RNA interference is induced in the glassy winged sharpshooter <i>Homalodisca vitripennis</i> by actin dsRNA

Rosa, Cristina; Kamita, Shizuo G.; Falk, Bryce W.

doi:10.1002/ps.3253

Cited by 28 publications

(23 citation statements)

References 54 publications

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“…A colony of H. vitripennis was maintained at the University of California-Davis Contained Research Facility (CRF) in cages containing a mixture of host plants as previously described [20]. Eight day-old adult insects (males and females) were collected in three different pools of ∼50 insects each.…”

Section: Methodsmentioning

confidence: 99%

“…H . vitripennis is a very economically important pest of a wide range of plants including Citrus spp., grapes ( Vitis vinifera ), and almonds ( Prunus dulcis ) [20]. This insect also serves as a very robust vector of the bacterium Xylella fastidiosa, the causal agent of Pierce’s disease of grapevines and citrus variegated chlorosis disease [21].…”

Section: Introductionmentioning

confidence: 99%

“…In a few highly established model organisms, such as Drosophila , hairpin constructs can be used to overexpress dsRNA in particular tissues and at specific developmental stages [24]. Similar strategies have been tested against adult H. vitripennis [20], [25] and other insects [26]. However, robust systemic RNAi is absent in insects [27] because they lack a functional RNA dependent RNA polymerase (RdRP) that amplifies small RNAs.…”

Section: Introductionmentioning

confidence: 99%

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Sequencing and De Novo Assembly of the Transcriptome of the Glassy-Winged Sharpshooter (Homalodisca vitripennis)

et al. 2013

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BackgroundThe glassy-winged sharpshooter Homalodisca vitripennis (Hemiptera: Cicadellidae), is a xylem-feeding leafhopper and important vector of the bacterium Xylella fastidiosa; the causal agent of Pierce’s disease of grapevines. The functional complexity of the transcriptome of H. vitripennis has not been elucidated thus far. It is a necessary blueprint for an understanding of the development of H. vitripennis and for designing efficient biorational control strategies including those based on RNA interference.ResultsHere we elucidate and explore the transcriptome of adult H. vitripennis using high-throughput paired end deep sequencing and de novo assembly. A total of 32,803,656 paired-end reads were obtained with an average transcript length of 624 nucleotides. We assembled 32.9 Mb of the transcriptome of H. vitripennis that spanned across 47,265 loci and 52,708 transcripts. Comparison of our non-redundant database showed that 45% of the deduced proteins of H. vitripennis exhibit identity (e-value ≤1−5) with known proteins. We assigned Gene Ontology (GO) terms, Kyoto Encyclopedia of Genes and Genomes (KEGG) annotations, and potential Pfam domains to each transcript isoform. In order to gain insight into the molecular basis of key regulatory genes of H. vitripennis, we characterized predicted proteins involved in the metabolism of juvenile hormone, and biogenesis of small RNAs (Dicer and Piwi sequences) from the transcriptomic sequences. Analysis of transposable element sequences of H. vitripennis indicated that the genome is less expanded in comparison to many other insects with approximately 1% of the transcriptome carrying transposable elements.ConclusionsOur data significantly enhance the molecular resources available for future study and control of this economically important hemipteran. This transcriptional information not only provides a more nuanced understanding of the underlying biological and physiological mechanisms that govern H. vitripennis, but may also lead to the identification of novel targets for biorationally designed control strategies.

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Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Sequencing and De Novo Assembly of the Transcriptome of the Glassy-Winged Sharpshooter (Homalodisca vitripennis)

et al. 2013

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“…Alternative, efficient, pathogen-specific, environmentally friendly, and safe approaches to control these diseases would lead to long-term sustainability of crop systems. Strategies to control X. fastidiosa currently in development, based on genomics-derived knowledge and the production of transgenic plants, focus on either the pathogen (e.g., Chatterjee et al 2008c;Dandekar et al 2012) or the vector (such as RNAi to impact insect development, not discussed here; Rosa et al 2012), or on both partners and their interactions during insect transmission . We limit our discussion to these approaches, although it should be mentioned that genomic data has provided great insights into the biology, ecology, evolution, and taxonomy of X. fastidiosa, much of which has been useful to devise other disease management strategies and for detection and quarantine purposes.…”

Section: Disease Management Strategiesmentioning

confidence: 99%

Genomic Insights into Xylella fastidiosa Interactions with Plant and Insect Hosts

Retchless

Labroussaa

Shapiro

et al. 2014

Genomics of Plant-Associated Bacteria

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“…Microinjection of a synthetic dsRNA solution into the hemolymph has been demonstrated to induce RNAi in hemipterans, such as whitefly ( B . tabaci) [21], pea aphid ( Acyrthosiphon pisum ) [22,23], glassy winged sharpshooter ( Homalodiscus vitripennis ) [24], and for dipterans, such as the fruit fly ( Drosophila melanogaster ) [25]. Initially, microinjection was the prefered mode of delivery because it was not known whether RNAi was cell-autonomous and/or systemic among taxonomically different groups of insect groups [12,26–28].…”

Section: Introductionmentioning

confidence: 99%

Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs

et al. 2017

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Control of the whitefly Bemisia tabaci (Genn.) agricultural pest and plant virus vector relies on the use of chemical insecticides. RNA-interference (RNAi) is a homology-dependent innate immune response in eukaryotes, including insects, which results in degradation of the corresponding transcript following its recognition by a double-stranded RNA (dsRNA) that shares 100% sequence homology. In this study, six whitefly ‘gut’ genes were selected from an in silico-annotated transcriptome library constructed from the whitefly alimentary canal or ‘gut’ of the B biotype of B. tabaci, and tested for knock down efficacy, post-ingestion of dsRNAs that share 100% sequence homology to each respective gene target. Candidate genes were: Acetylcholine receptor subunit α, Alpha glucosidase 1, Aquaporin 1, Heat shock protein 70, Trehalase1, and Trehalose transporter1. The efficacy of RNAi knock down was further tested in a gene-specific functional bioassay, and mortality was recorded in 24 hr intervals, six days, post-treatment. Based on qPCR analysis, all six genes tested showed significantly reduced gene expression. Moderate-to-high whitefly mortality was associated with the down-regulation of osmoregulation, sugar metabolism and sugar transport-associated genes, demonstrating that whitefly survivability was linked with RNAi results. Silenced Acetylcholine receptor subunit α and Heat shock protein 70 genes showed an initial low whitefly mortality, however, following insecticide or high temperature treatments, respectively, significantly increased knockdown efficacy and death was observed, indicating enhanced post-knockdown sensitivity perhaps related to systemic silencing. The oral delivery of gut-specific dsRNAs, when combined with qPCR analysis of gene expression and a corresponding gene-specific bioassay that relates knockdown and mortality, offers a viable approach for functional genomics analysis and the discovery of prospective dsRNA biopesticide targets. The approach can be applied to functional genomics analyses to facilitate, species-specific dsRNA-mediated control of other non-model hemipterans.

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RNA interference is induced in the glassy winged sharpshooter Homalodisca vitripennis by actin dsRNA

Abstract: The findings indicate that RNAi is induced in post-embryonic leafhoppers by dsRNA. The present system can be used to screen potential gene-silencing targets that can be used for reducing the vector competence of H. vitripennis and other leafhoppers.

Cited by 28 publications

References 54 publications

Sequencing and De Novo Assembly of the Transcriptome of the Glassy-Winged Sharpshooter (Homalodisca vitripennis)

Sequencing and De Novo Assembly of the Transcriptome of the Glassy-Winged Sharpshooter (Homalodisca vitripennis)

Genomic Insights into Xylella fastidiosa Interactions with Plant and Insect Hosts

Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs

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