Abstract

Simple SummaryThe Chinese white pine beetle, Dendroctonus armandi, is a destructive pest of coniferous forests in the middle Qinling Mountains of China. The short neuropeptide F (sNPF) and short NPF receptor (sNPFR) genes have important roles in a broad range of physiological and behavioral processes. However, the function of sNPF signaling pathway in regulating the food intake of Dendroctonus armandi has been unclear. In this study, we cloned and characterized cDNAs encoding sNPF and sNPF receptor in D. armandi and analyzed their spatiotemporal expression patterns, as well as changes in expression after starvation. Addtionally, sNPF and sNPFR knockdown in beetles using RNA interference significantly increased mortality and reduced their food intake and body weight through changes of a biosynthesis and metabolism pathway. From these results, we conclude that sNPF signaling pathway is important in the feeding control of D. armandi.The short neuropeptide F (sNPF) is an essential signaling molecule that is evolutionarily conserved and involved in a broad range of physiological functions in the invertebrates, by interacting with sNPF receptors, which belong to G protein-coupled receptors (GPCR). However, the function of sNPF in regulating the food intake of Dendroctonus armandi has been unclear. In this study, we cloned and characterized cDNAs encoding sNPF and sNPF receptor in the D. armandi and made bioinformatics predictions on the deduced amino acid sequences. They had a high degree of similarity to that of Dendroctonus ponderosa. Quantitative real-time reverse transcription PCR (qRT-PCR) revealed that the transcript levels of both sNPF and sNPFR varied across developmental stages and body parts. In addition, the sNPF and sNPFR expression levels were upregulated in starved beetles, and the expression levels recovered after re-feeding. Furthermore, RNAi knockdown by the injection of sNPF and sNPFR dsRNA into beetles significantly increased mortality and reduced their food intake and body weight, and also caused decrease of glycogen and free fatty acid and increase of trehalose. These results indicate that sNPF signaling pathway plays an important role in the regulation of food intake and provides a potential molecular target for the eco-friendly control strategies of this pest.

Highlights

  • Neuropeptides are essential signaling molecules that are evolutionarily conserved and involved in a broad range of physiological functions

  • When the Drosophila short neuropeptide F (sNPF) receptor was overexpressed, this led to an augmentation in both body size and food uptake, while RNA interference (RNAi) mediated knockdown of the receptor resulted in smaller animals [12]

  • The phylogenetic tree of sNPF (Figure 2) and sNPFR (Figure S2) indicated that proteins were clustered with the Coleoptera group with D. ponderosae

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Summary

Introduction

Neuropeptides are essential signaling molecules that are evolutionarily conserved and involved in a broad range of physiological functions. When the Drosophila sNPF receptor was overexpressed, this led to an augmentation in both body size and food uptake, while RNA interference (RNAi) mediated knockdown of the receptor resulted in smaller animals [12]. This positive correlation between sNPF signaling and feeding behavior was demonstrated in several other insect species. We identified the full-length sNPF and sNPFR cDNAs from D. armandi and performed gene expression analysis and RNA interference experiments These results provide an important step forward to the improvement of eco-friendly pest control strategies of bark beetles

Insect Sample Preparation
Total RNA Isolation and cDNA Synthesis
Analysis of cDNA Sequences
RT-qPCR
Survival Test and Body Weight Measurement
Statistical Analysis
Sequence Characteristics and Bioinformatics Analysis
Starvation and Re-Feeding Assays
Effect of dsRNA Treatment on Mortality and Body Weight
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