Technique to deliver dsRNA in Bemisia tabaci nymphs and its validation through silencing of juvenile hormone esterase gene

Abstract

AbstractRNA interference (RNAi) is a technique employed to investigate the functions of various genes in whitefly and other insects. The effectiveness of the RNAi technique depends upon the delivery of dsRNA into the insects and it has been achieved through microinjection, oral feeding, egg soaking, transfection, topical application, spraying with nanoparticles, leaf‐mediated dsRNA feeding or plant‐mediated/ transgenic plants. In Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae), most of the studies on RNAi were conducted on the adult through either injection or oral feeding. There is a need to develop a system to deliver dsRNA in whitefly nymphs because the first instar nymphs after hatching find a suitable place on the leaf and feed on the same place till adult emergence. So, we evaluated nine different techniques using different sponges, tubes, and, Petri plates for feeding nymphs. The detached leaf survived for maximum number of days (8.25 days) with minimum nymphal mortality (12.01%) in the Whitefly Nymphal Feeding System‐1 (WNFS‐1) treatment; therefore, WNFS‐1 was selected for the delivery of dsRNA to B. tabaci nymphs. In WNFS‐1, blue food dye was used to confirm that nymphs were being fed, and the results were evidenced by blue‐coloured bodies. The juvenile hormone esterase (jhe) gene was silenced in order to successfully deliver dsRNA to nymphs using WNFS‐1 and resulting in much higher nymphal mortality (68.36%). Therefore, WNFS‐1 is a reliable method of delivering dsRNA for the purpose of silencing the genes in the nymphal stages of whiteflies. The WNFS system may also be used extensively to evaluate any chemistry (insecticides, hormones, biopesticides, etc.) in nymphs of whitefly.