Plant and Aphid Partners of Poleroviruses: Role in Virus Transmission by Aphids?

Comité de lecture : trueConférence invitée : falseDate de début de l'événement : 2011-07-11Date de fin de l'évenement : 2011-07-14Date de validation : Tue Aug 13 15:11:30 CEST 2013Diffusion de la pièce jointe : Publique, PubliqueIdentifiant : 200587Langue du titre : engNombre de consultation de la notice : 77Nombre de téléchargements de la pièce jointe : 8Pays de l'événement : BRAPublic visé : ScientifiqueType de communication avec actes : Présentation oraleType d'événement : SymposiumPoleroviruses are phloem limited viruses strictly transmitted by aphids in a circulative and non propagative manner. Virions are acquired by aphids when ingesting sap from infected plants. Virus particles cross the gut epithelium and the accessory salivary gland cells before being released, together with saliva, into the plant during a subsequent feed. This highly specific transcytosis mechanism relies on the presence of virus receptors on the surface of the aphid cells. We developed several approaches to identify virus partners in the plant and in the aphid to analyse their role in virus transmission by the vector. By screening different aphid cDNA libraries using a yeast two hybrid system, only few candidates were able to bind virus structural proteins. Among them, we found two nuclear proteins (GAR1 and ALY) which may not be the true virusreceptors but could be considered as virus-sensors. An Ephrin receptor-like protein was also found to interact with the viral proteins. Involvement of these candidates in virus transport through the aphid needs to be analyzed by developing in the insect RNAi-based techniques. These experiments are in progress. We also looked for plant virus-partners and identified several phloem proteins able to bind purified virions in vitro. We showed that these proteins could stimulate virus transmission by aphids when added together with purified virus to the aphid diet (Bencharki et al. 2010, M.P.M.I., 23: 799). By developing a yeast two hybrid system using a phloem specific cDNA library, we identified five additional proteins able to bind viral proteins. Among them, we found ALY proteins already identified as aphid virus-partners suggesting that orthologous plant and aphid proteins could be implicated in the virus cycle. So far, a direct implication of these proteins in aphid transmission has not been observed and experiments are on going to analyze their functions

In Vitro Acquisition of Specific Small Interfering RNAs Inhibits the Expression of Some Target Genes in the Plant Ectoparasite Xiphinema index

Xiphinema index is an important plant parasitic nematode that induces direct damages and specifically transmits the Grapevine fanleaf virus, which is particularly harmful for grapevines. Genomic resources of this nematode species are still limited and no functional gene validation technology is available. RNA interference (RNAi) is a powerful technology to study gene function and here we describe the application of RNAi on several genes in X. index. Soaking the nematodes for 48 h in a suspension containing specific small interfering RNAs resulted in a partial inhibition of the accumulation of some targeted mRNA. However, low reproducible silencing efficiency was observed which could arise from X. index silencing pathway deficiencies. Indeed, essential accustomed proteins for these pathways were not found in the X. index proteome predicted from transcriptomic data. The most reproducible silencing effect was obtained when targeting the piccolo gene potentially involved in endo-exocytosis of synaptic molecules. This represents the first report of gene silencing in a nematode belonging to the Longidoridae family

Specific companion cell profiling to investigate plant virus long distance movement

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Impact of Mutations in Arabidopsis thaliana Metabolic Pathways on Polerovirus Accumulation, Aphid Performance, and Feeding Behavior

During the process of virus acquisition by aphids, plants respond to both the virus and the aphids by mobilizing different metabolic pathways. It is conceivable that the plant metabolic responses to both aggressors may be conducive to virus acquisition. To address this question, we analyze the accumulation of the phloem-limited poleroviru