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Intron-containing hairpin RNA interference vector for OBP8 show promising mortality in peach potato aphid
Authors:Afroz  Amber  Aslam  Safeena  Rashid  Umer  Malik  Muhammad Faheem  Zeeshan  Nadia  Khan  Muhammad Ramzan  But  Muhammad Qasim Shahzad  Khan  Sabaz Ali
Institution:1.Department of Biochemistry and Biotechnology, University of Gujrat, Gujrat, Punjab, Pakistan
;2.Department of Zoology, Faculty of Sciences, University of Gujrat, Gujrat, Punjab, Pakistan
;3.National Institute of Genomics and Advanced Biotechnology, NARC Islamabad, Islamabad, Pakistan
;4.Biotechnology Program, COMSATS University Islamabad-Abbottabad Campus, University Road, Abbottabad, 22060, Pakistan
;
Abstract:

Myzus persicae is a devastating pest affecting potato production. Intron-containing hairpin RNA (ihpRNA) silenced the odorant-binding protein 8 (OBP8) for enhanced protection against Myzus persicae in potatoes. OBP8 was isolated from M. persicae, sequenced, with the allotment of GenBank ID. 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-OBP8 (7 µg/µL) oral feeding resulted in a 69% mortality, and 58% OBP8 reduced expression 8D post-feeding compared to control. Golden Gate cloning is used for RNA interference taking advantage of type IIs restriction enzyme Eco31I. ihpRNA-OBP8 introduced by agroinfiltration in Solanum tuberosum. Transgenic S. tuberosum fed Myzus show 57.6% mortality and 49% reduction in OBP8 expression 8D post-feeding, compared to control. This work proves OBP8 as promising ihpRNA targets in potato and related crops for whom Myzus is a destructive pest.

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