Efficacy of arginine kinase as a promising RNAi target in Aphis gossypii genome as revealed through aphid bioassay on field-grown transgenic cotton plants

The cotton aphid Aphis gossypii is a major agricultural pest of cotton that causes substantial damage to the crop not only by sucking sap but also through virus transmission. Globally adopted traditional and contemporary approaches to control aphid infestation have certain limitations and are hazardous to human health. RNA interference (RNAi) technology has unfolded its potential as an effective crop protection strategy against various pests. In this study, we adopted plant-mediated RNAi strategy to enhance aphid resistance in cotton by targeting arginine kinase (AK), which is a crucial enzyme responsible for energy homeostasis in insects. We selected a 312bp dsRNA fragment containing eight siRNAs and showing optimum GC content, Hb index, and stable secondary structure based on computational prediction studies. The binary construct expressing dsRNA was used to transform local cotton variety MNH886 and four transgenic lines were obtained in the T1 generation. Out of the four T1 transgenic cotton lines, dsA-7 exhibited the highest aphid mortality (73.3%), whereas, dsA-1, dsA-3 and dsA-6 revealed 60%, 61%, and 66.6% aphid mortality, respectively, in comparison to 13.3% mortality in the mock control cotton line. Moreover, significant knockdown in mRNA expression of AK was observed in aphids fed dsA-7 which was 79%. In comparison, 54%, 47%, and 45% downregulation was recorded in aphids which fed on dsA-6, dsA-3, and dsA-1 transgenic cotton lines, respectively. These results revealed that plant-mediated downregulation of aphid RNA induced significant RNA interference in A. gossypii which resulted in considerable aphid mortality and led to plant protection against aphids.