Identification of RNAi efficiency‐related gene in the green peach aphid, Myzus persicae (Hemiptera), based on comparative transcriptomics

Abstract BACKGROUND RNA interference (RNAi) is a promising strategy for aphid control; however, its practical application has been limited by low efficiency in aphids. To address this, we combined transcriptomic analysis with an ‘RNAi of RNAi’ approach to identify genes associated with RNAi efficiency in green peach aphid Myzus persicae . RESULTS Using the artificial diet‐mediated dsRNA delivery method, we observed that ingestion of both dsMpHunchback and dsfusion (a dsRNA targeting three genes) induced RNAi effects. These included significant gene silencing and reduced nymph production at 24, 36, and 48 h post‐feeding compared to dsGFP. Expression profiling of core RNAi machinery revealed that samples from 36 and 48 h post‐feeding were critical for RNAi efficiency, prompting their selection for transcriptome sequencing. Weighted gene co‐expression network analysis (WGCNA) and analysis of Differentially Expressed Genes (DEGs) were employed to elucidate the RNA‐seq data. Subsequent validation through RNAi of RNAi assay demonstrated functional roles for genes encoding basic region leucine zipper ( gene11325 ), cathepsin B‐like ( gene12476 ), glucosidase KIAA1161‐like ( gene6489 ), and piggyBac transposable element‐derived protein 3‐like ( gene7736 ). Notably, gene11325 and gene6489 were associated with RNAi efficiency, whereas g ene12476 and gene7736 appeared to exert compensatory effects that counteracted the RNAi effects induced by dsMpHunchback and dsfusion. CONCLUSION These findings highlight the functional diversity of genes influencing RNAi efficiency in green peach aphid, enhancing our understanding of RNAi mechanisms and establishing a foundation for optimizing RNAi‐based aphid control strategies. © 2025 Society of Chemical Industry.