Intron-Containing Hairpin RNA Interference Vector for OBP8 Show Promising Mortality in Peach Potato Aphid

Abstract

\n Myzus persicae is a devastating pest affecting potato production. RNA Interference technology is used against essential odorant binding protein 8 (OBP8) to enhance protection against Myzus persicae in potato. Gene was isolated, sequenced and GenBank IDs were allotted and ERNAi was used to design siRNA targets from OBP8 with no off-targets. Multiple Sequence Alignment show M. persicae OBP8 resemblance with Acyrthosiphon pisum, Rhopalosiphum maidis, Aphis fabae, and Sitobion avenae. DsRNA (7 µg/µl) oral acquisition had resulted in 69% mortality and 58% reduction in OBP8 expression 8D post dsRNA feeding in comparison to control. Golden Gate (GG) cloning based RNAigg is used for RNA interference taking advantage of type IIs restriction enzyme Eco31I. Agro infiltration assay was used for introduction of intron-containing hairpin RNA (ihpRNA) in Solanum tuberosum. Aphids feeding on transgenic S. tuberosum show 57.6% mortality and 49% reduction in OBP8 expression 8d post-feeding in comparison to control. This work proves OBP8 as promising ihpRNA targets in potato and related crops for whom Myzus is a devastating target.