Habrobracon hebetor is a parasitoid wasp capable of infesting many lepidopteran larvae. It uses venom proteins to immobilize host larvae and prevent host larval development, thus playing an important role in the biocontrol of lepidopteran pests. To identify and characterize its venom proteins, we developed a novel venom collection method using an artificial host (ACV), i.e., encapsulated amino acid solution in paraffin membrane, allowing parasitoid wasps to inject venom. We performed protein full mass spectrometry analysis of putative venom proteins collected from ACV and venom reservoirs (VRs) (control). To verify the accuracy of proteomic data, we also collected venom glands (VGs), Dufour’s glands (DGs) and ovaries (OVs), and performed transcriptome analysis. In this paper, we identified 204 proteins in ACV via proteomic analysis; compared ACV putative venom proteins with those identified in VG, VR, and DG via proteome and transcriptome approaches; and verified a set of them using quantitative real-time polymerase chain reaction. Finally, 201 ACV proteins were identified as potential venom proteins. In addition, we screened 152 and 148 putative venom proteins identified in the VG transcriptome and the VR proteome against those in ACV, and found only 26 and 25 putative venom proteins, respectively, were overlapped with those in ACV. Altogether, our data suggest proteome analysis of ACV in combination with proteome–transcriptome analysis of other organs/tissues will provide the most comprehensive identification of true venom proteins in parasitoid wasps.

Bactrocera dorsalis (Hendel) (B. dorsalis) is an important agricultural invasive pest that causes significant economic losses in tropical and subtropical fruit and vegetable crops. In this study, the proteins related to the sense of smell and taste of B. dorsalis, such as OBP, PBP, OR, IR, SNMP and CSP, were screened based on B. dorsalis transcriptome data. By integrating the compounds that were reported to be attractive to B. dorsalis, similar compounds of hydrocarbon compounds were obtained. Molecular docking was used to predict the binding between the similar compounds and the OBP, PBP, OR, IR, SNMP and CSP proteins. Network pharmacology was used to screen the potentially attractive compounds, and ecological experiments with B. dorsalis were finally conducted to verify the effect of these potentially attractive compounds on B. dorsalis. The results showed that the G protein-coupled receptor [BR: KO04030] and ion channel [BR: KO04040] pathways were closely related to the odor tropism of B. dorsalis. A total of 84 compounds, such as mitemcinal, exemestane and midecamycin, have potential binding effects on the B. dorsalis odor receptor proteins. The results of the ecological experiments showed that 1 mg/mL and 0.1 mg/mL 19-norandrostenedione, 1 mg/mL progesterone compounds was significantly attractive to B. dorsalis males, while 0.1 mg/mL exemestane was significantly attractive to B. dorsalis females. In this study, network pharmacology technology was used to discover the potential attractive compounds for B. dorsalis, which is important for the development and subsequent prevention and control of B. dorsalis. It can provide a reference in improving the success rates of clinical trials of new pest control products and in reducing the time and cost of drug development.

Zeugodacus cucuribitae (Coquillett) (Z. cucuribitae) is a global extremely invasive quarantine pest which has a wide host range of fruits and vegetables. At present, there are a few control measures for Z. cucuribitae, and deltamethrin and avermectin are commonly used. Among the hosts of Z. cucuribitae, Luffa acutangular, Luffa cylindrica, Sechium edule, Brassica oleracea var. botrytis, Musa nana, and Fragaria × ananassa are non-favored hosts. However, it is still not clear why these hosts are non-favored and whether there are any repellent components of Z. cucuribitae in these hosts. In this study, the components of these six hosts were collected from the literature, and the genes of odor and chemical sensation were determined from the genome of Z. cucuribitae. After the potential relationships between these components and genes were determined by molecular docking methods, the KEGG and GO enrichment analysis of these genes was conducted, and a complex network of genes vs. components vs. Kegg pathway vs. GO terms was constructed and used to select the key components for experiments. The results show that oleanolic acid (1 mg/mL, 0.1 mg/mL, and 0.01 mg/mL), rotenone (1 mg/mL, 0.1 mg/mL, and 0.01 mg/mL), and beta-caryophyllene oxide (1 mg/mL, 0.1 mg/mL, and 0.01 mg/mL) had a significant repellent effect on Z. cucuribitae, and three components, rotenone (1 mg/mL and 0.1 mg/mL), echinocystic acid (1 mg/mL, 0.1 mg/mL, and 0.01 mg/mL), and beta-caryophyllene oxide (1 mg/mL, and 0.1 mg/mL) had significant stomach toxicity in Z. cucuribitae. Furthermore, a complex signaling pathway was built and used to predict the effect of these components on Z. cucuribitae. These components probably play roles in the neuroactive ligand–receptor interaction (ko04080) and calcium signaling (ko04020) pathways. This study provides a reference for the prevention and control of Z. cucuribitae and a scientific reference for the rapid screening and development of new pest control drugs.