Cyantraniliprole resistance in Tuta absoluta: selection, comparative transcriptomes, and nanocarrier-mediated RNAi studies

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The tomato leaf miner Tuta absoluta (Meyrick) is an important invasive tomato pest that causes serious economic losses worldwide.Cyantraniliprole, a second-generation anthranilic diamide insecticide, has been widely used against insect pests, including T. absoluta.In this study, a cyantraniliprole-resistant strain (CyanRS) exhibiting 23.9-fold resistance was established through continuous selection with cyantraniliprole over eight consecutive generations.Additionally, we employed the Illumina RNA-seq to obtain comparative transcriptomes, followed by nanocarrier-mediated RNAi to investigate the functional role of key P450 genes linked to cyantraniliprole resistance in T. absoluta.Transcriptomic results showed that a total of 2435 differentially expressed genes, of these, 1197 (49.15%) were up-regulated, and 1238 (50.84%) were down-regulated among CyanRS and (susceptible strain) SS of T. absoluta.GO analysis indicated that most genes were enriched in cellular processes, metabolic processes, biological regulation, catalytic activity, binding, cellular anatomical entities, and intracellular terms.Most of these DEGs were identified in numerous KEGG pathways, such as oxidative phosphorylation, carbon metabolism, drug metabolism, and cytochrome P450.RT-qPCR results confirmed the overexpression of four P450 genes (CYP338A1, CYP6B5, CYP307A1, and CYP314A1), aligning with the RNA-seq data.Moreover, the cytochrome P450 activity was significantly higher in CyanRS compared to SS of T. absoluta.Nanocarrier-mediated RNAi of the CYP338A1, CYP6B5, CYP307A1, and CYP314A1 decreased the mRNA expression levels and P450 enzyme activity, ultimately increasing the sensitivity of CyanRS to cyantraniliprole, with higher mortality rates as compared to the controls.Overall, these results showed that several P450s, especially the 20E synthesis pathway genes, were linked with cyantraniliprole resistance in T. absoluta.This study provides in-depth knowledge to understand the molecular basis of cyantraniliprole resistance in this key invasive pest.

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