A New RoVe Method to Purify Endophytic Fungi from Medicinal Plants Western Ghats of Karnataka India

Author(s):  
Ankala Basappa Vedamurthy ◽  
Rohit Shankar Mane

Endophytic fungi from medicinal plants were hardly ever reported as compared to those from soil and marine sources. The present results associate the presence of endophytic fungi in medicinal plants by isolating them from three medicinal plants i.e. Bombax ceiba, Aloe vera, and Ximenia americana. In the present research investigation, surface sterilization method and media were standardized, and 32 endophytic fungi were isolated from three medicinal plants. We assessed the competence of three different surface sterilization methods and four media for isolation of endophytic fungi. The RoVe method used was more effective in eliminating epiphytic microorganisms. Therefore by using new method we have isolated total of 32 fungal endophytes and those were belongs to Aspergillus nomius (63.20%), Aspergillus niger (41.60%), Thielaviopsis basicola (38.33%), Fusarium oxysporum (33.20%), Pestalotiopsis inflexa (27.20%), Nigrospora sphaerica (20%), Alternaria alternata (15.30%) and Phomopsis archeri (20%). This is the first report of successful isolation of endophytic fungi from the said medicinal plants, for using newly formulated surface sterilization method.

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 140 ◽  
Author(s):  
Ahmed Mohamed Aly Khalil ◽  
Saad El-Din Hassan ◽  
Sultan M. Alsharif ◽  
Ahmed M. Eid ◽  
Emad El-Din Ewais ◽  
...  

Endophytic fungi are widely present in internal plant tissues and provide different benefits to their host. Medicinal plants have unexplored diversity of functional fungal association; therefore, this study aimed to isolate endophytic fungi associated with leaves of medicinal plants Ephedra pachyclada and evaluate their plant growth-promoting properties. Fifteen isolated fungal endophytes belonging to Ascomycota, with three different genera, Penicillium, Alternaria, and Aspergillus, were obtained from healthy leaves of E. pachyclada. These fungal endophytes have varied antimicrobial activity against human pathogenic microbes and produce ammonia and indole acetic acid (IAA), in addition to their enzymatic activity. The results showed that Penicillium commune EP-5 had a maximum IAA productivity of 192.1 ± 4.04 µg mL−1 in the presence of 5 µg mL−1 tryptophan. The fungal isolates of Penicillium crustosum EP-2, Penicillium chrysogenum EP-3, and Aspergillus flavus EP-14 exhibited variable efficiency for solubilizing phosphate salts. Five representative fungal endophytes of Penicillium crustosum EP-2, Penicillium commune EP-5, Penicillium caseifulvum EP-11, Alternaria tenuissima EP-13, and Aspergillus flavus EP-14 and their consortium were selected and applied as bioinoculant to maize plants. The results showed that Penicillium commune EP-5 increased root lengths from 15.8 ± 0.8 to 22.1 ± 0.6. Moreover, the vegetative growth features of inoculated maize plants improved more than the uninoculated ones.


2021 ◽  
Author(s):  
Mohammad Magdy El-Metwally ◽  
Atia Mohamed Eisa ◽  
Amal AI Mekawey ◽  
Samy F. Mahmoud ◽  
Yasser El Halmouch

Abstract One of hidden mine of antibiotics is endophytic fungi especially that inhibited medicinal plants. In this regard, leaves, stems, fruits and bulbs of some commonly medicinal plants growing in Northwestern coast of Egypt were subjected for isolation of endophytic fungi with screening study of its antimicrobial activity. Practically, more than one hundred (101) endophytic fungal species isolated from Scorpiurus muricatus, Mellilotus indicus, Lotus polyphyllos, Ononis vaginalis, Nicotiana glauca, Lycium europaeum, Asphodelus aestivus, Echium angustifolium,Fagonia cretica, Pancratium maritimum,and Carduus getulus were tested in vitro for their antimicrobial activities against E. coli, Pseudomonas argenosa, Staphylococcus aureus, Bacillus subtilus, Candida albicanus, Candida glabrata, Penicillium expansum, Aspergillus flavus. The values of applied diversity indices revealed significant differences in presence, absence and abundance among endophytic fungal isolates. To the best of our knowledge, the present study is the first to report of Alternaria pluriseptata as endophytic species with most dominant and most active in its anitimicrobial activity among the isolated species. Seven distinctive groups were revealed from the two-way cluster analysis showing the intensity of antimicrobial activity against tested pathogens: Twenty-five percent of the isolates (26 strains assembled together in group V) exhibited no antimicrobial activity against all tested pathogens while six percent (6 isolates) assembled in group VII revealed high antimicrobial activity against five pathogens.


2018 ◽  
Vol 10 (4) ◽  
pp. 1286-1290
Author(s):  
Sushma K.S ◽  
Jayashankar M. ◽  
Mohammed Ali Saeed ◽  
Vinu A.K.

Twenty fungal endophytes were isolated from different medicinal plants of Biligirirangana Hill, Chamarajanagar dist. Phytochemical analysis of ethyl acetate extract showed positive results for tanins, flavonoids, steroids, terpenoids, glycosides, saponins and alkaloids except saponin and flavonoid tests. The crude extracts of the fungal endophytes were tested against two gram positive and two gramnegetive bacteria for its antibacterial activity. The highest zone of inhibition was produced by Fusarium sps (AB9)35mm. All the crude extracts were found to be effective against Escherichia coli, Bacillus subtilis and Staphylococcus aureus rather in Pseudomonas aerogenosa. Among them the highest inhibition zone was produced in Penicillium sp (AB11) 24mm, Cladosporium sp(AB3) 21mm, and Aspergillus sp(AB12) 30mm. Therefore, endophytic fungi can be a good source to inhibit the growth of harmful pathogens.  


2021 ◽  
Vol 24 (2) ◽  
pp. 66
Author(s):  
Indriati Ramadhani ◽  
Hasnadiazahra Rohadi ◽  
Yeni Yuliani ◽  
Muhammad Ilyas

An assemblage of endophytic fungi was isolated from Moringa oleifera Lam. collected from Lombok island, West Nusa Tenggara Province, Indonesia. Fungal endophytes were isolated using surface sterilization methods with slight modification. Forty-six selected endophytic fungal strains were isolated from the leaves, petioles, and stems of M. oleifera. The fungal strains identification through morphological observation and ITS rDNA-based molecular analysis showed that fungal endophytes were associated with host plants belonging to the taxa Alternaria, Cladosporium, Colletotrichum (Glomerella), Corynespora, Curvularia (Cochliobolus), Fusarium, Mucor, Ochrocladosporium, Phomopsis (Diaporthe), and Trametes. In this study, endophytic Phomopsis dominating the obtained strains, whereas 26% (12/46) strains were isolated from host plant samples.


2021 ◽  
Vol 12 (6) ◽  
pp. 7584-7595

Medicinal plants are a promising host for endophytic fungi to produce secondary metabolites relevant for food and health. In this study, we evaluate antioxidant activity to determine the species of endophytic fungi isolated from Syzygium aqueum. Endophytic fungi were isolated from leaf stalks through surface sterilization. The fungi’s isolate was identified with morphology and molecular analysis (ITS-rDNA). The pure fungi strain was cultivated on PDB media for 4 weeks, and metabolites were extracted using ethyl acetate. The crude extract of endophytic fungi was examined for its antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH). The pure compound was isolated using the chromatography method, and its structure was determined using spectroscopy analysis involving NMR 1D and 2D. In total, four obtained endophytic fungi were isolated from leaf stalks. The fungi with good antioxidant activity (IC50 59.2 µg/mL) were identified as Beltrania rhombica. The characteristics of the pure compound are white-yellowish powder with IC50 44.2 µg/mL. Based on spectroscopy analysis, the pure compound was identified as 3-(hydroxyl(3,4,5-trihydroxylphenyl)methyl)-3,4-dihydro-2H-pyran-4,5,6-triol.


2021 ◽  
Vol 948 (1) ◽  
pp. 012037
Author(s):  
N D Rahayu ◽  
N Sukarno ◽  
S Listiyowati ◽  
M Rafi ◽  
S Mursidawati ◽  
...  

Abstract Geodorum densiflorum is an ornamental and medicinally important orchid. The medicinal metabolites are produced by endophytic fungi associated with orchid tissue, particularly flowers. However, there is no report of the endophytic fungi from the G. densiflorum flowers. This research aimed to study the endophytic fungi from different parts of G. densiflorum flower. Fungal isolation was carried out from sepal, petal, labium, stigma, and anther using surface sterilization method. The fungi were identified by combining morphological and molecular characteristics of ITS rDNA region. The results showed that each flower organ had different species of endophytic fungi. A total of seven species of endophytic fungi were obtained; four species were successfully identified by molecular identification and three species based on morphology. The four species, namely Hypomontagnella barbarensis, Aspergillus oryzae, Curvularia pseudobranchyspora, and Nigrospora chinensis, while the three species, namely Gonatobotrys sp., Humicola sp., Aspergillus section Nigri. The labium inhabited by Curvularia pseudobranchyspora, Nigrospora chinensis, Aspergillus section Nigri. Three species isolated from petals, namely Hypomontagnella barbarensis, Gonatobotrys sp., and Aspergillus oryzae. The sepal is inhabited by Humicola sp. There were no endophytic fungi in stigma and anther. This indicates that each flower part is a unique habitat of endophytic fungi.


PeerJ ◽  
2020 ◽  
Vol 8 ◽  
pp. e8309 ◽  
Author(s):  
Sareh Hatamzadeh ◽  
Kamran Rahnama ◽  
Saeed Nasrollahnejad ◽  
Khalil Berdi Fotouhifar ◽  
Khodayar Hemmati ◽  
...  

L-asparaginase is an important anticancer enzyme that is used in the first line treatment of acute lymphoblastic leukemia. This study was conducted to isolate L-asparaginase-producing endophytic fungi from medicinal plants of family Asteraceae. Seven healthy medicinal plants from family Asteraceae were selected for the isolation of endophytic fungi using standard surface sterilization techniques. A total of 837 isolates belonging to 84 species were comprised of the stem (55.6%), leaf (31.1%), root (10.6%) and flower (2.7%). Initial screening of L-asparaginase-producing endophytes was performed by qualitative plate assay on modified Czapex dox’s agar medium. L-asparaginase activity of fungal endophytes was quantified by the nesslerization method. Identification of endophytic fungi was performed using both morphological characteristics and phylogenetic analyses of DNA sequence data including ribosomal DNA regions of ITS (Internal transcribed spacer) and LSU (partial large subunit rDNA), TEF1 (Translation Elongation Factor) and TUB (β-tubulin). Of the 84 isolates, 38 were able to produce L-asparaginase and their L-asparaginase activities were between 0.019 and 0.492 unit/mL with Fusarium proliferatum being the most potent. L-asparaginase-producing endophytes were identified as species of Plectosphaerella, Fusarium, Stemphylium, Septoria, Alternaria, Didymella, Phoma, Chaetosphaeronema, Sarocladium, Nemania, Epicoccum, Ulocladium and Cladosporium. This study showed that endophytic fungi from Asteraceae members have a high L-asparaginase-producing potential and they can be used as an alternative source for production of anticancer enzymes.


2020 ◽  
Author(s):  
Neha Kapoor ◽  
Abu Mushtaque ◽  
Lokesh Gambhir

Abstract Endophytic fungi have been in the spotlight as a reservoir of novel agents with diverse bioactivities. Similarity in chemical diversity with the host plant makes them an amenable target for industrial interventions. A wide range of compounds as secondary metabolites and enzymes are manufactured inside the endophytic fungal factory. However, utilization of endophytic fungi as industrially imperative enzyme producers has been a scarce event. The present study was conducted to bio-prospect the fungal endophytes present in the high altitude medicinal plants of Uttarakhand, as industrially imperative enzyme producers. A total of 58 different endophytic isolates were obtained from Pinus sabiniana, Cinnamomum tamala, Cinnamomum verum, Ocimum tenuiflorum and Rhododendron arboreum. Endophytic fungal colonization was highest, 31%, in Pinus sabiniana. The pure isolates were further explored for the production of amylases, cellulases, proteases and L-asparaginase. Out of 58 isolates, 40 isolates exhibited potent enzyme productivity. #7PSSTB isolate was considered as superlative contender on account of its relatively higher production of all the three enzymes viz. amylases, cellulases, proteases. Partial purification of #7PSSTB extract showed compelling enzymatic activity corroborating the existence of exogenous enzyme in the extract. Interestingly, #9 RASTB, #11 RASTB and #17 RASTB exhibited the production of therapeutically imperative L-asparaginase enzyme. The present study puts the spotlight on endophytic diversity in the high altitude medicinal plants as a source of enzymes of industrial interest. Production of L-asparaginase paves the way of pharmaceutical intervention to explore anti-oncogenic effects in the endophytic fungal repository of high altitude regions.


2019 ◽  
Vol 20 (5) ◽  
Author(s):  
NANI RADIASTUTI ◽  
DWI NINGSIH SUSILOWATI ◽  
HUSHSHILA A BAHALWAN

Abstract. Radiastuti N, Bahalwan HA, Susilowati DN. 2019. Phylogenetic study of endophytic fungi associated with Centella asiatica from Bengkulu and Malaysian accessions based on the ITS rDNA sequence. Biodiversitas 20:  1248-1258. Centella asiatica is one of the medicinal plants which is known to be symbiotic with various endophytic fungi. The purpose of this research was to determine diversity of culturable fungal endophyte from C. asiatica. Identification was conducted using molecular phylogenetic analysis based on the ITS rDNA sequence. The result showed that obtained 145 isolates endophytic fungi (from stolons, leaves, roots, petioles) were grouped into 18 morphotaxa (Bengkulu) and 23 morphotaxa (Malaysian). The fungal endophytic were identified as Aspergillus austroafricanus, Aspergillus oryzae, Acrocalymma vagum, Ceratobasidium cornigerum, Ceratobasidium sp., Chaetomium globosum, Colletotrichum karstii, C. gigasporium, C. tabaci, Colletotrichum siamense, Eutypella sp., Fusarium solani, F. oxysporum, F. falciforme, F. keratoplasticum, F. striatum, Fusarium sp., Penicillium capsulatum, Phoma multirostrata, Perenniporia tephropora, Perenniporia sp., Phanerochaete chrysosporium, Phanerochaete stereoides, Phyllosticta capitalensis, Phomopsis asparagi, Peroneutypa scoparia, Phialemoniopsis sp., Mycochaetophora gentinae, Talaromyces sp., Earliella scabrosa, and Trichaptum sp. based on molecular phylogenetic analysis. Fusarium (Nectriaceae) were the most found of fungal endophytes in both C. asiatica Bengkulu and Malaysian accession. The majority strain are including Ascomycetes and Basidiomycetes. The identification of endophytic fungi from medicinal plants is needed as a preliminary study to determine the potential of endophytic fungi producing bioactive compounds.


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