Aims: The present study was carried out to determine the diversity of endophytic fungi that colonize the leaves of Psidium guajava, and to evaluate their antagonistic activity against Fusarium oxysporum f.sp. cubense and Mycosphaerella fijiensis which are the two main phytopathogens of banana plants. Place and Duration of Study: The research was carried out at Microbiology Laboratory, Faculty of Sciences, University of Yaoundé I and Microbiology Laboratory, Faculty of Biotechnologies, University of Agronomic Sciences and Veterinary Medicine Bucharest, between April 2018 and February 2020. Methodology: Fragments of surface sterilized leaves of Psidium guajava were inoculated on Potato Dextrose Agar supplemented with chloramphenicol. The isolated and purified endophytic fungi were identified based on their macroscopic and microscopic characters using a mycological atlas as guide. The non-sporulating isolates were identified by comparing the ITS regions of their DNA to those of known fungi registered in the GenBank database. The antagonistic activity of the endophytic fungi isolated against Fusarium oxysporum and Mycosphaerella fijiensis was screened using dual culture method. Results: A total of 28 endophytic fungal were isolated from the leaves of Psidium guajava corresponding to a colonization frequency of 33.33%. These isolates were identified as: Aspergillus sp., Botryosphaeria sp., Fusarium sp., Neoscytalidium sp., Xylaria sp., Phyllosticta capitalensis, Cercospora apii, Xylaria longipes, Phomopsis sp., Phomopsis asparagi, Aspergillus versicolor, Pallidocercospora thailandica, and Xylaria grammica that belonged to the Deuteromycota and Ascomycota divisions. These endophytic fungi inhibited the growth of Fusarium oxysporum f.sp. cubense and Mycosphaerella fijiensis with the percentage inhibition varying respectively from 23.25% to 73.52% and from 21.36% to 100%. The species Botryosphaeria sp., Phomopsis sp., Phomopsis asparagi, and Xylaria longipes exhibited the greatest activity. Conclusion: The leaves of Psidium guajava have a fairly varied diversity of endophytic fungi. These endophytic fungi can serve as potential biological control agents against Panama and Sigatoka diseases of banana and also would produce secondary metabolites with antifungal properties.
This study evaluated the antibacterial efficacy of methanolic extracts of isolated endophytic fungi from stem barks and leaves of Alstonia boonei De Wild and Greenwayodendron suaveolens (Engl. and Diels) Verdc. subsp. Suaveolens against Klebsiella pneumoniae ATCC 43816, Haemophilus influenzae ATCC 49247, Pseudomonas aeruginosa ATCC 27853, and Escherichia coli ATCC 35218, responsible for causing pneumonia. The endophytic fungi were isolated and characterized in the Potato Dextrose Agar (PDA), Sabouraud Dextrose Agar (SDA), and Czapek Dox Agar (CDA) media. The fungi and their methanolic extracts were tested for in vitro antibacterial potential by antagonistic assay for endophytic fungi against bacterial pathogens and microdilution method. The phytochemical screening of extracts was carried out according to the colorimetric and precipitation methods to reveal the presence of secondary metabolites. The results showed that 24 macroscopically and microscopically distinct endophytic fungi were isolated, identified, and stored. These endophytic fungi possessed antibacterial activity against the selected bacterial strains with inhibition zones ranging from 7.00 to 25.00 mm. The endophytic fungi GS15 and AB24 have presented the inhibitions zones of 20.33 mm and 25.00 mm, respectively, and these were better than the ones obtained for Levofloxacin®. The endophytes with inhibition zones greater than 10 mm were used for extraction of their secondary metabolites. The endophytic fungi extracts showed antibacterial activity with the minimum inhibitory concentrations (MICs) ranging from 6.25 × 10−4 to 2 × 10−2 g/L and the minimum bactericidal concentrations (MBCs) ranging from 2.5 × 10−3 to 2 × 10−2 g/L. The endophytic fungi GS15 extract was the most effective extract; it showed bactericidal effects on the tested bacterial strains. The phytochemical screening of the extracts revealed the presence of secondary metabolites classes, responsible for causing the obtained antibacterial activity. Thus, the endophytic fungi methanolic extracts from A. boonei and G. suaveolens have the potential to inhibit the growth of bacteria responsible for nosocomial pneumonia.
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