Abstract: Insufficient aroma of tobacco leaves has always been a major factor limiting the quality of tobacco leaves and their industrial availability. In order to develop a new source of spices, the R59 strain with a sweet fruit aroma was isolated from healthy leaves of tobacco collected from Lichuan, Hubei province. This strain was identified as Aureobasidium melanogenum. In this study, eleven aroma components were detected in the fermentation broth. Among them, phenylethyl alcohol, phenylacetaldehyde, and isoamyl acetate, which possessed fruity arama, were produced more than 100 μg/mL. After fermentation medium optimization, the production of phenylacetaldehyde and isoamyl acetate by the R59 strain was increased significantly when cultured in 30 g/L glucose, 15 g/L corn steep liquor at pH 5.0. Spraying of the fermentation broth onto cut tobacco at a dosage of 10 μL/g could effectively improve its coordination, aroma and aftertaste. Due to its low fermentation cost and simple addition process, the R59 strain can be used as an aroma source to significantly improve the quality of cigarette products.