Azadirachtin downregulates the expression of the CREB gene and protein in the brain and directly or indirectly affects the cognitive behavior of the Spodoptera litura fourth-instar larvae.

Abstract

Azadirachtin has the potential to be used for pest control. Nevertheless, few studies have investigated the effects of azadirachtin on the cognitive behavior of pests. In this study, expression of the cAMP response element-binding protein (CREB) and its gene were studied via a series of experiments in Spodoptera litura larvae treated with azadirachtin. RNA-Seq analysis of S. litura larvae treated with azadirachtin was undertaken. According to Kyoto Encyclopedia of Genes and Genomes analysis, the top 20 enriched pathways included neuroactive ligand-receptor interaction pathways, with seven significantly differentially expressed genes including CREB. Quantitative real time polymerase chain reaction (qRT-PCR) results indicated that the CREB gene was expressed during all developmental stages of S. litura, but relative expression of the CREB gene was significantly downregulated after treatment with azadirachtin. Grayscale statistical analysis also showed that expression levels of protein kinase A (PKA), extracellular signal-regulated kinase (ERK) and CREB proteins were significantly downregulated after treatment with azadirachtin. Moreover, RNA interference results showed that the effect of azadirachtin on the cognitive behavior of S. litura was consistent with that seen after interfering with CREB. In addition, larval selectivity to addictive odor sources was reduced, and the initial reaction time was increased. This study clarified that azadirachtin can affect the cognitive behavior of S. litura and treatment with azadirachtin resulted in a downregulation of gene and protein expression of CREB and its pathway proteins. © 2020 Society of Chemical Industry.