Expression, affinity, and binding mode analysis of antennal-binding protein X in the variegated cutworm Peridroma saucia (Hübner).

The variegated cutworm Peridroma saucia (Hübner) is a worldwide pest that causes serious damage to many crops. Odorant-binding proteins (OBPs) are small soluble proteins involved in the first step of odorant reception. In moths, antennal-binding protein Xs (ABPXs) represent a main subfamily of classic OBPs. However, their functions remain unclear. Here, we cloned the ABPX gene from the antennae of P. saucia. RT-qPCR and western-blot analyses showed that PsauABPX is antenna-predominant and male-biased. Further temporal expression investigation indicated that the expression of PsauABPX started 1 day before eclosion and reached the highest 3 days after eclosion. Next, fluorescence binding assays revealed that recombinant PsauABPX had high binding affinities with P. saucia female sex pheromone components Z11-16: Ac and Z9-14: Ac. Then, molecular docking, molecular dynamics simulation, and site-directed mutagenesis were employed to identify key amino acid residues involved in the binding of PsauABPX to Z11-16: Ac and Z9-14: Ac. The results demonstrated that Val-32, Gln-107 and Tyr-114 are essential for the binding to both sex pheromones. This study not only give us insight into the function and binding mechanism of ABPXs in moths, but could also be used to explore novel strategies to control P. saucia.

GOBP1 from the Variegated Cutworm Peridroma saucia (Hübner) (Lepidoptera: Noctuidae) Displays High Binding Affinities to the Behavioral Attractant (Z)-3-Hexenyl acetate.

The variegated cutworm Peridroma saucia (Hübner) is a worldwide pest that causes serious damage to many crops. To recognize sex pheromones and host plant volatiles, insects depend on olfactory chemoreception involving general odorant-binding proteins (GOBPs). In this study, PsauGOBP1 was cloned from the adult antennae of P. saucia. RT-qPCR and Western-blot analysis showed that PsauGOBP1 was specifically and equally expressed in the adult antennae of both females and males. Fluorescence competitive-binding assays with sex pheromones and host plant volatiles demonstrated that PsauGOBP1 bound to six host plant volatiles: (Z)-3-hexenyl acetate (KD = 4.0 ± 0.1 µM), citral (KD = 5.6 ± 0.4 µM), farnesol (KD = 6.4 ± 0.6 µM), nonanal (KD = 6.8 ± 0.3 µM), (Z)-3-hexen-1-ol (KD = 8.5 ± 0.6 µM), and benzaldehyde (KD = 9.4 ± 0.5 µM). Electroantennogram recordings with the six host plant volatiles indicated that (Z)-3-hexenyl acetate elicited the strongest responses from both male and female antennae. Further bioassays using Y-tube olfactometers showed that (Z)-3-hexenyl acetate was attractive to adult P. saucia of both sexes. These results suggest that PsauGOBP1 might be involved in detecting host plant volatiles and that (Z)-3-hexenyl acetate might serve as a potential attractant for the biological control of P. saucia.

Identification of Candidate Chemosensory Receptors in the Antennae of the Variegated Cutworm, Peridroma saucia Hübner, Based on a Transcriptome Analysis.

Insect chemoreception, including olfaction and gustation, involves several families of genes, including odorant receptors (ORs), ionotropic receptors (IRs), and gustatory receptors (GRs). The variegated cutworm Peridroma saucia Hübner (Lepidoptera: Noctuidae) is a worldwide agricultural pest that causes serious damage to many crops. To identify such olfactory and gustatory receptors in P. saucia, we performed a systematic analysis of the antennal transcriptome of adult P. saucia through Illumina sequencing. A total of 103 candidate chemosensory receptor genes were identified, including 63 putative ORs, 10 GRs, 24 IRs, and 6 ionotropic glutamate receptors (iGluRs). Phylogenetic relationships of these genes with those from other species were predicted, and specific chemosensory receptor genes were analyzed, including ORco, pheromone receptors (PRs), sugar receptors, CO2 receptors, and IR co-receptors. RT-qPCR analyses of these annotated genes revealed that 6 PRs were predominantly expressed in male antennae; 3 ORs, 1 GR, 2 IRs, and 2 iGluRs had higher expression levels in male than in female antennae; and 14 ORs, 1 GR, and 3 IRs had higher expression levels in female than in male antennae. This research increases the understanding of olfactory and gustatory systems in the antennae of P. saucia and facilitates the discovery of novel strategies for controlling this pest.