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Delivery of dsRNA through topical feeding for RNA interference in the citrus sap piercing‐sucking hemipteran, Diaphorina citri
Author(s) -
Killiny Nabil,
Kishk Abdelaziz
Publication year - 2017
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/arch.21394
Subject(s) - diaphorina citri , rna interference , nymph , biology , rna silencing , gene silencing , instar , hemiptera , rna , botany , gene , genetics , larva
RNA interference (RNAi) is a powerful means to study functional genomics in insects. The delivery of dsRNA is a challenging step in the development of RNAi assay. Here, we describe a new delivery method to increase the effectiveness of RNAi in the Asian citrus psyllid Diaphorina citri . Bromophenol blue droplets were topically applied to fifth instar nymphs and adults on the ventral side of the thorax between the three pairs of legs. In addition to video recordings that showed sucking of the bromophenol blue by the stylets, dissected guts turned blue indicating that the uptake was through feeding. Thus, we called the method topical feeding . We targeted the abnormal wing disc gene ( awd ), also called nucleoside diphosphate kinase ( NDPK ), as a reporter gene to prove the uptake of dsRNA via this method of delivery. Our results showed that dsRNA‐ awd caused reduction of awd expression and nymph mortality. Survival and lifespan of adults emerged from treated nymphs and treated adults were affected. Silencing awd caused wing malformation in the adults emerged from treated nymphs. Topical feeding as a delivery of dsRNA is highly efficient for both nymphs and adults. The described method could be used to increase the efficiency of RNAi in D. citri and other sap piercing‐sucking hemipterans.