Lawsone (2-hydroxy-1, 4-napthoquinone),also known as hennotannic acid, is an orange red dyeused as a popular skin and hair colorant. The dye isproduced in the leaves ofLawsonia inermisL, oftenreferred to as the ‘‘henna’’ tree. In this study, we reportthe production of lawsone by an endophytic fungus,Gibberella moniliformisisolated from the leaf tissues ofLawsonia inermis.The fungus produced the orange-reddye in potato dextrose agar and broth, independent of thehost tissue. Presence of lawsone was confirmed spectrometrically using HPLC and ESI-MS/MS analysis.The fragmentation pattern of lawsone was identical toboth standard lawsone and that extracted from planttissue. This is a first report of lawsone being produced byan endophytic fungus, independent of the host tissue.The study opens up interesting questions on the possiblebiosynthetic pathway through which lawsone is produced by the fungus.