Systemic RNAi in the small hive beetle<i>Aethina tumida</i>Murray (Coleoptera: Nitidulidae), a serious pest of the European honey bee<i>Apis mellifera</i>

Powell, Michelle E.; Bradish, Hannah M; Gatehouse, John A.; Fitches, Elaine

doi:10.1002/ps.4365

Cited by 51 publications

(47 citation statements)

References 42 publications

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“…Not surprisingly, due to the lack of sequence similarity, dsSK also showed no effects. A similar observation in A. mellifera was reported by Powell et al ., who demonstrated that injections into honeybees of 50 ng of dsRNAs Laccase 2 and vacuolar ‐ATPase V‐type subunit A , designed to target Aethina humidity , similarly had no effect on bee survival and did not induce the suppression of either of the target genes. An alignment of A. mellifera and A. tumida Laccase 2 and V‐ATPase subunit A mRNAs indicated sequence identities of 74% and 68%, respectively.…”

Section: Discussionsupporting

confidence: 82%

“…Stability studies of dsRNA provided proof that the dsRNA of the target genes was not degraded significantly during 48 h. Our results agree with those of Li et al ., which indicated that three dsRNAs targeting different sites within a gene encoding vacuolar ATP synthase subunit E in Nilaparvata lugens were found to be stable in 0.1 g mL –1 sucrose solution at 22 h. The Ap_ST1 dsRNAs (sugar transporter gene) from an aphid kept for a week at room temperature was not degraded, thus again demonstrating stability . However, 1 μg of V‐ATPase subunit dsRNA targeting A. tumida in 10 μL of 50% sucrose solution after 8 h is completely degraded . These contrasting results demonstrate the importance of evaluating the stability of specific dsRNAs under the appropriate conditions.…”

Section: Discussionmentioning

confidence: 99%

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Targeting the potassium ion channel genes SK and SH as a novel approach for control of insect pests: efficacy and biosafety

Alshukri

Astarita

Al‐Esawy

et al. 2019

Pest Management Science

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BACKGROUND Potassium ion channels play a critical role in the generation of electrical signals and thus provide potential targets for control of insect pests by RNA interference. RESULTS Genes encoding the small conductance calcium‐activated potassium channel (SK) and the voltage‐gated potassium channel (SH) were knocked down in Tribolium castaneum by injection and oral delivery of dsRNA (dsTcSK and dsTcSH, respectively). Irrespective of the delivery mechanism a dose‐dependent effect was observed for knockdown (KD) of gene expression and insect mortality for both genes. Larvae fed a 400 ng dsRNA mg–1 diet showed significant gene (P < 0.05) knockdown (98% and 83%) for SK and SH, respectively, with corresponding mortalities of 100% and 98% after 7 days. When injected (248.4 ng larva–1), gene KD was 99% and 98% for SK and SH, causing 100% and 73.4% mortality, respectively. All developmental stages tested (larvae, early‐ and late‐stage pupae and adults) showed an RNAi‐sensitive response for both genes. LC50 values were lower for SK than SH, irrespective of delivery method, demonstrating that the knockdown of SK had a greater effect on larval mortality. Biosafety studies using adult honeybee Apis mellifera showed that there were no significant differences either in expression levels or mortality of honeybees orally dosed with dsTcSK and dsTcSH compared to control‐fed bees. Similarly, there was no significant difference in the titre of deformed wing virus, used as a measure of immune suppression, between experimental and control bees. CONCLUSION This study demonstrates the potential of using RNAi targeting neural receptors as a technology for the control of T. castaneum. © 2019 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

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Section: Discussionsupporting

confidence: 82%

Section: Discussionmentioning

confidence: 99%

Targeting the potassium ion channel genes SK and SH as a novel approach for control of insect pests: efficacy and biosafety

Alshukri

Astarita

Al‐Esawy

et al. 2019

Pest Management Science

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“…The level of sensitivity to RNAi by microinjection and oral exposure can be variable among insects within the same order. For example, in the Coleoptera order, D. virgifera virgifera , T. castaneum , Leptinotarsa decemlineata , Meligethes aeneus , Rhynchophorus ferrugineus , C. puncticollis , C. brunneus and A. planipennis are sensitive to both methods of dsRNA delivery, whereas Anthonomus grandis , Anthonomus eugenii and Aethina tumida are recalcitrant to RNAi by oral exposure . These studies demonstrate the importance of evaluating the RNAi sensitivity by microinjection as well as by feeding in the target insect species since it is not possible to predict the RNAi effectiveness based on the RNAi response found in other insects, even in closely related species.…”

Section: Discussionmentioning

confidence: 68%

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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“…V‐ATPase seems to be involved in growth and body formation in various orders of insects. Phenotypes of deformity induced by RNAi of V‐ATPases have been reported among several orders of insects including L. migratoria , A. tumida , and P. maidis (Li & Xia, ; Yao et al ., ; Powell et al ., ) (Fig. ).…”

Section: Discussionmentioning

confidence: 93%

“…To our knowledge, functions of V-ATPases in insects have been studied only in species belonging to six orders (Orthoptera, Thysanoptera, Hemiptera, Coleoptera, Lepidoptera, Diptera) by RNA interference (RNAi), which can inhibit gene expression in a sequence specific manner by introduction of double-stranded RNA (dsRNA) (Miller et al, 2012;Miyata et al, 2014). Knockdown of V-ATPase causes lethal effect on various species suggesting this molecule is essential in vital physiological activities (Baum et al, 2007;Whyard et al, 2009;Gu et al, 2011;Upadhyay et al, 2011;Li & Xia, 2012;Khan et al, 2013;Yao et al, 2013;Badillo-Vargas et al, 2015;Jin et al, 2015;Rebijith et al, 2016;Powell et al, 2017). Therefore, V-ATPase has been proposed as a putative target molecule for the development of in planta dsRNA system to control insect pests (Baum et al, 2007;Kola et al, 2015).…”

Section: Introductionmentioning

confidence: 99%

Systemic RNAi of V‐ATPase subunit B causes molting defect and developmental abnormalities in Periplaneta fuliginosa

et al. 2018

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The vacuolar (H )-ATPases (V-ATPases) are ATP-driven proton pumps with multiple functions in many organisms. In this study, we performed structural and functional analysis of vha55 gene that encodes V-ATPase subunit B in the smokybrown cockroach Periplaneta fuliginosa (Blattodea). We observed a high homology score of the deduced amino acid sequences between 10 species in seven orders. RNAi of the vha55 gene in P. fuliginosa caused nymphal/nymphal molting defects with incomplete shedding of old cuticles, growth inhibition, as well as bent and wrinkled cuticles of thoraxes and abdominal segments. Since growth inhibition caused by vha55 RNAi did not interfere in the commencement of cockroach molting, molting timing and body growth might be controlled by independent mechanism. Our study suggested V-ATPases might be a good candidate molecule for evolutionary and developmental studies of insect molting.

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Systemic RNAi in the small hive beetleAethina tumidaMurray (Coleoptera: Nitidulidae), a serious pest of the European honey beeApis mellifera

Abstract: This is the first study to show evidence for systemic RNAi in A. tumida in response to injected dsRNA, but further research is required to develop methods to induce RNAi effects via ingestion. © 2016 Crown copyright. Pest Management Science © 2016 Society of Chemical Industry.

Cited by 51 publications

References 42 publications

Targeting the potassium ion channel genes SK and SH as a novel approach for control of insect pests: efficacy and biosafety

Targeting the potassium ion channel genes SK and SH as a novel approach for control of insect pests: efficacy and biosafety

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

Systemic RNAi of V‐ATPase subunit B causes molting defect and developmental abnormalities in Periplaneta fuliginosa

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