Silencing abnormal wing disc gene of the Asian citrus psyllid, Diaphorina citri disrupts adult wing development and increases nymph mortality

PLoS One. 2013 May 29;8(5):e65392. doi: 10.1371/journal.pone.0065392. Print 2013.

Abstract

Huanglongbing (HLB) causes considerable economic losses to citrus industries worldwide. Its management depends on controlling of the Asian citrus Psyllid (ACP), the vector of the bacterium, Candidatus Liberibacter asiaticus (CLas), the causal agent of HLB. Silencing genes by RNA interference (RNAi) is a promising tool to explore gene functions as well as control pests. In the current study, abnormal wing disc (awd) gene associated with wing development in insects is used to interfere with the flight of psyllids. Our study showed that transcription of awd is development-dependent and the highest level was found in the last instar (5(th)) of the nymphal stage. Micro-application (topical application) of dsRNA to 5(th) instar of nymphs caused significant nymphal mortality and adult wing-malformation. These adverse effects in ACP were positively correlated with the amounts of dsRNA used. A qRT-PCR analysis confirmed the dsRNA-mediated transcriptional down-regulation of the awd gene. Significant down-regulation was required to induce a wing-malformed phenotype. No effect was found when dsRNA-gfp was used, indicating the specific effect of dsRNA-awd. Our findings suggest a role for awd in ACP wing development and metamorphosis. awd could serve as a potential target for insect management either via direct application of dsRNA or by producing transgenic plants expressing dsRNA-awd. These strategies will help to mitigate HLB by controlling ACP.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Amino Acid Sequence
  • Animals
  • Citrus / microbiology
  • Citrus / parasitology
  • Gene Expression Regulation, Developmental
  • Hemiptera / genetics*
  • Hemiptera / growth & development
  • Hemiptera / microbiology
  • Host-Parasite Interactions
  • Insect Control / methods
  • Insect Proteins / chemistry
  • Insect Proteins / genetics*
  • Insect Proteins / metabolism
  • Insect Vectors / genetics
  • Insect Vectors / growth & development
  • Insect Vectors / microbiology
  • Models, Molecular
  • Molecular Sequence Data
  • Nucleoside-Diphosphate Kinase / chemistry
  • Nucleoside-Diphosphate Kinase / genetics*
  • Nucleoside-Diphosphate Kinase / metabolism
  • Nymph / genetics*
  • Nymph / growth & development
  • Nymph / metabolism
  • Plant Diseases / microbiology
  • Plant Diseases / parasitology
  • Protein Conformation
  • RNA Interference*
  • Reverse Transcriptase Polymerase Chain Reaction
  • Rhizobiaceae / physiology
  • Sequence Homology, Amino Acid
  • Wings, Animal / growth & development
  • Wings, Animal / metabolism*

Substances

  • Insect Proteins
  • Nucleoside-Diphosphate Kinase

Grants and funding

Ibrahim El-Shesheny was supported with a scholarship from the Egyptian government. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.