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

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 (d...

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Autores principales: Suzuki, Takeshi, Nunes, Maria Andreia, España, María Urizarna, Namin, Hooman Hosseinzadeh, Jin, Pengyu, Bensoussan, Nicolas, Zhurov, Vladimir, Rahman, Tawhid, De Clercq, Rebecca, Hilson, Pierre, Grbic, Vojislava, Grbic, Miodrag
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5507529/
https://www.ncbi.nlm.nih.gov/pubmed/28704448
http://dx.doi.org/10.1371/journal.pone.0180654
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author Suzuki, Takeshi
Nunes, Maria Andreia
España, María Urizarna
Namin, Hooman Hosseinzadeh
Jin, Pengyu
Bensoussan, Nicolas
Zhurov, Vladimir
Rahman, Tawhid
De Clercq, Rebecca
Hilson, Pierre
Grbic, Vojislava
Grbic, Miodrag
author_facet Suzuki, Takeshi
Nunes, Maria Andreia
España, María Urizarna
Namin, Hooman Hosseinzadeh
Jin, Pengyu
Bensoussan, Nicolas
Zhurov, Vladimir
Rahman, Tawhid
De Clercq, Rebecca
Hilson, Pierre
Grbic, Vojislava
Grbic, Miodrag
author_sort Suzuki, Takeshi
collection PubMed
description 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.
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spelling pubmed-55075292017-07-25 RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing Suzuki, Takeshi Nunes, Maria Andreia España, María Urizarna Namin, Hooman Hosseinzadeh Jin, Pengyu Bensoussan, Nicolas Zhurov, Vladimir Rahman, Tawhid De Clercq, Rebecca Hilson, Pierre Grbic, Vojislava Grbic, Miodrag PLoS One Research Article 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. Public Library of Science 2017-07-12 /pmc/articles/PMC5507529/ /pubmed/28704448 http://dx.doi.org/10.1371/journal.pone.0180654 Text en © 2017 Suzuki et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Suzuki, Takeshi
Nunes, Maria Andreia
España, María Urizarna
Namin, Hooman Hosseinzadeh
Jin, Pengyu
Bensoussan, Nicolas
Zhurov, Vladimir
Rahman, Tawhid
De Clercq, Rebecca
Hilson, Pierre
Grbic, Vojislava
Grbic, Miodrag
RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing
title RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing
title_full RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing
title_fullStr RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing
title_full_unstemmed RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing
title_short RNAi-based reverse genetics in the chelicerate model Tetranychus urticae: A comparative analysis of five methods for gene silencing
title_sort rnai-based reverse genetics in the chelicerate model tetranychus urticae: a comparative analysis of five methods for gene silencing
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5507529/
https://www.ncbi.nlm.nih.gov/pubmed/28704448
http://dx.doi.org/10.1371/journal.pone.0180654
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