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<i>Ceratocystis paradoxa</i>, the causal agent of stem-bleeding disease of the coconut palm, causes great losses to the global coconut industry. As the mechanism of pathogenicity of <i>C. paradoxa</i> has not been determined, an exogenous gene marker was introduced into the fungus. In this study, pCT74-sGFP, which contains the green fluorescent protein (GFP) gene, and the hygromycin B resistance gene as a selective marker, was used as an expression vector. Several protoplast release buffers were compared to optimize protoplast preparation. The plasmid pCT74-sGFP was successfully transformed into the genome of <i>C. paradoxa</i>, which was verified using polymerase chain reaction and green fluorescence detection. The transformants did not exhibit any obvious differences from the wild-type isolates in terms of growth and morphological characteristics. Pathogenicity tests showed that the transformation process did not alter the virulence of the X-3314 <i>C. paradoxa</i> strain. This is the first report on the polyethylene glycol-mediated transformation of <i>C. paradoxa</i> carrying a &#8216;reporter&#8217; gene GFP that was stably and efficiently expressed in the transformants. These findings provide a basis for future functional genomics studies of <i>C. paradoxa</i> and offer a novel opportunity to track the infection process of <i>C. paradoxa</i>.