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siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes
INRA, UMR 1355 Institut Sophia Agrobiotech, Interactions Plantes-Nematodes, Sophia Antipolis F-06903, France
Université de Nice Sophia Antipolis, Sophia Antipolis F-06903, France
CNRS, UMR 7254 Institut Sophia Agrobiotech, Interactions Plantes-Nematodes, Sophia Antipolis F-06903, France
Present address: INRA, UMR 1163 Biotechnologie des Champignons Filamenteux, Université Aix-Marseille, Marseille F-13288, France.
* Author to whom correspondence should be addressed.
Received: 10 May 2012; in revised form: 18 June 2012 / Accepted: 26 June 2012 / Published: 10 July 2012
Abstract: Expanding genomic data on plant pathogens open new perspectives for the development of specific and environment friendly pest management strategies based on the inhibition of parasitism genes that are essential for the success of infection. Identifying such genes relies on accurate reverse genetics tools and the screening of pathogen knock-down phenotypes. Root-knot nematodes are major cosmopolitan crop pests that feed on a wide range of host plants. Small interfering RNAs (siRNAs) would provide a powerful tool for reverse genetics of nematode parasitism genes provided that they could (1) target genes expressed in inner tissues of infective nematodes and (2) target genes expressed during parasitism. In this study, we show that siRNAs can access inner tissues of the infective juveniles during soaking and accumulate in the esophagus, amphidial pouches and related neurons of the nematode. We provide evidence that siRNAs can trigger knock-down of the parasitism gene Mi-CRT, a calreticulin gene expressed in the esophageal glands of Meloidogyne incognita. Mi-CRT knock-down in infective juveniles affected nematode virulence. However, Mi-CRT knock-down was not persistent after plant infection, indicating that siRNA-mediated RNAi is best suited for functional analysis of genes involved in pre-parasitic stages or in the early steps of infection.
Keywords: Meloidogyne incognita; plant parasitic nematode; RNA interference; RNAi; root-knot nematode; RKN; short interfering RNA; siRNA
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Arguel, M.-J.; Jaouannet, M.; Magliano, M.; Abad, P.; Rosso, M.-N. siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes. Genes 2012, 3, 391-408.
Arguel M-J, Jaouannet M, Magliano M, Abad P, Rosso M-N. siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes. Genes. 2012; 3(3):391-408.
Arguel, Marie-Jeanne; Jaouannet, Maëlle; Magliano, Marc; Abad, Pierre; Rosso, Marie-Noëlle. 2012. "siRNAs Trigger Efficient Silencing of a Parasitism Gene in Plant Parasitic Root-Knot Nematodes." Genes 3, no. 3: 391-408.