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

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

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.

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Chitin Synthase / genetics
  • Cloning, Molecular
  • Coleoptera / genetics*
  • DNA Primers / genetics
  • Insect Control / methods*
  • Laccase / genetics
  • Larva
  • Molecular Sequence Data
  • Phenotype*
  • RNA Interference*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Sequence Alignment

Substances

  • DNA Primers
  • Laccase
  • Chitin Synthase
  • chitin synthase 2