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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 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.

Keywords: biopesticide, chitin synthase 2, laccase 2, systematic RNAi
Abbreviations: Bt, Bacillus thuringiensis; CHS, chitin synthase; dsRNA, double-stranded RNA; Lac, laccase; RNAi, RNA interference; RT-PCR, reverse transcriptase polymerase chain reaction; qRT-PCR, quantitative reverse transcriptase polymerase chain reaction

Correspondence: ^a*analiza.alves@pioneer.com, ^bmarce@ksu.edu, ^crichard.beeman@ars.usda.gov, ^djfoster1@unl.edu, ^ebsiegfried1@unl.edu, *Corresponding author
Associate Editor: Brad Coates was editor of this paper.

Received: 24 September 2008 | Accepted: 22 April 2009 | Published: 24 September 2010

Copyright: This is an open access paper. We use the Creative Commons Attribution 3.0 license that permits unrestricted use, provided that the paper is properly attributed.  

ISSN: 1536-2442 | Volume 10, Number 162

Alves AP, Lorenzen MD, Beeman RW, Foster JE, Siegfried BD. 2010. RNA interference as a method for target-site screening in the western corn rootworm, Diabrotica virgifera virgifera. Journal of Insect Science 10:162, available online: insectscience.org/10.162