Screening And Identification Of Odor Receptor Genes Of Paratrioza Sinica Yang&Li

Paratrioza sinica Yang&Li is one of the important pests of Lycium barbaru,it causes immediate damage by sucking the sap of leaf and twig in the stages of adult and nymph,leading to the weak tree.Its overwintering adults carry Aceri macrodonis Keifer through the winter,the latter also can cause serious damage to the medlar.In order to carry out the control of olfactory gene targets in P.sinica,and to understand its olfactory recognition mechanism.The attractiveness and repellency ofplant volatile compounds at different concentrations to adults of P.sinica using Y-tube olfactory behavior test and field experiments were analyzed,the transcriptome was sequenced,the odorant receptor gene was screened and identified,and its gene sequence was cloned and analyzed by bioinformatics.By q RT-PCR,the expression levels of two OR genes in the head,chest and abdomen of P.sinica were analyzed.The results are as follows:1.Octanal(0.01 μL/μL)has a very significant attracting effect on female P.sinica.1-octen-3-ol(1 μL/μL)and nonanal(0.001 μL/μL)have significant attractiveness to female adults;significant to males.Nonanal(0.01 μL/μL,0.001μL/μL)has attracting effect,nonanal(1 μL/μL)and jasmone(0.01 μL/μL)has repelling effect,3-hexen-1-ol,(Z)-(1 μL/μL)is effective against P.sinica.males are more repellent than females.With a high degree of attraction,1-octanol always repels both male and female adults of P.sinica.Among the 11 odorants in the field,nonanal(0.1 μL/μL)and octanal(0.01 μL/μL)have a higher trapping rate than the other treatments for adults.It has lower selection rate for linalool and foliol.Based on the results of the indoor and outdoor adults selection of odorants,nonanal and octanal have a significant attraction to the adults,but linaloola and foliol have a significant repellent response to adults.2.There were 48 647 unigenes with the total length of 54 114 993 bp,average length of 1 112 bp and an N50 of 1 780 bp.Through functional annotation based on seven databases,we found the most unigenes were annotated in NR database.According to GO database,more unigenes were annotated as cellular process、metabolic process,cell,cell part,binding and catalytic activity.In KEGG database,it is found that the most annotated pathway is signal transduction.By further screening,10 olfactory receptors(ORs)were identified.3.The Psin OR15 and Psin OR2 a gene sequences of P.sinica were cloned.The analysis revealed that the Psin OR15 gene has a full length of 749 bp,open reading frame of 237 aa,encodes 250 amino acids,a molecular weight of 28.73 KD,and an isoelectric point of 8.21.It is a hydrophilic protein with two transmembrane regions.And,the Psin OR2 a gene has a full length of 623 bp,the open reading frame is 138 aa,it encodes 117 amino acids,the molecular weight is 13.54 KD,and the isoelectric point is 9.87.It belongs to hydrophobin and may have a transmembrane region.There is no signal peptide in both of them.The secondary structure of Psin OR15 and Psin OR2 a proteins is dominated by α helix,and the tertiary structure is mainly formed by α helix and random coil winding and folding.4.The Psin OR15 and Psin OR2 a genes of P.sinica are expressed in different tissues of female and male adults.Among them,the expression of Psin OR15 gene was the highest in the breasts of female and male adults,followed by the head.It was slightly expressed in the abdomen,and the expression level in the breasts of males was higher than that of females.The expression level of Psin OR2 a gene in different tissues of female and male adults of P.sinica was not significantly different.