Differentially Protein Analysis Between Wild Type And ?veA Deletion Mutant Of Eurotium Cristatum

veA gene as an important regulatory factor,conservative in many fungi,especially in the ascomycetes,plays a key role in coordinating secondary metabolism and differentiation processes such as asexual or sexual sporulation in fungus.The Eurotium cristatum as a good material to study the mechanism of spore production is the predominate germ in the fungus growing process of Fuzhuan brick tea and it can produce not only sexual spore but also asexual spore.Under the condition of low osmotic pressure,wild type generates only ascospores and ?veA deletion mutant generates only conidia.veA gene regulate both sexual spores positively and asexual spore production negatively.So far,the mechanism of spore production research is still in its infancy.Protein is the executor of the life.The mechanism of spore production research has very important significance from the aspects of proteomics.In the studies,the protein which were from wild type and ?veA deletion mutant of Eurotium cristatum were focused on 24-cm immobilized pH gradient(linear pH: 4-7)strip(IPGS),and then IPGS was electrophoresised on the sodium dodecyl sulfate polyacrylamide gel(SDS-PAG).After SDS-PAG was stained with silver staining and coomassie blue R-250,differential proteins were detected by ImageMaster 2D platinum on the basis of difference multiple >2,and P<0.05.These proteins were identified by MALDI-TOF-MS and the search of NCBI database,and then were analyzed online through bioinformatics such as GO and KEGG.Results are as follows:1)Protein concentration of wild type and ?veA deletion mutant respectively: 3.208?g/?L ? 3.411?g/?L.Differentially protein of wild type and ?veA deletion mutant by 2-DE respectively: 1378±53?1732±94.2)Differentially expressed proteins were analyzed for significant down-or upregulation,which was calculated by twice folds,a total of 193 unique proteins were identified,of which there were 77 up-regulated significantly differential proteins in wild type and 97 up-regulated in ?veA deletion mutant.Seventysix differentially expressed protein spots identified successfully by mass spectrometry,the results show that there were 23 down-regulated significantlydifferential proteins and 53 up-regulated in ?veA deletion mutant relative to wild type.3)The results of Gene Ontology show that from the perspective of the function of molecular,ion binding and oxidoreductase activity play the dominant role;from the perspective of the biological proces,small molecule metabolic process and biosynthetic process play the dominant role;from the perspective of the cellular component,cytoplasm and intracellular play the dominant role.The results of KEGG pathway show that there were nine important metabolic pathways involved in differentially protein related to spore production.These protein were related to sugar metabolism,biosynthesis of amino acids and biological oxidation,as well as some unknown protein.4)The protein play an important role in spore production including four important proteins probably related to certain metabolic pathways: tubulin,protein phosphatase and eukaryotic translation initiation factor up-regulated in ?veA deletion mutant.The function and regulation mechanism of these protein need further verification related to spore production and development in Eurotium cristatum.