Development of efficient RNAi in <i>Nezara viridula</i> for use in insecticide target discovery

Riga, Maria; Denecke, Shane; Livadaras, Ioannis; Geibel, Sven; Nauen, Ralf; Vontas, John

doi:10.1002/arch.21650

Cited by 21 publications

(18 citation statements)

References 39 publications

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“…Also, we identified suitable target genes for oral delivery. Previously, the core genes of the RNAi mechanism were found to be expressed in N. viridula, 33 and more recently, Riga et al 29 have demonstrated RNAi functionality by microinjection of gene-specific dsRNA in this species.…”

Section: Discussionmentioning

confidence: 99%

“…28 In SGSB, recently, it has been demonstrated that microinjection-mediated delivery of dsRNA targeting essential genes led to significant mortality in the nymphs. 29 The main objectives of this study were to first identify suitable target genes for oral delivery and then to evaluate the RNAi response of SGSB to orally delivered dsRNA. It is critical to identify the RNAi efficacy of orally delivered dsRNA in order to develop biopesticides based on this technology.…”

Section: Introductionmentioning

confidence: 99%

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RNAi‐mediated mortality in southern green stinkbug Nezara viridula by oral delivery of dsRNA

Sharma

Christiaens

Taning

et al. 2020

Pest Management Science

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BACKGROUND: The southern green stinkbug, Nezara viridula (Hemiptera: Pentatomidae), is an important emerging polyphagous pest infesting soybean in the United States, Brazil and Argentina. The indiscriminate use of synthetic insecticides to control stinkbugs has limited the effectiveness of current management strategies. Alternatively, RNA interference (RNAi) has emerged as a novel mode of action to control pests in an eco-friendly manner. RESULTS: Here, we assessed the potential of RNAi technology by oral delivery of double-stranded RNA (dsRNA) for the control of N. viridula. Initially, ten candidate genes were tested by microinjection assay to select the best target genes for oral delivery. Seven genes resulted in more than 90% mortality after microinjection. To evaluate RNAi efficacy by oral delivery of dsRNA, five genes were tested by feeding the insects on gene-specific dsRNA mixed with an artificial diet. Significant mortality of 43% and 45% was observed after 14 days of treatment with ds⊍Cop and dsvATPase A, respectively. To elucidate the lower RNAi efficacy via oral delivery of dsRNA, ex vivo dsRNA degradation in the saliva and the midgut juice was performed, which indicated that the reduced RNAi efficacy is accompanied by a rapid degradation of dsRNA by digestive secretions. CONCLUSION: This study proves that RNAi can be triggered by orally delivered dsRNA in N. viridula and can be exploited to control this economically important pest. The reduced stability of dsRNA in saliva and midgut that was observed indicates a need to further improve RNAi efficacy, for example by use of specific formulations.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

RNAi‐mediated mortality in southern green stinkbug Nezara viridula by oral delivery of dsRNA

Sharma

Christiaens

Taning

et al. 2020

Pest Management Science

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“…Post-transcriptional gene silencing also known as RNA interference (RNAi) was first elucidated in 1998 [5] and has since then been widely used as a tool in the study of gene function. In the hemipteran group, RNAi has been used to study the role of genes in insect development/reproduction of several species, such as Oncopeltus fasciatus [6,7], Nezara viridula ( [8], Diaphorina citri [9], Halyomorpha halys [10], among others. RNAi is a highly conserved mechanism among eukaryotic organisms, in which the messenger RNA (mRNA) is cleaved by the RNAi machinery, leading to the inactivation of gene expression [11].…”

Section: Introductionmentioning

confidence: 99%

RNAi and CRISPR/Cas9 as Functional Genomics Tools in the Neotropical Stink Bug, Euschistus heros

Cagliari

Smagghe

Zotti

et al. 2020

Insects

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The Neotropical brown stink bug, Euschistus heros, is one of the most important stink bug pests in leguminous plants in South America. RNAi and CRISPR/Cas9 are important and useful tools in functional genomics, as well as in the future development of new integrated pest management strategies. Here, we explore the use of these technologies as complementing functional genomic tools in E. heros. Three genes, abnormal wing disc (awd), tyrosine hydroxylase (th) and yellow (yel), known to be involved in wing development (awd) and the melanin pathway (th and yel) in other insects, were chosen to be evaluated using RNAi and CRISPR/Cas9 as tools. First, the genes were functionally characterized using RNAi knockdown technology. The expected phenotype of either deformed wing or lighter cuticle pigmentation/defects in cuticle sclerotization was observed for awd and th, respectively. However, for yel, no obvious phenotype was observed. Based on this, yel was selected as a target for the development of a CRISPR/Cas9 workflow to study gene knockout in E. heros. A total of 719 eggs were injected with the Cas9 nuclease (300 ng/µL) together with the sgRNA (300 ng/µL) targeting yel. A total of six insects successfully hatched from the injected eggs and one of the insects showed mutation in the target region, however, the phenotype was still not obvious. Overall, this study for the first time provides a useful CRISPR/Cas9 gene editing methodology to complement RNAi for functional genomic studies in one of the most important and economically relevant stink bug species.

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“…Hemiptera genes have been successfully silenced through this method (Christiaens & Smagghe, 2014;Bansal et al, 2016;Ghosh et al, 2017;Lu et al, 2017;Mogilicherla et al, 2018, Riga et al, 2019.…”

Section: Discussionmentioning

confidence: 99%

Tyramine receptor drives olfactory response to (E)-2-decenal in the stink bugHalyomorpha halys

Finetti

Pezzi

Civolani

et al. 2020

Preprint

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In insects, the tyramine receptor 1 (TAR1) has been shown to control several physiological functions, including olfaction. We investigated the molecular and functional profile of the Halyomorpha halys type 1 tyramine receptor gene (HhTAR1) and its role in olfactory functions of this pest. Molecular and pharmacological analyses confirmed that the HhTAR1 gene codes for a true TAR1. The RT-qPCR analysis revealed that HhTAR1 is expressed mostly in adult brain and antennae as well as in early development stages (eggs, 1st and 2nd instar nymphs). In particular, among the antennomeres that compose a typical H. halys antenna, HhTAR1 was more expressed in flagellomeres. Scanning electron microscopy (SEM) investigation revealed the type and distribution of sensilla on adult H. halys antennae: both flagellomeres appear rich in trichoid and grooved sensilla, known to be associated with olfactory functions. Through a RNAi approach, topically delivered HhTAR1 dsRNA induced a 50 % gene downregulation after 24 h in H. halys 2nd instar nymphs. An innovative behavioral assay revealed that HhTAR1 RNAi-silenced 2nd instar nymphs were less susceptible to the alarm pheromone component (E)-2 decenal as compared to control. These results provide critical information concerning the TAR1 role in olfaction regulation, especially alarm pheromone reception, in H. halys. Furthermore, considering the emerging role of TAR1 as target of biopesticides, this work paves the way for further investigation on innovative methods for controlling H. halys.

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Development of efficient RNAi in Nezara viridula for use in insecticide target discovery

Cited by 21 publications

References 39 publications

RNAi‐mediated mortality in southern green stinkbug Nezara viridula by oral delivery of dsRNA

RNAi‐mediated mortality in southern green stinkbug Nezara viridula by oral delivery of dsRNA

RNAi and CRISPR/Cas9 as Functional Genomics Tools in the Neotropical Stink Bug, Euschistus heros

Tyramine receptor drives olfactory response to (E)-2-decenal in the stink bugHalyomorpha halys

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