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.
Highlights
With the growing world population there is increasing need of increased crop production
RNA extraction, and Complementary DNA (cDNA) synthesis was followed by Polymerase Chain Reaction (PCR) and Tm for odorant binding protein 8 (OBP8) (465 bp) was found 54oC (Fig 1B)
In this study new pRNAiBAB7ihp is made, for making silencing OBP8 gene in M. persicae for the 1st time in one restriction ligation cloning step. intron-containing hairpin RNA (ihpRNA) constructed based on Golden Gate (GG) cloning method, efficiently suppressed the OBP8 genes and caused 57.6% reduced expression
Summary
With the growing world population there is increasing need of increased crop production. Hemipteran insects like Myzus persicae (Green peach aphid) adapt to varying environmental conditions by switching between the primary and secondary hosts for its survival; responsible for great crop loss (Liu et al 2020). It is a pest affecting more than 40 plant families for parthenogenetic and sexual reproduction either by direct feeding damage to plants, or by indirect transmittance by 100 plant viruses such as cucumber mosaic virus, tobacco rattle virus (Faisal et al 2019; Bwye et al 1997; Mulot et al 2016). Type IIs restriction enzyme is used for at both sense and antisense orientations for ihpRNA of OBP8 gene in single tube
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