| Summary: | <i>Calamus castaneus</i> is a common rattan palm species in the tropical forests of Peninsular Malaysia and is noticeable by the yellow-based spines that cover the stems. This study aimed to determine the prevalence of fungal endophytes within <i>C. castaneus</i> spines and whether they inhibit the growth of fungal pathogens. Twenty-one genera with 40 species of fungal endophytes were isolated and identified from rattan palm spines. Based on molecular identification, the most common isolates recovered from the spines were <i>Colletotrichum</i> (<i>n =</i> 19) and <i>Diaporthe</i> spp. (<i>n =</i> 18), followed by <i>Phyllosticta</i> spp., <i>Xylaria</i> sp., <i>Trichoderma</i> spp., <i>Helminthosporium</i> spp., <i>Penicillium</i> spp., <i>Fusarium</i> spp., <i>Neopestalotiopsis</i> spp., <i>Arthrinium</i> sp., <i>Cyphellophora</i> sp., <i>Cladosporium</i> spp., <i>Curvularia</i> sp., <i>Bionectria</i> sp., and <i>Acremonium</i> spp. Non-sporulating fungi were also identified, namely <i>Nemania primolutea</i>, <i>Pidoplitchkoviella terricola</i>, <i>Muyocopron laterale</i>, <i>Acrocalymma fici</i>, <i>Acrocalymma medicaginis</i>, and <i>Endomelanconiopsis endophytica</i>. The isolation of these endophytes showed that the spines harbor endophytic fungi. Most of the fungal endophytes inhibited the growth of several plant pathogenic fungi, with 68% of the interactions resulting in mutual inhibition, producing a clear inhibition zone of <2 mm. Our findings demonstrate the potential of the fungal endophytes from <i>C. castaneus</i> spines as biocontrol agents.
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