Rohitukine, a chromone alkaloid, has gained considerable international attention in recent years becauseof its novel semi-synthetic derivative, flavopiridol and P-276-00. Both these molecules are in advancedstages of clinical development and trial for cancer treatment. Recently, flavopiridol was approved as anorphan drug for treatment of chronic lymphocytic leukemia cancer. The natural occurrence of rohitukineis restricted to only four plant species, Amoora rohituka and Dysoxylum binectariferum (both from theMeliaceae family) and from Schumanniophyton magnificum and Schumanniophyton problematicum (bothfrom the Rubiaceae family). Recently, an endophytic fungi isolated from D. binectariferum was reportedto produce rohitukine in culture. In this study, we report the production of rohitukine and its subsequentattenuation by endophytic fungi, Fusarium oxysporum (MTCC-11383), Fusarium oxysporum (MTCC-11384)and Fusarium solani (MTCC-11385), all isolated from D. binectariferum and Gibberella fujikuroi (MTCC-11382)isolated from Amoora rohituka. The fungal rohitukine which was analyzed by HPLC, LC–MS andLC–MS/MS was identical to reference rohitukine and that produced by the plant. The rohitukine contentin the mycelial samples ranged from 192.78�g to 359.55 �g 100 g−1of dry weight of and in broth itranged from 14.10 to 71.90 �g 100 ml−1. In all the fungal cultures, the production declined from first tofourth sub-culture. Studies are underway to unravel the mechanism by which the fungi produce the hostmetabolite in culture.