Differential Expression Of Diapause-associated Proteins In Exorista Civilis (Rondani.)

Exorista civilis Rondani is affiliated with Diptera:Tachinidae,is a kind of enemy heavenly enemy.It has a wide range of parasitic parasites and is a dominant natural enemy of important crop pests such as armyworm and grasshopper.And Exorista civilis is an important regulatory factor affecting the population of armyworm and grasshopper.At present,the research on the Exorista civilis includes biological characteristics,multi-host selectivity,ISSR analysis of different geographical locations,flight capability,rejuvenation and propagation,and diapause induction.Exorista civilis are used as parasitic natural enemies to study their large scale propagation and storage.Research on the diapause control mechanism of natural enemies in sheds can increase the shelf-life of the product to extend the control of pests.In this paper,the differential expression of diapause-associated proteins was investigated using the iTRAQ proteomics technique and bioinformatics analysis method.A preliminary investigation was conducted combining with multidisciplinary and differential proteomics information,the quantitative and functional expression of the diapause-associated proteins was investigated,and the differential proteins associated with diapause were found.The first chapter of this thesis mainly introduces the research progress of diapauses and the proteomics,differential proteomics,research progress,technical level and practical application of drosophila,and clarifies the necessity of using proteomics to study the diapause mechanism Exorista civilis.The second chapter of this thesis introduces the overall idea of using iTRAQ technology and describes each step in detail.After the sample acquisition,protein quantification,protein extraction and detection,enzyme release salt,iTRAQ label,and mass spectrometry detection,then using Proteome Discoverer2.1 software to retrieve the data of the uniprot_taxonomy₄3755.fasta database and combining with bioinformatics to analyze the protein functions.The final results showed that the number of total proteins was 1055,and the number of peptides was 6233.The total number of secondary spectra identified was 22,315 by the iTRAQ technology.The 1892 proteins were annotated by KEGG and classified into six major categories based on function: cellular processes,environmental information processing,genetic information processing,human diseases,metabolism,and organic systems;the protein was annotated by the GO system function to the 1393 protein and was divided into organisms Process,molecular function,and cellular components are three major types;711 proteins are annotated in the COG system and are occupied by two major categories of translation and post-translational modification.Among the 1055 proteins identified,there were 95 different proteins,of which 24 were up-regulated proteins,and 71 were down-regulated proteins.The overall level of the differential protein was down to down expression,which was closely related to the low metabolism in the diapaused Exorista civilis.Through the functional annotation of the GO system combined with the pathway enrichment of KEGG,the differential proteins of diapauses were analyzed.It was found that there was an up-regulation of protein expression associated with the cold resistance of diapause insects in diapauses.Shock protein,which also further proves that heat shock protein is a chaperone protein of insect diapause.In addition,we found related with energy metabolism and lipid metabolism,amino acid metabolism and carbohydrate metabolic pathway showed different levels of up or down,explain the source of the Exorista civilis chasing energy supply in the process of Exorista civilis diapause: on the one hand is to reduce its energy consumption,such as low level of carbohydrate metabolism,on the other hand is to explain the activation of reserve substances the energy,such as fatty acid degradation.