Isolation, Identification and Molecular Characterization of Endophytic Fungi from the leaves of Coelogyne species, and their role as an Antimicrobial agent

Author(s):  
Akila Shree J ◽  
Krishnaveni C

Endophytic fungi from Orchids believed to have an essential symbiotic relationship with the plant for both the germination of seeds and the development of young plantlets. Endophytes are microorganisms which live in the intercellular spaces of healthy host tissues without causing apparent symptoms. Endophytic fungi isolated from the medicinal plants are used for the development of drugs as they act as the source of bioactive compounds. This study has been designed to isolate the fungal endophytes from the leaves of three epiphytic orchid species (Coelogyne nervosa, A. Rich., Coelogyne mossiae, Rolf and Coelogyne cristata, Lindl) from the same genus Coelogyne, collected from Ooty flora, Coonoor. About five different endophytes were identified and their morphological characters were studied. Coelogyne cristata showed maximum colonization frequency. Among the five species, Fusarium species showed antibacterial activity against the gram-positive bacteria Bacillus Subtilis, a pathogen found in soil, water and food. Fungal genomic DNA isolated for molecular identification from the four fungal species.

2016 ◽  
Vol 14 (1) ◽  
pp. 149-156 ◽  
Author(s):  
Hoang Kim Chi ◽  
Le Huu Cuong ◽  
Tran Thi Nhu Hang ◽  
Nguyen Dinh Luyen ◽  
Tran Thi Hong Ha ◽  
...  

In recent years, a considerable number of studies on the role of microbes in agarwood production have been carried out in plants of the species Aquilaria. Based on the fact that there is a relationship between the microorganisms residing inside the plant and the agarwood formation, we isolated and characterized endophytic fungi associated with A. crassna samples collected from Southern Vietnam. Morphological identification and DNA barcoding analysis of the fungal endophytic isolates indicated that they were classified at least into three groups of diverse genera: Geotrichum, Fusarium and Colletotrichum belonging to families Dipodascaceae, Nectriaceae and Glomerellaceae, respectively.  Noteworthy, Geotrichum candium strain SHTr1 isolated from a dark colored woody sample of agarwood was able to produce a fruity odor and exhibited a slight antimicrobial activity against the test bacterium Staphylococcus aureus. Another fungal isolate, Fusarium verticillioides SHTr3’s, showed a moderate antimicrobial activity against a test Gram positive bacteria Bacillus subtillis and S. aureus with MIC values at 50 μg.mL-1. At 200 μg.mL-1, the ethyl acetate extracts of fungal isolates F. verticillioides SHTr3 and Colletotrichum truncatum SHTrHc7 were found to have comparable scavenging abilities on DPPH-free radicals with 53.87 and 71.82%, respectively. The present results contribute to a depiction of a diverse fungal endophytic community in Vietnamese agarwood plant A. crassna and provide important information for further understanding of the role of endophytic fungi in agarwood formation and therapeutic applications of host plants in general.


2020 ◽  
Vol 10 (4) ◽  
pp. 502-512
Author(s):  
Channabasava ◽  
Govindappa Melappa

Background: Many biologically important bioactive compounds have been identified in Calophyllum tomentosum extract. Hence, we aimed to identify different endophytic fungal species from different parts of the plants and their bioactive compounds. Objective: The study includes identification and characterization of bioactive compounds in the extracts of endophytic fungal species of Calophyllum tomentosum. Methods: Different parts were used to assess the different endophytic fungi. Two different solvents were used: extracts and qualitative solvents. TLC and GC-MS were employed to identify the bioactive compounds. The bioactive compounds producing endophytes were identified by using18S rRNA. Results: In total, 13 different fungal species were identified from different parts of the Calophyllum tomentosum. The C. tomentosum leaf part showed seven different fungal endophytes from the barks (four) and stems (two). The methanol extract of Fusarium species (stem), Cladosporium species (bark) and hexane extract of Alternaria species (bark) showed a higher amount of coumarins in qualitative methods and TLC. Based on the above results, for further GC-MS and molecular identification studies, we selected three endophytes. In the GC-MS analysis, the methanol extract of Fusarium species (stem) showed psoralen, and furocoumarin (imperatonin or ammidin); Cladosporium species (bark) showed coumarin (2H-1-benzopyran-2-one), coumaric acid (3-benzofurancarboxylic acid), hynecromone (coumarin 4), 4-hydroxy-9-(3-methyl-2-butyl)furo(3,2-g)chloronen-7- one; and hexane extract of Alternaria species (bark) showed coumaric acid (3-benzofurancarboxylic acid). The three fungal endophytes were identified as Fusarium equiseti, Cladosporium uredinicola and Alternaria alternata from 18S rRNA analysis. Conclusions: Based on the above results, the three endophytic fungal species confirm the presence of coumarins and therefore, can be used for the production of coumarins instead of the plant.


2002 ◽  
Author(s):  
◽  
Kumindra Devrajh Ramsunder

Fusarium species produce toxic mycotoxins that are known to exert adverse health effects in humans and animals. No attempts have been made to establish mycotoxin-producing capabilities of isolates of Fusarium species from bananas exhibiting symptoms of crown rot. Crown rot is one of the most serious post harvest problems in banana and the disease is caused by different fungal species, principally Fusarium species. Banana, which is of great economic significance in growing countries (i.e. Costa Rica, Cameroon, Ecuador) is seriously affected by crown rot and is a major cause of fruit loss


2020 ◽  
Vol 62 (1-2) ◽  
pp. 175-186
Author(s):  
M. F. Vianna ◽  
S. Pelizza ◽  
M. L. Russo ◽  
S. Rodriguez ◽  
A. C. Scorsetti

The diversity of fungal endophytes is poorly known and particularly in the case of Nicotiana tabacum, the literature is limited. The present study assessed and compared the diversity and distribution of endophytic fungi between different organs of tobacco plants. We calculated the relative frequency and rates of colonisation and of isolation of endophytic fungi in roots, stems and leaves, as well as the Shannon–Wiener and Simpson diversity indexes. Similarities between assemblages from the studied organs were also analysed. A total of 1588 endophytic fungal strains assigned to 31 morphospecies were isolated. The highest diversity of endophytes was found in leaves, being Fusarium graminearum and Alternaria botrytis the most common fungal species. This study provides information on the distribution of fungal endophytes inhabiting leaves, stems, and roots of N. tabacum and thus can serve as a starting point for increasing our comprehension on the interactions in which these fungi are involved.


Author(s):  
M. Chaithra ◽  
S. Vanitha ◽  
A. Ramanathan ◽  
V. Jegadeeshwari ◽  
V. Rajesh ◽  
...  

Aim: To identify cocoa associated endophytic fungi through morphological and molecular techniques. Place and Duration of Study: Department of Plant Pathology, TNAU, Coimbatore, Tamil Nadu from April 2018 to November 2019. Methodology: Endophytic fungal isolates were isolated from different parts of cocoa using Petrini method. Isolated endophytic fungal strains were grown in Malt extract broth and total genomic DNA was isolated and amplified using universal primers ITS1F and ITS4R. Amplified rDNA was visualized and documented. Results: Morphological and molecular characterization of six endophytic fungi revealed that they are from four different taxa viz., Lasiodiplodia pseudotheobromae PAK-7, Arthrinium rasikravindrae P11, Arthrinium rasikravindrae P12, Diaporthe sp. Vef-3, Lasiodiplodia theobromae TN-R-3,  Colletotrichum sp. TN-9-2 belonging to four different families viz., Botryosphaeriaceae, Apiosporaceae, Diaporthaceae and Glomorellaceae under Phylum Ascomycota. Conclusion:  The present study indicates the distribution and diversity of fungal endophytes in different plant parts of the cocoa tree in south India.


2006 ◽  
Vol 52 (5) ◽  
pp. 427-435 ◽  
Author(s):  
Mysore V Tejesvi ◽  
Basavanna Mahesh ◽  
Monnanda S Nalini ◽  
Harishchandra S Prakash ◽  
Kukkundoor R Kini ◽  
...  

Endophytic fungi represent an interesting group of microorganisms associated with the healthy tissues of terrestrial plants. They represent a large reservoir of genetic diversity. Fungal endophytes were isolated from the inner bark segments of ethnopharmaceutically important medicinal tree species, namely Terminalia arjuna, Crataeva magna, Azadirachta indica, Holarrhena antidysenterica, Terminalia chebula, and Butea monosperma (11 individual trees), growing in different regions of southern India. Forty-eight fungal species were recovered from 2200 bark segments. Mitosporic fungi represented a major group (61%), with ascomycetes (21%) and sterile mycelia (18%) the next major groups. Species of Fusarium, Pestalotiopsis, Myrothecium, Trichoderma, Verticillium, and Chaetomium were frequently isolated. Exclusive fungal taxa were recovered from five of the six plant species considered for the study of endophytic fungi. Rarefaction indices for species richness indicated the highest expected number of species for bark segments were isolated from T. arjuna and A. indica (20 species each) and from C. magna (18 species).Key words: bark, endophytes, medicinal trees, southern India.


Author(s):  
Biplab Bagchi ◽  

Aerial tissues of the woody lianas- Combretum sp. was selected and assessed for study of endophytic fungal diversity from three forest areas of West Medinipur and Jhargram districts of West Bengal. In monsoon study it was observed that out of 225 various tissue segments ,165 segments had been colonized by endophytic fungi Combretum roxburghii and 209 endophytic isolates were isolated from them. Average colonization frequency (CF)was 73.32 percent. Highest CF was in plant of Belpahari (76%) and in petiole (82.66%). Out of isolated fungal genera Pestalotiopsis sp. was highest in number. Diplodia sp., Beltrania sp., Chaetomium sp., Fusarium sp., Arthrinium sp. were also greater in number. Dominance index was highest in Belpahari (0.1547). Simpson’s diversity was maximum in Godapiasal (0.8693) and Shannon-Wiener index was also highest (2.152) in plant of Godapiasal. Highest diversity of endophytic fungi was in plants of Chilkigarh. Isolated endophytic fungi were Lasiodiplodia sp., Diplodia sp., Fusarium sp., Chaetomium sp., Arthrinium sp., Aspergillus sp., Pestalotiopsis sp. etc. Simpson’s diversity and Shannon-Wiener index were maximum in plants of Godapiasal. The study of antimicrobial activity of isolated fungal endophytes indicated that few plugs showed antimicrobial activity against few pathogenic bacteria. Among all isolates tested only seven showed antimicrobial activity. Aspergillus sp. and one unidentified fungi showed the antimicrobial activity against three bacteria i. e., Bacillus cereus, Escherechia coli and Vibrio cholera and maximum inhibition zone was observed against E. coli, diameter of inhibition zone is 1.4 cm. Other fungi showed less antimicrobial activity. Diameter of zone of inhibition varied (0.7 – 1.4) and this variation might be due to the difference of bioactive compound produced or the varied concentrations or amount degree of same compound.


2021 ◽  
Vol 18 (2) ◽  
pp. 263-269
Author(s):  
Jawed Shaikh ◽  
Ashfaque Mehboob Khan ◽  
Mirza Mushtaq Vaseem Baig

Fungal endophytesare colonized in different part of the plants and play important role in survival of plants in stressful habitat. In search of potential endophytic fungito produce bioactive metabolites inthis study we investigate thediversity of endophytic fungi associated with leaves of the Acacia nilotica plant. Twenty-six endophytic fungi were subjected to morphologicaland molecular identification with internal transcribes spacer (ITS) region sequenced. All 26 endophytic fungi were divided into nine genera Chaetomium, Amesia, Ovatospora, Penicillium, Phialemonium, Colletotrichum, Crinipellis, Acrophialophora, Cribbea. Most of them belonged to the phylum Ascomycota only one belonging to the phylum Basidiomycota. This study shows that Acacia leaves inhabitant by diverse group of endophytic fungi. The biodiversity analysis showed Chaetomium sp. Being dominant with the highest colonization frequency (26.9%). One of the Chaetomium sp. showed sequence similarity of93% with the species reported earlier, Further investigationsarein needed to harness the bioactive compounds.


2018 ◽  

<p>Plant-associated microorganisms, especially endophytic fungi, represent an untapped resource for the discovery of biologically active natural products. The objectives of this study were to isolate, identify endophytic fungi, and produce their bioactive metabolites from the leaves of two varieties of Egyptian artichoke namely: French Hyrious and Egyptian Baladi. In addition, assess of their total antioxidant capacity (TAC), total phenolic content (TPC) and total flavonoid content (TFC). The results of this novel study show a total of 35 endophytic fungal species belonging to 14 genera were isolated from both artichoke leaves with gross total counts of colonizing endophytic fungi ranged from 71 to 123 cfu which is matching 78.89% to 136.67% of colonization frequency. All taxa recovered were assigned to Ascomycetes. In addition, there is high species richness and diversity indices of endophytic filamentous fungi in the leaves Baladi Artichoke as compared to its French rival. &nbsp;<em>Alternaria alternata</em> were found to be the most frequently isolated dominant species. The TAC, TPC and TFC of the fungal cultures ranged from 163 to 681 mgAAE/gDW, 10.38 to 40.30 mgGAE/ gDW, and 13.92 to 173.55 mgQE/gDW, respectively. Furthermore, LC-ESI-MS/MS confirmed the presence of 1,3-dicaffeoylquinic acid and 1,5-dicaffeoylquinic acid in the methanolic extract of <em>A. alternata.</em> Hence, this novel study suggested that the metabolites produced by endophytic fungi associated with Egyptian artichoke could be explored as an economic and potential natural resources with diverse pharmaceutical and biological activities.</p>


2015 ◽  
Vol 2015 ◽  
pp. 1-8 ◽  
Author(s):  
Yougen Wu ◽  
Sisay Girmay ◽  
Vitor Martins da Silva ◽  
Brian Perry ◽  
Xinwen Hu ◽  
...  

We hypothesize that the fungal endophytes of noni may possibly play a role in its overall pharmacological repertoire, especially since the perceived efficacy of the fruit in ethnomedicinal use is associated with the fermented juice. The foremost goal of this study is to explore the role of endophyte-derived secondary metabolites in the purported anticancer properties of noni. To that end, culturable endophytic fungi resident within the healthy leaves and fruit of the plant were isolated and identified by molecular sequence analysis of the 5.8S gene and internal transcribed spacers (ITS). Purified organisms were subjected toin vitrofermentation in malt extract broth for 8 weeks under anaerobic conditions at room temperature (25°C), in order to simulate the conditions under which traditional fermented noni juice is prepared. The cytotoxic potential of organic extracts derived from the fermented broths of individual endophytes was then tested against three major cancers that afflict humans. Twelve distinct endophytic fungal species were obtained from the leaves and 3 from the fruit. Three of the leaf endophytes inhibited the growth of human carcinoma cell lines LU-1 (lung), PC-3 (prostate), and MCF-7 (breast) with IC50values of ≤10 μg/mL.


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