Gustatory receptors (GRs) are crucial in the peripheral coding of the non-volatile compounds in insects, and thus play important roles in multiple behaviors including feeding, mating, and oviposition. However, little research has been done on GRs in lepidopteran pests. In the current work with Spodoptera litura, an important worldwide crop’s pest, a candidate fructose GR gene (SlitGR8) was cloned in full length, and its spatial and temporal expression profiles were determined by quantitative real-time PCR (qPCR). It revealed that SlitGR8 was highly expressed in antennae of both male and female adults, as well as in larva of first, fifth and sixth instar. Functional analyses were further conducted using the Xenopus oocyte system. SlitGR8 responded specifically to D-fructose among 12 tested sugar compounds. In addition, the behavioral assay demonstrated that both female and male moths could respond with proboscis extension behavior to D-fructose applied onto the antenna, but females showed higher sensitivity than males. The results provide an important base for further elucidation of molecular mechanisms of gustation, and a potential target for development of feeding interfering technique in S. litura.

The olfactory system in insects are crucial for recognition of host plants and oviposition sites. General odorant binding proteins (GOBPs) are thought to be involved in detecting odorants released by host plants. Orthaga achatina (Lepidoptera: Pyralidae) is one of the most serious pests of camphor trees, Cinnamomum camphora (L.) Presl, an important urban tree species in southern China. In this study, we study the GOBPs of O. achatina. Firstly, two full-length GOBP genes (OachGOBP1 and OachGOBP2) were successfully cloned according to transcriptome sequencing results, and real-time quantitative PCR measurements showed that both GOBP genes were specifically expressed in the antennae of both sexes, proposing their important roles in olfaction. Then, both GOBP genes were heterologous expressed in Escherichia coli and fluorescence competitive binding assays were conducted. The results showed that OachGOBP1 could bind Farnesol (K_i = 9.49 μM) and Z11-16: OH (K_i = 1.57 μM). OachGOBP2 has a high binding affinity with two camphor plant volatiles (Farnesol, K_i = 7.33 μM; α-Phellandrene, K_i = 8.71 μM) and two sex pheromone components (Z11-16: OAc, K_i = 2.84 μM; Z11-16: OH, K_i = 3.30 μM). These results indicate that OachGOBP1 and OachGOBP2 differ in terms of odorants and other ligands. Furthermore, key amino acid residues that bind to plant volatiles were identified in GOBPs using 3-D structure modeling and ligand molecular docking, predicting the interactions between the GOBPs and the host plant volatiles.