Correction: Protocols for the delivery of small molecules to the two-spotted spider mite, Tetranychus urticae

Suzuki, Takeshi; España, María Urizarna; Nunes, Maria Andréia; Zhurov, Vladimir; Dermauw, Wannes; Osakabe, Masahiro; Leeuwen, Thomas Van; Grbić, Miodrag; Grbić, Vojislava

doi:10.1371/journal.pone.0190025

Cited by 7 publications

(7 citation statements)

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“…of 513 bp (DNA sequences used for the generation of dsRNA-TuCOPB2-A and dsRNA-TuCOPB2-B are shown in Fig. 1A)) and delivered them at a concentration of 160 ng/µL to mites using the soaking method 32,34 . As a control, we used the dsRNA-NC (382 bp in length) that has complementarity against the non-transcribed intergenic region of the T. urticae genomic scaffold 12, Fig.…”

Section: Resultsmentioning

confidence: 99%

“…www.nature.com/scientificreports/ dsRNA oral delivery. dsRNA was delivered to mites using the soaking method described by Suzuki et al 32,34 . Briefly, fifty newly-emerged adult female mites were soaked in 50 μL of dsRNA solution (160 ng/μL, unless stated otherwise; 0.1% v/v Tween 20) at 20 °C for 24 h. Subsequently, mites were washed in 100 μL of double distilled water and were transferred onto either bean leaf or bean leaf discs placed on water-soaked cotton kept at 26 °C, 16/8 h light/dark photoperiod, and 50% RH.…”

Section: Preparation Of 32 P-labeled Dsrna Fragments a 1 µL Aliquot mentioning

confidence: 99%

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Environmental RNA interference in two-spotted spider mite, Tetranychus urticae, reveals dsRNA processing requirements for efficient RNAi response

Bensoussan

Dixit

Tabara

et al. 2020

Sci Rep

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Comprehensive understanding of pleiotropic roles of RNAi machinery highlighted the conserved chromosomal functions of RNA interference. The consequences of the evolutionary variation in the core RNAi pathway genes are mostly unknown, but may lead to the species-specific functions associated with gene silencing. The two-spotted spider mite, Tetranychus urticae, is a major polyphagous chelicerate pest capable of feeding on over 1100 plant species and developing resistance to pesticides used for its control. A well annotated genome, susceptibility to RNAi and economic importance, make T. urticae an excellent candidate for development of an RNAi protocol that enables high-throughput genetic screens and RNAi-based pest control. Here, we show that the length of the exogenous dsRNA critically determines its processivity and ability to induce RNAi in vivo. A combination of the long dsRNAs and the use of dye to trace the ingestion of dsRNA enabled the identification of genes involved in membrane transport and 26S proteasome degradation as sensitive RNAi targets. Our data demonstrate that environmental RNAi can be an efficient reverse genetics and pest control tool in T. urticae. In addition, the species-specific properties together with the variation in the components of the RNAi machinery make T. urticae a potent experimental system to study the evolution of RNAi pathways.

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Section: Resultsmentioning

confidence: 99%

Section: Preparation Of 32 P-labeled Dsrna Fragments a 1 µL Aliquot mentioning

confidence: 99%

Environmental RNA interference in two-spotted spider mite, Tetranychus urticae, reveals dsRNA processing requirements for efficient RNAi response

Bensoussan

Dixit

Tabara

et al. 2020

Sci Rep

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“…Previously, Kwon et al 30,31 identified TuCOPB2 as a sensitive RNAi target following leaf-disc mediated delivery of dsRNA. To test the ability of dsRNA-TuCOPB2 to induce RNAi-associated phenotypic changes, we designed two dsRNA-TuCOPB2 fragments (fragment A of 308 bp and B of 513 bp ( Figure 1A)) and delivered them to mites using the soaking method 32,34 . As a control, we used the dsRNA-NC (382 bp in length) that has complementarity against the non-transcribed intergenic region of the T. urticae genomic scaffold 12, Figure 1A.…”

Section: Tucopb2 Is a Sensitive Target For Rnaimentioning

confidence: 99%

“…Fifty newly-emerged adult female mites were soaked in 50 μL of dsRNA solution (160 ng/μL, unless stated otherwise; 0.1% v/v Tween 20) at 20°C for 24 hours 34 . Subsequently, mites were washed in 100 μL of double distilled water and were transferred onto either bean leaf or bean leaf discs placed on watersoaked cotton kept at 26°C, 16/8 h light/dark photoperiod and 50% RH.…”

Section: Dsrna Oral Deliverymentioning

confidence: 99%

Environmental RNA interference in two-spotted spider mite,Tetranychus urticae, reveals dsRNA processing requirements for efficient RNAi response

Bensoussan

Dixit

Tabara

et al. 2020

Preprint

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Comprehensive understanding of pleiotropic roles of RNAi machinery highlighted the conserved chromosomal functions of RNA interference. The consequences of the evolutionary variation in the core RNAi pathway genes are mostly unknown, but may lead to the species-specific functions associated with gene silencing. The two-spotted spider mite, Tetranychus urticae, is a major polyphagous chelicerate pest capable of feeding on over 1,100 plant species and developing resistance to pesticides used for its control. A well annotated genome, susceptibility to RNAi and economic importance, make T. urticae an excellent candidate for development of an RNAi protocol that enables high-throughput genetic screens and RNAibased pest control. Here, we show that the length of the exogenous dsRNA critically determines its processivity and ability to induce RNAi in vivo. A combination of the long dsRNAs and the use of dye to trace the ingestion of dsRNA enabled the identification of genes involved in membrane transport and 26S proteasome degradation as sensitive RNAi targets. Our data demonstrate that environmental RNAi can be an efficient reverse genetics and pest control tool in T. urticae. In addition, the species-specific properties together with the variation in the components of the RNAi machinery make T. urticae a potent experimental system to study the evolution of RNAi pathways.

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“…Phytophagous mites pierce parenchymatic plant cells using stylets to suck their nutrients, and cause severe chlorosis leading to a reduction in crop yield ( Park and Lee, 2002 ; Farouk and Osman, 2011 ; Bensoussan et al, 2016 ). T. urticae is a model within chelicerate herbivores with its genome sequenced and a broad range of tools and protocols developed ( Grbic et al, 2011 ; Cazaux et al, 2014 ; Suzuki et al, 2017 ). Besides, mite ability to feed on Arabidopsis thaliana and the wide available toolkits for this plant species have provided an outstanding opportunity for functional studies of plant-mite interaction ( Santamaria et al, 2012 , 2015a , 2017 ; Zhurov et al, 2014 ).…”

Section: Introductionmentioning

confidence: 99%

Arabidopsis Kunitz Trypsin Inhibitors in Defense Against Spider Mites

et al. 2018

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Tetranychus urticae (two-spotted spider mite) is a striking example of polyphagy among herbivores with an extreme record of pesticide resistance and one of the most significant pests in agriculture. The T. urticae genome contains a large number of cysteine- and serine-proteases indicating their importance in the spider mite physiology. This work is focused on the potential role of the Kunitz trypsin inhibitor (KTI) family on plant defense responses against spider mites. The molecular characterization of two of these genes, AtKTI4 and AtKTI5, combined with feeding bioassays using T-DNA insertion lines for both genes was carried out. Spider mite performance assays showed that independent KTI silencing Arabidopsis lines conferred higher susceptibility to T. urticae than WT plants. Additionally, transient overexpression of these inhibitors in Nicotiana benthamiana demonstrated their ability to inhibit not only serine- but also cysteine-proteases, indicating the bifunctional inhibitory role against both types of enzymes. These inhibitory properties could be involved in the modulation of the proteases that participate in the hydrolysis of dietary proteins in the spider mite gut, as well as in other proteolytic processes.

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Correction: Protocols for the delivery of small molecules to the two-spotted spider mite, Tetranychus urticae

Cited by 7 publications

References 1 publication

Environmental RNA interference in two-spotted spider mite, Tetranychus urticae, reveals dsRNA processing requirements for efficient RNAi response

Environmental RNA interference in two-spotted spider mite, Tetranychus urticae, reveals dsRNA processing requirements for efficient RNAi response

Environmental RNA interference in two-spotted spider mite,Tetranychus urticae, reveals dsRNA processing requirements for efficient RNAi response

Arabidopsis Kunitz Trypsin Inhibitors in Defense Against Spider Mites

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