RNA interference as a method for target-site screening in the western corn rootworm, Diabrotica virgifera virgifera

Authors

Analiza P. Alves, University of Nebraska-LincolnFollow
Richard W. Beeman, USDA-ARS-CGAHRFollow
Marce D. Lorenzen, USDA-ARS-CGAHRFollow
John E. Foster, University of Nebraska-LincolnFollow
Blair D. Siegfried, University of Nebraska-LincolnFollow

Date of this Version

2010

Citation

Journal of Insect Science 162, 2010

Comments

U.S. Government work

Abstract

To test the efficacy of RNA interference (RNAi) as a method for target-site screening in Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) larvae, genes were identified and tested for which clear RNAi phenotypes had been identified in the Coleopteran model, Tribolium castaneum. Here the cloning of the D. v. vergifera orthologs of laccase 2 (DvvLac2) and chitin synthase 2 (DvvCHS2) is reported. Injection of DvvLac2-specific doublestranded RNA resulted in prevention of post-molt cuticular tanning, while injection of DvvCHS2- specific dsRNA reduced chitin levels in midguts. Silencing of both DvvLac2 and DvvCHS2 was confirmed by RT-PCR and quantitative RT-PCR. As in T. castaneum, RNAi-mediated gene silencing is systemic in Diabrotica. The results indicate that RNAi-induced silencing of D. v. vergifera genes provides a powerful tool for identifying potential insecticide targets.