Odorant receptor (OR) is crucial for insects to detect and recognize external chemical cues closely related to their survival.  The insect OR forms a heteromeric complex composed of a ligand-specific receptor and a ubiquitously odorant receptor coreceptor (Orco).  This study used the CRISPR/Cas9 technique to knock out (KO) Orco and reveal its essential role in acting on OR-meditated olfactory behavior in a critical invasive agricultural pest, the fall armyworm (FAW), Spodoptera frugiperda.  Electroantennogram (EAG) results suggested that the Orco mutants of both male and female moths severely reduced their electrophysiological responses to the eight tested plant volatiles and two sex pheromones.  However, the Orco gene played distinct roles in mating behavior between sexes: the mating behavior was fully disrupted in mutated males but not in mutated females.  The oviposition result indicated that the Orco KO females displayed reduced egg laying by 24.1% compared with the mated wild type (WT) females.  Overall, these results strongly suggest that Orco is an excellent target for disrupting FAW’s normal behavior and provides a feasible pest control approach.

Pea aphid, Acyrthosiphon pisum, is a serious pest of many different leguminous plants, and it mainly relies on its odorant receptors (Ors) to discriminate among host species.  However, less is known about the role that Ors play in the host plant location.  In this study, we identified a novel conserved odorant receptor clade by phylogenetic analysis, and conducted the functional analysis of ApisOr23 in A. pisum.  The results showed that the homologous Ors from A. pisum, Aphis glycines and Aphis gossypii share 94.28% identity in amino acid sequences.  Moreover, conserved motifs were analyzed using the annotated homologous Or23 from eight aphid species, providing further proof of the high conservation level of the Or23 clade.  According to the tissue expression pattern analysis, ApisOr23 was mainly expressed in the antennae.  Further functional study using a heterologous Xenopus expression system revealed that ApisOr23 was tuned to five plant volatiles, namely trans-2-hexen-1-al, cis-2-hexen-1-ol, 1-heptanol, 4´-ethylacetophenone, and hexyl acetate.  Among them, trans-2-hexen-1-al, which is one of the main volatile organic compounds released from legume plants, activated the highest response of ApisOr23.  Our findings suggest that the conserved Or23 clade in most aphid species might play an important role in host plant detection.

Lipoxygenases (LOXs) are a group of non-heme, iron-containing enzymes and extensively involved in plant growth and development, ripening and senescence, stress responses, biosynthesis of regulatory molecules and defense reaction. In our previous study, 18 LOXs in melon genome were screened and identified, and five 13-LOX genes (CmLOX08, CmLOX10, CmLOX12, CmLOX13 and CmLOX18) were predicted to locate in chloroplast. Phylogenetic analysis result showed that the five genes have high homology with jasmonic acid (JA) biosynthesis-related LOXs from other plants. In addition, promoter analysis revealed that motifs of the five genes participate in gene expression regulated by hormones and stresses. Therefore, we analyzed the expressions of the five genes and LOX activity in leaves of four-leaf stage seedlings of oriental melon cultivar Yumeiren under abiotic stress: wounding, cold, high temperature and hydrogen peroxide (H2O2), and signal molecule treatments: methyl jasmonate (MeJA), abscisic acid (ABA) and salicylic acid (SA). Real time qPCR revealed that wounding and H2O2 induced the expressions of all the five genes. Only CmLOX08 was induced by cold while only CmLOX13 was suppressed by high temperature. ABA induced the expressions of CmLOX10 and CmLOX12 while inhibited CmLOX13 and CmLOX18. MeJA increased the 3 genes expressions except CmLOX08 and CmLOX13, whereas SA decreased the effect, apart from CmLOX12. All the abiotic stresses and signal molecules treatments increased the LOX activity in leaves of oriental melon. In summary, the results suggest that the five genes have diverse functions in abiotic stress and hormone responses, and might participate in defense response. The data generated in this study will be helpful in subcellular localization and transgenic experiment to understand their precise roles in plant defense response.