Strigolactones (SLs) stimulate seed germination of root parasitic weeds of the Orobanchaceae, resulting in yield losses of important cereals, legumes and vegetables in many areas of the world. SLs also induce hyphal branching of arbuscular mycorrhizal (AM) fungi to promote beneficial symbiosis for plants. SLs endogenously function as regulators of plant architecture and development as a class of plant hormones. So far, more than 30 SLs have been identified in root exudates of various plants while the biological significance of the structural diversification of SLs is still unclear. Fabaceae plants have been reported to produce orobanchol, which is one of major SLs and orobanchol-derived SLs. For instance, medicaol was isolated from the root exudates of Medicago truncatula (barrel medic) and has a unique seven-membered A-ring. Fabacyl acetate identified from the root exudates of Pisum sativum (pea) has an epoxide group and an acetoxy group. Several other Fabaceae plants including Vigna unguiculata (cowpea) and pea produce orobanchyl acetate, which has an acetoxy group. Although the biosynthetic pathway to orobanhol has been revealed, the biosynthetic pathway and biological roles of these orobanchol-derived SLs remain unknown. We report the identification of 2-oxo-glutarate dependent dioxygenases and BAHD acyltransferases responsible for the conversion of orobanchol to these orobanchol derivatives in Fabaceae plants. Candidate genes were selected by transcriptome analysis and co-expression analysis. The recombinant proteins of the candidate genes were expressed in E. coliand the enzyme activity was characterized. These findings provide the molecular basis behind the structural diversity of SLs.