JXB Advance Access originally published online on July 22, 2009
Journal of Experimental Botany 2009 60(14):4041-4050; doi:10.1093/jxb/erp237
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RESEARCH PAPER |
Tobacco rattle virus mediates gene silencing in a plant parasitic root-knot nematode
INRA-UNSA-CNRS, UMR 1064, Interactions Plantes-Microorganismes et Santé Végétale, 400, route des Chappes, BP 167, F-06903 Sophia Antipolis, France
To whom correspondence should be addressed. E-mail: rosso{at}sophia.inra.fr
Root-knot nematodes (RKNs) are sedentary biotrophic parasites that induce the differentiation of root cells into feeding cells that provide the nematodes with the nutrients necessary for their development. The development of new control methods against RKNs relies greatly on the functional analysis of genes that are crucial for the development of the pathogen or the success of parasitism. In the absence of genetic transformation, RNA interference (RNAi) allows for phenotype analysis of nematode development and nematode establishment in its host after sequence-specific knock-down of the targeted genes. Strategies used to induce RNAi in RKNs are so far restricted to small-scale analyses. In the search for a new RNAi strategy amenable to large-scale screenings the possibility of using RNA viruses to produce the RNAi triggers in plants was tested. Tobacco rattle virus (TRV) was tested as a means to introduce double-stranded RNA (dsRNA) triggers into the feeding cells and to mediate RKN gene silencing. It was demonstrated that virus-inoculated plants can produce dsRNA and siRNA silencing triggers for delivery to the feeding nematodes. Interestingly, the knock-down of the targeted genes was observed in the progeny of the feeding nematodes, suggesting that continuous ingestion of dsRNA triggers could be used for the functional analysis of genes involved in early development. However, the heterogeneity in RNAi efficiency between TRV-inoculated plants appears as a limitation to the use of TRV-mediated silencing for the high-throughput functional analysis of the targeted nematode genes.
Key words: Reverse genetics, RNA interference, virus-induced gene silencing
* Present address: CEA, iBiTecS, Gif sur Yvette, F-91191, France.
Present address: INRA, Université Bordeaux 2, UMR 1090, Génomique, Diversité et Pouvoir Pathogène, Domaine de la Grande Ferrade, F-33883 Villenave-D'ornon, France.
Present address: Laboratory of Nematology, Graduate School for Experimental Plant Sciences, Wageningen University, 6709 PD Wageningen, The Netherlands.
Received 15 May 2009; Revised 26 June 2009 Accepted 6 July 2009