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

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    Author
    Vyas, Meenal
    Raza, Amir
    Ali, Muhammad Yousaf
    Ashraf, Muhammad Aleem
    Mansoor, Shahid
    Shahid, Ahmad Ali
    Brown, Judith K.
    Affiliation
    Univ Arizona, Sch Plant Sci
    Issue Date
    2017-01-03
    
    Metadata
    Show full item record
    Publisher
    PUBLIC LIBRARY SCIENCE
    Citation
    Knock down of Whitefly Gut Gene Expression and Mortality by Orally Delivered Gut Gene-Specific dsRNAs 2017, 12 (1):e0168921 PLOS ONE
    Journal
    PLOS ONE
    Rights
    © 2017 Vyas et al. This is an open access article distributed under the terms of the Creative Commons Attribution License
    Collection Information
    This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at repository@u.library.arizona.edu.
    Abstract
    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 a, Alpha glucosidase 1, Aquaporin 1, Heat shock protein 70, Trehalasel, and Trehalose transported. 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 a 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.
    Note
    Open Access Journal
    ISSN
    1932-6203
    DOI
    10.1371/journal.pone.0168921
    Version
    Final published version
    Sponsors
    USDA [6402-21310-003-10S]; Higher Education Commission of Pakistan-International Research Support Initiative Program
    Additional Links
    http://dx.plos.org/10.1371/journal.pone.0168921
    ae974a485f413a2113503eed53cd6c53
    10.1371/journal.pone.0168921
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