Effect Of Mitochondrion Proteins Expression Of Colletotrichum Gloeosporioides In Response Toborate Based On Comparative Proteomic Analysis

This study is based on a comparative proteomics,using two-dimensional electrophoresis(2-DE)and MALDI-TOF-TOF-MS echnology to study the impact of mitochondrion proteins of Colletotrichum gloeosporioides and Alternaria alternata under normal condition and borate stress.And quantitative real-time PCR assay was used to investigated the correlation between transcription and translation.Firstly,obtain different protein maps by 2-DE which different concentration of potassium tetraborate(0 mM,5 mM and 10 mM)were treated on anthracnose conidium at 48 h and 72 h.Then by means of MALDI-TOF-TOF-MS echnology,analysis and identified of differentially expressed proteins.Sixteen proteins were selected from the differential expression proteins(DPEs)and twenty DPEs to analyse variation of expression quantity of gene.The results are as follows:(1).It was detected about 600 protein spots by 2-DE and PDQuest software(Ver8.0,Bio-Rad)in the mitochondrial map of Colletotrichum gloeosporioides,At the same time,115 differentially expressed protein spots were analyzed.By MALDI-TOF-TOF-MS 58 protein spots were successfully identified.It was detected about 600 protein spots by 2-DE and PDQuest software(Ver8.0,Bio-Rad)in the mitochondrial map of Alternaria alternata,at the same time,110 differentially expressed protein spots were analyzed.By MALDI-TOF-TOF-MS 56 protein spots were successfully identified.(2).Through the expression analysis of 58 differentially expressed mitochondria protein spots in Colletotrichum gloeosporioides,12 protein spots were significantly up-regulated.25 protein spots were significantly down-regulated and 21 protein spots were both significantly up-regulated,also significantly down-regulated.The expression of 58 differentially expressed mitochondrial protein spots in Alternaria alternata,11 protein spots were significantly up-regulated.14 protein spots were significantly down-regulated and 31 protein spots were both significantly up-regulated,also significantly down-regulated.(3).The functional analysis of 57 different protein spots of mitochondria protein in Colletotrichum gloeosporioides was carried out,and then differentially expressed proteins can be divided into 8 functional types.They include electron transport chain(38%),Post-translational modification,protein turnover,and chaperones(15%),nucleotide transport and metabolism(13%),others and unknown(12%),membrane and transport(8%),amino acid transport and metabolism(7%),carbohydrate metabolism(5%),Signal transduction(2%).The functional analysis of 56 different protein spots of mitochondria protein in Alternaria alternata was carried out,and then differentially expressed proteins also can be divided into 8 functional types.The functional classification include electron transport chain(54%),Post-translational modification,protein turnover,and chaperones(16%),function unknown(12%),Tricarboxylic Acid Cycle(10%),amino acid transport and metabolism(3%),carbohydrate metabolism(3%),nucleotide transport and metabolism(2%),membrane and transport(2%).Among them,the largest functional groups of Colletotrichum gloeosporioides and Alternaria alternata were electron transport chain related proteins and Post-translational modification,protein turnover,and chaperones proteins.(4).KEGG pathway analysis was performed on 58 different proteins of Colletotrichum gloeosporioides by using KOBAS website.There were twelve differential proteins involved in 25 pathways,and then According to the function of these pathways were divided into carbohydrate metabolism related pathway,energy metabolism related pathway,amino acid metabolism related pathway,nucleoside metabolism related pathways and other metabolic pathways 5 categories pathway.The pathways which is the most differential proteins are involved are energy-related pathways.KEGG pathway analysis was performed on 56 different proteins of Alternaria alternata by using KOBAS website.There were fourteen differential proteins involved in 18 pathways,and then According to the function of these pathways were divided into carbohydrate metabolism related pathway,energy metabolism related pathway,amino acid metabolism related pathway,Metabolism of cofactors and vitamins pathways and other metabolic pathways 5 categories pathway.The pathways which is the most differential proteins are involved are energy-related pathways,also.(5).According to the different biological function classification of the expression proteins,Sixteen proteins and twenty proteins from the differential expression proteins of Colletotrichum gloeosporioides and Alternaria alternate,respectively,then to identify by quantitative real-time PCR.It is find that protein and gene expression had a good relativity but individual inconformity.So,it is necessary to detect on transcription after detection of proteins.