RESUMO
In this study, nine endophytic fungi capable of producing multiple phenolic compounds were screened and identified from 152 fungi isolated from pigeon pea in a natural habitat (Honghe, Yunnan Province, China). Talaromyces neorugulosus R-209 exhibited the highest potential for phenolic compound production. L-phenylalanine feeding was used to enhance phenolic compound production in T. neorugulosus R-209 cultures. Under the optimal feeding conditions (l-phenylalanine dose of 0.16 g/L and feeding phase of 6 days), the yields of genistein, apigenin, biochanin A, and cajaninstilbene acid increased by 15.59-fold, 7.20-fold, 25.93-fold, and 10.30-fold over control, respectively. T. neorugulosus R-209 fed with l-phenylalanine was found to be stable in the production of phenolic compounds during ten successive subcultures. Moreover, bioactivities of extracts of T. neorugulosus R-209 cultures were significantly increased by l-phenylalanine feeding. Overall, l-phenylalanine feeding strategy made T. neorugulosus R-209 more attractive as a promising alternative source for the production of health-beneficial phenolic compounds in the nutraceutical/medicinal industries.
Assuntos
Cajanus , Endófitos , Fenóis , Fenilalanina , Talaromyces , Talaromyces/metabolismo , Fenilalanina/metabolismo , Endófitos/metabolismo , Endófitos/isolamento & purificação , Fenóis/metabolismo , Cajanus/microbiologia , China , EcossistemaRESUMO
The secondary metabolites of the endophytic fungus Talaromyces malicola hosted in the arthropod Armadillidium vulgare were separated by silica gel column chromatography, gel column chromatography, and semi-preparative high-performance liquid chromatography. Eleven compounds(1-11) were obtained from the ethyl acetate fraction of the fermentation broth of T. malicola, and their structures were identified by NMR, HR-ESI-MS, UV, IR, and ECD. The 11 compounds were talarosesquiterpene A(1),(3ß,5α,6α,15α,22E)-5,6-epoxyergosta-8(14),22-diene-3,7,15-triol(2), vermistatin(3), hydroxyvermistatin(4), bercheminol A(5), penicillide(6), lunatinin(7), penipurdin A(8), emodin(9), BE-25327(10), and(-)-regiolone(11). Compound 1 was a new diaporol-type sesquiterpene. Compounds 2, 4-5, and 7-11 were isolated from Talaromyces for the first time.
Assuntos
Endófitos , Metabolismo Secundário , Talaromyces , Talaromyces/metabolismo , Talaromyces/química , Animais , Endófitos/química , Endófitos/metabolismo , Estrutura Molecular , Cromatografia Líquida de Alta Pressão , Espectroscopia de Ressonância MagnéticaRESUMO
AIMS: The immense therapeutic value of Valeriana jatamansi is attributed to the presence of bioactive secondary metabolites (valepotriates and sesquiterpenoids). Its over-exploitation in wild habitats resulted in extensive depletion, necessitating alternative approaches to produce its therapeutic metabolites. This study sought to assess the ability of endophytes of V. jatamansi to boost the biosynthesis of secondary metabolites in the leaf-cell suspension (LCS) culture of V. jatamansi. METHODS AND RESULTS: A total of 11 fungal endophytes were isolated from the rhizomes of V. jatamansi. Isolated endophytes were found to belong to phylum Ascomycota, Basidiomycota, and Mucoromycota. Supplementation of extracts of endophyte Phaeosphaeriaceae sp. VRzFB, Mucor griseocyanus VRzFD, Penicillium raistrickii VRzFK, and Penicillium sajarovii VRzFL in the LCS culture of V. jatamansi increased the fresh cell biomass by 19.6%-39.1% and dry cell biomass by 23.4%-37.8%. Most of the endophytes' extract could increase the content of valepotriates (26.5%-76.5% valtrate and 40.5%-77.9% acevaltrate) and sesquiterpenoids (19.9%-61.1% hydroxyl valerenic acid) in LCS culture. However, only two endophytes, Irpex lacteus VRzFI and Fusarium oxysporum VRzFF, could increase the sesquiterpenoids acetoxy valerenic acid (36.9%-55.3%). In contrast, some endophytes' extracts caused negative or no significant effect on the cell biomass and targeted metabolites. Increased secondary metabolites were corroborated with increased expression of iridoid biosynthesis genes in LCS culture. Production of H2O2 and lipid peroxidation was also varied with different endophytes indicating the modulation of cellular oxidative stress due to endophyte elicitors. CONCLUSIONS: The results suggest the distinct effect of different fungal endophytes-extract on LCS culture, and endophytes can serve as biotic elicitors for increasing the secondary metabolite production in plant in vitro systems.
Assuntos
Endófitos , Folhas de Planta , Sesquiterpenos , Valeriana , Endófitos/metabolismo , Sesquiterpenos/metabolismo , Valeriana/microbiologia , Valeriana/metabolismo , Folhas de Planta/microbiologia , Fungos/metabolismo , Ascomicetos/metabolismo , Rizoma/microbiologia , Penicillium/metabolismo , Metabolismo SecundárioRESUMO
The present study revealed the consequences of the interaction of a widely used bioinsecticide and endophyte Metarhizium anisopliae with the hazardous mycotoxin zearalenone (ZEN) as a pure substance and with ZEN as a native component of a crude Fusarium extract. In the environment, microorganisms encounter a mixture of metabolites secreted by other organisms living in the same area, not single substances. The obtained results suggest that M. anisopliae, exposed to a variety of active substances produced by Fusarium graminearum, is able to eliminate ZEN. Within 14 days, M. anisopliae biotransformed 90.8% and 85.8% of ZEN as a pure substance and ZEN as a native component of the F. graminearum extract from Rice Medium (E-Fg-RM), respectively, through reduction predominantly to α-epimers of zearalenols and zearalanols, considered more estrogenic than ZEN, which can raise concerns. Compared to pure ZEN, E-Fg-RM significantly affected the production of Metarhizium secondary metabolites by increasing the destruxins amount by approximately 20-25% and reducing the swainsonine content by 96.2%. All these findings provide a possible picture of the interaction of M. anisopliae with ZEN in the wild, mainly as a result of the use of crude extract from Fusarium, which contained a mixture of different metabolites.
Assuntos
Endófitos , Fusarium , Metarhizium , Zearalenona , Fusarium/metabolismo , Zearalenona/metabolismo , Metarhizium/metabolismo , Endófitos/metabolismo , Micotoxinas/metabolismoRESUMO
BACKGROUND: Antimicrobial resistance has emerged as a major global health threat, necessitating the urgent development of new antimicrobials through innovative methods to combat the rising prevalence of resistant microbes. With this view, we developed three novel nanoconjugates using microbial natural pigment for effective application against certain pathogenic microbes. RESULTS: A natural red pigment (RP) extracted from the endophyte Monascus ruber and gamma rays were applied to synthesize RP-ZnO, RP-CuO, and RP-MgO nanoconjugates. The synthesized nanoconjugates were characterized by different techniques to study their properties. The antimicrobial potential of these nanoconjugates was evaluated. Moreover, the antibiofilm, protein leakage, growth curve, and UV light irradiation effect of the synthesized nanoconjugates were also studied. Our results confirmed the nano-size, shape, and stability of the prepared conjugates. RP-ZnO, RP-CuO, and RP-MgO nanoconjugates showed broad antimicrobial potential against the tested bacterial and fungal pathogens. Furthermore, the RP-ZnO nanoconjugate possessed the highest activity, followed by the RP-CuO against the tested microbes. The highest % inhibition of biofilm formation by the RP-ZnO nanoconjugate. Membrane leakage of E. coli and S. aureus by RP-ZnO nanoconjugate was more effective than RP-MgO and RP-CuO nanoconjugates. Finally, UV light irradiation intensified the antibiotic action of the three nanoconjugates and RP-ZnO potential was greater than that of the RP-MgO, and RP-CuO nanoconjugates. CONCLUSION: These findings pave the way for exploiting the synthesized nanoconjugates as potential materials in biomedical applications, promoting natural, green, and eco-friendly approaches.
Assuntos
Monascus , Nanoconjugados , Monascus/metabolismo , Nanoconjugados/química , Biofilmes/efeitos dos fármacos , Pigmentos Biológicos/química , Fermentação , Cobre/química , Cobre/farmacologia , Endófitos/metabolismo , Endófitos/química , Óxido de Zinco/química , Óxido de Zinco/farmacologia , Testes de Sensibilidade Microbiana , Óxido de Magnésio/química , Óxido de Magnésio/farmacologia , Antibacterianos/farmacologia , Antibacterianos/química , Escherichia coli/efeitos dos fármacosRESUMO
Seaweed endophytes are a rich source of microbial diversity and bioactive compounds. This review provides a comprehensive analysis of the microbial diversity associated with seaweeds and their interaction between them. These diverse bacteria and fungi have distinct metabolic pathways, which result in the synthesis of bioactive compounds with potential applications in a variety of health fields. We examine many types of seaweed-associated microorganisms, their bioactive metabolites, and their potential role in cancer treatment using a comprehensive literature review. By incorporating recent findings, we hope to highlight the importance of seaweed endophytes as a prospective source of novel anticancer drugs and promote additional studies in this area. We also investigate the pharmacokinetic and pharmacodynamic profiles of these bioactive compounds because understanding their absorption, distribution, metabolism, excretion (ADMET), and toxicity profiles is critical for developing bioactive compounds with anticancer potential into effective cancer drugs. This knowledge ensures the safety and efficacy of proposed medications prior to clinical trials. This study not only provides promise for novel and more effective treatments for cancer with fewer side effects, but it also emphasizes the necessity of sustainable harvesting procedures and ethical considerations for protecting the delicate marine ecology during bioprospecting activities.
Assuntos
Antineoplásicos , Bactérias , Endófitos , Fungos , Alga Marinha , Alga Marinha/química , Endófitos/metabolismo , Endófitos/química , Antineoplásicos/farmacologia , Antineoplásicos/farmacocinética , Humanos , Fungos/efeitos dos fármacos , Fungos/metabolismo , Bactérias/efeitos dos fármacos , Bactérias/metabolismo , Neoplasias/tratamento farmacológico , Animais , BiodiversidadeRESUMO
Fungal phytopathogens represent a large and economically significant challenge to food production worldwide. Thus, the application of biocontrol agents can be an alternative. In the present study, we carried out biological, metabolomic, and genetic analyses of three endophytic isolates from nodules of Chamaecytisus albus, classified as Pseudomonas chlororaphis acting as antifungal agents. The efficiency of production of their diffusible and volatile antifungal compounds (VOCs) was verified in antagonistic assays with the use of soil-borne phytopathogens: B. cinerea, F. oxysporum, and S. sclerotiorum. Diffusible metabolites were identified using chromatographic and spectrometric analyses (HPTLC, GC-MS, and LC-MS/MS). The phzF, phzO, and prnC genes in the genomes of bacterial strains were confirmed by PCR. In turn, the plant growth promotion (PGP) properties (production of HCN, auxins, siderophores, and hydrolytic enzymes, phosphate solubilization) of pseudomonads were bioassayed. The data analysis showed that all tested strains have broad-range antifungal activity with varying degrees of antagonism. The most abundant bioactive compounds were phenazine derivatives: phenazine-1-carboxylic acid (PCA), 2-hydroxy-phenazine, and diketopiperazine derivatives as well as ortho-dialkyl-aromatic acids, pyrrolnitrin, siderophores, and HCN. The results indicate that the tested P. chlororaphis isolates exhibit characteristics of biocontrol organisms; therefore, they have potential to be used in sustainable agriculture and as commercial postharvest fungicides to be used in fruits and vegetables.
Assuntos
Antifúngicos , Pseudomonas chlororaphis , Antifúngicos/farmacologia , Antifúngicos/química , Pseudomonas chlororaphis/metabolismo , Ascomicetos/metabolismo , Fusarium/efeitos dos fármacos , Fusarium/metabolismo , Botrytis/efeitos dos fármacos , Endófitos/metabolismo , Metabolômica/métodos , Compostos Orgânicos Voláteis/farmacologia , Compostos Orgânicos Voláteis/metabolismo , Compostos Orgânicos Voláteis/química , Fenazinas/metabolismo , Fenazinas/farmacologia , Fenazinas/química , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controleRESUMO
The natural environment has been significantly impacted by human activity, urbanization, and industrialization, leading to changes in living organisms and their adaptation to harsh conditions. Species, including plants, adapt to these changes by creating mechanisms and modifications that allow them to survive in harsh environments. Also, endophytes, microorganisms that live inside plants, can support plant growth and defense mechanisms in these conditions by synthesizing antimicrobial secondary metabolites. What is more, endophytes produce bioactive metabolites, including alkaloids, amines, and peptides, which play a crucial role in the relationship between endophytes and their host organisms. Endophytes themselves benefit from this by creating a stable environment for their survival and development. The aim of this review is to gain insight into endophytic bioactive metabolites from chosen synanthropic ruderal plants. Industrial activities release pollutants like heavy metals, by-products, and waste, which challenge living organisms and require adaptation. Synanthropic plants, where endophytes are abundant, are particularly valuable for their bioactive compounds, which are used in agriculture and medicine. This review presents, among others, endophytes of herbaceous ruderal plants from central Europe-Chelidonium majus L., Urtica dioica L., Plantago lanceolata L., Matricaria chamomilla L., Equisetum arvense L., Oenothera biennis L., Silybum marianum L., and Mentha piperita L.
Assuntos
Bactérias , Endófitos , Endófitos/metabolismo , Endófitos/fisiologia , Bactérias/metabolismo , Europa (Continente) , Plantas/microbiologia , Plantas/metabolismoRESUMO
Two endophytes from the same Ginkgo biloba host were isolated and cultured separately. Three new eremophilane sesquiterpenoids (1-3), three new furan derivates (6, 8-9), one new polyketide (10), and four known compounds (4, 5, 7, 11) from Paraphaeosphaeria sp. and two new 10-membered macrolides (12-13), a new liner polyketide (14), a new benzofuran (15), and six known compounds (16-21) from Nigrospora oryzae were isolated. The structures of the isolated compounds were determined by spectroscopic methods, NMR calculations, and ECD calculations. The compounds 3-7, 9-10, 12, and 14-17 showed significant antiphytopathogenic effects against mycotoxigenic Alternaria sp. comparable to the activity of nystatin (positive control). Compounds 2, 6, 8, 9, and 18 indicated inhibitions against phytopathogen Fusarium asiaticum with MICs < 10 µg/mL. In addition, the compounds with weak antifungal activities from two endophytes were mixed to test their antifungal activity. The results showed that the metabolites from two endophytes had synergistic antifungal effects, and the beneficial interactions between natural products can induce more antifungal effects against plant pathogens than that of single compounds.
Assuntos
Ascomicetos , Endófitos , Ginkgo biloba , Doenças das Plantas , Endófitos/química , Endófitos/metabolismo , Ginkgo biloba/microbiologia , Ginkgo biloba/química , Ascomicetos/química , Ascomicetos/efeitos dos fármacos , Ascomicetos/metabolismo , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Estrutura Molecular , Fusarium/efeitos dos fármacos , Alternaria/efeitos dos fármacos , Alternaria/química , Fungicidas Industriais/farmacologia , Fungicidas Industriais/química , Antifúngicos/farmacologia , Antifúngicos/química , Testes de Sensibilidade MicrobianaRESUMO
BACKGROUND: Pseudomonas eucalypticola, a new species of the P. fluorescens group that generates most Pseudomonas-based biocontrol agents, has not been found in any plants other than Eucalyptus dunnii leaves. Except for antagonism to the growth of a few fungi, its features in plant growth promotion and disease control have not been evaluated. Here, we identified a similar species of P. eucalypticola, 1021Bp, from endophyte cultures of healthy leaves of English boxwood (Buxus sempervirens 'Suffruticosa') and investigated its antifungal activity, plant growth promotion traits, and potential for boxwood blight control. RESULTS: Colorimetric or plate assays showed the properties of 1021Bp in nitrogen fixation, phosphate solubilization, and production of indole-3-acetic acid (IAA) and siderophores, as well as the growth suppression of all five plant fungal pathogens, including causal agents of widespread plant diseases, gray mold, and anthracnose. Boxwood plant leaves received 87.4% and 65.8% protection from infection when sprayed with cell-free cultural supernatant (CFS) but not the resuspended bacterial cells at 108-9/mL of 1021Bp at one and seven days before inoculation (dbi) with boxwood blight pathogen, Calonectria pseudonaviculata, at 5 × 104 spores/mL. They also received similarly high protection with the 1021Bp cell culture without separation of cells and CFS at 14 dbi (67.5%), suggesting a key role of 1021Bp metabolites in disease control. CONCLUSIONS: Given the features of plant growth and health and its similarity to P. eucalypticola with the P. fluorescens lineage, 1021Bp has great potential to be developed as a safe and environmentally friendly biofungicide and biofertilizer. However, its metabolites are the major contributors to 1021Bp activity for plant growth and health. Application with the bacterial cells alone, especially with nonionic surfactants, may result in poor performance unless survival conditions are present.
Assuntos
Doenças das Plantas , Folhas de Planta , Pseudomonas , Doenças das Plantas/microbiologia , Doenças das Plantas/prevenção & controle , Pseudomonas/genética , Pseudomonas/crescimento & desenvolvimento , Pseudomonas/metabolismo , Folhas de Planta/microbiologia , Antibiose , Ácidos Indolacéticos/metabolismo , Fungos/crescimento & desenvolvimento , Fungos/genética , Fungos/classificação , Fungos/efeitos dos fármacos , Sideróforos/metabolismo , Endófitos/metabolismo , Endófitos/genética , Desenvolvimento Vegetal , Agentes de Controle Biológico , Antifúngicos/farmacologia , Antifúngicos/metabolismoRESUMO
Grass endophytic fungi have garnered increasing attention as a prolific source of bioactive metabolites with potential application across various fields, including pharmaceticals agriculture and industry. This review paper aims to synthesize knowledge on the diversity, isolation, and bioactivity of metabolites produced by grass endophytic fungi. Additionally, this approach aids in the conservation of rare and endangered plant species. Advanced analytical techniques such as high-performance liquid chromatography, liquid chromatograpy-mass spectrometry and gas chromatography are discussed as critical tools for metabolite identification and characterization. The review also highlights significant bioactive metabolites discovered to date, emphasizing their antimicrobial, antioxidant, and insecticidal activities and plant growth regulation properties. Besides address the challenges and future prospects in harnessing grass endophytic fungi for sustainable biotenological applications. By consolidating recent advancements and identifying agaps in the current research, this paper provides a comprehensive overview of the potential grass endophytic fungi as a valuable resource for novel bioactive compounds.
Assuntos
Produtos Biológicos , Endófitos , Fungos , Poaceae , Endófitos/metabolismo , Fungos/metabolismo , Poaceae/microbiologia , Produtos Biológicos/metabolismo , Antioxidantes/metabolismo , Anti-Infecciosos/metabolismo , Inseticidas/metabolismoRESUMO
Glycyrrhizin (GL) from licorice alleviates intracerebral hemorrhage (ICH) injuries by interacting with high-mobility group box (HMGB) 1, an inflammatory factor. We found that GL is bioconverted by endophyte coexisting with licorice and succeeded in isolating two derivatives. The aim of this study was to identify the compound with more potent HMGB1 inhibitory activity inspired by these GL derivatives. We took advantage of a ketone introduced by an endophyte at the C-3 position and attempted methyl esterification at the C-30 position because it was suggested that the water or lipid solubility of the molecule plays an important role. Among three derivatives synthesized, the product that is both ketonized and esterified showed more potent HMGB1 inhibitory activity than GL in macrophages and significantly improved adverse events occurred in ICH in vivo. These results suggest that modification of the hydrophilicity of GL, particularly at the C-3 and C-30 positions, enhances the HMGB1 inhibitory activity.
Assuntos
Glycyrrhiza , Ácido Glicirrízico , Proteína HMGB1 , Ácido Glicirrízico/farmacologia , Ácido Glicirrízico/química , Ácido Glicirrízico/metabolismo , Proteína HMGB1/metabolismo , Proteína HMGB1/antagonistas & inibidores , Animais , Glycyrrhiza/química , Camundongos , Masculino , Células RAW 264.7 , Hemorragia Cerebral/tratamento farmacológico , Hemorragia Cerebral/microbiologia , Hemorragia Cerebral/metabolismo , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Endófitos/química , Endófitos/metabolismo , Relação Estrutura-Atividade , Fungos/metabolismo , Fungos/efeitos dos fármacosRESUMO
Alternaria species (Deuteromycetes, Ascomycota) as ubiquitous fungi and prolific producers of a variety of toxic compounds are a part of microbiomes of plants, humans, and animals, mainly causing disease, allergic reactions, and toxicosis. However, some species have also been reported as endophytic microorganisms with highly bioactive metabolites. Our previous results indicate that potentially endophytic Alternaria species from Cupressaceae produce bioactive metabolites that possibly contribute to plant holobiont's health. Here, a possible mechanism behind this bioactivity is elucidated. As some endophytic fungi are reported to produce cytotoxic taxane diterpenoids, eight potentially endophytic Alternaria isolates from our collection were analyzed for the presence of the key genes of the paclitaxel (Taxol) biosynthetic pathway, i.e., taxadin synthase (ts), 10-deacetylbaccatin III-10-O-acetyltransferase (dbat), and C-13-phenylpropanoid side-chain CoA acyltransferase (bapt). The presence of all genes, i.e., ts, dbat, and bapt, was detected by PCR in six isolates and dbat and bapt in two isolates. Chemical analyses of the fermentation broths by TLC and HPLC chromatography and IR spectroscopy indicated the synthesis of the final product, i.e., paclitaxel. So, we introduce the synthesis of taxane diterpenoids as a possible mechanism by which Alternaria occupies the plant niches and protects the plant holobiont in the presence of competing microorganisms.
Assuntos
Alternaria , Vias Biossintéticas , Taxoides , Alternaria/genética , Alternaria/metabolismo , Taxoides/metabolismo , Vias Biossintéticas/genética , Endófitos/metabolismo , Endófitos/genética , Endófitos/isolamento & purificação , Endófitos/classificação , Hidrocarbonetos Aromáticos com Pontes/metabolismo , Diterpenos/metabolismo , Paclitaxel/biossíntese , Proteínas Fúngicas/genética , Proteínas Fúngicas/metabolismo , Genômica , FilogeniaRESUMO
Previous studies have reported the functional role, biochemical features and synthesis pathway of podophyllotoxin (PTOX) in plants. In this study, we employed combined morphological and molecular techniques to identify an endophytic fungus and extract PTOX derivatives. Based on the analysis of ITS sequences and the phylogenetic tree, the isolate was classified as Penicillium herquei HGN12.1C, with a sequence identity of 98.58%. Morphologically, the HGN12.1C strain exhibits white colonies, short-branched mycelia and densely packed hyphae. Using PacBio sequencing at an average read depth of 195×, we obtained a high-quality genome for the HGN12.1C strain, which is 34.9 Mb in size, containing eight chromosomes, one mitochondrial genome and a GC content of 46.5%. Genome analysis revealed 10 genes potentially involved in PTOX biosynthesis. These genes include VdtD, Pinoresinollariciresinol reductase (PLR), Secoisolariciresinol dehydrogenase (SDH), CYP719A23, CYP71BE54, O-methyltransferase 1 (OMT1), O-methyltransferase 3 (OMT3), 2-ODD, CYP71CU and CYP82D61. Notably, the VdtD gene in fungi shares functional similarities with the DIR gene found in plants. Additionally, we identified peltatin, a PTOX derivative, in the HGN12.1C extract. Docking analysis suggests a potential role for the 2-ODD enzyme in converting yatein to deoxypodophyllotoxin. These findings offer invaluable insights into the synthesis mechanism of PTOX in fungi, shedding light on the relationship between host plants and endophytes.
Assuntos
Vias Biossintéticas , Genoma Fúngico , Penicillium , Filogenia , Podofilotoxina , Podofilotoxina/biossíntese , Podofilotoxina/análogos & derivados , Penicillium/genética , Penicillium/metabolismo , Vias Biossintéticas/genética , Endófitos/genética , Endófitos/metabolismo , Análise de Sequência de DNA , Composição de Bases , GenômicaRESUMO
BACKGROUND: To adapt to constantly changing environments, ancient gymnosperms have coevolved with diverse endophytic fungi that are essential for the fitness and adaptability of the plant host. However, the effect of sex on plant-endophyte interactions in response to environmental stressors remains unknown. RNA-seq integrated with ITS analysis was applied to reveal the potential mechanisms underlying the sex-specific responses of Taxus mairei to ultraviolet (UV)-B radiation. RESULTS: Enrichment analysis suggested that sex influenced the expression of several genes related to the oxidation-reduction system, which might play potential roles in sex-mediated responses to UV-B radiations. ITS-seq analysis clarified the effects of UV-B radiation and sex on the composition of endophytic fungal communities. Sex influenced various secondary metabolic pathways, thereby providing chemicals for T. mairei host to produce attractants and/or inhibitors to filter microbial taxa. Analysis of fungal biomarkers suggested that UV-B radiation reduced the effect of sex on fungal communities. Moreover, Guignardia isolate #1 was purified to investigate the role of endophytic fungi in sex-mediated responses to UV-B radiation. Inoculation with spores produced by isolate #1 significantly altered various oxidation-reduction systems of the host by regulating the expression of APX2, GST7 NCED1, ZE1, CS1, and CM1. CONCLUSION: These results revealed the roles of endophytic fungi in sex-mediated responses to UV-B radiation and provided novel insights into the sex-specific responses of Taxus trees to environmental stressors. Video Abstract.
Assuntos
Metabolismo Secundário , Taxus , Raios Ultravioleta , Taxus/microbiologia , Endófitos/genética , Endófitos/metabolismo , Fungos/genética , Fungos/classificação , Fungos/efeitos da radiação , Fungos/metabolismo , MicrobiotaRESUMO
Papaver genus, commonly known as popies, is a valuable source of alkaloids used in medicine, including papaverine, morphine, codeine, and thebaine. We isolated six endophytic fungal isolates producing morphinan alkaloids from four Papaver species growing in Kurdistan Province, Iran. To do this, a 1:1 mixture of methanol and chloroform was used to extract fungal cultures. The contents of morphinan alkaloids in the extracts were subsequently determined using phase high-performance liquid chromatography. Among the morphinan alkaloid-producing fungal isolates, IRAN 4653C had the highest yield giving 23.06 (mg/g) morphine and 2.03 (mg/g) codeine when grown in potato dextrose liquid medium. The identity of this isolate was examined and recognized as a new fungal species named as Pithoascus kurdistanesis sp. nov. based on multi-gene phylogenetic analyses of ITS, TEF-1α, and TUB2 sequences data and morphological features. The morphinan-producing endophytic fungus and the isolated Pithoascus species from Papaver are being reported for the first time. Accordingly, this fungus shows promise as a new source of valuable compounds which is illustrated and introduced here as a new Microascaceae member belonging to Pithoascus from Kurdistan Province, Iran. Moreover, the morphinan productivity of P. kurdistanesis was further validated by gas chromatography-mass spectrometry (GC-MS).
Assuntos
Endófitos , Morfina , Papaver , Filogenia , Endófitos/metabolismo , Endófitos/genética , Papaver/microbiologia , Papaver/metabolismo , Ascomicetos/metabolismo , Ascomicetos/genética , Ascomicetos/isolamento & purificação , Irã (Geográfico) , AlcaloidesRESUMO
There is interest in studying microbes that colonize maize silks (style tissue, critical for reproduction) including the fungal pathogen Fusarium graminearum (Fg) and its interactions with the microbiome and biocontrol agents. In planta imaging of these interactions on living silks using confocal fluorescence microscopy would provide key insights. However, newly discovered microbes have unknown effects on human health, and there are regulatory requirements to prevent the release of fluorescently tagged microbes into the environment. Therefore, the microbe infection, colonization, and interaction stages on silks prior to microscopy must be contained. At the same time, silk viability must be maintained and experiments conducted that are biologically relevant (e.g. silks should remain attached to the cob), yet the silk tissue must be accessible to the researcher (i.e. not within husk leaves) and allow for multiple replicates. Here we present methods that meet these five contrasting criteria. We tested these methods using Fg and four silk-derived bacterial endophytes. The endophytes were previously known to have anti-Fg activity in vitro, but in planta observations were lacking. In Method 1, a portion of the tip of a cob was dissected, and silks remained attached to the cob in a Petri dish. The cob was placed on a water agar disc to maintain hydration. DsRed-tagged bacteria and GFP-tagged Fg were inoculated onto the silks and incubated, allowing the two microbes to grow towards one another before staining with propidium iodide for confocal microscopy. A variation of the protocol was presented in Method 2, where detached silk segments were placed directly on water agar where they were inoculated with bacteria and Fg to promote dense colonization, and to allow for many replicates and interventions such as silk wounding. The bacterial endophytes were successfully observed colonizing Fg hyphae, silk trichomes, and entering silks via cut ends and wounds. These protocols can be used to study other silk-associated microbes including several globally important fungal pathogens that enter maize grain through silks.
Assuntos
Fusarium , Microscopia de Fluorescência , Zea mays , Fusarium/patogenicidade , Zea mays/microbiologia , Microscopia de Fluorescência/métodos , Microbiota , Interações Microbianas , Endófitos/metabolismo , Microscopia Confocal/métodos , Doenças das Plantas/microbiologia , Bactérias/genéticaRESUMO
Endophytic fungi have been shown to synthesize bioactive secondary metabolites, some of which promote plant growth through various mechanisms. In our previous study, endophytic fungi were isolated from mango trees (Mangifera indica L.). The present study examined fifty endophytic fungal isolates for mineral solubilization activity, ammonia production, and siderophore production. It was shown that these isolates could produce phytohormones indole-3-acetic acid and gibberellic acid, as well as inhibit plant pathogens, specifically Colletotrichum gloeosporioides and Lasiodiplodia theobromae. The results showed that all the isolated fungal endophytes exhibited various activities. Based on the findings, two fungal endophytes-Aureobasidium pullulans CY.OS 13 and Aspergillus tamarii CY.OS 144-were selected for dual inoculation in chili plants under pot-scale conditions to investigate their potential to improve growth-related traits such as seed germination, shoot and root length, biomass, and chlorophyll content. Seed treated with A. pullulans CY.OS 13 and/or A. tamarii CY.OS 144 showed a significant (p < 0.05) increase in seed germination and growth parameters of chili plants grown under pot-scale conditions. Particularly, chili plants whose seeds were injected with a combination of the two selected endophytic fungi showed the highest plant development traits. Therefore, the selected endophytic fungi have the potential to be used as biofertilizers, especially when combined. They could eventually replace chemical fertilizers because they are environmentally friendly, beneficial to humans, and can even promote sustainable agriculture.
Assuntos
Colletotrichum , Endófitos , Ácidos Indolacéticos , Mangifera , Reguladores de Crescimento de Plantas , Endófitos/isolamento & purificação , Endófitos/metabolismo , Mangifera/microbiologia , Reguladores de Crescimento de Plantas/metabolismo , Ácidos Indolacéticos/metabolismo , Colletotrichum/crescimento & desenvolvimento , Giberelinas/metabolismo , Raízes de Plantas/microbiologia , Raízes de Plantas/crescimento & desenvolvimento , Sideróforos/metabolismo , Desenvolvimento Vegetal , Germinação , Ascomicetos/crescimento & desenvolvimento , Ascomicetos/metabolismo , Ascomicetos/isolamento & purificação , Fungos/isolamento & purificação , Fungos/classificação , Fungos/metabolismo , Fungos/crescimento & desenvolvimento , Sementes/microbiologia , Sementes/crescimento & desenvolvimento , Aspergillus/crescimento & desenvolvimento , Aspergillus/metabolismo , Aspergillus/isolamento & purificação , Clorofila/metabolismo , Amônia/metabolismo , Capsicum/microbiologia , Capsicum/crescimento & desenvolvimento , BiomassaRESUMO
Epothilones are one of the common prescribed anticancer drugs for solid tumors, for their exceptional binding affinity with ß-tubulin microtubule, stabilizing their disassembly, causing an ultimate arrest to the cellular growth. Epothilones were initially isolated from Sornagium cellulosum, however, their extremely slow growth rate and low yield of epothilone is the challenge. So, screening for a novel fungal endophyte dwelling medicinal plants, with higher epothilone productivity and feasibility of growth manipulation was the objective. Aspergillus niger EFBL-SR OR342867, an endophyte of Latania loddegesii, has been recognized as the heady epothilone producer (140.2 µg/L). The chemical structural identity of the TLC-purified putative sample of A. niger was resolved from the HPLC, FTIR and LC-ESI-MS/MS analyses, with an identical molecular structure of the authentic epothilone B. The purified A. niger epothilone B showed a resilient activity against MCF-7 (0.022 µM), HepG-2 (0.037 µM), and HCT-116 (0.12 µM), with selectivity indices 21.8, 12.9 and 4, respectively. The purified epothilone B exhibited a potential anti-wound healing activity to HepG-2 and MCF-7 cells by ~ 54.07 and 60.0%, respectively, after 24 h, compared to the untreated cells. The purified epothilone has a significant antiproliferative effect by arresting the cellular growth of MCF-7 at G2/M phase by ~ 2.1 folds, inducing the total apoptosis by ~ 12.2 folds, normalized to the control cells. The epothilone B productivity by A. niger was optimized by the response surface methodology, with ~ 1.4 fold increments (266.9 µg/L), over the control. The epothilone productivity by A. niger was reduced by ~ 2.4 folds by 6 months storage as a slope culture at 4 °C, however, the epothilone productivity was slightly restored with ethylacetate extracts of L. loddegesii, confirming the plant-derived chemical signals that partially triggers the biosynthetic genes of A. niger epothilones. So, this is the first report emphasizing the metabolic potency of A. niger, an endophyte of L. loddegesii, to produce epothilone B, that could be a new platform for industrial production of this drug.
Assuntos
Antineoplásicos , Aspergillus niger , Endófitos , Epotilonas , Cicatrização , Epotilonas/farmacologia , Epotilonas/biossíntese , Epotilonas/química , Epotilonas/metabolismo , Humanos , Endófitos/metabolismo , Endófitos/química , Aspergillus niger/efeitos dos fármacos , Aspergillus niger/metabolismo , Antineoplásicos/farmacologia , Antineoplásicos/química , Cicatrização/efeitos dos fármacos , Células MCF-7 , Células Hep G2 , Ciclo Celular/efeitos dos fármacos , Linhagem Celular Tumoral , Proliferação de Células/efeitos dos fármacosRESUMO
Co-cultivation is a powerful emerging tool for awakening biosynthetic gene clusters (BGCs) that remain transcriptionally silent under artificial culture conditions. It has recently been used increasingly extensively to study natural interactions and discover new bioactive metabolites. As a part of our project aiming at the discovery of structurally novel and biologically active natural products from mangrove endophytic fungi, an established co-culture of a strain of Phomopsis asparagi DHS-48 with another Phomopsis genus fungus DHS-11, both endophytes in mangrove Rhizophora mangle, proved to be very efficient to induce the production of new metabolites as well as to increase the yields of respective target metabolites. A detailed chemical investigation of the minor metabolites produced by the co-culture of these two titled fungal strains led to the isolation of six alkaloids (1-6), two sterols (7, 8), and six polyketides (9-14). In addition, all the compounds except 8 and 10, as well as three new metabolites phomopyrazine (1), phomosterol C (7), and phomopyrone E (9), were not present in discrete fungal cultures and only detected in the co-cultures. The structures were elucidated on the basis of spectroscopic analysis, and the absolute configurations were assumed by electronic circular dichroism (ECD) calculations. Subsequently, the cytotoxic, immunosuppressive, and acetylcholinesterase inhibitory properties of all the isolated metabolites were determined in vitro. Compound 8 exhibited moderate inhibitory activity against ConA-induced T and LPS-induced B murine splenic lymphocytes, with IC50 values of 35.75 ± 1.09 and 47.65 ± 1.21 µM, respectively.