Antennal Responses and Odorant-Binding Protein 7 Binding of Rhoptroceros cyatheae (Selandriidae: Rhopographus) to Volatile Organic Compounds from Alsophila spinulosa

Rhoptroceros cyatheae (Hymenoptera: Selandriidae) is a dominant herbivorous pest of Alsophila spinulosa in southwestern China, including Guizhou and Sichuan provinces. Infestation by this pest impairs spore reproduction of A. spinulosa and reduces the photosynthetic capacity of host plants. However, the chemosensory genes of R. cyatheae have not been reported, and the molecular basis of antennal detection of host volatile organic compounds (VOCs) is poorly understood. This study aims to screen and identify bioactive VOCs potentially involved in host searching behavior of R. cyatheae, analyze antennal VOC detection patterns, and explore the in vitro binding characteristics of an odorant-binding protein (OBP) involved in olfactory recognition, thereby providing a preliminary theoretical basis for the green management of R. cyatheae. Dynamic headspace sampling, gas chromatography-mass spectrometry (GC-MS), and gas chromatography-electroantennography (GC-EAD) were used to measure antennal electrophysiological responses of R. cyatheae to volatiles from its host A. spinulosa. Y-tube olfactometer assays were conducted to evaluate behavioral responses. For RcyaOBP7, fluorescence competitive binding assays, structural modeling, and molecular docking were integrated to investigate its in vitro binding characteristics with nine selected bioactive VOCs. Nine A. spinulosa volatiles were identified that elicited antennal electrophysiological responses in R. cyatheae, and the sawfly showed behavioral orientation to these VOCs, confirming that its antennae can detect host VOCs. In vitro binding assays showed that RcyaOBP7 exhibited strong binding affinity to p-ethylacetophenone, suggesting its potential involvement in antennal olfactory recognition of this volatile. Specific VOCs released by A. spinulosa are among the signaling molecules detected by the antennae of R. cyatheae. In vitro findings indicate that RcyaOBP7 binds specifically to p-ethylacetophenone, suggesting a possible role in antennal olfactory recognition and behaviors such as host location. However, in vivo functional validation and field trials under ecologically relevant conditions are needed to confirm these roles. This study characterizes the in vitro binding properties of RcyaOBP7 and provides a basis for further research on green management strategies for R. cyatheae based on antennal olfactory signals.