Differential Expression Of Olfactory Genes From Batocera Horsfieldi(Hope) Adult Made On Host Plant And Its Semiochemicals In Host

Longicorn has the habits of complementing nutrition and its adult living free and has a long operating life.Furthermore,longicorn performed a preference on different hosts.Batocera horsfieldi(Hope)is a significant pest on timber forest,economic forest and protection forest,as it is widespread,hardly controlled,serious harm and hosts various.Besides,B.horsfieldi is an elusive insect that has a strong stress resistance,and it could also make a devastating loss on forest.So it is important to harmless control the insect by making the most use of its preference as the forestry protection in the new development era.For those reasons,Headspace Solid Phase Micro-extractio(HS-SPME),Gas Chromatograph-Mass Spectrometry(GC-MS),Electroantennogram Responses(EAG),Gel Chromatography(Sephadex G75),Fluorescence Binding Assays(1-NPN),Reverse Transcriptase PCR(RT-PCR),Quantitative PCR(RT-qPCR)and Illumina HiSeq were conducted to investigate the differential expression of olfactory genes from B.horsfieldi adult made on host plant and its semiochemicals in host systematically,and explored the genetic selection mechanism and locating mechanism of B.horsfieldi adult made on host plant.The main results were follows:(1)The transcriptome library of B.horsfieldi contained 137485 transcripts and 69214 Unigenes;the mean number of transcript was 1142 bp,the mean number of Unigene was 1983 bp.And 41636 Unigenes were exhibited in NR library,14895 Unigenes were exhibited in NT library,19287 Unigenes were exhibited in KO library,33442 Unigenes were exhibited in SwissProt library,34687 Unigenes were exhibited in PFAM library,35321 Unigenes were exhibited in GO library,20582 Unigenes were exhibited in KOG library.The results of NR comment showed that 72.0%Unigenes of B.horsfieldi had similarities with Tribolium castaneum and Dendroctonus ponderosae.Transcriptome gene function classification of B.horsfieldi declared that:5 mainly function classification were exhibited in GO library,they were cellular process,metabolic process,single-organism process,binding and catalyticactivity respectively,and they contained 20912,19086,17202,21477 and 15823 Unigenes respectively;20585 Unigenes were exhibited in 26 groups of KOG library,and posttranslational modification,protein turnover and chaperones totally contained 1977,General function prediction only contained 3285,signal transductionmechanisms contained 3053,all above its contained 40.39%Unigenes totally;19287 Unigenes were exhibited in 5 function classification of KEGG library,they were cellular process(6793),environmental informationprocessing(6255),genetic information processing(3038),metabolism(3852)and organismal system(3508).Furthermore,the transcriptome library of B.horsfieldi also showed that when those insects were in different developmental stage,Unigenes expressed difference,including olfactory genes.(2)For those 10 kinds of olfactory genes expression,CSP2 gene expressed most,and CSP1,OBP1 and Minus-C OBP3 gene expressed least in B.horsfieldi parasitize on Populus tomentosa;OBP2 gene expressed most(CSP2 gene next),and CSP1,Minus-C OBP3 and Minus-C OBP4 gene expressed least in B.horsfieldi parasitize on J.regia,B.horsfieldi parasitize on P.tomentosa(except male adults)expressed more than those insects parasitize on J.regia significantly.And in all olfactory genes,CSP2 gene of male adult parasitize on P.tomentosa expressed most,CSP1 gene of larva parasitize on P.tomentosa expressed least.Besides,the olfactory genes of adults expressed more than larva usually,and male and female performed difference.Furthermore,when adults were fed by host plant(except female adults fed by J.regia),OBP2,OBP3,CSP1,CSP2,Minus-C OBP1 and Minus-C OBP3 gene expressed more than before significantly,and OBP1?OBP3 and CSP2 gene played the an important role,but the effect of Minus-C OBP3 and Minus-C OBP4 gene was small;in addition,when male adults were fed by J.regia,olfactory genes expressed significant more than before,it might be declared male adults performed prefer to J.regia;when female adults were fed by J.regia and Eucalyptus robusta,olfactory genes expressed significant less than before,it might be declared female adults performed anorexia to J.regia.(3)12 kinds of macromolecular crude extract protein(pro.I-XII,Protein I-XII)were found in antenna of B.horsfieldi.When those twelve kinds of macromolecular crude extract protein were isolated and purified by Sephadex G75,a purified protein(pur-pro.I,purified protein ?)was found,and it could combine with fluorescent probe(N-phenyl-1-naphthylamine,1-NPN)too.The relative molecular weight of pur-pro.? was 15.2 KDa approximately,and it was pro.?(Protein ?),when contrasted with all kinds of macromolecular crude extract protein.All above,it declared that the isolated and purified protein pur-pro.I was one of the odorant binding proteins of B.horsfieldi.(4)The volatile components in B.horsfieldi host plant Juglans regia L.(bark and leaf),Rosa cymosa Tratt.and Rosay multiflora Thunb.were mainly terpene,hydrocarbon,aromatic compounds,alcohols,aldehydes,ketones,esters and quinones and so on.And the relative content of terpene,alcohols and aldehydes were higher,the relative content of ketones,hydrocarbon and aromatic compounds were lower in host plant.The volatile components in host plant J.regia bark were 9 species 36 kinds,and 8 species 42 kinds in J.regia leaf,5 species 16 kinds in R.cymosa,7 species 23 kinds in R.multiflora.Besides,the bark and leaf of J.regia all contained 4 species 13 kinds of volatile components,and the relative content of terpene was highest in bark and leaf,43.371%and 78.948%respectively;the relative content of hydrocarbon was lowest in bark,1.043%;the relative content of ketones was lowest in leaf,0.009%.R.cymosa and R.multiflora all contained 3 species 8 kinds of volatile components,and the relative content of aldehydes was highest in R.multiflora,81.387%;the relative content of alcohols was highest in R.cymosa,55.519%;he relative content of aromatic hydrocarbons was highest in R.multiflora,0.679%;the relative content of phenols was highest in R.cymosa,0.747%.(5)For those 24 kinds of selected host volatile components,not all of them had effect on B.horsfieldi significantly.Those volatile components that had significant EAG effect on adults were:2-pentylfuran,1-octen-3-ol,terpinolene,2-ethylfuran,linalool(except male adults B.horsfieldi),salicylaldehyde,?-ocimene,(2E,4E)-hepta-2,4-dienal,p-cymene,eucalyptol,1-caryophyllene,(-)-?-pinene,guaiacol and 5-hydroxy-1,4-naphthoquinone.Besides,the results also showed that host volatile components had different effect on adults significantly,adults performed different preferences.And 1-octen-3-ol had most EAG effect on male adults significantly;salicylaldehyde and linalool had most EAG effect on female adults significantly.Furthermore,both male and female performed preferences for the same volatile component and their isomers.And the EAG response had a positive correlation with the concentration of volatile compounds in some degree.