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Viruses 2016, 8(11), 316; doi:10.3390/v8110316

Comparative Analysis of RNAi-Based Methods to Down-Regulate Expression of Two Genes Expressed at Different Levels in Myzus persicae

1
Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France
2
INRA, UMR BGPI INRA-CIRAD-SupAgro, CIRAD TA-A54/K, Campus International de Baillarguet, 34398 Montpellier, France
3
Plant Protection Department, Shiraz University, Shiraz, Iran
These authors contributed equally to this work.
*
Author to whom correspondence should be addressed.
Academic Editor: Ralf Georg Dietzgen
Received: 4 August 2016 / Revised: 9 November 2016 / Accepted: 11 November 2016 / Published: 19 November 2016
(This article belongs to the Special Issue Molecular Plant Virus—Insect Vector Interactions)
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Abstract

With the increasing availability of aphid genomic data, it is necessary to develop robust functional validation methods to evaluate the role of specific aphid genes. This work represents the first study in which five different techniques, all based on RNA interference and on oral acquisition of double-stranded RNA (dsRNA), were developed to silence two genes, ALY and Eph, potentially involved in polerovirus transmission by aphids. Efficient silencing of only Eph transcripts, which are less abundant than those of ALY, could be achieved by feeding aphids on transgenic Arabidopsis thaliana expressing an RNA hairpin targeting Eph, on Nicotiana benthamiana infected with a Tobacco rattle virus (TRV)-Eph recombinant virus, or on in vitro-synthesized Eph-targeting dsRNA. These experiments showed that the silencing efficiency may differ greatly between genes and that aphid gut cells seem to be preferentially affected by the silencing mechanism after oral acquisition of dsRNA. In addition, the use of plants infected with recombinant TRV proved to be a promising technique to silence aphid genes as it does not require plant transformation. This work highlights the need to pursue development of innovative strategies to reproducibly achieve reduction of expression of aphid genes. View Full-Text
Keywords: gene silencing; functional validation; aphids gene silencing; functional validation; aphids
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MDPI and ACS Style

Mulot, M.; Boissinot, S.; Monsion, B.; Rastegar, M.; Clavijo, G.; Halter, D.; Bochet, N.; Erdinger, M.; Brault, V. Comparative Analysis of RNAi-Based Methods to Down-Regulate Expression of Two Genes Expressed at Different Levels in Myzus persicae. Viruses 2016, 8, 316.

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