| Summary: | In solving the issue of basal stem rot diseases caused by Ganoderma, an investigation of <i>Scytalidium parasiticum</i> as a biological control agent that suppresses Ganoderma infection has gained our interest, as it is more environmentally friendly. Recently, the fungal co-cultivation has emerged as a promising method to discover novel antimicrobial metabolites. In this study, an established technique of co-culturing <i>Scytalidium parasiticum</i> and <i>Ganoderma boninense</i> was applied to produce and induce metabolites that have antifungal activity against <i>G. boninense</i>. The crude extract from the co-culture media was applied to a High Performance Liquid Chromatography (HPLC) preparative column to isolate the bioactive compounds, which were tested against <i>G. boninense</i>. The fractions that showed inhibition against <i>G. boninense</i> were sent for a Liquid Chromatography-Time of Flight-Mass Spectrometry (LC-TOF-MS) analysis to further identify the compounds that were responsible for the microbicidal activity. Interestingly, we found that eudistomin I, naringenin 7-O-beta-D-glucoside and penipanoid A, which were present in different abundances in all the active fractions, except in the control, could be the antimicrobial metabolites. In addition, the abundance of fatty acids, such as oleic acid and stearamide in the active fraction, also enhanced the antimicrobial activity. This comprehensive metabolomics study could be used as the basis for isolating biocontrol compounds to be applied in oil palm fields to combat a <i>Ganoderma</i> infection.
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