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Abstract
Comprehensive understanding of pleiotropic roles of RNAi machinery highlighted the conserved chromosomal functions of RNA interference. The consequences of the evolutionary variation in the core RNAi pathway genes are mostly unknown, but may lead to the species-specific functions associated with gene silencing. The two-spotted spider mite, Tetranychus urticae, is a major polyphagous chelicerate pest capable of feeding on over 1100 plant species and developing resistance to pesticides used for its control. A well annotated genome, susceptibility to RNAi and economic importance, make T. urticae an excellent candidate for development of an RNAi protocol that enables high-throughput genetic screens and RNAi-based pest control. Here, we show that the length of the exogenous dsRNA critically determines its processivity and ability to induce RNAi in vivo. A combination of the long dsRNAs and the use of dye to trace the ingestion of dsRNA enabled the identification of genes involved in membrane transport and 26S proteasome degradation as sensitive RNAi targets. Our data demonstrate that environmental RNAi can be an efficient reverse genetics and pest control tool in T. urticae. In addition, the species-specific properties together with the variation in the components of the RNAi machinery make T. urticae a potent experimental system to study the evolution of RNAi pathways.
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1 The University of Western Ontario, Department of Biology, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884)
2 Tokyo University of Agriculture and Technology, Department of Applied Biological Science, Fuchu, Japan (GRID:grid.136594.c); Tokyo University of Agriculture and Technology, Institute of Global Innovation Research, Fuchu, Japan (GRID:grid.136594.c)
3 Tokyo University of Agriculture and Technology, Graduate School of Bio-applications and Systems Engineering, Koganei, Japan (GRID:grid.136594.c)
4 Tokyo University of Agriculture and Technology, Department of Applied Biological Science, Fuchu, Japan (GRID:grid.136594.c); Tokyo University of Agriculture and Technology, Graduate School of Bio-applications and Systems Engineering, Koganei, Japan (GRID:grid.136594.c); Tokyo University of Agriculture and Technology, Institute of Global Innovation Research, Fuchu, Japan (GRID:grid.136594.c)
5 Bayer AG, Division Crop Science, Research and Development, Monheim, Germany (GRID:grid.420044.6) (ISNI:0000 0004 0374 4101)
6 The University of Western Ontario, Department of Biology, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Instituto de Ciencias de la Vid y el Vino, Logrono, Spain (GRID:grid.481584.4); University of Belgrade, Department of Biology, Belgrade, Serbia (GRID:grid.7149.b) (ISNI:0000 0001 2166 9385)
7 The University of Western Ontario, Department of Biology, London, Canada (GRID:grid.39381.30) (ISNI:0000 0004 1936 8884); Instituto de Ciencias de la Vid y el Vino, Logrono, Spain (GRID:grid.481584.4)