Fusarium equiseti is a pathogenic fungus of plant root rot, and there are few studies on the biocontrol strains of plant wilt caused by F. equiseti. Hence, we conducted a screening and antimicrobial characterization study of marine-origin biocontrol fungi from water samples of the Yap Trench. A new Talaromyces strain DYM25 was screened from water samples of the Yap Trench in the western Pacific Ocean, and its potential as a biocontrol agent against Fusarium wilt of cucumber was studied. 18S rRNA and ITS gene sequencing verified that strain DYM25 belongs to the genus Talaromyces. The growth of F. equiseti was inhibited by strain DYM25 through the antibiosis effect. A preliminary test was first conducted to examine the bioactive stability of filtered DYM25 broth against F. equiseti under various conditions, including high temperature, UV light, alkaline environment, and the presence of metal ions, which indicated its potential as a bio-control agent. The results of the pot experiment showed that F. equiseti caused cucumber wilt, which could be mitigated using the fermentation broth of strain DYM25 (52.9 %). On the other hand, the alkaloid chromogenic reaction showed that the alkaloid salts present in the crude n-butanol extracts were most likely the major components that might have an antimicrobial effect. Therefore, Talaromyces sp. DYM25 represents a new species that can be used as a novel biocontrol agent against cucumber wilt.