Analysis Of Related Protein About The Dispause For Over Wintering Larvae Of Ostrinia Furnacalis

In this study, the Asion corn borer Ostrinia furnacalis (Guenée)5th instar larvaewere as the research object,using two-dimendional electrophoresis (2-DE) and massspectrometry identification combined with bioinformatics to study the process of theAsion corn borer protein abundance level in diapause from the proteome level andexplore the function of differentially expressed proteins in this period, to provides newtheoretical basis for the study of the molecular mechanisms of Asian corn borer indiapause.The plasma in hemolymph is responsible for the energy-delivering and materialtransportation, wound healing,and temperature regulation,the plasma proteins is thekey members to perform these functions.The preparation of the hemolymph proteinfrom larvae is crucial important to the two-dimensional electrophoresis.In this studycombined hemolymph characteristics with former protein extraction methods,compared TCA/acetone extraction method,the direct extraction method and PEGextraction method,by SDS-PAGE and2-D analyzed,we found that the PEG extractionmethod is the best method,it can remove most kinds of interference impurities andeasily dissolved,the SDS-PAGE electrophoresis has the largest number of strips,theseparated effect is the best,the two dimendional electrophoresisthe has the largestnumber of spots,874,and distributed widely,resolution high,repeatability excellent,theseparate effect is the best,these results lay the theoretic foundation for the analysis ofthe Asian corn borer related proteins in diapause.Collected the Asian corn borer larvae hemolymph in diapause and thehemolymph from the control group, obtain the plasma proteins using the PEGextraction,conduct two-dimensional electropresis, analysis the map using silverstaining and the software imageMasterTM2D Platinum6.0, the results show thatexpression abundance changes obove3times having53protein spots in diapauseconditions,including43upregulated,7downregulated,2specific expressioned and1spot disapperd.Combined with MALDI-TOF/TOF MS,11spots were identified from20spots,including heat shock70kDa protein,small heat shock protein, triosephosphateisomerase,phosphoglycerate mutase,arginine kinase,glutathionetransferase sepia and aliphatic nitrilase,putative.The protein function analysis based on the informations provided by theUniProtKB database and the NCBI database.Results show that heat shock70kDaprotein and small heat shock protein play the role of molecular chaperones,maintainthe cell structure,protect cells,involved in the process of apoptosis,these are critical forAsian corn borer surviving the harsh environment.Trioephosphate isomerase andphosphoglycerate mutase are important enzymes in the glycolysis,is essential foreffective energy obtained,glycolysis is the polysaccharide degradation’s commonway,energy is very important in the anaerobic,its metabolites can be other waysintermediate product.Arginine kinase is a kind of phosohagen kinase invertebrates,which played the role of energy storage and energy buffer.Energy iscrucial to maintenance life and against the cold environment in Asian corn borerdiapause.Glutathioneglutathione transferse and the nitriase is a detoxificationenzyme,helping insects against harmful substances.Above all,these studies provide a new theoretical basis for the study of themolecular mechanisms of Asian corn borer in diapause,and provid the experimentalbasis for revealing the relationship of cold hardiness and disaster for the Asian cornborer larvae,aim at preventing the Asian corn borer effectively.