scholarly journals Chaenomeles speciosa fruit endophytic fungi isolation and characterization of their antimicrobial activity and the secondary metabolites composition

Author(s):  
Yurii V. Lykholat ◽  
Nina O. Khromykh ◽  
Oleh O. Didur ◽  
Oksana A. Drehval ◽  
Tetyana V. Sklyar ◽  
...  

Abstract Background Endophytes promote the survival of the host plants affected by unfavorable environment. To confirm the endophytes role in Chaenomeles speciosa pathogenic resistance, fungal isolates were derived from both fruit peel and pulp, and presumptively identified using macroscopic and microscopic techniques. Antifungal potential of the isolates was assayed by dual culture method and agar disc method against Alternaria alternata, Fusarium culmorum, and Fusarium oxysporum strains. Two most active fungal isolates were cultured in Czapek's liquid medium to obtain metabolites. The crude ethyl acetate extracts of metabolites were characterized for antibacterial activity against Basillus subtilis and Erwinia dissolvens, and for component composition by GC–MS technique. Results Nine fungal endophytic isolates were derived from the peel and pulp of C. speciosa fruits and tentatively attributed to Penicillium spp. (seven isolates), Aspergillus spp. (one isolate), Mucor spp. (one isolate). Two fungal isolates, one each of the fruit peel and pulp, were designated as Penicillium sp. I and Penicillium sp. II respectively, and selected for further research. Both isolates showed similar activity against A. alternata; however, Penicillium sp. I activity against F. culmorum and F. oxysporum exceeded the activity of Penicillium sp. II. Cultural medium ethyl acetate extracts of both endophytes exhibited higher antibacterial activity against Gram-positive B. subtilis, while mycelium extracts were more active against Gram-negative E. dissolvens. In general, Penicillium sp. I antibacterial activity was higher in cultural medium extracts, while activity of Penicillium sp. II dominated in mycelium extracts. GC–MS analysis of the fungal metabolites component composition revealed the identity of 27 and 17 compounds, respectively in the ethyl acetate extracts of Penicillium sp. I and Penicillium sp. II cultural medium. Basic compounds produced by the first isolate, were represented by 3-Furanacetic acid, 4-hexyl-2,5-dihydro-2,5-dioxo, Diisooctyl phthalate, 11-Hexadecyn-1-ol, and Propanedioic acid, dihydroxy. At the same time, Phthalic acid diisooctyl ester and other phthalates constituted the main part of the second isolate metabolites, followed by Hexadecanoic acid, Eicosyl isopropyl ether, and 4-Butoxy-2-butanone at a lower content. Conclusions The findings showed that the antimicrobial potential of Chaenomeles fruits endophytic fungi is promising and deserves further investigation.

2020 ◽  
Vol 55 (3) ◽  
pp. 207-214
Author(s):  
MM Haque ◽  
N Sultana ◽  
SMT Abedin ◽  
N Hossain ◽  
SE Kabir

The fatty acid analysis and biological activity of n-hexane, dichloromethane, ethyl acetate and methanol extracts of Nyctanthes arbor-tristis L. flowers are reported. Five fatty acids namely palmitic (44.15%), stearic (19.34%), arachidic (15.06%), behenic (9.77%) and lignoceric (11.69%) acids were identified. From cytotoxicity test, the LC50 values (the median lethal concentration) for n-hexane, dichloromethane, ethyl acetate and methanol extracts as well as for standard vincristine sulphate were found 7.05, 4.67, 3.14, 5.53 and 0.50 μg/ml, respectively. Antibacterial activity results of different extracts were compared with standard antibiotic ciprofloxacin and tetracycline by measuring the zone of inhibition diameter in millimeter. The dichloromethane and ethyl acetate extracts showed significant antibacterial activity. From antioxidant activity test, IC50 values (50% inhibitory concentration) of n-hexane, dichloromethane, ethyl acetate, methanol extracts and ascorbic acid were found to be 291.92 mg/ml, 45.74 μg/ml, 21.86 μg/ml, 64.30 μg/ml and 3.98 μg/ml, respectively. Bangladesh J. Sci. Ind. Res.55(3), 207-214, 2020


2016 ◽  
Vol 11 (9) ◽  
pp. 1934578X1601100
Author(s):  
Sakon Monggoot ◽  
Jariya Burawat ◽  
Patcharee Pripdeevech

A total of 17 endophytic fungal isolates were obtained from the leaves of Mentha cordifolia Opiz (Lamiaceae). Seven isolates were identified to the level of genus by using taxonomically relevant morphological traits. Colletotrichum and Phomopsis species were dominant among these strains. All strains were separated from M. cordifolia leaf for the first time. The ethyl acetate extracts of all endophytic fungi were tested for antibacterial activity against Salmonella typhimurium TISTR1166 and Pseudomonas aeruginosa TISTR781. Most endophytes exhibited antibacterial activity. Ustilago sp. MFLUCC15-1024 presented the highest inhibition zone diameter with a MIC of 31.25 μg/mL against the tested pathogens. The chemical composition of the ethyl acetate extract of this strain was investigated using gas chromatography-mass spectrometry. Twenty-one components were identified. 2-Phenylethanol (38.7%), E-ligustilide (12.4%), α-eudesmol (10.2%), β-vetivone (4.6%), β-ylangene (3.7%) and verbanol (3.4%) were the major components of the extract. The strong antibacterial activity of Ustilago sp. MFLUCC15-1024 ethyl acetate extract may be attributed to the presence of a high concentration of bioactive compounds including phenyl ethyl alcohol, E-ligustilide and α-eudesmol. The results indicate that there is high diversity of endophytic fungi in M. cordifolia leaf, and that Ustilago sp. MFLUCC15-1024 strain could be an excellent resource of natural antibacterial compounds.


2018 ◽  
Vol 1 (2) ◽  
Author(s):  
A. Kwaji ◽  
H. M. Adamu ◽  
I. Y. Chindo ◽  
R. Atiko

The present study is aimed at the isolation and characterization of betulin from Entada africana.  A dichloromethane soluble portion of the stem bark methanol/acetone (1:1 v/v) extract was subjected to gradient elution using ethyl acetate in hexane (5 – 30 %) on an open column. A pure compound was obtained with Rf = 0.61 in hexane/ethyl acetate (8:2 v/v) after repeated washing and recrystallization from methanol and coded Enac1. The pure compound was analyzed using IR, 1H & 13C NMR and GC-MS. Clinical isolates of Escherichia coli, Klebsiella pneumoniae, Salmonella typhi and Staphylococcus aureus were used to assess the antibacterial activity of the pure compound while its preliminary Cytotoxicity was evaluated using brine shrimp nauplii. Based on the spectroscopic data obtained and in comparison with literature, the isolated compound was identified as betulin. The minimum inhibitory concentration (MIC) of betulin ranged between 62.50 - 250.00 µg/mL for all the four bacterial isolates in the study while its fifty percent lethal concentration (LC50) was 10.00 µg/mL. Significant Cytotoxicity with moderate antibacterial activity was observed. The study therefore justifies the existence of bioactive compounds in the stem bark of Entada africana and its use in traditional medicine.


2020 ◽  
Vol 6 (1) ◽  
pp. 26-32
Author(s):  
Mulya Sari ◽  
Nampiah Sukarno ◽  
Irmanida Batubara ◽  
Rohani Cinta Badia Br Ginting

Endophytic fungi isolated from red ginger (Zingiber officinale) can inhibit growth of Fusarium oxysporum, but the ability of the fungi to control Candida albicans  as human pathogen has not been reported. The aims of this research were to study the mechanism of ten endophytic fungi isolates derived from red ginger to control C. albicans in vitro using dual culture methode and fungal extract, and to determine fungal bioactive chemical groups produced by the fungi. Three out of ten isolates tested, Talaromyces assiutensis JMa 7, T. assiutensis JMbt 3, and Curvularia affinis JMbt 9 inhibited growth of C. albicans with inhibition zones were 4.0 mm, 4.9 mm, and 11.3 mm, respectively. The cultures of the three potential endophytic fungi were extracted by maceration method using 3 solvents i.e ethyl acetate, n-hexane and ethanol. The three isolates were grown in PDB separately for 21 days incubation. At harvest, the culture filtrate was extracted by ethyl acetate and n-hexane, while fungal mycelia were extracted by all the three solvents. Ethyl acetate extracts obtained from culture filtrate of all the three fungal isolates consistently inhibited C. albicans with inhibition zones were 2.0-3.8 mm. For n-hexane extract, however, only Talaromyces assiutensis JMbt 3 that had positive effect with inhibition zone was 2.0 mm. All extracts from mycelia did not have any effects on C. albicans. The ethyl acetate extract of T. assiutensis JMbt 3 was analysed to determine its chemical groups using visible color on thin layer chromatography (TLC). The results showed that the bioactive compounds was terpenoids, and antioxidant.


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


Author(s):  
Nenengsiti Silfi Ambarwati ◽  
Amarila Malik ◽  
Ageng Tri Listari ◽  
Nirwana Nirwana ◽  
Berna Elya ◽  
...  

Objectives: The emergence of new infections and increase in bacterial drug resistance has created a serious need for the expansion of new antibacterial agents from natural sources. The study was carried out to evaluate the antibacterial activity of fractions of ethyl acetate extract of Garcinia latissima Miq. fruits.Methods: The fractionation was done using a silica gel column and organic solvents as the eluent, i.e., n-hexane, ethyl acetate, and methanol. All fractions were assayed for antibacterial activity, which was done by performing disc diffusion for growth inhibition against Bacillus subtilis and Pseudomonas aeruginosa. In addition, the growth inhibition activity was also examined by performing bioautography assay using pre-coated silica gel 60 GF 254 plates as the stationary phase. Fractions A-F were eluted using n-hexane:chloroform (1:4), while Fractions G-K were used ethyl acetate:dichloromethane (4:1) as the mobile phase. The plate was visualized by ultraviolet at λ 254 nm and 366 nm, while the other one was contacted with the inoculated agar medium to observe zone inhibition. Further, the minimum inhibitory concentration (MIC) value was determined by performing microdilution.Results: The result showed that the antibacterial activity of all fractions was more active at inhibiting the growth of B. subtilis than P. aeruginosa, mainly for Fractions H and J. However, the strongest antibacterial activity was showed by Fractions H and J against B. subtilis, MIC 312.5 μg/mL (lower than reference, which is erythromycin antibiotic (25 μg/mL), followed by Fraction D against B. subtilis MIC 625 μg/mL, Fraction K against P. aeruginosa MIC 625 μg/mL, whereas Fractions C, E, and G against B. subtilis, and Fraction E against P. aeruginosa also showed low MIC values (1.250 μg/mL).Conclusions: The results indicated that fractions of G. latissima Miq. fruit ethyl acetate extracts possessed antibacterial activity. The most active fraction that inhibited the growth of B. subtilis was shown by Fractions H and J; these fractions have the potential to be developed as new antibacterial agents.


2021 ◽  
Vol 21 (1) ◽  
Author(s):  
Ziling Mao ◽  
Weihao Zhang ◽  
Chunyin Wu ◽  
Hao Feng ◽  
Yuanhang Peng ◽  
...  

Abstract Background Eucalyptus bacterial wilt caused by Ralstonia solanacearum is an important eucalyptus disease. Endophytic fungi, an important source of natural active substances, provide a new breakthrough for the control of plant diseases. Results In the present study, 80 endophytic fungal isolates were obtained from the healthy branches and fruits of Eucalyptus exserta. Fifteen distinct isolates (MK120854-MK120868) were selected for further taxonomic identification through morphological trait assessments and internal transcribed spacer (ITS) region-rRNA gene sequence analysis. Thirteen genera, namely, Phyllosticta, Penicillium, Eutypella, Purpureocillium, Talaromyces, Lophiostoma, Cladosporium, Pestalotiopsis, Chaetomium, Fusarium, Gongronella, Scedosporium and Pseudallescheria, were identified on the basis of their morphological characteristics. Members of the genus Phyllosticta were the primary isolates, with a colonization frequency (CF) of 27.5 %. Most of the fungal isolates displayed antibacterial activity. The crude extracts obtained from Lophiostoma sp. Eef-7, Pestalotiopsis sp. Eef-9 and Chaetomium sp. Eef-10 exhibited strong inhibition on the test bacteria, and Lophiostoma sp. Eef-7 was further cultured on a large scale. Three known compounds, scorpinone (1), 5-deoxybostrycoidin (2) and 4-methyl-5,6-dihydro-2 H-pyran-2-one (3), were isolated from the endophytic fungus Lophiostoma sp. Eef-7 associated with E. exserta. The structures of these compounds were elucidated by analysis of 1D and 2D NMR and HR-ESI-MS spectra and a comparison of their spectral data with published values. Compounds 1 and 2 showed weak antimicrobial activity against Ralstonia solanacearum. Conclusions Endophytic fungi from Eucalyptus exserta may represent alternative sources of antimicrobial agents. Lophiostoma sp. Eef-7 can produce 2-azaanthraquinone derivatives and shows weak antibacterial activity against Ralstonia solanacearum.


2019 ◽  
Vol 2 (1) ◽  
pp. 07-12
Author(s):  
Sony Eka Nugraha ◽  
Suryadi Achmad ◽  
Erly Sitompul

North Sumatera is one of the central areas of purple passion fruit production in Indonesia. Processing passion fruit into beverage products (passion fruit juice) produces peel  has not been utilized. The use of passion fruit skin needs to be studied so that it can be useful as a raw material for antibacterial drug preparations. The aim of this study was to determine the phytochemical constituent screening  and antibacterial activity of ethyl acetate fraction of purple passion fruit peel against Staphylococcus aureus and Escherichia coli. Simplicia and ethyl acetate fraction were determinated  its phytochemical properties.  The extraction process by percolation method using ethanol  96% and continue to fractionation process by liquid liquid extraction method using n-hexane and ethyl acetate. The antibacterial activity were tested  against Staphylococcus aureus and Escherichia coli using agar diffusion method with paper discs. The result showed  that  the simplicia characteristic of passion fruit peel  were water content of  8.64%, water soluble extract of  31.69%, ethanol soluble extract of 13.02%, ash total of 7.89%, and insoluble ash in acid of 0.816%. The phytochemical screening simplicia and ethyl acetate fraction showed the presence of flavonoids, glycosides, saponins and tannins. The antibacterial activity test showed that the ethyl acetate fraction has an effective inhibition at the concentration of 100 mg/ml against Staphylococcus aureus and Escherichia coli, it showed dose dependent manner. The ethyl acetate fraction of passion fruit peel (Passiflora edulis Sims) has an antibacterial activity  on Staphylococcus aureus and Escherichia coli


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