin K and 6-epi-ophiobolin K, which may help the fungus to colonise and occupy the substratum as well as survive in natural environments.
Asunto(s)
Antifúngicos/química , Antimaláricos/química , Antineoplásicos/química , Aspergillus/química , Sesterterpenos/química , Antifúngicos/aislamiento & purificación , Antimaláricos/aislamiento & purificación , Antineoplásicos/aislamiento & purificación , Asteraceae/microbiología , Línea Celular Tumoral , HumanosRESUMEN
The endophytic fungal community associated with the native cactus Opuntia humifusa in the United States was investigated and its potential for providing antifungal compounds. A hundred-eight endophytic fungal isolates were obtained and identified by molecular methods into 17 different taxa of the genera Alternaria, Aureobasidium, Biscogniauxia, Cladosporium, Cryptococcus, Curvularia, Diaporthe, Epicoccum, Paraconiothyrium, Pestalotiopsis and Phoma. The most frequent species associated with O. humifusa were Alternaria sp. 3, Aureobasidium pullulans and Diaporthe sp. The fungal community of O. humifusa had a high richness and diversity; additionally, the species richness obtained indicates that the sample effort was enough to recover the diversity pattern obtained. Six extracts of endophytes showed antifungal properties and (1)H NMR analyses of the extracts of Alternaria sp. 5 Ohu 8B2, Alternaria sp. 3 Ohu 30A, Cladosporium funiculosum Ohu 17C1 and Paraconiothyrium sp. Ohu 17A indicated the presence of functional groups associated with unsaturated fatty-acid olefinic protons and fatty acid methylene and methyl protons. GC-FID analysis of these extracts confirmed the presence of a mixture of different fatty acids. The (1)H NMR analyses of Biscogniauxia mediterranea Ohu 19B extracts showed the presence of aromatic compounds. From the extract of B. mediterranea we isolated the compound 5-methylmellein that displayed moderate antifungal activity against the phytopathogenic fungi Phomopsis obscurans. Our results suggest that native medicinal cacti of the United States can live symbiotically with rich and diverse endophytic communities and may be a source of bioactive molecules, including those able to inhibit or control plant disease pathogens.
Asunto(s)
Antifúngicos/metabolismo , Biodiversidad , Productos Biológicos/farmacología , Hongos/clasificación , Hongos/genética , Opuntia/microbiología , Antifúngicos/química , Antifúngicos/aislamiento & purificación , Productos Biológicos/química , Productos Biológicos/aislamiento & purificación , Análisis por Conglomerados , ADN de Hongos/química , ADN de Hongos/genética , ADN Espaciador Ribosómico/química , ADN Espaciador Ribosómico/genética , Hongos/química , Hongos/aislamiento & purificación , Espectroscopía de Resonancia Magnética , Pruebas de Sensibilidad Microbiana , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN , Estados UnidosRESUMEN
We surveyed the diversity and capability of producing bioactive compounds from a cultivable fungal community isolated from oligotrophic soil of continental Antarctica. A total of 115 fungal isolates were obtained and identified in 11 taxa of Aspergillus, Debaryomyces, Cladosporium, Pseudogymnoascus, Penicillium and Hypocreales. The fungal community showed low diversity and richness, and high dominance indices. The extracts of Aspergillus sydowii, Penicillium allii-sativi, Penicillium brevicompactum, Penicillium chrysogenum and Penicillium rubens possess antiviral, antibacterial, antifungal, antitumoral, herbicidal and antiprotozoal activities. Bioactive extracts were examined using (1)H NMR spectroscopy and detected the presence of secondary metabolites with chemical shifts. Our results show that the fungi present in cold-oligotrophic soil from Antarctica included few dominant species, which may have important implications for understanding eukaryotic survival in cold-arid oligotrophic soils. We hypothesize that detailed further investigations may provide a greater understanding of the evolution of Antarctic fungi and their relationships with other organisms described in that region. Additionally, different wild pristine bioactive fungal isolates found in continental Antarctic soil may represent a unique source to discover prototype molecules for use in drug and biopesticide discovery studies.
Asunto(s)
Bioprospección , Frío Extremo , Hongos/aislamiento & purificación , Microbiota , Microbiología del Suelo , Aedes/efectos de los fármacos , Animales , Regiones Antárticas , Antiinfecciosos/aislamiento & purificación , Antiinfecciosos/toxicidad , Productos Biológicos/aislamiento & purificación , Productos Biológicos/toxicidad , Citotoxinas/aislamiento & purificación , Citotoxinas/toxicidad , Hongos/química , Hongos/clasificación , Humanos , Insecticidas/aislamiento & purificación , Insecticidas/toxicidad , Lactuca/efectos de los fármacos , Células MCF-7RESUMEN
The endophytic fungal assemblages associated with Smallanthus sonchifolius (Poepp.) H. Rob. and Smallanthus uvedalius (L.) Mack. ex Small growing in vitro autotrophic cultures and in the greenhouse were identified and evaluated for their ability to produce bioactive compounds. A total of 25 isolates were recovered that were genetically closely related to species of the genera Bionectria , Cladosporium , Colletotrichum , Fusarium , Gibberella , Hypocrea , Lecythophora , Nigrospora , Plectosphaerella , and Trichoderma . The endophytic assemblages of S. sonchifolius presented a greater diversity than the group isolated from S. uvedalius and demonstrated the presence of dominant generalist fungi. Extracts of all fungi were screened against the fungal plant pathogens. Ten extracts (41.6%) displayed antifungal activities; some of them had a broad antifungal activity. The phylotypes Lecythophora sp. 1, Lecythophora sp. 2, and Fusarium oxysporum were isolated from in vitro autotrophic cultures and displayed antifungal activity. The presence of bioactive endophytic fungi within S. sonchifolius and S. uvedalius suggests an ecological advantage against pathogenic attacks. This study revealed reduced numbers of endophytes in association with both Smallanthus species in controlled cultivation conditions compared with the endophytic communities of hosts collected in the wild environments. Even as reduced endophytic communities, these fungi continue to provide chemical protection for the host.
Asunto(s)
Antifúngicos/farmacología , Ascomicetos/química , Asteraceae/microbiología , Hongos/efectos de los fármacos , Ascomicetos/clasificación , Ascomicetos/efectos de los fármacos , Ascomicetos/genética , Biodiversidad , ADN Espaciador Ribosómico/genética , Endófitos/química , Endófitos/clasificación , Endófitos/genética , Filogenia , ARN Ribosómico 5.8S/genéticaRESUMEN
Conyza canadensis (L.) Cronquist syn. (horseweed) is a problematic and invasive weed with reported allelopathic properties. To identify the phytotoxic constituents of the aerial parts, a systematic bioactivity-guided fractionation of the dichloromethane extract was performed. Three active enyne derivatives, (2Z,8Z)-matricaria acid methyl ester, (4Z,8Z)-matricaria lactone, and (4Z)-lachnophyllum lactone, were identified. The lactones inhibited growth of the monocot Agrostis stolonifera (bentgrass) and the dicot Lactuca sativa (lettuce) at 1 mg mL(-1), while the (2Z,8Z)-matricaria acid methyl ester was less active. In a dose-response screening of the lactones for growth inhibitory activity against Lemna paucicostata , (4Z)-lachnophyllum lactone was the most active with an IC50 of 104 µM, while the (4Z,8Z)-matricaria lactone was less active (IC50 of 220 µM). In a fungal direct bioautography assay, the two lactones at 10 and 100 µg/spot inhibited growth of the plant pathogenic fungi Colletotrichum acutatum , Colletotrichum fragariae , and Colletotrichum gloeosporioides . In a dose-response screening of the lactones against six different plant pathogenic fungi, (4Z,8Z)-matricaria lactone was more active than the commercial fungicide azoxystrobin on Col. acutatum , Col. fragariae , and Col. gloeosporioides at 30 µM and about as active as the commercial fungicide captan against Col. gloeosporioides , while (4Z)-lachnophyllum lactone was less active.