4.6 Article

Feeding-based RNA interference of a trehalose phosphate synthase gene in the brown planthopper, Nilaparvata lugens

Journal

INSECT MOLECULAR BIOLOGY
Volume 19, Issue 6, Pages 777-786

Publisher

WILEY
DOI: 10.1111/j.1365-2583.2010.01038.x

Keywords

RNA interference; double-stranded RNA; trehalose phosphate synthase; feeding; Nilaparvata lugens

Funding

  1. National Basic Research Program of China [2010CB126200]
  2. National Natural Science Foundation of China [30930061]
  3. National Science & Technology Pillar Program [2008BADA5B04]

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The brown planthopper, Nilaparvata lugens, is the most devastating rice insect pest to have given rise to an outbreak in recent years. RNA interference (RNAi) is a technological breakthrough that has been developed as a powerful tool for studying gene function and for the highly targeted control of insect pests. Here, we examined the effects of using a feeding-based RNAi technique to target the gene trehalose phosphate synthase (TPS) in N. lugens. The full-length cDNA of N. lugens TPS (NlTPS) is 3235 bp and has an open reading frame of 2424 bp, encoding a protein of 807 amino acids. NlTPS was expressed in the fat body, midgut and ovary. Quantitative real-time PCR (qRT-PCR) analysis revealed that NlTPS mRNA is expressed continuously with little change during the life of the insect. Efficient silencing of the TPS gene through double-stranded RNA (dsRNA) feeding led to rapid and significant reduction levels of TPS mRNA and enzymatic activity. Additionally, the development of N. lugens larvae that had been fed with the dsRNA was disturbed, resulting in lethality, and the cumulative survival rates dropped to 75.56, 64.44, 55.56 and 40.00% after continuous ingestion of 0.5 mu g/mu l dsRNA for 2, 4, 7 and 10 days, respectively. These values were significantly lower than those of the insects in the control group, suggesting that NlTPS dsRNA may be useful as a means of insect pest control.

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