Tobacco rattle virus mediates gene silencing in a plant parasitic root-knot nematode

• Published in: Journal of experimental botany Vol. 60; no. 14; pp. 4041 - 4050
• Main Authors: Dubreuil, G, Magliano, M, Dubrana, M.P, Lozano, J, Lecomte, P, Favery, B, Abad, P, Rosso, M.N
• Format: Journal Article
• Language: English
• Published: Oxford Oxford University Press 01.01.2009; Oxford University Press (OUP)
• Subjects: Animals; Biological and medical sciences; Botanics; caenorhabditis-elegans; cell; Double stranded RNA; early development; expression; Fundamental and applied biological sciences. Psychology; Gene silencing; Gene Targeting - methods; Genes; genetic transformation; Genetic Vectors - genetics; Genetic Vectors - metabolism; host-parasite relationships; hosts; ingestion; interference; Life Sciences; meloidogyne-incognita; Nematoda; Nematoda - genetics; Nematoda - virology; Nematodes; new methods; Nicotiana - parasitology; Parasitism; Parasitism and symbiosis; phenotype; Phytopathology. Animal pests. Plant and forest protection; Plant Diseases - parasitology; Plant gall; Plant parasitic nematodes; Plant physiology and development; Plant roots; Plant Roots - parasitology; Plant Viruses - genetics; Plant Viruses - metabolism; Plants; progeny; protein; Research Papers; Reverse genetics; RNA; RNA Interference; root-knot nematodes; screening; small interfering RNA; Tobacco rattle virus; transgenic plants; troponin-c gene; vector; Vegetal Biology; virus-induced gene silencing; Viruses; virus-induced gene silencing; RNA interference; Reverse genetics; Plant pathogen; Root; Meloidogyne; Virus; Gene silencing; Tobravirus; Pest; Botany; Helmintha; Genetics; Nemathelminthia; Tobacco rattle virus; Invertebrata; Nematoda; Parasitic plant; RNA INTERFERENCE; RELATION HOTE-PARASITE; VIRUS-INDUCED GENE SILENCING; REVERSE GENETICS
• ISSN: 0022-0957; 1460-2431
• DOI: 10.1093/jxb/erp237

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.