| Plodia interpunctella（Lepidoptera:Pyralidae）is one of the major storage pests with global distribution,it is urgent to develop green and efficient prevention and control measures for it.P.interpunctella adults perceive odorant molecules in the environment,search and locate suitable spawning places.But the olfactory recognition mechanism of P.interpunctella to perceive the key volatiles in its spawning place remains to be unclear.This paper adopted the P.interpunctella as subject of study,used transcriptome sequencing and bioinformatics analysis to screen for odorant receptors（ORs）of the P.interpunctella,and used the methods of analytical chemistry,molecular biology,electrophysiology and behavior to conduct a preliminary study on the ORs-mediated olfactory recognition mechanism of the female P.interpunctella towards the oviposition site and obtained the following experimental results:（1）Among the volatiles of food,50 volatiles were able to elicit antennal responses in P.interpunctella mated females.In this paper,52 volatiles from 9 host food of P.interpunctella were taken as odor stimuli,and the antennae potential responses of unmated females,mated females and males were examined by EAG.The results showed that mated females responded to larger values（≤-0.119 m V）of acetophenone,hexanoic acid,nonanal and decanal;at the same time,the EAG responses of mated females to Decanal,acetophenone,octanal and decane were significantly greater than those of unmated females（p<0.05）.（2）Through transcriptome sequencing and bioinformatics analysis,31 full-length Pint ORs genes were screened out,among which 17 genes were biased or highly expressed in the female antennae.we sequenced the transcriptomes including antennae of P.interpunctella larvae,adult females and males,adult mouthparts,thorax,abdomen,legs and wings,screened a total of 56 Pint ORs using bioinformatics analysis,of which 31 were full-length;based on the FPKM values of full-length Pint ORs in different tissues,the analysis revealed that 17 of them were biased or highly expressed in female antennae.（3）Pint OR23 and Pint OR26 specifically recognize acetophenone which is able to significantly attract mated females.In this study,a total of five full-length Pint ORs including Pint OR5,Pint OR11,Pint OR18,Pint OR23 and Pint OR26,and the expression of these five genes in female antennae was higher than that in other tissues verified by RT-PCR.The oocyte-bipolar voltage clamp system was used to screen the binding relationship between five Pint ORs and candidate volatiles.The results showed that Pint OR11 could recognize hexanal and pentanal;Pint OR26 could recognize acetophenone,benzyl alcohol and hexanol;Pint OR23 had a relatively wide recognition spectrum,recognizing six volatiles such as acetophenone,benzaldehyde,benzyl alcohol and undecane,which showed the largest electrophysiological response to acetophenone(（mean value was 305.1±16.08 n A）,the higher the concentration of acetophenone,the larger the response value(1×10-4m M～1m M).In order to investigate the specific function of acetophenone on P.interpunctella a"Y"type olfactometer behavior test was conducted.The results showed that acetophenone was significantly attractive to mated females when the concentration of acetophenone was 1μg/μL（p<0.05）.In conclusion,acetophenone is able to cause a large EAG response in mated females and has significant attraction to mated females.This volatiles can provide application guidance for the development of efficient and green trapping agents for females;In the process of identifying host storage food,Pint OR26 and Pint OR23 highly expressed in the antennae of females can specifically recognize acetophenone.Therefore,Pint OR26 and Pint OR23 can be used as potential molecular targets and action sites to provide a theoretical basis for the production of effective interfering agents. |
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