Advanced search
1 file | 2.53 MB Add to list

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

(2017) PLOS ONE. 12(7).
Author
Organization
Abstract
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 (TuVAT-Pase), 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.
Keywords
C-ELEGANS, INTERFERENCE, INSECT, GENOME, DSRNA, ARABIDOPSIS, EUKARYOTES, GENERATION, SEQUENCE, ACIDS

Downloads

  • 8567016.pdf
    • full text
    • |
    • open access
    • |
    • PDF
    • |
    • 2.53 MB

Citation

Please use this url to cite or link to this publication:

MLA
Suzuki, Takeshi et al. “RNAi-based Reverse Genetics in the Chelicerate Model Tetranychus Urticae: A Comparative Analysis of Five Methods for Gene Silencing.” PLOS ONE 12.7 (2017): n. pag. Print.
APA
Suzuki, Takeshi, Nunes, M. A., España, M. U., Namin, H. H., Jin, P., Bensoussan, N., Zhurov, V., et al. (2017). RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing. PLOS ONE, 12(7).
Chicago author-date
Suzuki, Takeshi, Maria Andreia Nunes, Maria Urizarna España, Hooman Hosseinzadeh Namin, Pengyu Jin, Nicolas Bensoussan, Vladimir Zhurov, et al. 2017. “RNAi-based Reverse Genetics in the Chelicerate Model Tetranychus Urticae: A Comparative Analysis of Five Methods for Gene Silencing.” Plos One 12 (7).
Chicago author-date (all authors)
Suzuki, Takeshi, Maria Andreia Nunes, Maria Urizarna España, Hooman Hosseinzadeh Namin, Pengyu Jin, Nicolas Bensoussan, Vladimir Zhurov, Tawhid Rahman, Rebecca De Clercq, Pierre Hilson, Vojislava Grbic, and Miodrag Grbic. 2017. “RNAi-based Reverse Genetics in the Chelicerate Model Tetranychus Urticae: A Comparative Analysis of Five Methods for Gene Silencing.” Plos One 12 (7).
Vancouver
1.
Suzuki T, Nunes MA, España MU, Namin HH, Jin P, Bensoussan N, et al. RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing. PLOS ONE. 2017;12(7).
IEEE
[1]
T. Suzuki et al., “RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing,” PLOS ONE, vol. 12, no. 7, 2017.
@article{8567526,
  abstract     = {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 (TuVAT-Pase), 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.},
  articleno    = {e0180654},
  author       = {Suzuki, Takeshi and Nunes, Maria Andreia and España, Maria Urizarna and Namin, Hooman Hosseinzadeh and Jin, Pengyu and Bensoussan, Nicolas and Zhurov, Vladimir and Rahman, Tawhid and De Clercq, Rebecca and Hilson, Pierre and Grbic, Vojislava and Grbic, Miodrag},
  issn         = {1932-6203},
  journal      = {PLOS ONE},
  keywords     = {C-ELEGANS,INTERFERENCE,INSECT,GENOME,DSRNA,ARABIDOPSIS,EUKARYOTES,GENERATION,SEQUENCE,ACIDS},
  language     = {eng},
  number       = {7},
  pages        = {22},
  title        = {RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing},
  url          = {http://dx.doi.org/10.1371/journal.pone.0180654},
  volume       = {12},
  year         = {2017},
}

Altmetric
View in Altmetric
Web of Science
Times cited: