Diversity and biological activities of endophytic fungi associated with Catharanthus roseus.
ABSTRACT: BACKGROUND:The present study involves diversity and bioactivity of the endophytic fungal community from Catharanthus roseus inhabiting the coastal region. This study has been conducted hypothesizing that the microbial communities in the coastal regions would tolerate a range of abiotic stress such as salinity, humidity, temperature and soil composition, and it may produce new metabolites, which may possess bioactive property. Therefore in the current study, the cytotoxicity and free radical scavenging potential of the fungal organic extracts have been investigated. Moreover, the apoptotic and the antioxidant potential of the fungus that exhibited the best activity in preliminary screening has also been demonstrated. RESULTS:Twenty endophytic fungal isolates were obtained from different parts of the plant, and identified using internal transcribed spacer region analysis. Based on the colonization frequency, the dominant genera were found to be Colletotrichum, Alternaria and Chaetomium with colonization frequency % of 8.66, 7.00 and 6.33, respectively. It was observed that the species diversity and richness was the highest in bark followed by leaf and stem regions of the plant. On screening the fungal ethyl acetate extracts for cytotoxicity against the HeLa cells, the Chaetomium nigricolor extract exhibited potent cytotoxic activity of 92.20% at 100??g?mL-?1 concentration. Comparison between the different organic extracts (ethyl acetate, chloroform, dichloromethane and hexane) of Chaetomium nigricolor mycelial and culture filtrate, it was observed that the mycelial as well the culture filtrate ethyl acetate extracts and the culture filtrate hexane extract showed significant cytotoxic potential against the HeLa and MCF-7 cells, respectively. The apoptotic- and mitochondrial membrane depolarisation-induction potential of the Chaetomium nigricolor ethyl acetate extract has also been demonstrated in this study. Further the screening of antioxidant potential of the ethyl acetate fungal extracts using DPPH scavenging assay showed that Chaetomium nigricolor extract exhibited potential activity with a significant EC50 value of 22??g?mL-?1. The ethyl acetate extract of Chaetomium nigricolor also exhibited superoxide radical scavenging potential. CONCLUSION:These results indicated that diverse endophytic fungal population inhabits Catharanthus roseus. One of the fungal isolate Chaetomium nigricolor exhibited significant cytotoxic, apoptotic and antioxidant potential.
Project description:BACKGROUND:The endophytes of medicinal plants, such as Justicia adhatoda L., represent a promising and largely underexplored domain that is considered as a repository of biologically active compounds. OBJECTIVES:The aim of present study was isolation, identification, and biological evaluation of endophytic fungi associated with the J. adhatoda L. plant for the production of antimicrobial, antioxidant, and cytotoxic compounds. MATERIALS AND METHODS:Endophytic fungi associated with the J. adhatoda L. plant were isolated from healthy plant parts and taxonomically characterized through morphological, microscopic, and 18S rDNA sequencing methods. The screening for bioactive metabolite production was achieved using ethyl acetate extracts, followed by the optimization of different parameters for maximum production of bioactive metabolites. Crude and partially purified extracts were used to determine the antimicrobial, antioxidant, and cytotoxic potential. RESULTS:Out of six endophytic fungal isolates, Chaetomium sp. NF15 showed the most promising biological activity and was selected for detailed study. The crude ethyl acetate extract of NF15 isolate after cultivation under optimized culture conditions showed promising antimicrobial activity, with significant inhibition of the clinical isolates of Staphylococcus aureus (87%, n=42), Pseudomonas aeruginosa (> 85%, n = 41), and Candida albicans (62%, n = 24). CONCLUSIONS:The present study confirms the notion of selecting endophytic fungi of medicinal plant Justicia for the bioassay-guided isolation of its bioactive compounds, and demonstrates that endophytic fungus Chaetomium sp. NF15 could be a potential source of bioactive metabolites.
Project description:Jatropha curcas L., a perennial plant grown in tropics and subtropics is popularly known for its potential as biofuel. The plant is reported to survive under varying environmental conditions having tolerance to stress and an ability to manage pest and diseases. The plant was explored for its endophytic fungi for use in crop protection. Endophytic fungi were isolated from leaf of Jatropha curcas, collected from New Delhi, India. Four isolates were identified as Colletotrichum truncatum, and other isolates were identified as Nigrospora oryzae, Fusarium proliferatum, Guignardia cammillae, Alternaria destruens, and Chaetomium sp. Dual plate culture bioassays and bioactivity assays of solvent extracts of fungal mycelia showed that isolates of Colletotrichum truncatum were effective against plant pathogenic fungi Fusarium oxysporum and Sclerotinia sclerotiorum. Isolate EF13 had highest activity against S. sclerotiorum. Extracts of active endophytic fungi were prepared and tested against S. sclerotiorum. Ethyl acetate and methanol extract of C. truncatum EF10 showed 71.7% and 70% growth inhibition, respectively. Hexane extracts of C. truncatum isolates EF9, EF10, and EF13 yielded oil and the oil from EF10 was similar to oil of the host plant, i.e., J. curcas.
Project description:Bioactive natural compounds, isolated from fungal endophytes, play a promising role in the search for novel drugs. They are an inspiring source for researchers due to their enormous structural diversity and complexity. During the present study fungal endophytes were isolated from a well-known medicinal shrub, Berberis aristata DC. and were explored for their antagonistic and antioxidant potential. B. aristata, an important medicinal shrub with remarkable pharmacological properties, is native to Northern Himalayan region. A total of 131 endophytic fungal isolates belonging to eighteen species and nine genera were obtained from three hundred and thirty surface sterilized segments of different tissues of B. aristata. The isolated fungi were classified on the basis of morphological and molecular analysis. Diversity and species richness was found to be higher in leaf tissues as compared to root and stem. Antibacterial activity demonstrated that the crude ethyl acetate extract of 80% isolates exhibited significant results against one or more bacterial pathogens. Ethyl acetate extract of Alternaria macrospora was found to have potential antibacterial activity. Significant antioxidant activity was also found in crude ethyl acetate extracts of Alternaria alternata and Aspergillus flavus. Similarly, antagonistic activity of the fungal endophytes revealed that all antagonists possessed inhibition potential against more than one fungal pathogen. This study is an important step towards tapping endophytic fungal diversity for bioactive metabolites which could be a step forward towards development of novel therapeutic agents.
Project description:Endophytic fungi have been recognized as a potential source of bioactive secondary metabolites. The endophytic Trichoderma species were isolated from Vinca plants (Vinca major, Vinca herbacea, and Vinca minor), found in Iran and screened for antimicrobial and anti-proliferative activity. Based on morphological and phylogenetic analyses, four fungal species were identified: T. asperellum, T. brevicompactum, T. koningiopsis, and T. longibrachiatum. In addition, endophytic fungi bioactivity of methanol and ethyl acetate extracts (7.8-250 ?gml-1) were assessed against a panel of pathogenic fungi and bacteria and IC80 was calculated. Data showed that both methanol and ethyl acetate extracts from all endophytic isolates had significant cytotoxic effects against the model target fungus Pyricularia oryzae. Further research indicated that they had significant antimicrobial bioactivity against the human pathogenic bacteria Staphylococcus aureus and Escherichia coli, and plant pathogenic bacteria Ralstonia solanacearum and Clavibacter michiganensis as well. According to the bioactivity results, crude ethyl acetate extract of T. koningiopsis VM115 isolate was determined for TLC and GC-MS analysis. An antifungal compound was isolated from ethyl acetate extract of T. koningiopsis VM115 based on bioassay guided fractionation. The 1H-NMR and 13C-NMR spectroscopic data showed that the compound was trichodermin, which exhibited strong fungicidal effects against P. oryzae, Aspergillus fumigatus, and Botrytis cinera with MICs of 31.25 ?g ml-1 through in vitro antifungal tests. GC-MS analysis identified six classes of volatile compound produced by T. koningiopsis VM115 (alcohols, esters, pyrones (lactones), acids, furanes and lipids). 6-n-pentyl-6H-pyran-2-one (6PP) was identified as one of the most abundant metabolites in this research. These results indicate that the fungal endophytes from Vinca plants had antibacterial and cytotoxic activities; evidence that endophytes are a good source of biological activity and compounds. This work is the first report of Trichodermin production by T. koningiopsis species.
Project description:Endophytic fungal occurrences were studied in aerial regions of <i>Digitaria bicornis</i> and <i>Paspalidium flavidum</i> by three isolation methods: potato dextrose agar (PDA), malt extract agar (MEA), and moist blotters. Seventy species of 29 genera of endophytic fungi in <i>D. bicornis</i> and 71 species of 30 genera in <i>P. flavidum</i> were documented. Endophytic fungal communities were grouped into 40 and 43 anamorphic ascomycetes (21 and 23 genera) and 20 teleomorphic ascomycetes (6 and 7 genera) in <i>D. bicornis</i> and <i>P. flavidum,</i> respectively. PDA supported the expression of larger number of fungal communities than MEA and MB; and <i>P. flavidum</i> hosted more number of endophytic fungi than <i>D. bicornis</i>. Seasons played an important role in supporting the assemblage of fungal endophytes. Endophytic fungal species richness and assemblages in plant regions were determined for alpha, beta, and gamma diversities. The ethyl acetate followed by methanolic extracts of certain fungal species showed good antagonistic and antibacterial activities. Among fungal endophytes, <i>Curvularia protuberata</i> and <i>Penicillium citrinum</i> exhibited high antagonistic and antibacterial activities. The high-resolution orbitrap liquid chromatography-mass spectrometry of ethyl acetate crude extracts of <i>C. protuberata</i> and <i>P. citrinum</i> revealed the presence of antifungal and antimicrobial, besides a host of compounds in the extracts. The present study indicated that grass endophytes are the sources of compounds with antimicrobial and other pharmacological activities.
Project description:The present study was carried out to determine the free radical scavenging potential of culture filtrate of Streptomyces sp. AM-S1. Antioxidant activity of culture filtrate, lyophilized culture filtrate and ethyl acetate extract of Streptomyces sp. AM-S1 was determined by various in vitro assays such as ferric reducing power assay, phosphomolybdenum reduction, DPPH and ABTS radical scavenging activities. The results revealed that the culture filtrate of Streptomyces sp. AM-S1 effectively scavenged DPPH (IC50 90.2 ?l/ml) and ABTS (IC50 13.2 ?l/ml) radicals in a concentration dependent manner. In all the assays, ethyl acetate extract registered higher antioxidant activity when compared with the lyophilized culture filtrate (LCF). In addition, ethyl acetate extract (1123.4 ?mole Fe(II)/mg extract) exhibited higher ferric reducing activity than the standard BHA (814.4 ?mole Fe(II)/mg extract). Further works are needed on the isolation and identification of antioxidant molecules from the ethyl acetate extract of Streptomyces sp. AM-S1 culture filtrate.
Project description:Schinus terebinthifolius Raddi is a medicinal plant widely used for the treatment of various diseases. The secondary metabolites responsible for the pharmacological properties can be produced directly by the plant or by endophytic fungi. The objective of this study was to evaluate the diversity of endophytic fungi of different parts of S. terebinthifolius and to identify chemical compounds produced by endophytes and their antioxidant and antibacterial activities. For this, fruits, stem bark and roots were dried, ground and placed in fungal growth medium. The selected endophytes were grown and subjected to extraction with ethyl acetate. DPPH, FRAP, ?-carotene bleaching and antimicrobial assays were performed. The phylogenetic tree was elaborated, encompassing 15 different species. The fungal extracts showed hydroxybenzoic acids and 1-dodecanol as predominant compounds. All fungal extracts exhibited antioxidant activity. The fungal extracts exhibited bactericidal and bacteriostatic activities against Gram-positive and Gram-negative bacterial ATCC strains and against methicillin-resistant nosocomial bacteria. Among the 10 endophytic fungi evaluated, the extract of the fungus Ochrocladosporium elatum showed higher phenolic content and exhibited higher antioxidant and antibacterial activities in all tests. Together, the results increase the known diversity of S. terebinthifolius endophytic fungi, secondary metabolites produced and their antioxidant and antibacterial activities.
Project description:Hypersaline environments are known to support diverse fungal species from various orders. The production of secondary metabolites is one of the strategies that fungi adopt to thrive under such extreme environments, bringing up the stress tolerance response. Some such unique secondary metabolites also exhibit clinical significance. The increasing prevalence of drug resistance in cancer therapy demands further exploration of these novel bioactive compounds as cancer therapeutics. In the present study, a total of 31 endophytic fungi harboring inside red, green, and brown marine algae have been isolated and identified. The maximum likelihood analysis and diversity indices of fungal endophytes revealed the phylogenetic relationship and species richness. The genus <i>Aspergillus</i> was found to be the dominating fungus, followed by <i>Cladosporium</i> spp. All the isolated endophytic fungal extracts were tested for their cytotoxicity against HeLa and A431 cancer cell lines. Nine isolates were further analyzed for their cytotoxic activity from the culture filtrate and mycelia extract. Among these isolates, <i>Biscogniauxia petrensis</i> showed potential cytotoxicity with CC<sub>50</sub> values of 18.04 and 24.85 μg/ml against HeLa and A431 cells, respectively. Furthermore, the media and solvent extraction optimization revealed the highest cytotoxic active compounds in ethyl acetate extract from the potato dextrose yeast extract broth medium. The compound-induced cell death <i>via</i> apoptosis was 50-60 and 45% when assayed using propidium iodide-live/dead and loss of mitochondrial membrane potential assay, respectively, in HeLa cells. Four bioactive fractions (bioassay-based) were obtained and analyzed using chromatography and spectroscopy. This study reports, for the first time, the cytotoxic activity of an endophytic fungal community that was isolated from marine macro-algae in the Rameswaram coastal region of Tamil Nadu, India. In addition, <i>B. petrensis</i> is a prominent apoptotic agent, which can be used in pharmaceutical applications as a therapeutic.
Project description:The marine ecosystem is an extraordinary reserve of pharmaceutically important, bioactive compounds even in this "synthetic age". Marine algae-associated endophytic fungi have gained prominence as an important source of bioactive compounds. This study was conducted on secondary metabolites of Chaetomium globosum-associated with marine green alga Chaetomorpha media from the Konkan coastline, India. Its ethyl acetate extract (CGEE) exhibited an IC50 value of 7.9?±?0.1 µg/mL on MCF-7 cells. CGEE exhibited G2M phase cell cycle arrest, ROS production and MMP loss in MCF-7 cells. The myco-components in CGEE contributing to the cytotoxicity were found by Gas Chromatography/Mass Spectrometry analyses. Chrysin, a dihydroxyflavone was one of the forty-six myco-components which is commonly found in honey, propolis and passionflower extracts. The compound was isolated and characterized as fungal chrysin using HPLC, UV-Vis spectroscopy, LC-MS, IR and NMR analyses by comparing with standard chrysin. The purified compound exhibited an IC50 value of 49.0?±?0.6 µM while that of standard chrysin was 48.5?±?1.6 µM in MCF-7 cells. It induced apoptosis, G1 phase cell cycle arrest, MMP loss, and ROS production. This is the first report of chrysin from an alternative source with opportunities for yield enhancement.
Project description:Soil is considered an extensively explored ecological niche for microorganisms that produce useful biologically active natural products suitable for pharmaceutical applications. The current study aimed at investigating biological activities and metabolic profiles of three fungal strains identified from different desert sites in Saudi Arabia. Soil fungal isolates were collected from AlQasab, Tabuk, and Almuzahimiyah in Saudi Arabia and identified. Furthermore, their antibacterial activity was investigated against Staphylococcus aureus, Enterococcus faecalis, Klebsiella pneumonia, and Escherichia coli in blood, nutrient, and Sabouraud dextrose agars. Moreover, fungal extracts were evaluated on cell viability/proliferation against human breast carcinoma and colorectal adenocarcinoma cells. To identify the biomolecules of the fungal extracts, High-performance liquid chromatography HPLC-DAD coupled to analytical LC-QTOF-MS method was employed for fungal ethyl acetate crude extract. Identified fungal isolates, Chaetomium sp. Bipolaris sp. and Fusarium venenatum showed varied inhibitory activity against tested microbes in relation to crude extract, microbial strain tested, and growth media. F. venenatum showed higher anticancer activity compared to Chaetomium sp. and Bipolaris sp. extracts against four of the tested cancer cell lines. Screening by HPLC and LC/MS-QTOF identified nine compounds from Chaetomium sp. and three from Bipolaris sp. however, for F. venenatum extracts compounds were not fully identified. In light of the present findings, some biological activities of fungal extracts were approved in vitro, suggesting that such extracts could be a useful starting point to find compounds that possess promising agents for medical applications. Further investigations to identify exact biomolecules from F. venenatum extracts are needed.