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Double-stranded RNA Oral Delivery Methods to Induce RNA Interference in Phloem and Plant-sap-feeding Hemipteran Insects
Published on: May 4, 2018
Double-stranded RNA in the biological control of grain aphid (Sitobion avenae F.)
1Institute of Crop Sciences/The National Key Facility for Crop Gene Resource and Genetic Improvement, Chinese Academy of Agricultural Sciences (CAAS), 12 Zhongguanchun South Street, Beijing, 10081, China.
Abstract:
Grain aphid (Sitobion avenae F.) is the most dominant and destructive pest of wheat, which causes significant yield loss of cereal plants each year by inflicting damage both through the direct effects of feeding and by vectoring debilitating plant viruses. In this study, we performed de novo transcriptome sequencing of grain aphid via Roche 454 GS-FLX pyrosequencing. A total of 1,106,696 reads were obtained and assembled into 32,277 unigenes, of which 25,389, 21,635, and 16,211 unigenes matched the Nt, Nr, and Swiss-Prot databases, respectively. Functional annotation of these unigenes revealed not only the presence of genes that encode the key components of RNAi machinery such as Dicer and Argonaute but also the genes encoding the TAR RNA binding protein (TRBP) and the SID-1 protein, which function in assisting the RNA-induced silencing complex (RISC) formation in microRNA (miRNA) pathway and mediating a systemic RNA interference (RNAi) effect though a cellular uptake mechanism. Furthermore, among a set of 66 unigenes selected for a double-stranded RNA (dsRNA) artificial diet assay, four novel effective RNAi targets, which led to high mortality of aphids due to the down-regulation of the expression of the respective target gene, were identified. Moreover, the expansion of systemic RNAi effect in grain aphid was observed by adding the fluorescently labeled dsRNA in an artificial diet assay.
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