RNA interference as a method for target-site screening in the Western corn rootworm, Diabrotica virgifera virgifera.
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.
Department of Entomology, University of Nebraska - Lincoln, Lincoln, NE 68583-0816, USA. analiza.alves@pioneer.com
Analiza P
AP
Alves
Marcé D
MD
Lorenzen
Richard W
RW
Beeman
John E
JE
Foster
Blair D
BD
Siegfried
D000595:Amino Acid Sequence; D000818:Animals; D002687:Chitin Synthase / Q000235:genetics; D003001:Cloning, Molecular; D001517:Coleoptera / Q000235:genetics*; D017931:DNA Primers / Q000235:genetics; D007300:Insect Control / Q000379:methods*; D042845:Laccase / Q000235:genetics; D007814:Larva; D008969:Molecular Sequence Data; D010641:Phenotype; D034622:RNA Interference; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
D017931:DNA Primers; D042845:Laccase; D002687:Chitin Synthase; C400821:chitin synthase 2