Expression Patterns Of Cold Hardiness Genes In Larvae Of Anoplophora Glabripennis During Different Overwintering Stages

Anoplophora glabripennis is an important forest borer pest in China and an important quarantine object in the world.There are a wide range of hosts,larvae mainly damage and overwintering in the host moth.Low temperature in winter have direct effects on the growth,development,reproduction and expansion of overwintering insects,therefore also affect the number distribution of insect populations in the next year.Therefore,the cold tolerance of insects has become an important and decisive condition for their survival and reproduction.At present,A.glabripennis has gradually spread to high latitudes and altitudesand,we predicted that is related to its cold resistance enhancement.In our study,we used the transcriptome sequencing technologies to explore the expression of cold tolerance genes and alternative splicing events of the A.glabripennis larvea in different overwintering stages to explore the regulations of cold tolerance genes and alternative splicing events and reveal the factors and the adaptation mechanism of its cold tolerance.The main results are as follows:1.All the genetic information at transcription level of A.glabripennis was obtained.A total of 822365302 of clean data passed the Illumina quality filter after transcriptome sequencing of fifteen samples.A total of 22,319 gene were annoted,including 3,909 gene annotations to COG database,8,103 gene annotations to GO database,and 4,581 gene annotations to KEGG database.1,518 genes are annotated in all databases.2.The transcriptional differential information of longhorned beetle larvae in different overwintering stages was determined.The analysis of 5 clustering subsets of 9 clusters of significantly differentially expressed genes showed that the variation trend of gene expression in 5 subsets was different.The main annotation functions of 2,4,6,7 and 8 clusters of genes were as follows:glycoside hydrolytic enzyme protein,odor binding protein,antimicrobial peptide,serine protease,enzyme related to energy and carbohydrate metabolism,shsp21.3.The expression rules of the target genes related to cold tolerance were screened out according to the differentially expressed genes.The expression patterns of 9 genes which were selected from 5 subclusters,mainly composed of glycoside hydrolytic enzyme family genes,antimicrobial peptide genes,serine protease genes,histoprotease genes,cysteine protease genes,and heat-induced protein HSP21 genes,were verified.These genes showed different trends after quantitative expression,indicating that the genes involved in regulating cold tolerance of longhorns glabra larvae were related to metabolic process,cell components,immune system,environmental process and signal transmission.4.The variation of variable shear events in the larvae of A.glabripennis during different overwintering stages was determined.The mutuallye xclusive exon(MXE)events occurred the most frequently in the larvae of A.glabripennis during the whole overwintering period.The AS events of A.glabripennis larvae in different wintering stages was affected by the ambient temperature.This study reveals that cold environment can induce A.glabripennis larvae produce more AS,the changed form of AS is an important way of A.glabripennis larvae to adapt more environment changes.