Global climate change is pushing insects into colder regions. Understanding their cold tolerance is important for predicting population dynamics. During overwintering, Streltzoviella insularis larvae activate the AMPK signaling pathway. This suggests that energy metabolism plays a key role under cold stress. In this study, we used enzyme activity assays, LC-MS-based targeted metabolomics, and transcriptome sequencing. We focused on six key enzymes in glycolysis and the TCA cycle. We also measured related metabolites and regulatory genes. Hexokinase (HK) and citrate synthase (CS) activities were highly sensitive to temperature. HK increased then markedly decreased; CS was significantly downregulated. Pyruvate kinase (PK), isocitrate dehydrogenase (IDH), and α-ketoglutarate dehydrogenase (KGD) showed trends that matched changes in larval cold tolerance, exhibiting an up–down–up expression trend. Glycolytic metabolites (glucose-6-phosphate, fructose-6-phosphate, 1,6-fructose-diphosphate, phosphoenolpyruvic acid) peaked at −10 °C. TCA intermediates (citrate, acetyl-CoA, α-ketoglutaric acid, and isocitrate) were more abundant at 0–4 °C. Pyruvate increased significantly. PYR content showed a significant increase followed by a decrease, peaking at 0 °C. It was converted into lactate and acetyl-CoA. ATP levels dropped and then increased, reaching their lowest level at 0 °C. These results suggest a shift from aerobic to mixed aerobic–anaerobic metabolism. Transcriptome data showed differential expression of key metabolic genes such as phosphoenolpyruvate carboxykinase, phosphoglycerate kinase, and ATP synthase subunit beta. These gene changes supported the trends in enzymes and metabolites. Our findings reveal a coordinated metabolic and transcriptional response to cold. This provides a basis for understanding the cold adaptation and potential range expansion of S. insularis.
The Russian olive (Elaeagnus angustifolia), which functions as a “dead-end trap tree” for the Asian long-horned beetle (Anoplophora glabripennis) in mixed plantations, can successfully attract Asian long-horned beetles for oviposition and subsequently kill the eggs by gum. This study aimed to investigate gum secretion differences by comparing molecular and metabolic features across three conditions—an oviposition scar, a mechanical scar, and a healthy branch—using high-performance liquid chromatography and high-throughput RNA sequencing methods. Our findings indicated that the gum mass secreted by an oviposition scar was 1.65 times greater than that secreted by a mechanical scar. Significant differences in gene expression and metabolism were observed among the three comparison groups. A Kyoto Encyclopedia of Genes and Genomes annotation and enrichment analysis showed that an oviposition scar significantly affected starch and sucrose metabolism, leading to the discovery of 52 differentially expressed genes and 7 differentially accumulated metabolites. A network interaction analysis of differentially expressed metabolites and genes showed that EaSUS1, EaYfcE1, and EaPGM1 regulate sucrose, uridine diphosphate glucose, α-D-glucose-1P, and D-glucose-6P. Although the polysaccharide content in the OSs was 2.22 times higher than that in the MSs, the sucrose content was lower. The results indicated that the Asian long-horned beetle causes Russian olive sucrose degradation and D-glucose-6P formation. Therefore, we hypothesized that damage caused by the Asian long-horned beetle could enhance tree gum secretions through hydrolyzed sucrose and stimulate the Russian olive’s specific immune response. Our study focused on the first pair of a dead-end trap tree and an invasive borer pest in forestry, potentially offering valuable insights into the ecological self-regulation of Asian long-horned beetle outbreaks.
Elaeagnus angustifolia L. can attract adult Asian longhorned beetle (ALB), Anoplophora glabripennis (Motschulsky), and kill their offspring by gum secretion in oviposition scars. This plant has the potential to be used as a dead-end trap tree for ALB management. However, there is a limited understanding of the attraction ability and biochemical defense response of E. angustifolia to ALB. In this study, we conducted host selection experiments with ALB and then performed physiological and biochemical assays on twigs from different tree species before and after ALB feeding. We analyzed the differential metabolites using the liquid chromatograph–mass spectrometer method. The results showed that ALB’s feeding preference was E. angustifolia > P.× xiaohei var. gansuensis > P. alba var. pyramidalis. After ALB feeding, the content of soluble sugars, soluble proteins, flavonoids, and tannins decreased significantly in all species. In three comparison groups, a total of 492 differential metabolites were identified (E. angustifolia:195, P.× xiaohei var. gansuensis:255, P. alba var. pyramidalis:244). Differential metabolites were divided into overlapping and specific metabolites for analysis. The overlapping differential metabolites 7-isojasmonic acid, zerumbone, and salicin in the twigs of three tree species showed upregulation after ALB feeding. The specific metabolites silibinin, catechin, and geniposide, in E. angustifolia, significantly increased after being damaged. Differential metabolites enriched in KEGG pathways indicated that ALB feeding activated tyrosine metabolism and the biosynthesis of phenylpropanoids in three tree species, with a particularly high enrichment of differential metabolites in the flavonoid biosynthesis pathway in E. angustifolia. This study provides the metabolic defense strategies of different tree species against ALB feeding and proposes candidate metabolites that can serve as metabolic biomarkers, potentially offering valuable insights into using E. angustifolia as a control measure against ALB.