Trascriptome Sequencing Of The Yellow Peach Moth,Conogethes Punctiferalis(Guenée)(Lepidoptera:Crambidae) And Functional Analysis Of The Pheromone Binding Protein CpunPBP1

The olfaction system is essential for insect locating hosts,spouses and looking for mates.Among them,some physiological and behavioral experiments have shown that OBPs is the first step in insect olfactory sense mechanisms biochemical reactions.The yellow peach moth(YPM),Conogethes punctiferalis(Guenee)is one of the most important pest of agriculture and forestry in China during recent years.Further study on OBPs in the yellow peach moth could provide us new methods to control this pest through interring their olfaction perception.In this study,we obtained the antennae transcriptome data of C.punctiferalis using Illumina HiSeqTM 2500 sequencing and several olfactory related genes were identified by bioinformatics analysis method.We use quantitative real-time PCR method to verify sequencing data and the ordrant functional of C.punctiferalis are researched.The main results are as follows:1.Using transcriptome sequencing we obtained 101366 transcripts with total length of 116001455 bp and average length is 1144 by,a total of 47109 unigenes were generated by paired-end joining and gap-filling.Based on sequence similarity comparison against the five databases(NR,Swiss-Prot,GO,COG and KEGG)using Blastx,a total of 18990 unigenes were annotated.2.We used bioinformatics analysis method to discover several olfactory related genes,including 15 odorant binding protein(OBP)genes,46 odorant receptor(OR)genes and 7 ionotropic receptors(IRs).3.When FDR and p-value was less than or equal to 0.05 as the threshold,the antennal transcriptomes of male and female C.punctiferalis had 1308 significant differential trascripts,with 759 up-regulated genes and 549 down-regulated genes,and including 30BPs and 70rs.We choose olfactory genes unigenes to detect their expression between male and female antennae using RT-qPCR.The result revealed that the real time PCR results of are consistene with the DEG data.4.A novel PBP cDNA(CpunPBP1)was cloned,expressed and purification from C.punctiferalis.The binding affinity of CpunPBP1 with 15 odorant compounds were measured by using a fluorescence probe 1-NPN with the fluorescence competitive binding experiments.The results showed that CpunPBP1 had the capability of binding 2 pheromone chemicals(cis-10-hexadecenal and hexadecanoyl),with binding constants of 7.32 and 9.39 ?mol/L respectively.In addition,CpunPBPl could bind with 8 plant volatiles.Of which,camphene had the strongest binding capability,with the binding constant of 3.76 ?mol/L.