Binding properties of olfactory proteins to host volatiles, free fatty acids and cuticular hydrocarbons in the termite Reticulitermes aculabialis.

As eusocial insects prevalent in tropical and subtropical regions, termites are characterized by highly organized behaviors and exceptional adaptability, rooted in caste differentiation and chemical communication. These traits make them excellent models for studying insect social structures and ecological interactions. Investigating how termites use chemical signals to perceive and respond to their environment provides insights into their coordination and adaptation within complex ecosystems. This study delved into the chemosensory mechanisms of Reticulitermes aculabialis, examining the interactions of four olfactory proteins with 70 ligands, including host volatiles, cuticular hydrocarbons (CHCs), and free fatty acids (FFAs). Molecular docking simulations revealed varied affinities of the olfactory proteins for long-chain hydrocarbons (n-C to n-C), suggesting a nuanced chemical communication system through specific hydrocarbon detection. RacuCSP1 and RacuCSP2 exhibited specific binding to linoleic acid and undecanoic acid, respectively, highlighting the significance of FFAs in the physiological and behavioral processes of termites. The four olfactory proteins showed a strong affinity for longifolene in fluorescence competitive binding experiments. Notably, RacuOBPs exhibited unique affinities for terpenoid volatiles such as β-lonone and neocembrene, while RacuCSPs specifically bound with terpenoids like 3-carene, myrtenol, α-pinene oxide and β-pinene indicating their critical roles in host detection. Behavioral observations following gene silencing revealed that RacuOBP5 was essential for recognizing longifolene and α-lonone recognition, while RacuCSP1 was key for detecting α-pinene in termites. These findings enhance our understanding of the termite chemosensory system and offer insights for developing precise pest management strategies.