Development of Sprayable long hairpin dsRNA Encapsulated with layered double hydroxide nanoparticles against Bemisia tabaci and Pectinophora gossypiella

RNA interference is an eco-friendly pest control mechanism regulating gene expression at a post-transcriptional level in eukaryotes and offers a promising alternative to conventional chemical pesticides, which face challenges such as resistant development and non-target toxicity. The sprayable gene-specific double-stranded RNA (dsRNA) will suppress the insecticide detoxifying genes (Acetylcholine esterase (AChE), orcokinin (Orc), sex lethal protein, Ecdysone receptor (ECR) genes (B. tabaci Asia-1, PgCadl, Cadherin aIIeles (rl9 and r20), rl5A and rl5B, rl4 aIIele of the pink boIIworm cadherin gene (PgCadl), rl3PgCadl, PgABCA2, PgCadl aIIeles (rl-r20), and PgABCC2) of white fly and pink bollworm. Initially, this study will screen differences in insecticidal activity across various open reading frames of target genes using similarly sized (approximately 300 bp) dsRNAs. The optimal length of dsRNA will be determined by preparing samples ranging from 100 to 700 bp. Different formulations of dsRNA spray shielded with layered double hydroxide nanoparticles will be used against different target genes. The effects of dsRNA on non-target organisms (NTOs) will be evaluated against honey bees, Apis mellifera, A.cerana, and a natural enemy, Orius laevigatus. The optimal length of hairpin dsRNA will be capsulated with LDH and harmless to NTOs, and then it will be sprayed on Bemisia tabaci and Pectinophora gossypiella infesting lab- and greenhouse-cultivated cotton plants. This study will lead to a significant reduction in pink bollworm and white fly populations compared to the control efficacy of different synthetic chemical insecticides.