RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing

  1. Suzuki, T. 25
  2. Nunes, M.A. 12
  3. España, M.U. 2
  4. Namin, H.H. 2
  5. Jin, P. 2
  6. Bensoussan, N. 2
  7. Zhurov, V. 2
  8. Rahman, T. 27
  9. Clercq, R.D. 4
  10. Hilson, P. 346
  11. Grbic, V. 2
  12. Grbic, M. 28
  1. 1 Centro de Citricultura Sylvio Moreira, Instituto Agronômico de Campinas, Cordeirópolis, São Paulo, Brazil
  2. 2 University of Western Ontario
    info

    University of Western Ontario

    London, Canadá

    ROR https://ror.org/02grkyz14

  3. 3 Ghent University
    info

    Ghent University

    Gante, Bélgica

    ROR https://ror.org/00cv9y106

  4. 4 Department of Plant Systems Biology, VIB, Ghent, Belgium
  5. 5 Tokyo University of Agriculture and Technology
    info

    Tokyo University of Agriculture and Technology

    Tokio, Japón

    ROR https://ror.org/00qg0kr10

  6. 6 Institut Jean Pierre Bourgin
    info

    Institut Jean Pierre Bourgin

    Versalles, Francia

    ROR https://ror.org/01wqd6v19

  7. 7 Saskatoon Research Centre, Agriculture and Agri-Food Canada, Saskatoon, SK, Canada
  8. 8 Universidad de La Rioja
    info

    Universidad de La Rioja

    Logroño, España

    ROR https://ror.org/0553yr311

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Revista:
PLoS ONE

ISSN: 1932-6203

Año de publicación: 2017

Volumen: 12

Número: 7

Tipo: Artículo

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DOI: 10.1371/JOURNAL.PONE.0180654 SCOPUS: 2-s2.0-85023186041 WoS: WOS:000405649600039 GOOGLE SCHOLAR

Otras publicaciones en: PLoS ONE

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Resumen

RNA interference (RNAi) can be used for the protection against agricultural pests through the silencing of genes required for pest fitness. To assess the potential of RNAi approaches in the two-spotted spider mite, Tetranychus urticae, we compared 5 methods for the delivery of double-stranded RNA (dsRNA). These methods include mite feeding on either (i) leaves floating on a dsRNA solution, (ii) dsRNA-expressing plants, (iii) artificial diet supplemented with dsRNA, or (iv) dsRNA-coated leaves, and (v) mite soaking in a dsRNA solution. In all cases, the gene targeted for method validation was the Vacuolar-type H+-ATPase (TuVATPase), encoding a constitutively expressed ATP-driven proton pump located in the membrane. Down-regulation of TuVATPase increased mortality and/or reduced fecundity in all methods, but with variable efficiency. The most efficient methods for dsRNA delivery were direct soaking of mites in the dsRNA solution and mite feeding on dsRNA-coated leaves that mimics dsRNA application as a sprayable pesticide. Both resulted in a dark-body phenotype not observed in mites treated with a control dsRNA. Although with lower efficiency, dsRNA designed for TuVATPase silencing and expressed in transgenic Arabidopsis plants impacted the fitness of mites feeding on these plants. RNAi may thus be a valuable strategy to control spider mite populations, either as a sprayable pesticide or through transgenic crops. This comparative methodological study focusing on the induction of RNAi-based gene silencing in T. urticae paves the way for reverse genetics approaches in this model chelicerate system and prepares large-scale systematic RNAi screens as a first step towards the development of specific RNA-based pesticides. Such alternative molecules may help control spider mites that cause significant damages to crops and ornamental plant species, as well as other chelicerates detrimental to agriculture and health. © 2017 Suzuki et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.