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Validation of RNA interference in western corn rootworm Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) adults.
Pest Manag Sci. 2012 Apr; 68(4):587-91.PM

Abstract

BACKGROUND

RNA interference (RNAi) is commonly used in insect functional genomics studies and usually involves direct injection of double-stranded RNA (dsRNA). Only a few studies have involved exposure to dsRNAs through feeding. For western corn rootworm (Diabrotica virgifera virgifera) larvae, ingestion of dsRNA designed from the housekeeping gene, vacuolar ATPase (vATPase) triggers RNAi causing growth inhibition and mortality; however, the effect of dsRNA feeding on adults has not been examined. In this research, WCR adults were fed with vATPase-dsRNA-treated artificial diet containing a cucurbitacin bait, which is a proven feeding stimulant for chrysomelid beetles of the subtribe Diabroticina to which rootworms belong.

RESULTS

Real-time PCR confirmed suppression of vATPase expression and western blot analysis indicated reduced signal of a protein that cross-reacted with a vATPase polyclonal antiserum in WCR adults exposed to artificial diet treated with dsRNA and cucurbitacin bait. Continuous feeding on cucurbitacin and dsRNA-treated artificial diet resulted in more than 95% adult mortality within 2 weeks while mortality in control treatments never exceeded 20%.

CONCLUSIONS

This research clearly demonstrates the effect of RNAi on WCR adults that have been exposed to dsRNA by feeding and establishes a tool to screen dsRNAs of potential target genes in adults. This technique may serve as an alternative to target screening of larvae which are difficult to maintain on artificial diets.

Authors+Show Affiliations

Department of Entomology, University of Nebraska, Lincoln, NE 68583-0816, USA.No affiliation info available

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

22500293

Citation

Rangasamy, Murugesan, and Blair D. Siegfried. "Validation of RNA Interference in Western Corn Rootworm Diabrotica Virgifera Virgifera LeConte (Coleoptera: Chrysomelidae) Adults." Pest Management Science, vol. 68, no. 4, 2012, pp. 587-91.
Rangasamy M, Siegfried BD. Validation of RNA interference in western corn rootworm Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) adults. Pest Manag Sci. 2012;68(4):587-91.
Rangasamy, M., & Siegfried, B. D. (2012). Validation of RNA interference in western corn rootworm Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) adults. Pest Management Science, 68(4), 587-91.
Rangasamy M, Siegfried BD. Validation of RNA Interference in Western Corn Rootworm Diabrotica Virgifera Virgifera LeConte (Coleoptera: Chrysomelidae) Adults. Pest Manag Sci. 2012;68(4):587-91. PubMed PMID: 22500293.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Validation of RNA interference in western corn rootworm Diabrotica virgifera virgifera LeConte (Coleoptera: Chrysomelidae) adults. AU - Rangasamy,Murugesan, AU - Siegfried,Blair D, PY - 2012/4/14/entrez PY - 2012/4/14/pubmed PY - 2012/6/23/medline SP - 587 EP - 91 JF - Pest management science JO - Pest Manag Sci VL - 68 IS - 4 N2 - BACKGROUND: RNA interference (RNAi) is commonly used in insect functional genomics studies and usually involves direct injection of double-stranded RNA (dsRNA). Only a few studies have involved exposure to dsRNAs through feeding. For western corn rootworm (Diabrotica virgifera virgifera) larvae, ingestion of dsRNA designed from the housekeeping gene, vacuolar ATPase (vATPase) triggers RNAi causing growth inhibition and mortality; however, the effect of dsRNA feeding on adults has not been examined. In this research, WCR adults were fed with vATPase-dsRNA-treated artificial diet containing a cucurbitacin bait, which is a proven feeding stimulant for chrysomelid beetles of the subtribe Diabroticina to which rootworms belong. RESULTS: Real-time PCR confirmed suppression of vATPase expression and western blot analysis indicated reduced signal of a protein that cross-reacted with a vATPase polyclonal antiserum in WCR adults exposed to artificial diet treated with dsRNA and cucurbitacin bait. Continuous feeding on cucurbitacin and dsRNA-treated artificial diet resulted in more than 95% adult mortality within 2 weeks while mortality in control treatments never exceeded 20%. CONCLUSIONS: This research clearly demonstrates the effect of RNAi on WCR adults that have been exposed to dsRNA by feeding and establishes a tool to screen dsRNAs of potential target genes in adults. This technique may serve as an alternative to target screening of larvae which are difficult to maintain on artificial diets. SN - 1526-4998 UR - https://www.unboundmedicine.com/medline/citation/22500293/Validation_of_RNA_interference_in_western_corn_rootworm_Diabrotica_virgifera_virgifera_LeConte__Coleoptera:_Chrysomelidae__adults_ L2 - https://doi.org/10.1002/ps.2301 DB - PRIME DP - Unbound Medicine ER -