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Identification of a Resin Detoxification P450 in Dendroctonus ponderosae
AuthorGorzalski, Andrew J.
AdvisorTittiger, Claus R.
Biochemistry and Molecular Biology
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Monoterpene hydroxylation plays a critical role in resin detoxification in Dendroctonus ponderosae. Hydroxylation of (+)- and (-)-a-pinene to myrtenol and trans-verbenol was seen in both larvae and female mountain pine beetle. The hydroxylation activity was highest in the head, midgut, and fat body of fed female MPB. The expression of five potential resin detoxification P450s identified from functional genomic research was analyzed in a monoterpene saturated environment. Expression of CYP6DH2 was significantly induced in the presence of monoterpene vapors. The full ORF of CYP6DH2 was determined using RACE. A paralog, CYP6DH1, was identified and is 97.7% identical to CYP6DH2. Further expression analysis determined CYP6DH2 to be significantly upregulated by all monoterpenes, but no effect was seen on the expression of CYP6DH1. When exposed to monoterpene vapors, expression of CYP6DH2 was highest in the head, midgut, and carcass. Expression during feeding was highest in the hindgut for both genes and sexes. Recombinant CYP6DH2 hydroxylated (+)-a-pinene to myrtenol. These results suggest that CYP6DH2 is a resin detoxification P450. Further experiments are necessary to determine the role CYP6DH1 plays in MPB.