In solving the issue of basal stem rot diseases caused by Ganoderma, an investigation of Scytalidium parasiticum 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 Scytalidium parasiticum and Ganoderma boninense was applied to produce and induce metabolites that have antifungal activity against G. boninense. 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 G. boninense. The fractions that showed inhibition against G. boninense 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 Ganoderma infection.

DisclosureThe authors declare no conflicts of interest. All the experiments undertaken in this study comply with the current laws of the country where they were performed.The objective of this study was to assess the interactions between Scytalidium parasiticum and Ganoderma boninense, the causal agent of basal stem rot in oil palm (Elaeis guineensis).When compared with S. ganodermophthorum and S. sphaerosporum, S. parasiticum (Sp) showed greater inhibition toward all Ganoderma isolates during in vitro co-culturing. At the Downloaded by [University of Sussex Library] at 05:14 28 June 2016 2 interaction zone, coiling of host hyphae, formation of short lateral enlarged contact structures, and production of potential appressorium-like organs were observed in Sp on G. boninense.These were followed by the degradation, shrinkage, and deformation of G. boninense mycelia.Sp also reduced mycelial survival and fruiting body regeneration of G. boninense. Nonvolatile metabolites from Sp suppressed the growth of G. boninense. Our results show that Sp could be a necrotrophic mycoparasite of G. boninense. Nursery experiments revealed that Sp was non-pathogenic to oil palm seedlings, and it could suppress Ganoderma infection and reduce disease severity. Sp also increased the height of palms in the positive control with non-Ganoderma-inoculated rubber wood block and Sp inoculum compared to similar control without Sp. Leaf area was significantly greater in the G. boninense G8 inoculated palms when Sp was present compared to without Sp. These results show that Sp might be a potential biocontrol candidate against BSR of oil palm.

Basal stem rot (BSR) disease caused by Ganoderma boninense basidiomycetous fungus is the most economically important disease in oil palms in South East Asia. Unfortunately, there is no single most effective control measure available. Tremendous efforts have been directed in incorporation of environmentally friendly biocontrol approaches in minimizing BSR disease. This study investigated the performance of two potential biocontrol agents (BCAs), AAT0115 and AAB0114 strains recovered from oil palm on suppression of BSR in planta, and also assessed their plant-growth-promoting (PGP) performance. ITS rRNA-sequence phylogeny discriminated the two ascomycetous Talaromyces apiculatus (Ta) AT0115 and Clonostachys rosea (Cr) AAB0114 biocontrol species with PGP characteristics. In vitro studies have demonstrated both Ta and Cr are capable of reducing linear mycelial growth of G. boninense. Inoculation of individual Cr and Ta—as well as Cr+Ta consortium—induced a significant increment in leaf area and bole girth of oil-palm seedlings five months post-inoculation (MPI) under nursery conditions. At five months post-inoculation, shoot and root biomass, and nutrient contents (nitrogen, phosphorus, potassium, calcium, magnesium and boron) were significantly higher in Ta-inoculated seedlings compared to control treated with non-Ta-inoculated maize. Chlorophyll and carotenoids contents in rapidly growing oil-palm seedlings challenged with Cr, Ta or a combination of both were not negatively affected. Cr, Ta and Cr+Ta consortium treated seedlings had 4.9–60% BSR disease reduction compared to the untreated control. Co-inoculation of Cr and Ta resulted in increased BSR control efficiencies by 18–26% (compared with Cr only) and 48–55% (compared with Ta only). Collectively, Cr and Ta, either individually or in consortium showed potential as BSR biocontrol agents while also possess PGP traits in oil palm.

Basal stem rot disease of oil palm caused by Ganoderma boninense is one of the most devastating diseases in oil palm plantation resulting in low yield, loss of palm stands and shorter replanting cycle. To-date, there is no effective treatment for Ganoderma infected palms. Control measures, either chemical or cultural approaches, show varying degrees of effectiveness. The application of biological control agents which is environmental-friendly could be an attractive solution to overcome the problem. Earlier, we had isolated a mycoparasite, Scytalidium parasiticum, from the basidiomata of Ganoderma boninense. In vitro assay and nursery experiment showed that this fungus could suppress Ganoderma infection and reduce disease severity. However, metabolites which might contribute to the antagonistic or mycoparasitic effect remain unknown. In the current study, optimization of fungal sample processing, extraction, and analytical procedures were conducted to obtain metabolites from the maize substrate colonized by mycoparasitic ascomycetous Scytalidium parasiticum. This technique capable of producing sexual spores in sac-like organs. Untargeted metabolomics profiling was carried out by using Liquid Chromatography Time of Flight Mass Spectrometry (LC-ToF-MS). We found that S. parasiticum in both liquid-and solid-state cultivation gave higher metabolite when extracted with 60% methanol with 1% formic acid in combination with homogenisation methods such as ultrasonication and grinding. The findings from this study are useful for optimisation of metabolite extraction from other fungi-Ganoderma-plant interactions.

A mycoparasite, Scytalidium parasiticum sp. nov., isolated from the basidiomata of Ganoderma boninense causing basal stem rot of oil palm in Johor, Malaysia, is described and illustrated. It is distinct from other Scytalidium species in having smaller asci and ascospores (teleomorphic stage), longer arthroconidia (anamorphic stage), hyaline to yellowish chlamydospores, and producing a fluorescent pigment. The phylogenetic position of S. parasiticum was determined by sequence analyses of the internal transcribed spacers and the small-subunit ribosomal RNA gene regions. A key to identify Scytalidium species with teleomorphic stage is provided.

The influence of different medium components (glucose, sucrose, and fructose) on the growth of different Ganoderma isolates and species was investigated using mixture design. Ten sugar combinations based on three simple sugars were generated with two different concentrations, namely 3.3% and 16.7%, which represented low and high sugar levels, respectively. The media were adjusted to either pH 5 or 8. Ganoderma isolates (two G. boninense from oil palm, one Ganoderma species from coconut palm, G. lingzhi, and G. australe from tower tree) grew faster at pH 8. Ganoderma lingzhi proliferated at the slowest rate compared to all other tested Ganoderma species in all the media studied. However, G. boninense isolates grew the fastest. Different Ganoderma species were found to have different sugar preferences. This study illustrated that the mixture design can be used to determine the optimal combinations of sugar or other nutrient/chemical components of media for fungal growth.

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The presence of entomopathogenic <em>Beauveria bassiana</em> on Pleasing Fungus Beetle <em>Episcapha quadrimacula</em> has been reported on fruiting bodies of <em>Ganoderma boninense</em> for the first time in Malaysia.