A fluorescent base wye (Yt base) was isolated from Torulopsis utilis tRNAPhe. The structure was established as 4,9-dihydro-4,6-dimethyl-9-oxo-1H-imidazo[1,2-alpha]purine based on ultraviolet (uv), nuclear magnetic resonance (NMR), and mass spectra, and by direct comparison with synthetic material. The nucleoside, wyosine (Yt), was isolated from purified tRNAPhe by enzymatic degradations followed by column and thin-layer chromatographies. The structure of wyosine is proposed as 3-ribofuranosyl-4,9-dihydro-4,6-dimethyl-9-oxo-1H-imidazol[1,2-alpha]purine (the ribosyl group is attached to the N-9 position of guanine nucleus) on the basis of comparison with model compounds.