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Characterizing the mechanism of action of double-stranded RNA activity against western corn rootworm (Diabrotica virgifera virgifera LeConte).

ABSTRACT: RNA interference (RNAi) has previously been shown to be effective in western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) larvae via oral delivery of synthetic double-stranded RNA (dsRNA) in an artificial diet bioassay, as well as by ingestion of transgenic corn plant tissues engineered to express dsRNA. Although the RNAi machinery components appear to be conserved in Coleopteran insects, the key steps in this process have not been reported for WCR. Here we characterized the sequence of events that result in mortality after ingestion of a dsRNA designed against WCR larvae. We selected the Snf7 ortholog (DvSnf7) as the target mRNA, which encodes an essential protein involved in intracellular trafficking. Our results showed that dsRNAs greater than or equal to approximately 60 base-pairs (bp) are required for biological activity in artificial diet bioassays. Additionally, 240 bp dsRNAs containing a single 21 bp match to the target sequence were also efficacious, whereas 21 bp short interfering (si) RNAs matching the target sequence were not. This result was further investigated in WCR midgut tissues: uptake of 240 bp dsRNA was evident in WCR midgut cells while a 21 bp siRNA was not, supporting the size-activity relationship established in diet bioassays. DvSnf7 suppression was observed in a time-dependent manner with suppression at the mRNA level preceding suppression at the protein level when a 240 bp dsRNA was fed to WCR larvae. DvSnf7 suppression was shown to spread to tissues beyond the midgut within 24 h after dsRNA ingestion. These events (dsRNA uptake, target mRNA and protein suppression, systemic spreading, growth inhibition and eventual mortality) comprise the overall mechanism of action by which DvSnf7 dsRNA affects WCR via oral delivery and provides insights as to how targeted dsRNAs in general are active against insects.

SUBMITTER: Bolognesi R

PROVIDER: S-EPMC3469495 | biostudies-literature | 2012

REPOSITORIES: biostudies-literature

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Characterizing the mechanism of action of double-stranded RNA activity against western corn rootworm (Diabrotica virgifera virgifera LeConte).

Bolognesi Renata R Ramaseshadri Parthasarathy P Anderson Jerry J Bachman Pamela P Clinton William W Flannagan Ronald R Ilagan Oliver O Lawrence Christina C Levine Steven S Moar William W Mueller Geoffrey G Tan Jianguo J Uffman Joshua J Wiggins Elizabeth E Heck Gregory G Segers Gerrit G

PloS one 20121011 10

RNA interference (RNAi) has previously been shown to be effective in western corn rootworm (WCR, Diabrotica virgifera virgifera LeConte) larvae via oral delivery of synthetic double-stranded RNA (dsRNA) in an artificial diet bioassay, as well as by ingestion of transgenic corn plant tissues engineered to express dsRNA. Although the RNAi machinery components appear to be conserved in Coleopteran insects, the key steps in this process have not been reported for WCR. Here we characterized the seque ...[more]

PMID: 23071820

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Project description:The western corn rootworm, Diabrotica virgifera virgifera (LeConte) (Coleoptera: Chrysomelidae), is an economically important pest of corn in the northern United States. Some populations have developed resistance to management strategies including transgenic corn that produces insecticidal toxins derived from the bacterium Bacillus thuringiensis (Bt). Knowledge of insect dispersal is of critical importance for models of resistance evolution. Larval density affects survival in the field, and stress from crowding often affects facultative long-distance dispersal of adult insects. In this study, we used laboratory flight mills to characterize western corn rootworm flight performance as a function of larval rearing density. Larvae were reared under three densities and the resulting adult females were either allowed to fly voluntarily for 22 h or forced to fly specified durations. For both experiments we also measured lifetime fecundity following flight. The three rearing densities placed differential levels of stress on individuals, as evidenced by decreased survival to adulthood and decreased size of adults at greater rearing density. When larvae were reared under crowded conditions the resulting females were more likely to engage in flight activity, including long uninterrupted flights lasting >10 min, than those reared under low density conditions. Flight and egg production are both energy intensive processes. However, we found no evidence in either voluntary or forced flight experiments of a tradeoff between flight activity and female fecundity. The results suggest that females emerging from high density populations in cornfields are more likely to disperse and disperse farther than those emerging from low density populations. These results are important because they imply that variation in population density in the landscape will affect dispersal, which may in turn require computer models of resistance evolution to incorporate multiple dispersal rates arising from varying larval densities among fields.

Dataset Information

Characterizing the mechanism of action of double-stranded RNA activity against western corn rootworm (Diabrotica virgifera virgifera LeConte).

Publications

Characterizing the mechanism of action of double-stranded RNA activity against western corn rootworm (Diabrotica virgifera virgifera LeConte).

Similar Datasets

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