RNAi-based gene silencing through dsRNA injection or ingestion against the African sweet potato weevil<i>Cylas puncticollis</i>(Coleoptera: Brentidae)

Prentice, Katterinne; Christiaens, Olivier; Pertry, Ine; Bailey, Ana M.; Niblett, C. L.; Ghislain, Marc; Gheysen, Godelieve; Smagghe, Guy

doi:10.1002/ps.4337

Cited by 94 publications

(88 citation statements)

References 45 publications

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“…The degradation in C. puncticollis gut secretions happens faster than in C. brunneus . Comparing the oral RNAi results presented here and by Prentice et al 38,. we observed that C. brunneus is more sensitive to oral RNAi than C. puncticollis .…”

Section: Discussionsupporting

confidence: 63%

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RNA interference: a promising biopesticide strategy against the African Sweetpotato Weevil Cylas brunneus

Christiaens

Prentice

Pertry

et al. 2016

Sci Rep

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The African sweetpotato weevil Cylas brunneus is one of the most devastating pests affecting the production of sweetpotatoes, an important staple food in Sub-Saharan Africa. Current available control methods against this coleopteran pest are limited. In this study, we analyzed the potential of RNA interference as a novel crop protection strategy against this insect pest. First, the C. brunneus transcriptome was sequenced and RNAi functionality was confirmed by successfully silencing the laccase2 gene. Next, 24 potential target genes were chosen, based on their critical role in vital biological processes. A first screening via injection of gene-specific dsRNAs showed that the dsRNAs were highly toxic for C. brunneus. Injected doses of 200ng/mg body weight led to mortality rates of 90% or higher for 14 of the 24 tested genes after 14 days. The three best performing dsRNAs, targeting prosα2, rps13 and the homolog of Diabrotica virgifera snf7, were then used in further feeding trials to investigate RNAi by oral delivery. Different concentrations of dsRNAs mixed with artificial diet were tested and concentrations as low as 1 μg dsRNA/ mL diet led to significant mortality rates higher than 50%.These results proved that dsRNAs targeting essential genes show great potential to control C. brunneus.

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Section: Discussionsupporting

confidence: 63%

“…we observed that C. brunneus is more sensitive to oral RNAi than C. puncticollis . This could be related to the stronger dsRNA degradation capacity that was observed in C. puncticollis compared to C. brunneus 38.…”

Section: Discussionmentioning

confidence: 98%

RNA interference: a promising biopesticide strategy against the African Sweetpotato Weevil Cylas brunneus

Christiaens

Prentice

Pertry

et al. 2016

Sci Rep

Self Cite

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“…Similar to the results from previous studies of the cotton boll weevil and the African sweet potato weevil (Baum et al, ; Macedo et al, ; Prentice et al, ), our data show that robust gene silencing can be achieved when naked dsRNAs are delivered through injections, but not through oral ingestion. A recent study in the cotton boll weevil showed that midgut nucleases degraded dsRNA and precluded orally delivered dsRNA from achieving strong RNAi (Almeida Garcia et al, ).…”

Section: Discussionsupporting

confidence: 91%

“…RNA interference has been shown to be effective in inducing gene silencing in the cotton boll weevil Anthonomus grandis (Coleoptera: Curculionidae) and the African sweet potato weevil Cylas puncticollis (Coleoptera: Brentidae) (Gillet et al, ; Macedo et al, ; Prentice et al, ). Interestingly, unlike western corn rootworm, dsRNAs delivered through ingestion were not as effective as those delivered through injections in eliciting strong RNAi responses in either weevil species (Baum et al, ; Macedo et al, ; Prentice et al, ). A recent study in cotton boll weevil showed degradation by midgut nucleases was major cause for the lack of a strong RNAi response following oral delivery of dsRNA (Almeida Garcia et al, ).…”

Section: Introductionmentioning

confidence: 99%

Lethal RNA interference response in the pepper weevil

Taylor

Pinheiro

et al. 2019

J Applied Entomology

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The pepper weevil, Anthonomus eugenii Cano (Coleoptera: Curculionidae) is an important pest of peppers (Capsicum spp.) in the southern United States, Mexico, and Central America. Current management practices rely on chemical control of adults although grower options are limited. A more integrated strategy to manage the pepper weevil infestations may benefit from measures that can control this pest specifically such as RNA interference (RNAi). We cloned and sequenced transcript fragments of two possible essential genes of the pepper weevil, Snf7 and V‐ATPase‐A. We treated pepper weevil females with the double‐stranded RNAs (dsRNAs) of these genes and found that orally delivered dsRNA of either gene was not effective in eliciting strong RNAi response or causing mortality. In contrast, Snf7 and V‐ATPase‐A dsRNAs delivered through injections produced robust gene silencing and led to more than 90% mortality within 12 days. Our results suggest that the pepper weevil possesses the molecular machinery needed for a robust RNAi response and that Snf7 and V‐ATPase‐A genes likely encode essential functions in this insect. Moreover, RNAi‐mediated silencing of essential genes can potentially be used to control this pest if strong RNAi response can be induced by orally delivered dsRNA.

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“…We demonstrated significant reduction in the transcript levels of all genes tested 3 days after dsRNA injection, suggesting that SLW adults possess a functional RNAi machinery and systemic RNAi response. Previous studies in weevils Cylas puncticollis and Cylas brunneus reported that injection of dsRNA for the same genes resulted in high lethality . Although gene expression was not evaluated in these previous works, the insect mortality was presumed to be due to gene knockdown.…”

Section: Discussionmentioning

confidence: 99%

Delivery of gene‐specific dsRNA by microinjection and feeding induces RNAi response in Sri Lanka weevil, Myllocerus undecimpustulatus undatus Marshall

Pinheiro

Taylor

et al. 2019

Pest Management Science

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BACKGROUND RNA interference (RNAi) is widely used in entomological research for functional analysis of genes and is being considered as a new tool for insect pest management. Sri Lanka weevil (SLW) is a highly polyphagous pest of agronomically important plants, but currently only a few control methods are available for this insect. RESULTS In the present study, we evaluated the stability of candidate reference genes β‐ACT, α‐TUB, EF1‐α, RPL12 and GAPDH, and identified EF1‐α as the most reliable for gene expression normalization. Four target genes involved in different cellular processes, including Prosα2, RPS13, Snf7 and V‐ATPase A were selected to evaluate whether RNAi response in SLW adults can be triggered by microinjection and oral feeding of their double‐stranded RNAs (dsRNAs). Three days after injection of the dsRNAs for the target genes, their transcript levels were significantly reduced (up to 91.4%) when compared to the control. Additionally, weevils fed with the target dsRNAs showed significant decreases in gene transcript levels and significant mortality was observed in insects treated with Prosα2 and Snf7 dsRNAs (78.6 to 92.7%). CONCLUSION Our data demonstrate that microinjection and feeding of dsRNA produce a strong RNAi response in SLW, indicating that RNAi‐based strategies could be explored to develop a selective and environmentally safe control method against SLW. © 2019 Society of Chemical Industry

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RNAi-based gene silencing through dsRNA injection or ingestion against the African sweet potato weevilCylas puncticollis(Coleoptera: Brentidae)

Cited by 94 publications

References 45 publications

RNA interference: a promising biopesticide strategy against the African Sweetpotato Weevil Cylas brunneus

RNA interference: a promising biopesticide strategy against the African Sweetpotato Weevil Cylas brunneus

Lethal RNA interference response in the pepper weevil

Delivery of gene‐specific dsRNA by microinjection and feeding induces RNAi response in Sri Lanka weevil, Myllocerus undecimpustulatus undatus Marshall

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