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J Insect Sci. 2010;10:162. doi: 10.1673/031.010.14122.

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

Author information

1
Department of Entomology, University of Nebraska - Lincoln, Lincoln, NE 68583-0816, USA. analiza.alves@pioneer.com

Abstract

To test the efficacy of RNA interference (RNAi) as a method for target-site screening in Diabrotica virgifera virgifera LeConte (Coleptera: 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 double-stranded 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.

PMID:
21067417
PMCID:
PMC3395163
DOI:
10.1673/031.010.14122
[Indexed for MEDLINE]
Free PMC Article
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