Cloning and Expression of the Neuropeptide F and Neuropeptide F Receptor Genes and Their Regulation of Food Intake in the Chinese White Pine Beetle Dendroctonus armandi

Abstract

Neuropeptide F (NPF) is an important signaling molecule that acts as a neuromodulator to regulate a diversity of physiological and behavioral processes from vertebrates to invertebrates by interaction with NPF receptors, which are G protein-coupled receptors (GPCR). However, nothing is known about NPF in Chinese white pine beetle, Dendroctonus armandi, a destructive pest of natural and coniferous forests in the middle Qinling Mountains of China. We have cloned and characterized cDNAs encoding one NPF precursor and two NPF receptors in D. armandi and made bioinformatics predictions according to the deduced amino acid sequences. They were highly similar to that of Dendroctonus ponderosa. The transcription levels of these genes were different between larvae and adults of sexes, and there were significant differences among the different developmental stages and tissues and between beetles under starvation and following re-feeding states. Additionally, downregulation of NPF and NPFR by injecting dsRNA into beetles reduced their food intake, caused increases of mortality and decreases of body weight, and also resulted in a decrease of glycogen and free fatty acid and an increase of trehalose. These results indicate that the NPF signaling pathway plays a significant positive role in the regulation of food intake and provides a potential target for the sustainable management of this pest.

Identification of the Short Neuropeptide F and Short Neuropeptide F Receptor Genes and Their Roles of Food Intake in Dendroctonus armandi

“…Then, they were moved to the laboratory in plastic containers with moist paper and immediately stored at –80 °C until analysis. Logging slash of infested P. armandi was transported from the sample plot to a laboratory, where emerged adult beetles and larvae were collected and then reared in glass dishes (90 × 15 mm 2 ) using the meridic diet described by a previous report [ 27 ]. All conditions were controlled by an artificial climate cabinet under certain conditions of 25 ± 1 °C, 70% relative humidity (RH) and in the dark.…”

“…In addition, whole-body homogenates of each group were used to extract glycogen, trehalose, and free fatty acid. These three substances content levels were detected as previously described [ 27 ].…”

Roles of Krüppel Homolog 1 and Broad-Complex in the Development of Dendroctonus armandi (Coleoptera: Scolytinae)

“…RNA interference (RNAi) technology has now become a widely used tool to analyze the gene functions of Chinese white pine beetle ( Dendroctonus armandi Tsai and Li). Chen‘s team used RNAi to investigate and analyze the olfactory receptor coreceptor ( DarmOrco ), chemoreceptor ( DarmCSP2 ), aquaporins ( DaAqps ), antifreeze protein genes ( DaAFP ), Capa peptide receptors ( DaCapaRs ), neuropeptide F ( DaNPF ), and 3-hydroxy-3-methylglutaryl coenzyme A reductase genes ( HMGR ) in D. armandi ( Zhang et al, 2016 ; Li et al, 2018 ; Fu et al, 2019 , 2020 , 2021 ; Liu et al, 2021 ; Sun et al, 2021 ). Sun et al (2022) discovered that allatostatin C (PISCF/AST) and juvenile hormone acid O-methyltransferase ( JHAMT ) were major regulators of juvenile hormone synthesis in D. armandi after obtaining dsRNA technology via the L4440 vector construction.…”

Identification and functional characterization of the sulfakinin and sulfakinin receptor in the Chinese white pine beetle Dendroctonus armandi

“…Chinese white pine beetles were obtained from infested P. armandii at the Huoditang Experimental Forest Station, which is located on the southern slopes of the mid-Qinling Mountains, Shaanxi, China (33 °18′N, 108 °21′E), and reared on an artificial food source in a laboratory maintained at 25 ± 1 °C, 70% relative humidity (RH), and in the dark (Liu et al, 2021a).…”

“…Total RNA was isolated from three developmental stages of the Chinese white pine beetle (larvae, pupae, and adults from both sexes), and the concentration and quality were determined as described previously (Liu et al, 2021a). Total RNA for cDNA synthesis was prepared using the Fast King RT Reagent Kit with gDNA Eraser (Tiangen, China) according to the manufacturer protocol.…”