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Dryad

Data from: Ame-miR-2161 affects the survival and development of honeybee larvae through the juvenile hormone acid methyltransferase gene

Abstract

MicroRNAs (miRNAs), as endogenous small non-coding RNAs, play a role in regulating the growth and development processes of insects. However, there are still few reports on the coordinated regulation of honeybee development processes by miRNAs and hormones. In this study, the regulatory network of target genes of Apis mellifera miRNA-2161 (ame-miR-2161) was constructed. The association between ame-miR-2161 and the survival and development of workers larvae was analyzed. The results showed that ame-miR-2161 potentially targets 22 mRNAs, focusing on target gene juvenile hormone acid methyltransferase (Jhamt), which is a key rate-limiting enzyme in the final step of the juvenile hormone synthesis pathway. RT-qPCR revealed that the expression of ame-miR-2161 and Jhamt had the same trend during the larval stage. Dual-luciferase assays confirmed the targeting relationship between ame-miR-2161 and Jhamt. Overexpression of ame-miR-2161 increased the mRNA level of Jhamt, which led to significantly increased juvenile hormone (JH) titer in 4- to 6-day-old larvae, and the expression of the JH signaling downstream response gene Kr-h1 also gradually increased. Conversely, inhibition of ame-miR-2161 decreased the mRNA level of jhamt, significantly reduced JH titer in larvae, and markedly declined the expression of Kr-h1. This indicates that ame-miR-2161 positively regulates the expression of Jhamt. Further findings showed that overexpression of ame-miR-2161 improved larval survival rate, while inhibition of ame-miR-2161 reduced larval survival rate. Changes in ame-miR-2161 did not affect the pupation rate of larvae but influenced changes in larval body weight. The results suggest that ame-miR-2161 affects JH levels by positively regulating the expression of Jhamt, participating in the regulation of the survival and development of honeybee larvae. These findings provide important information for interpreting the regulation of miRNA on hormone levels and metamorphic development in honeybees.