Inoculation of Nicotiana tabacum with recombinant potato virus X induces RNA interference in the solenopsis mealybug, Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae)

Biotechnol Lett. 2015 Oct;37(10):2083-90. doi: 10.1007/s10529-015-1880-7. Epub 2015 Jun 19.

Abstract

Objectives: The chitin synthase 1 (CHS1) gene in Phenacoccus solenopsis (PsCHS1) was evaluated as a potential target of RNA interference (RNAi) by using Potato virus X (PVX) as a vector (recombinant PVX) for expressing RNAi triggering elements in Nicotiana tabacum L.

Results: RT-PCR analysis confirmed the expression of PsCHS1 in N. tabacum inoculated with recombinant-PVX-PsCHS1 (treated). RT- and multiplex-PCR further showed a reduction in mRNA levels of the target gene in mealybugs feeding on treated plants. Mortality in parent adults and emerging nymphs (21 and 29%) exposed to the treated plants was significantly higher (P < 0.05) than those exposed to uninoculated (-ve control) or inoculated with non-recombinant PVX (PVX-control). The number of surviving adults and the combined number of adults and nymphs (47 and 60%) was significantly (P < 0.05) lower on the treated plants than the -ve (76%) or PVX (74%) control. The visual observations verified the physical deformities in mealybugs exposed to the treated plants.

Conclusion: chitin synthase 1 is a potential RNAi target in P. solenopsis and the recombinant PVX can be used as a tool to evaluate candidate RNAi triggering elements in plants.

Keywords: Mealy bug; Phenacoccus solenopsis; Potato virus X; RNA interference; Recombinant virus; Solenopsis mealybug.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Chitinases / antagonists & inhibitors
  • Chitinases / genetics
  • Chitinases / metabolism*
  • Hemiptera / enzymology*
  • Hemiptera / physiology*
  • Multiplex Polymerase Chain Reaction
  • Nicotiana / parasitology*
  • Nicotiana / virology*
  • Nymph / physiology
  • Plants
  • Potexvirus / genetics
  • Potexvirus / growth & development*
  • RNA Interference*
  • Real-Time Polymerase Chain Reaction
  • Reverse Transcriptase Polymerase Chain Reaction
  • Survival Analysis

Substances

  • Chitinases