Methyl farnesoate (MF) plays hormonal regulatory roles in crustaceans. An epoxidated form of MF, known as juvenile hormone (JH), controls metamorphosis and stimulates reproduction in insects. To address the evolutionary significance of MF epoxidation, we generated mosquitoes completely lacking either of the two enzymes that catalyze the last steps of MF/JH biosynthesis and epoxidation, respectively: the JH acid methyltransferase (JHAMT) and the P450 epoxidase CYP15 (EPOX). jhamt-/- larvae lacking both MF and JH died at the onset of metamorphosis. Strikingly, epox-/- mutants, which synthesized MF but no JH, completed the entire life cycle. While epox-/- adults were fertile, the reproductive performance of both sexes was dramatically reduced. Our results suggest that although MF can substitute for the absence of JH in mosquitoes, it is with a significant fitness cost. We propose that MF can fulfill most roles of JH, but its epoxidation to JH was a key innovation providing insects with a reproductive advantage.

Department of Biological Sciences Biomolecular Science Institute Florida International University Miami FL 33199

Department of Biological Sciences Biomolecular Science Institute Florida International University Miami FL 33199;

Department of Biology Muhlenberg College Allentown PA 18104

Department of Chemistry and Biochemistry Florida International University Miami FL 33199

Department of Plant and Environmental Sciences University of Copenhagen Copenhagen DK 1871 Denmark

Department of Plants and Crops Faculty of Bioscience Engineering Ghent University Ghent B 9000 Belgium

Institute of Entomology Biology Centre Czech Academy of Sciences Ceske Budejovice 37005 Czech Republic

Institute of Parasitology Biology Centre Czech Academy of Sciences Ceske Budejovice 37005 Czech Republic

Institute of Zoology Slovak Academy of Sciences Bratislava 84506 Slovakia

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