The development of reliable processes for the synthesis of gold nanoparticles is an important aspect of current nanotechnology research. Recently, reports are published on the extracellular as well as intracellular biosynthesis of gold nanoparticles using microorganisms. However, these methods of synthesis are rather slow. In present study, rapid and extracellular synthesis of gold nanoparticles using a plant pathogenic fungus F. oxysporum f. sp. cubense JT1 (FocJT1) is reported. Incubation of FocJT1 mycelium with auric chloride solution produces gold nanoparticles in 60 min. Gold nanoparticles were characterized by UV-Vis spectroscopy, FTIR, and particle size analysis. The particles synthesized were of 22 nm sized, capped by proteins, and posed antimicrobial activity against Pseudomonas sp.