scholarly journals Endophytic fungal isolation from Blumea axillaris: Identification and biological activity of secondary metabolites

2021 ◽  
Vol 13 (2) ◽  
pp. 10953
Author(s):  
Nehru LAVANYA ◽  
Vellingiri MANON MANI ◽  
Nachimuthu SARANYA ◽  
Rajendran DEEPAKKUMAR ◽  
Kathirvel PREETHI

Medicinal plants are a wealthy source of natural medicinal properties and remain as base for new drug discoveries. Endophyte from the specific medicinal plants produce the analogous metabolites as that of the host plant. The metabolites from the endophytes comprise maximum therapeutic properties and have been extensively applied in treating various diseases and disorders. This study was focused on identification of the endophytic fungi from the medicinal plant Blumea axillaris and investigates the diversity of endophytic fungi from various explants of the same plant. The explants were cultured on potato dextrose agar and 6 endophytic fungi were successfully isolated from Blumea axillaris. They were identified morphologically and confirmed with molecular analysis as Xylaria arbuscula, Paraphoma radicina, Phomopsis phaseoli, Sordaria fimicola, Aspergillus amstelodami, Diaporthe eucalyptorum. The DNA sequences were analyzed by BLAST and the phylogenetic tree was constructed with neighbor joining method. The six isolates were subjected to antagonistic activity for the selection of potential strain and the bioactive strain Xylaria arbuscula was selected for the production of secondary metabolites by optimization. The parameters like pH, temperature, incubation period, carbon and nitrogen (organic and inorganic source) were optimized for secondary metabolite production. The fungal metabolite was extracted by solvent extraction method using polar and non-polar solvents like propanol, methanol, chloroform, acetone and ethyl acetate. To investigate the bioactivities of the fungal crude extract was subjected first for its antioxidant activity using DPPH radical scavenging method, followed by antimicrobial activity of methanolic (MeOH) extract of Xylaria arbuscula, that were also analyzed by the agar well-diffusion method against the clinical pathogens Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Escherichia coli, Pseudomonas aeruginosa, Klebsiella pnuemoniae, Proteus mirabilis, Aspergillus niger and Candida albicans.

2019 ◽  
Vol 3 (1) ◽  
pp. 10-18 ◽  
Author(s):  
Peter M. Eze ◽  
Joy C. Nnanna ◽  
Ugochukwu Okezie ◽  
Happiness S. Buzugbe ◽  
Chika C. Abba ◽  
...  

AbstractEndophytic fungi associated with Nigerian plants have recently generated significant interest in drug discovery programmes due to their immense potential to contribute to the discovery of new bioactive compounds. This study was carried out to investigate the secondary metabolites of endophytic fungi isolated from leaves ofNewbouldia laevis, Ocimum gratissimum, andCarica papayaThe plants were collected from Agulu, Anambra State, South-East Nigeria. Endophytic fungal isolation, fungal fermentation; and extraction of secondary metabolites were carried out using standard methods. The crude extracts were screened for antimicrobial activities using the agar well diffusion method, and were also subjected to high performance liquid chromatography (HPLC) analysis to identify their constituents. A total of five endophytic fungi was isolated, two fromN. laevis(NL-L1 and NL-L2), one fromO. gratissimum(SL-L1), and two fromC. papaya(PPL-LAC and PPL-LE2). In the antimicrobial assay, the extracts of NL-L2, SL-L1, and PPL-LE2 displayed mild antibacterial activity against both Gram negative and Gram positive test bacteria. PPL-LAC extract showed mild activity only againstS. aureus, while no antimicrobial activity was recorded for NL-L1 extract. All the endophytic fungal extracts showed no activity against the test fungiC. albicansandA. fumigatusHPLC analysis of the fungal extracts revealed the presence of ethyl 4-hydroxyphenyl acetate and ferulic acid in NL-L1; ruspolinone in NL-L2; protocatechuic acid, scytalone, and cladosporin in SL-L1; indole-3-acetic acid and indole-3-carbaldehyde in PPL-LE2; and indole-3-acetic acid in PPL-LAC. The findings of this study revealed the potentials possessed by these plants as source of endophytes that express biological active compounds. These endophytes hold key of possibilities to the discovery of novel molecules for pharmaceutical, agricultural and industrial applications.


F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 379 ◽  
Author(s):  
Dhurva Prasad Gauchan ◽  
Pratistha Kandel ◽  
Astha Tuladhar ◽  
Ashesh Acharya ◽  
Upendra Kadel ◽  
...  

Background: Endophytic fungi are largely underexplored in the discovery of natural bioactive products though being rich sources of novel compounds with promising pharmaceutical potential. In this study, Taxus wallichiana, which has huge medicinal value, was investigated for its endophytic diversity and capability to produce bioactive secondary metabolites by analyzing antioxidant, antimicrobial and cytotoxic properties. Methods: The endophytes were identified by ITS-PCR using genomic DNA samples. The secondary metabolites were extracted by solvent extraction method using ethyl acetate. The antioxidant activity was analyzed by Thin Layer Chromatography, Total Phenol Content (TPC), Total Flavonoid Content (TFC) and DPPH assay, and the antimicrobial activity was analyzed by agar-well diffusion method. Brine shrimp lethality assay was used to analyze the cytotoxicity of the fungal extracts. Results: Out of 16 different Taxus trees sampled from different locations of Dhorpatan, 13 distinctive endophytic fungi were isolated and grouped into 9 different genera: Bjerkandera, Trichoderma, Preussia, Botrytis, Arthrinium, Alternaria, Cladosporium, Sporormiella and Daldinia. The ethyl acetate extracts isolated from three endophytic fungi: Alternaria alternata, Cladosporium cladosporioides and Alternaria brassicae showed significant TPC values of 204±6.144, 312.3±2.147 and 152.7±4.958µg GAE/mg of dry extract, respectively, and TFC values of 177.9±2.911, 644.1±4.202 and 96.38±3.851µg RE/mg of dry extract, respectively. Furthermore, these three extracts showed a dose dependent radical scavenging activity with IC50 concentration of 22.85, 22.15 and 23.001 µg/ml, respectively. The extracts of C. cladosporioides and A. brassicae also showed promising antimicrobial activity against Escherichia coli, Staphylococcus aureus and Bacillus subtilis with a minimum inhibitory concentration of 250μg/ml for all bacteria. Both the samples showed cytotoxic property against shrimp nauplii with LC50 of 104.2 and 125.9µg/ml, respectively. Conclusions: The crude fungal extracts obtained from endophytes: A. alternata, C. cladosporioides and A. brassicae upon purification and further identification of the bioactive compounds can be a fascinating source for novel pharmaceutical agents.


2021 ◽  
Vol 948 (1) ◽  
pp. 012072
Author(s):  
W N Fadillah ◽  
N Sukarno ◽  
D Iswantini ◽  
M Rahminiwati ◽  
S Listiyowati

Abstract Marine sponges are associated with marine fungi. The associated fungi produce secondary metabolites for sponge survival in extreme habitats. Despite the important role of the associated fungi on their host, the research on marine fungi however has not been studied well. This study aimed to isolate sponge-associated marine fungi and analyze the potency of fungal secondary metabolites against Candida albicans. The sponge used was Clathria sp. collected from Pramuka Island, Indonesia. Fungal isolation used the direct inoculation method. Fungal identification was done by morphological and molecular characteristics of ITS rDNA region. Detection of anti-Candida used the well diffusion method. The isolate has typical morphological characteristics of the genus Gymnoascus with noduled chlamydospore and arthroconidia. The isolated fungus was identified as Gymnoascus udagawae based on morphological and molecular analysis. This is the first record of marine fungi G. udagawae from Indonesia. The ethyl acetate extract of fungal filtrate showed 1.4 cm inhibition diameter of 500 mg/μL extract. The inhibition is moderate category compared to that of clotrimazole a drug commonly used for candidiasis as the positive control with showed 2.8 cm inhibition diameter at 100 mg/μL. The fungus is a potential source of the secondary metabolite active against C. albicans.


Author(s):  
Smita Kishor Puri ◽  
Prasanna Vasantrao Habbu ◽  
Preeti Venkatrao Kulkarni ◽  
Venkatrao Hanumanthrao Kulkarni

Fungal endophytes are the microbes residing in internal tissues of the plant forming symbiotic, mutualistic, communalistic and trophobiotic relationship. Endophytes from medicinal plants are considered as essential source of secondary metabolites accompanied by interesting biological/pharmacological activities. In this study, an effort was made to isolate, characterize endophytic fungi from leaves of Andrographis paniculata and to screen the fungal fractions for in-vitro antioxidant and hepatoprotective activity. Two fungal endophytes (APLF-1 and APLF-2) from Andrographis paniculata were isolated and fermented to get chloroform (A1C, A2C), ethyl acetate (A1EA, A2EA) and n butanol (A1nB, A2nB) extracts. All the endophytic fractions of APLF-1 and APLF-2 were assayed for free radical scavenging properties against 2, 2-diphenyl-1-picrylhydrazyl (DPPH), hydroxyl radical and reducing power. Fractions of APLF-2 showed good scavenging activity compared to APLF-1 extracts. Further, A2EA and A2nB were evaluated for hepatoprotective activity against CCl4 induced hepatotoxicity. A2EA (50 mg/kg &100 mg/kg) and A2nB (50 mg/kg &100 mg/kg) reversed the elevated biochemical parameters with respect to CCl4 treated group (p<0.001). The LPO, SOD and CAT levels were also restored by A2EA and A2nB (100 mg/kg p.o). APLF-1 and APLF-2 were studied for rDNA sequencing by PCR technique. The endophytic fungi, APLF-1 and APLF-2 were identified as Diaporthe sp. A25 and Preussia sp. PPV3.6 respectively based on their morphology and molecular characterization. The presence of polyvalent secondary metabolites in A2EA and A2nB were confirmed by HPTLC analysis. Keyword: Endophytic fungi; Hepatoprotective; Antioxidant; Andrographis paniculata; Diaporthe sp. A25; Preussia sp. PPV3.6


2017 ◽  
Vol 5 (1) ◽  
pp. 15
Author(s):  
Salisu Abubakar ◽  
Rebeccah Wusa Ndana ◽  
Abolade Samuel Afolabi

The focus of this research work was to isolates and identify endophytic fungi from young leaves and stems (twig) of Azadirachta indica, family (Meliaceae) for the production and assay of the produced secondary metabolites. Altogether 126 segments were used, of which 63 segments each from leaves and stems (twig) tissues were screened, using modified surface sterilization techniques. A total of 12 species of endophytic fungi were purely isolated, Penicillium spp was randomly selected for the extraction and evaluation of its secondary metabolites. The eluent collected from column chromatography mixture of ethylacetate and n- hexane (50:50v/v) was phytochemically screened, and the results showed the presence of saponins, flavonoids and phenols. Based on disc diffusion method of sensitivity, the eluent possessed some degree of antibacterial and antifungal activities. In addition, in-vitro antioxidant potentiality of the eluent was also evaluated using 2, 2- Diphenyl-1-picrylhydrazyl radical (DPPH, Sigma Aldrich). Statistical package for social science (SPSS 3/93) software was used to analyze the results using one way Analysis of Variance (ANOVA), which revealed no significant difference, on the effect of concentrations of eluent on test organisms but the eluent showed significant difference on scavenging free radicals at a critical value (p >0.05).


Author(s):  
Shirly Kumala ◽  
Ng Vini Aprilia ◽  
Partomuan Simanjuntak

Objective: Colletotrichium capsici endophytic fungi isolated from Jamblang plant (Eugenia cumini  L).  The secondary metabolites of this plant has the potent antibacterial efficacy  as well as diarrheal and anti-diabetic. This research focussed on isolation of the endophytic microbes from branches of Jamblang plants and their secondary metabolites.Methods: Isolation of endophytes were performed in PDA(potato dextrose agar) using direct seed plant. Endophytic fungi isolates with strongest antimicrobial activity against the bacteria Staphylococcus aureus, Escherichia coli were fermented in Potato Dextrose Yeast (PDY) to produce  large scale of the metabolites.  Supernatant was extracted with ethyl acetate solvent. Ethyl acetate extract fractionated by column chromatography (SiO3, n-hexane- ethyl acetate = 50:1 ~ 1:1) and obtained three fractions. Further, agar diffusion method was performed to assess their anti-microbial activity.Results:  Antibacterial  test  results  indicated that fraction III had  the antibacterial  activity Staphylococcus aureus with  inhibition zone diameter  of 10.7 mm but no observed antibacterial activity against Escherichia coli.  Furthermore, identification  by GC-MS showed that compounds present in fraction III was mainly fatty acid and phenolic compounds.Conclusion:  In conclusion, secondary metabolites isolated from Jamblang plants branches contained predominantly fatty acid and phenol related compounds that could be responsible for its potent anti microbial activity. Keywords : Endophytic fungi, Jamblang (Eugenia cumini L.), antimicrobial activity


2016 ◽  
Vol 14 (4) ◽  
pp. 328-355 ◽  
Author(s):  
Hit Kishore Goswami ◽  
Kakali Sen ◽  
Radhanath Mukhopadhyay

AbstractSelective use of crude plant extracts has been the oldest ritual in ancient Indian Medicinal System ‘Ayurveda’, as well as in Traditional Chinese Medicine system for thousands of years. This has been well documented that herbal medicines of Chinese, Indian, Korean and Native American people had included bryophytes, lichens, lycophytes and ferns. Since antiquity, most of the ferns and fern allies have given many health benefits to ancient civilizations who had used them for food, tea and drugs. Modern approaches have combined multidisciplinary technologies and have specific chemical compounds extracted and identified for producing very particulate medicines from plant parts. Plants, which yield appreciable quality and quantity of polysaccharides, steroids, terpenoids, flavonoids, alkaloids and antibiotics are suitable for dragging out drugs for many ailments/diseases, including cancer treatments. Modern explorations on the functional activities of pteridophytes for human health by discovering specific compounds and their usage in medicines have widened the scope of pteridophytes by shaping these plants as a great boon for pharmaceutical companies and related industries. Even ‘fern weeds’, which invade our freshwater bodies and reduce the freshwater wealth of a lake, e.g. Azolla, Salvinia, Marsilea, Ceratopteris, etc. can be utilized to produce life saving drugs because they are reservoirs of very many organic compounds that are useful as medicines. Some of the fern genera have a few unique secondary metabolites, which have not been discovered in higher plants. Polyphenols are useful phytochemicals, which provide health benefits such as antioxidants. From experiments on screening of total polyphenol contents of 37 ferns and fern allies, Polystichum lepidocaulon and Polystichum polyblepharum were reported to have more than 13% of total polyphenols from dried materials of both fronds and rhizomes. In addition, fronds of Davallia mariesii and rhizomes of Cyrtomium fortune, Dicranopteris pedata, Athyrium niponicum and Dryopteris nipponensis showed more than 10% of total polyphenols from dried materials. High bioactivities of traditional medicinal ferns have been studied internationally to underscore their roles in medicine. These attempts have confirmed various bioactivities, such as antioxidant, antimicrobial, antiviral, anti-inflammatory, antitumor and anti-HIV, etc. The occurrence of antibiotic activity in the extracts of more than 200 species of pteridophytes has been shown to be of prime significance within the period of 1975–2015. The active substances in many cases were found to be antibacterial to penicillin-resistant Staphylococcus aureus, Mycobacterium phlei, Salmonella typhi, Vibrio cholera, and Pseudomonas aeruginosa. Dryopteris cochleata was active against both bacteria and fungi. Five other species of Dryopteris showed remarkable antibacterial activity. The ferns of ‘Adiantum group’ have been found to be particularly active against Gram-positive bacteria. The polypodiaceous ferns constitute a rich group of which Microsorum alternifolium, Leptochillus decurrens, Polypodium irioides, Pyrrosia mannii and Phymatodes ebenipes deserve special mention. Several thelypteroid, davallioid and athyrioid ferns, in addition to antibiotic activity have also been found to show most useful bioactivity for our life – the antioxidant activity. The latter superb biochemical quality of ferns alone makes most ferns of great advantage to human health. Lycophytes particularly Lycopodium clavatum and Equisetum hyemale and ferns (Dryopteris and Adiantums) have had constituted the backbone of Homeopathic medicines and now many more genera have been added to the network of modern medicinal approaches in the drug industry. These pteridophytes are indispensably integral parts of forests world over. A few of the aquatic ferns (Azolla, Salvinia) serve as excellent bio-fertilizers and bioremediation agents. Medicinal plants are under cultivation and cultured world over. Botanically, say a thousand years ago, these were wild and many of them were weeds. As pteridophytes have survived since Paleozoic, they have undergone series of disruptive adaptive changes of environment than any other vascular plants. These plants most likely, could withstand the tests of geological time on account of their being guarded with genetic capability of possessing many useful oils, phytochemicals (secondary metabolites) such as flavonoids, steroids, alkaloids, phenols, triterpenoid compounds, varieties of amino acids and fatty acids, which in turn offer inherent tolerance and defense system . Additionally, from evolutionary point of view majority of ferns have constituted carpet flora and have worked as ‘cradles’ in natural forests so as to nurture small animals particularly reptiles and mammals. Ferns are denominators of prevalent rich biodiversity in almost every part of the earth. Comparison of evolutionary adaptations and natural innovations illuminate the genetic basis for the development of organisms. It is emphasized that there should be good field stations just in the peripheral region of reserved forests with large green houses to function as ‘Fernariums/ Mossariums/ and/or Lichenariums’ to conserve and maintain rare, endangered and medicinally superlative species found in those areas/forests. Gene networks (DNA stretches) that retain similar wiring diagrams (some or many similar DNA sequences) among related, distantly related or even totally diverse organisms point to the ways in which regulatory regions of the genome have evolved. Indisputably, comparative genomics can help us in deciphering evolvability of gene network and conservation modes during vast geological journey in evolution. We need exhaustive genomics and multidimensional molecular genetic studies on pteridophytes so as to discover unique DNA sequences, which could turn the gates of modern medicine.


F1000Research ◽  
2020 ◽  
Vol 9 ◽  
pp. 379
Author(s):  
Dhurva Prasad Gauchan ◽  
Pratistha Kandel ◽  
Astha Tuladhar ◽  
Ashesh Acharya ◽  
Upendra Kadel ◽  
...  

Background: Endophytic fungi are largely underexplored in the discovery of natural bioactive products though being rich sources of novel compounds with promising pharmaceutical potential. In this study, Taxus wallichiana, which has huge medicinal value, was investigated for its endophytic diversity and capability to produce bioactive secondary metabolites by analyzing antioxidant, antimicrobial and cytotoxic properties. Methods: The endophytes were identified by ITS-PCR using genomic DNA samples. The secondary metabolites were extracted by solvent extraction method using ethyl acetate. The antioxidant activity was analyzed by Thin Layer Chromatography, Total Phenol Content (TPC), Total Flavonoid Content (TFC) and DPPH assay, and the antimicrobial activity was analyzed by agar-well diffusion method. Brine shrimp lethality assay was used to analyze the cytotoxicity of the fungal extracts. Results: Out of 16 different Taxus trees sampled from different locations of Dhorpatan, 13 distinctive endophytic fungi were isolated and grouped into 9 different genera: Bjerkandera, Trichoderma, Preussia, Botrytis, Arthrinium, Alternaria, Cladosporium, Sporormiella and Daldinia. The ethyl acetate extracts isolated from three endophytic fungi: Alternaria alternata, Cladosporium cladosporioides and Alternaria brassicae showed significant TPC values of 204±6.144, 312.3±2.147 and 152.7±4.958µg GAE/mg of dry extract, respectively, and TFC values of 177.9±2.911, 644.1±4.202 and 96.38±3.851µg RE/mg of dry extract, respectively. Furthermore, these three extracts showed a dose dependent radical scavenging activity with IC50 concentration of 22.85, 22.15 and 23.001 µg/ml, respectively. The extracts of C. cladosporioides and A. brassicae also showed promising antimicrobial activity against Escherichia coli, Staphylococcus aureus and Bacillus subtilis with a minimum inhibitory concentration of 250μg/ml for all bacteria. Both the samples showed cytotoxic property against shrimp nauplii with LC50 of 104.2 and 125.9µg/ml, respectively. Conclusions: The crude fungal extracts obtained from endophytes: A. alternata, C. cladosporioides and A. brassicae upon purification and further identification of the bioactive compounds can be a fascinating source for novel pharmaceutical agents.


Author(s):  
Kiran Kumar D. J. ◽  
Bhanupriya H. J.

Plant-derived substances have been used since antiquity for therapeutic and other purposes. The present study was undertaken to make a comparative analysis of antimicrobial and antioxidant properties of 8 medicinal plants, which are used in traditional medicine system. Methanolic extracts of Centella asiatica (leaves), Mentha piperita (leaves), Calotropis gigantean (leaves), Vitex nigundo (leaves), Bauhinia racemosus (leaves), Emblica officinalis (fruit), Bauhinia purpurea (leaves), Asperagus racemosus (stem) were tested for anti-microbial effects using the standard Agar Diffusion method. Antioxidant attributes of above extracts were studied by testing their ability to scavenge DPPH and ABTS radicles. Our results indicated that all tested extracts, with the exception of Bauhinia purpurea, were effective against E. Coli, while Candida albicans was only sensitive to Asperagus racemosus. Among tested extracts, Bauhinia racemosus was most potent in exhibiting radical scavenging activity. Our data provides rationale for exploitation of these plants for antimicrobial and antioxidant attributes.


2014 ◽  
Vol 5 (2) ◽  
Author(s):  
Widya Lestari

Endophytic fungi are a group of fungi whose part or whole life is in living plant tissue and usually does not harm the host. Endophytic fungi generally produce secondary metabolites that have beneficial biological activities such as anti-cancer compounds, antifungi or antibacterials. Control of plant diseases caused by Rigidoporus microporus and Ganoderma boninense can be done by using biological agents, namely by using endophytic fungi. This fungus has metabolite compounds that can inhibit the growth of Rigidoporus microporus and Ganoderma boninense. This study aims to determine what content is found in endophytic mushroom extracts isolated from rubber plants (Hevea brasiliensis). Endophytic fungi are extracted by maceration using methanol and ethyl acetate solvents. The extract was tested for antifungal activity using diffusion method. The content of secondary metabolites was tested by phytochemical test. The results of endophytic fungal extraction with methanol and ethyl acetate solvents resulted in antifungal activity with inhibitory zones in a row namely WL01: 2.19 and 2.15 mm Fungi Isolates on Ganoderma boninense while 3.09 mm WL02 against Rigidoporus microporus Based on the phytochemical test results, endophytic fungal extract with methanol and ethyl acetate solvents showed the presence of alkaloid, flavonoid, phenol and saponin. Keywords: endofit, fungi, hevea, secondary metabolites


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