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        1.
        2018.10 구독 인증기관·개인회원 무료
        We investigated the detoxification strategies of Helicoverpa armigera and Heliothis virescens, which allow them to feed successfully on cotton plants that produce toxic gossypol as a chemical defense compound. First, we tested CYP6AE14, a proposed candidate enzyme for gossypol detoxification, for its ability to detoxify gossypol. In incubation assays with gossypol and heterologously expressed CYP6AE14 no metabolites were detected. Our data show that CYP6AE14 is not directly involved in gossypol metabolism, at least under the assay conditions tested, but rather takes part in the general stress response of the herbivores to plant toxins. Second, we discovered that H. armigera and H. virescens excrete a large proportion (50%) of unmetabolized gossypol in the feces, but additionally metabolize gossypol by glycosylation. Analysis of larval feces revealed three monoglycosylated and up to five diglycosylated gossypol isomers when larvae fed on gossypol-supplemented diet. Based on their expression patterns we selected H. armigera candidate UGT genes and functionally expressed the respective proteins in insect cells. In enzymatic assays, we showed that UGT41B3 and UGT40D1 are capable of glycosylating gossypol mainly to a diglycosylated gossypol isomer that is characteristic for H. armigera and is absent in H. virescens feces. We offer novel insights into the detoxification mechanism of the plant defensive toxin, gossypol, by two generalist herbivores.