silencing of aphid genes by dsrna feeding from plants从植物蚜虫基因的沉默dsrna喂养.pdfVIP

Silencing of Aphid Genes by dsRNA Feeding from Plants 1,2. 1. 2 1 Marco Pitino , Alexander D. Coleman , Massimo E. Maffei , Christopher J. Ridout , Saskia A. Hogenhout1* 1 Department of Disease and Stress Biology, The John Innes Centre, Norwich Research Park, Norwich, United Kingdom, 2 Plant Physiology Unit, Department of Plant Biology, Innovation Centre, University of Turin, Turin, Italy Abstract Background: RNA interference (RNAi) is a valuable reverse genetics tool to study gene function in various organisms, including hemipteran insects such as aphids. Previous work has shown that RNAi-mediated knockdown of pea aphid (Acyrthosiphon pisum) genes can be achieved through direct injection of double-stranded RNA (dsRNA) or small-interfering RNAs (siRNA) into the pea aphid hemolymph or by feeding these insects on artificial diets containing the small RNAs. Methodology/Principal Findings: In this study, we have developed the plant-mediated RNAi technology for aphids to allow for gene silencing in the aphid natural environment and minimize handling of these insects during experiments. The green peach aphid M. persicae was selected because it has a broad plant host range that includes the model plants Nicotiana benthamiana and Arabidopsis thaliana for which transgenic materials can relatively quickly be generated. We targeted M. persicae Rack1, which is predominantly expressed in the gut, and M. persicae C002 (MpC002), which is predominantly expressed in the salivary glands. The aphids were fed on N. benthamiana leaf disks transiently producing dsRNA corresponding to these genes and on A. thaliana plants stably producing