Wednesday, December 12, 2001 -
D0573

Isolation and expression of a family IV P450 gene from insecticide resistant and susceptible western corn rootworms

Srinivas Parimi1, Blair D. Siegfried1, Mike E. Scharf2, Lance Meinke1, and Laurence D. Chandler3. (1) University of Nebraska, Department of Entomology, 202 PI Building, Lincoln, NE, (2) Purdue Univesity, Department of Entomology, Smith Hall, 105E, West Lafeyette, IN, (3) USDA ARS, Red River Valley Agricultural Research Center, P.O. Box 5677, Fargo, ND

Previous studies indicate the involvement of cytochrome P450 monooxygenases in metabolic resistance to organophosphate and carbamate resistance in western corn rootworms. Based on these results we isolated the P450 gene fragment CYP4AJ1 using degenerate PCR. Internal primers were designed from this gene fragment sequence and used to isolate full length cDNA following RACE (rapid amplification of cDNA ends) PCR. Using 5' and 3' RACE, full length gene was obtained which was approximately 1700 nucleotides long. A BLAST search of the full length cDNA sequence in the GENBANK revealed a high degree of similarity to other insect family IV P450 genes - T. castaneum (65%), M. sexta (62%), C. acinaciformis (58%), D. melanogaster and B. discoidalis (57%). Expression studies are under way with resistant and susceptible adults, in order to determine if there is an association of this gene with insecticide resistance.



Species 1: Coleoptera Chrysomelidae Diabrotica virgifera virgifera (western corn rootworm)
Keywords: Cytochrome P450 gene, Corn rootworm

The ESA 2001 Annual Meeting - 2001: An Entomological Odyssey of ESA