Orthosporin, a major component of the fermentation product of Lasiodiplodia theobromae - an endophytic fungus of Musa paradisiaca as a potential antimicrobial agent


  • Ugochukwu M. OKEZIE Nnamdi Azikiwe University, Department of Pharmaceutical Microbiology and Biotechnology, Awka, Anambra State (NG)
  • Peter M. EZE Nnamdi Azikiwe University, Department of Environmental Health Science, Faculty of Health Sciences and Technology, Awka, Anambra State (NG)
  • Festus B. C. OKOYE Nnamdi Azikiwe University, Department of Pharmaceutical and Medicinal Chemistry, Awka, Anambra State (NG)
  • Charles O. ESIMONE Nnamdi Azikiwe University, Department of Pharmaceutical Microbiology and Biotechnology, Awka, Anambra State (NG)




antimicrobial, antioxidant, antiviral, endophytic fungi, Lasiodiplodia theobromae, Musa paradisiaca, orthosporin


Endophytic fungi remain an unexplored reservoir of chemical diversity, and have become the primary focus is several bio-prospecting programs. This study was carried out to detect the constituents and bioactivity of the secondary metabolites of an endophytic Lasiodiplodia theobromae associated with Musa paradisiaca. Following standard protocols, the axenic fungus was isolated from healthy leaves of Musa paradisiaca harvested from Agulu, Anambra State, Nigeria. The isolated fungus was characterized using a standard taxonomic identification procedure involving the amplification and sequencing of the ITS region of the DNA. The fungus was subjected to solid state fermentation in rice medium, followed by extraction, chromatographic separation and chemical characterization of its secondary metabolites. The antimicrobial, antioxidant, antiviral and cytotoxic activities of fungal extract was evaluated using standard methods. At a 1 mg/mL, the fungal extract demonstrated inhibitory potentials against Bacillus subtilis, Pseudomonas aeruginosa and Escherichia coli with inhibition zone diameters that ranged from 3-7 mm. The extract, at 100 μg/mL, exhibited antioxidant effect having an IC50 of 65.8 µg/mL. The extract showed dose-dependent antiviral properties during the in vitro antiviral assay with 54% and 60% inhibition of reverse transcriptase observed at 0.5 and 1 mg/mL concentrations respectively. Following chromatographic separation and chemical analyses of the fungal metabolite, orthosporin was isolated. This widely known phytotoxic compound, together with other constituents of the fungal secondary metabolites, may have exhibited the observed bioactivities of the fungal extract recorded. Thus, our findings provide additional data on the potentials of endophytic fungi as producers of interesting bio-molecules.


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Abdullah HA, Mohanad KA, Muhammed AH, Adnan A, Zulfiqar A (2012). First report of grapevine dieback caused by Lasiodiplodia theobromae and Neoscytalidium dimidiatum in Basrah, southern Iraq. African Journal of Biotechnology 11(95):16165-16171. https://doi.org/10.5897/AJB12.010

Begoude D, Slippers B, Wingfield ML, Roux J (2009). Botryosphaeriaceae associated with Terminalia catappa in Cameroon, South Africa and Madagascar. Mycological Progress 9(1):101-123. https://doi.org/10.1007/s11557-009-0622-4

de Medeiros AG, Savi DC, Mitra P, Shaaban KA, Jha AK, Thorson JS, Rohr J (2018). Bioprospecting of Diaporthe terebinthifolii LGMF907 for antimicrobial compounds. Folia Microbiol 63(4):499-505. https://doi.org/10.1007/s12223-018-0587-2

Deanna AS, Michael GR, Stephen ES (2009). Dematiaceous fungi. Clinical Mycology. Chapter 14- (Second Edition), pp 329-354.

Deepika VB, Murali TS, Satyamoorthy K (2015). Modulation of genetic clusters for synthesis of bioactive molecules in fungal endophytes: A review. Microbiological Research 182:125-40. https://doi.org/10.1016/j.micres.2015.10.009

Ebada SS, Eze P, Okoye FBC, Esimone CO, Proksch P (2016). The fungal endophyte Nigrospora oryzae produces quercetin monoglycosides previously known only from plants. Chemistry Select 1(11):2767-2771. https://doi.org/10.1002/slct.201600478

Ebada SS, Edrada RA, Lin W, Proksch P (2008). Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates. Nature Protocols 3:1820-1831. https://doi.org/10.1038/nprot.2008.182.

Eze PM, Ojimba NK, Abonyi DO, Chukwunwejim CR, Abba CC, Okoye FBC, Esimone CO (2018). Antimicrobial activity of metabolites of an endophytic fungus isolated from the leaves of Citrus jambhiri (Rutaceae). Trop. J. Nat. Prod. Res. 2(3):145-149. http://dx.doi.org/10.26538/tjnpr/v2i3.9

Eze PM, Nnanna JC, Okezie U, Buzugbe HS, Abba CC, Chukwunwejim CR, … Esimone CO (2019). Screening of metabolites from endophytic fungi of some Nigerian medicinal plants for antimicrobial activities. The EuroBiotech Journal 3(1):10-18. https://doi.org/10.2478/ebtj-2019-0002

FAO (2005). Production Yearbook for FAOSTAT Data, Food and Agriculture Organization of the United Nations, Rome.

Feng CC, Chen GD, Zhao YQ, Xin SC, Li S, Tang JS, … Gao H (2014). New isocoumarins from a cold-adapted fungal strain Mucor sp. and their develop-mental toxicity to zebrafish embryos. Chemistry Biodiversity 11:1099-1108.

Hallock YF, Clardy J, Kenfield DS, Strobel G (1988). ‘De-O-methyldiaporthin, a phytotoxin from Drechslera siccans. Phytochemistry 27:3123-3125. https://doi.org/10.1016/0031-9422(88)80012-8

Harris JP, Mantle PG (2001). Biosynthesis of diaporthin and orthosporin by Aspergillus ochraceus. Phytochemistry 57(2):165-169. https://doi.org/10.1016/s0031-9422(01)00004-8

Hemingway RW, McGraw GW, Barras SJ (1977). Polyphenols in Ceratocystis minor infected Pinus taeda: fungal metabolites, phloem and xylem phenols. Journal of Agriculture and Food Chemistry 25:719-722.

Ichihara A, Hashimoto M, Hirai T, Takeda I, Sasamura Y, Sakamura S, … Tajimi A (1989). Structure, synthesis, and stereochemistry of (+)-orthosporin, a phytotoxic metabolite of Rhynchosporium Orthosporum. Chemistry Letters 18(8):1495-1498. https://doi.org/10.1246/cl.1989.1495

Jingtair S (2011). In: Postharvest Biology and Technology of Tropical and Subtropical Fruits. 1st Edition: Cocona to Mango. A volume in Woodhead Publishing Series in Food Science, Technology and Nutrition.

Ju ZR, Qin X, Lin XP, Wang JF, Kaliyaperumal K, Tian YQ, … Liu Y (2016). New phenyl derivatives from endophytic fungus Botryosphaeria sp. SCSIO KcF6 derived of mangrove plant ‘Kandelia candel’. Natural Product Research 30:192-198. https://doi.org/10.1080/14786419.2015.1050670

Kjer J, Debbab A, Aly AH, Proksch P (2010). Methods for isolation of marine-derived endophytic fungi and their bioactive secondary products. Nature Protocols 5:479-490. https://doi.org/10.1038/nprot.2009.233

Kumar RV, Venkatrajireddy G, Bikshapathi T, Reddy MK (2012). Antioxidant-the maximum expressed activity among 63 medicinal plants. Journal of Phytochemistry and Pharmacology 1:1-13.

Lee IK, Seok SJ, Kim WG, Yun BS (2006). Diaporthin and orthosporin from the fruiting body of Daldinia concentrica. Mycology 34(1):38-40. https://doi.org/10.4489/MYCO.2006.34.1.038

Liao HX, Shao TM, Mei RQ, Huang GL, Zhou XM, Zheng CJ, Wang CY (2019). Bioactive secondary metabolites from the culture of the mangrove-derived fungus Daldinia eschscholtzii HJ004. Marine Drugs 17(12):710. https://doi.org/10.3390/md17120710

Newman DJ, Cragg GM (2012). Natural products as sources of new drugs over the 30 years from 1981 to 2010. J. Nat. Prod. 75:311-335. https://doi.org/10.1021/np200906s

Okezie UM, Eze PM, Ajaghaku DL, Okoye FBC, Esimone CO (2015). Isolation and screening of secondary metabolites from endophytic fungi of Vernonia amygdalina and Carica papaya for their cytotoxic activity. Planta Medica 81-177. https://doi.org/10.1055/s-0035-1565554

Okezie UM, Eze PM, Okoye FBC, Ikegbunam MN, Ugwu MC, Esimone CO (2017). Secondary metabolites from an endophytic fungus of Vernonia amygdalina. African Journal of Pharmaceutical Research and Development. 9(1):24-26.

Okezie UM, Okoli OA, Ajaegbu EE, Okoye FBC, Esimone CO (2020). Antioxidant activities of extracts of endophytic fungi isolated from healthy leaves of Carica papaya. International Journal of Research Studies in Biosciences (IJRSB) 8(8):48-56. https://doi.org/10.20431/2349-0365.0808005

Okoye FBC, Nworu CS, Debbab A, Esimone CO, Proksch P (2015). Two new Cytochalasins from an endophytic fungus, KL-1.1 isolated from Psidium guajava leaves. Phytochemistry Letters 14:51-55. https://doi.org/10.1016/j.phytol.2015.09.004

Oladele E, Khokhar S (2011). Effect of domestic cooking on the polyphenolic content and antioxidant capacity of plantain (Musa paradisiaca). World Journal of Dairy and Food Sciences 6(2):189-194.

Rodriguez RJ, White JF, Arnold AE, Redman RS (2009). Fungal endophytes diversity and functional roles. New Phytologist 182:314-330. https://doi.org/10.1111/j.1469-8137.2009.02773.x

Rusman Y, Held BW, Blanchette RA, Wittlin S, Salomon CE (2015). Soudanones A–G: antifungal isochromanones from the Ascomycetous fungus Cadophora sp. isolated from an iron mine. Journal of Natural Products 78(6):1456-1460. https://doi.org/10.1021/acs.jnatprod.5b00204

Salvatore MM, Alves A, Andolf A (2020). Review secondary metabolites of Lasiodiplodia theobromae: distribution, chemical diversity, bioactivity, and implications of their occurrence. Toxins 12:457. https://doi.org/10.3390/toxins12070457

Tanapichatsakul C, Monggoot S, Gentekaki E, Pripdeevech P (2017). Antibacterial and antioxidant metabolites of Diaporthe spp. isolated from flowers of Melodorum fruticosum. Current Microbiology 75(4):476-483. https://doi.org/10.1007/s00284-017-1405-9

Teruhiko Y, Friedemann G (1999). Polyketides and other secondary metabolites including fatty acids and their derivatives. Comprehensive Natural Products Chemistry 1:117-138. https://doi.org/10.1016/B978-0-08-091283-7.00004-7

Ujam NT, Eze PM, Ejikeugwu C, Okoye FBC, Esimone CO (2020). Antimicrobial activity of metabolites of Lasiodiplodia theobromae isolated from Psiduim guajava. International Journal of Innovative Science, Engineering & Technology 7(2):123-129.

Uzor PF, Ebrahim W, Osadebe PO, Nwodo JN, Okoye FB, Müller WEG, … Proksch P (2015). Metabolites from Combretum dolichopetalum and its associated endophytic fungus Nigrospora oryzae-evidence for a metabolic partnership. Fitoterapia 105:147-150. https://doi.org/10.1016/j.fitote.2015.06.018

Wolfe K, Wu X, Liu RH (2003). Antioxidant activity of apple peels. Journal of Agricultural and Food Chemistry 51:609-614. https://doi.org/10.1021/jf020782a

Wu S, Chappell J (2008). Metabolic engineering of natural products in plants: tools of the trade and challenges for the future. Current Opinion in Biotechnology 19:145-152. https://doi.org/10.1016/j.copbio.2008.02.007

Zhang J (2014). Lasiodiplodia theobromae in citrus fruit (Diplodia Stem-End Rot). Academic Press. pp 309-335.

Zhang J, Swingle PP (2007) Effects of curing on green mold and stem-end rot of citrus fruit and its potential application under Florida Packing System. The American Phytopathological Society Publications. https://doi.org/10.1094/PD-89-0834



How to Cite

OKEZIE, U. M., EZE, P. M. ., OKOYE, F. B. C., & ESIMONE, C. O. (2022). Orthosporin, a major component of the fermentation product of Lasiodiplodia theobromae - an endophytic fungus of Musa paradisiaca as a potential antimicrobial agent. Notulae Scientia Biologicae, 14(2), 11084. https://doi.org/10.55779/nsb14211084



Research articles
DOI: 10.55779/nsb14211084

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