| Hyphantria cunea is a worldwide quarantine pest,because of its biological characteristics such as polyphagia,large appetite,surprisingly fecundity and excellent adaptability,once it invades,it is easy to break out.At present,Hyphantria cunea has become one of the important invasive species in China and caused widespread harm.In such a situation,measures should be taken in time to prevent and control Hyphantria cunea.Regulate the behavior of insects for the harmless biological control of pests is one of the current hot spots.In the various behaviors of insects,sensitive olfactory system plays a vital role,and a variety of olfactory related proteins are involved in the process of insect olfactory recognition,among which ORs play a central role.Thus,the functional study of ORs has become a core element in the insect olfactory mechanism and provides strategies for pest control.In the current work,based on the genome and transcriptome database,the study identified the OR genes of Hyphantria cunea,analyzed its homology and expression characteristics,furthermore,the functional characterization of Hcun ORs was analyzed.Based on the above,the study explored the electrophysiological and behavioral responses of Hyphantria cunea to the one of odor ligands(D-camphor).The main results are as following:1.By screening the annotation information of the genome and transcriptome database and necessary bioinformatics analysis,a total of 65 candidate Hcun OR genes were identified.2.A phylogenetic tree has been constructed with 485 ORs from 8 species of lepidopteran insects.Through the phylogenetic tree,ORco and three candidate PR of Hyphantria cunea were predicted,and found that most ORs have large functional differentiation,while individual ORs’ function is highly conserved.3.FPKM values were used to analyze the expression characteristics of Hcun OR genes among different tissues of adults,which indicate that most Hcun OR genes were predominantly expressed in the antennae.4.This study cloned the full-length sequences of Hcun ORco and 9 Hcun OR genes.The function of 8 Hcun ORs then was analyzed by Xenopus oocyte system and twoelectrode voltage clamp.Results showed that Hcun OR10 responds to 3 aromatic compounds,with the biggest response to benzaldehyde;Hcun OR14 responds to the 7odors,and the largest response to phenylacetaldehyde;Hcun OR15 responds to 2aromatic compounds,the largest response to eugenol;Hcun OR28 had no response to any test odors;Hcun OR42 reacts to nerolidol;Hcun OR46 reacts to 11 kinds of odors,among which,the highest reaction to D-camphor;Hcun OR47 is responsive to 20 kinds of odors,which are mainly aliphatic alcohols and aldehyde compounds,and has the greatest reaction to 1-nonanal;Hcun OR50 is responsive to 6 unsaturated short carbon chain compounds,has the greatest reaction to nerolidol.5.EAG and Y-tube were used to determine the antennal electrophysiological and behavioral responses of Hyphantria cunea to D-camphor,which indicated that 0.1 μg/μL and 1 μg/μL of D-camphor have no effect on Hyphantria cunea female and male adults.While the higher concentrations of D-camphor(10 μg/μL and 100 μg/μL)caused significant antennae response and have obvious repellent effect against Hyphantria cunea.Additionally,the antennae response and avoidance effect enhanced with the concentration of D-camphor increased.In summary,this study identified Hcun OR genes,analyzed its homology and expression characteristics,furthermore,first analyzed the odor ligands of 7 Hcun ORs,at the same time,found that a certain concentration of D-camphor has a significant avoidance effect on Hyphantria cunea,which lays an important foundation for clarifying the specific chemosensory mechanism and is of great significance for the future green prevention of Hyphantria cunea. |
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