Botryodiplodin (A Mycotoxin) detection in pathogenic Botryodiplodia theobromae isolated from diseased coconut fruits

Osayomore E. Ekhorutomwen(1), Chidi I. Nnamdi(2), Olalekan H. Shittu(3),

(1) Nigerian Institute for Oil Palm Research (NIFOR) & University of Benin, Benin, Edo State, Nigeria
(2) Nigerian Institute for Oil Palm Research (NIFOR)
(3) University of Benin, Benin, Edo State, Nigeria.
Corresponding Author


Botryodiplodia theobromae is a threat to crops because it produces botryodiplodin, that plays a role in the initial stages of plant infection, creating necrotic areas through which it can easily penetrate. In addition, the botryodiplodin produced is not easily detected during quarantine, and other techniques developed to detect botryodiplodin are not easily practicable for screening numerous samples. Hence, the need to develop an in-culture pigment formation method to identify and differentiate toxigenic and non-toxigenic pathogenic isolates of B. theobromae. In this study, to detect botryodiplodin produced by isolates of B. theobromae, PDA, CDA and modified CDA media were used. Only the modified CDA medium enhanced the detection of botryodiplodin produced by B. theobromae isolates due to the addition of glycine into the medium. The effect of modified CDA composition or formulation, sucrose, and glycine concentrations on botryodiplodin detection were also evaluated. Study on the effect of the modified CDA composition on the detection of botryodiplodin produced by isolates of B. theobromae revealed that only sucrose stimulated the detection of botryodiplodin in comparison with other ingredients in the modified medium. In addition, the results from the study also reveals that increasing sucrose and glycine concentrations directly enhanced botryodiplodin detection, with optimum concentration of sucrose and glycine for detecting botryodiplodin by isolates of B. theobromae established at 15 and 10 g/l respectively. Hence, there is no need to increase the concentration of both sucrose and glycine above these established concentrations when preparing an in-culture medium for screening B. theobromae isolates capable of producing botryodiplodin.


Botryodiplodia theobromae, coconut fruit, pathogenic, in-culture, botryodiplodin.


Alam S, Abbas HK, Sulyok M, Khambhati VH, Okunowo WO, & Shier WT (2022). Pigment produced by glycine-stimulated Macrophomina phaseolina is a (–)-botryodiplodin reaction product and the basis for an in-culture assay for (–)-Botryodiplodin Production. Pathogens, 11: 280-297.

Aldridge DC, Galt S, Giles D, & Turner WB (1971). Metabolites of Botryodiplodia theobromae. Journal of Chemical Society, 1971: 1623-1627.

Alves A, Crous PW, Correia A, & Phillips AJL (2008). Morphological and molecular data reveal cryptic speciation in Lasiodiplodia theobromae. Fungal Divers., 28: 1-13.

Bladt TT, Frisvad JC, Knudsen PB, & Larsen TO (2013). Anticancer and antifungal compounds from Aspergillus, Penicillium and other filamentous fungi. Molecules, 18: 11338-11376.

Dheepa R, Goplakrishnan C, Kamalakannan A, Nakkeeran S, Mahalingam CA, & Suresh (2018). Coconut Nut Rot Disease in India: Prevalence, Characterization of Pathogen and Standardization of Inoculation Techniques. International Journal of Current Microbiology and Applied Sciences, 7(2): 2046-2057.

Dunlap JR & Bruton DB (1986). Pigment biosynthesis by Macrophomina phaseolina: The glycine-specific requirement. Transactions of the British Mycology Society, 86: 111-115.

Ekhorutomwen OE, Udoh ME, & Omoregie KO (2019). A survey report on the incidence of fruit-rot and premature-nut-fall diseases of coconut in Badagry, Lagos State, Nigeria. NIFOR Annual Report, 58: 157 – 158

Ibrahim SR, Mohamed GA, Al Haidari RA, El-Kholy AA, Zayed MF, & Khayat MT (2018). Biologically active fungal depsidones: Chemistry, biosynthesis, structural characterization, and bioactivities. Fitoterapia, 129: 317-365.

Khambhati VH, Abbas HK, Sulyok M, Tomaso-Peterson M, & Shier WT (2020). First report of the production of mycotoxins and other secondary metabolites by Macrophomina phaseolina (Tassi) Goid. isolates from soybeans (Glycine max L.) symptomatic with charcoal rot disease. Journal of Fungi, 6: 332.

Machado AR, Pinho DB, & Pereira OL (2014). Phylogeny, identification, and pathogenicity of the Botryosphaeriaceae associated with collar and root rot of the biofuel plant Jatropha curcas in Brazil, with a description of new species of Lasiodiplodia. Fungal Divers., 67: 231-247.

Marques MW, Lima NB, Morais MA, Barbosa Souza MAG, Michereff SJ, Phillips AJL, & Câmara MPS (2013). Species of Lasiodiplodia associated with mango in Brazil. Fungal Divers., 61: 181-193.

McCurry PM & Abe K (1973). Stereochemistry and synthesis of the antileukemic agent botryodiplodin. Journal of the American Chemical Society, 95: 5824–5825.

Mohali S, Burgess TI, & Wingfield MJ (2005). Diversity and host association of the tropical tree endophyte Botryodiplodia theobromae revealed using simple sequence repeat markers. Phytopathology, 35: 385-396.

Moule Y & Darracq N (1984). Absence of DNA breaks during repair of DNA-protein cross-links induced by the mycotoxin botryodiplodin in mammalian cells. Carcinogenesis, 5: 1375-1377.

Moule Y, Decloitre F, & Hamon G (1981). Mutagenicity of the mycotoxin botryodiplodin in the Salmonella typhimurium/ microsomal activation test. Environmental Mutagenesis, 3: 287-291.

Moule Y, Douce C, Moreau S, & Darracq N (1981). Effects of the mycotoxin botryodiplodin on mammalian cells in culture. Chemical and Biological Interaction, 37: 155-164.

Phipps PM & Porter DM (1998). Collar rot of peanut caused by Botryodiplodia theobromae. Plant Diseases, 82: 1205-1209.

Punithalingam E (1980). Plant Diseases Attributed to Botryodiplodia theobromae. Pat. In: J. Cramer (eds). Bibliotheca Mycologica. Vaduz, Liechtenstein. Pp 71.

Punithalingam E (1976). Botryodiplodia theobromae. CMI Descriptions of Pathogenic Fungi and Bacteria, 519: 1-234.

Salvatore MM, Alves A, & Andolfi A (2020). Secondary metabolites of Botryodiplodia theobromae: distribution, chemical diversity, bioactivity, and implications of their occurrence. Toxins, 12: 457-286.

Shen W, Mao H, Huang Q, & Dong J (2015). Benzenediol lactones: A class of fungal metabolites with diverse structural features and biological activities. European Journal of Medicinal Chemistry, 97: 747-777.

Shier WT, Nelson J, Abbas HK, & Baird RE (2012). Root toxicity of the mycotoxin botryodiplodin in soybean seedlings. Toxicon, 60: 163.

Slippers B & Wingfield MJ (2007). Botryosphaeriaceae as endophytes and latent pathogens of woody plants: Diversity, ecology and impact. Fungal Biol. Rev., 21: 90 – 106.

Venugopal S & ChandraMohanan R (2006). Role of fungi in fruit rot and immature nut fall of coconut. Cord, 22(2): 1 – 8.

Viana FM, Uchoa CN, Freire FCO, Vieira IGP, Mendes FNP, & Saraiva HAO (2007). Tratamento do coco verde para exportação com ênfase no controle da podridão-basal-pós-colheita. Boletim de Pesquisa e Desenvolvimento. Fortaleza CE. Embrapa Agroindustria Tropical. pp. 1-29.

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