| Lonicera japonicaThunb(Caprifoliaceae)is perennial evergreen vine,the flower is a traditional medicine.The Xylotrechus grayii is a stem pest of honeysuckle.It is hidden darm.The use of chemical insecticides has no obvious effect on the prevention and control of natural calf and it also seriously affects the safety of honeysuckle medicinal materials.Therefore,people urgently need to develop a pollution-free control measure.In the long-term evolution,insects form highly sensitive and complex olfactory system.Insects use the olfactory system to sense and identify chemical information substances in the external environment to complete the search and location of the host plants and the spouses.The formation of insect olfactory signals involves the interaction of a series of related proteins such as odorant-binding proteins(OBPs)and chemosensory proteins(CSPs),elucidating the mechanism of insects' perception of volatile compounds in the environment,and the use of chemical pheromones for the control of insect pests such as disorientation and attraction.Provide theoretical basis.In this study,we used the bioinformatics method to identify the olfactoryassociated genes of the X.grayii adult antennae transcriptome,and analyzed the expression differences of CSPs in different tissues of the X.grayii.The olfactory sensory mechanism of the Xylotrechus grayii was initially revealed,which provides a theoretical basis for the scientific prevention and control of the X.grayii.The major results of this paper list as below:?.The X.grayii antennal transcriptome sequencing and bioinformatics:(1)The X.grayii antennal transcriptome sequencing received a total of 102535456 raw datas,removal of impurities obtained after 98460094 Clean reads,Q30 base percentage was not less than 90%,GC content was about 44%.After no reference genome was de novo assembled,127824 transcripts were obtained through Trinity splicing.Finally,90490 unigenes were screened out,of which 15281 were 1000 bp above length,with an average length of 710 nt and N50 length of 1416 nt.(2)functional annotation of the 90490 unigenes obtained,20651 of which are annotated to NR database,6648 unigenes annotations to KOG database,10292 unigenes annotations to KEGG database,16248 unigenes notes to GO database.?.Identification of the X.grayii olfactory genes:Using bioinformatics methodology analysis of transcriptome data,identified 34 odorant binding protein(OBPs)gene,15 chemosensory protein(CSPs)gene,60 odorant receptor(ORs)gene,4 sensory neuron membrane protein(SNMPs)gene.Phylogenetic tree was constructed from olfactory related genes of the X.grayii and other insect related genes,and the evolutionary relationship between olfactory related genes and other insect olfactory related genes was compared.?.The X.grayii has 4 chemosensory protein expression spectrum analysis:Using fluorescence quantitative PCR(qRT-PCR)to study 4 chemosensory protein gene in the X.grayii of female and male adults antennae(At),head(H),chest(C),abdomen(Ab),foot(F),the results showed that XgraCSP2 is mainly expressed in female and male antennaes,head,foots,expression of XgraCSP2 in abdomen of the male was higher than female,no expression in the chest.XgraCSP4 the highest expression in the male head,Secondly it is expressed in the antennae and chest,and has trace expression in the abdomen and feet.Females compared to males XgraCSP4 expressed more in the antenna than in the male,but the expression in the head was lower than that of the male.XgraCSP5 is expressed in a small quantity in female tissues,mainly in the male antennae,but microexpression in the head,chest,abdomen and foot of males.XgraCSP9 was mainly expressed in antennae,no expression in other tissues,there is a significant difference between the expression of female and male antennaes,the expression is 2.5 times of male female. |
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