title: Comparative Analysis of RNAi-Based Methods to Down-Regulate Expression of Two Genes Expressed at Different Levels in Myzus persicae.
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,ALYandEph, potentially involved in polerovirus transmission by aphids. Efficient silencing of onlyEphtranscripts, which are less abundant than those ofALY, could be achieved by feeding aphids on transgenicArabidopsis thalianaexpressing an RNA hairpin targetingEph, onNicotiana benthamianainfected with aTobacco rattle virus(TRV)-Eph recombinant virus, or on in vitro-synthesizedEph-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.
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. michael.mulot@colmar.inra.fr.
authors.forename: Michaël
authors.initials: M
authors.lastname: Mulot
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. sylvaine.boissinot@colmar.inra.fr.
authors.forename: Sylvaine
authors.initials: S
authors.lastname: Boissinot
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. baptiste.monsion@supagro.inra.fr.|INRA, UMR BGPI INRA-CIRAD-SupAgro, CIRAD TA-A54/K, Campus International de Baillarguet, 34398 Montpellier, France. baptiste.monsion@supagro.inra.fr.
authors.forename: Baptiste
authors.initials: B
authors.lastname: Monsion
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. rastgarmaryam@gmail.com.|Plant Protection Department, Shiraz University, Shiraz, Iran. rastgarmaryam@gmail.com.
authors.forename: Maryam
authors.initials: M
authors.lastname: Rastegar
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. gaclavijo@hotmail.com.
authors.forename: Gabriel
authors.initials: G
authors.lastname: Clavijo
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. david.halter@inra.fr.
authors.forename: David
authors.initials: D
authors.lastname: Halter
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. nicole.bochet@colmar.inra.fr.
authors.forename: Nicole
authors.initials: N
authors.lastname: Bochet
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. monique.erdinger@colmar.inra.fr.
authors.forename: Monique
authors.initials: M
authors.lastname: Erdinger
authors.affiliation: Université de Strasbourg, INRA, SVQV UMR-A 1131, 28 rue de Herrlisheim, Colmar, 68021 Strasbourg, France. veronique.brault@inra.fr.
authors.forename: Véronique
authors.initials: V
authors.lastname: Brault
mesh_terms: D000818:Animals; D001042:Aphids / Q000235:genetics* / Q000254:growth & development; D017360:Arabidopsis / Q000469:parasitology; D004772:Entomology / Q000379:methods*; D055785:Gene Knockdown Techniques / Q000379:methods*; D017344:Genes, Insect; D034622:RNA Interference; D014026:Nicotiana / Q000469:parasitology
keywords: aphids; functional validation; gene silencing