Electron Microscope And ITRAQ Protein Analysis Of T-CMS And Its Maintainer Line

In this study, in order to explore the mechanism of T-wheat cytoplasmic male sterility more deeply, T-CMS(T Ji 5418) and its maintainer line(Ji 5418) were choosen as the materials. Using iTRAQ technology the anther total protein to relatively quantitative analysis,and the differences in protein mass spectrum identification, GO annotation, COG comments,Pathway metabolic pathways, GO enrichment, Pathway enrichment analysis, combined with anther tapetum cell tem observation, to discuss tapetum and the dynamic changes of the mitochondria degradation.And clear the main metabolic process of male sterility.Then provide theoretical basis for the application of T male sterility.The results obtained are as follows:1. It is the cause of T wheat abortive that the anther tapetum cells dissolved too fast. It had shown that mononuclear period may not be the start period, but much of its period of abortive.And it also showed that tapetal cells endoplasmic reticulum may be associated with CMS. The main reason of the T- CMS abortive is that tapetum dissolved too fast, and it can’t continue to microspore function.2. Basic appraisal information differences in protein. The different protein relative to the molecular weight of the protein mainly between 10-100 kDa, peptide length distribution is mainly within the 12 amino acids of protein-based, 99.25% of the total difference in protein and with the increase in the number of protein and peptide length reduced peptide sequence coverage mainly between 5%-25%, the number of distribution-specific peptides with 10 amino acids within the protein-based, 97.49% of the total difference in protein.3. The number of different protein is slightly higher in binucleate stage than in monocytes. The results by i TRAQ relative quantification show that the difference of the total protein in monocytes 402, include regulated protein 222, down protein 180,less than binucleate stage 53, 17, 36,respectively.4. T-CMS is the result of karyoplasm interaction effects.It’s found by GO annotation,that differences between protein and found differences between proteins are mainlydistributed in cells(cells and cell membrane, mitochondria and chloroplasts.Differences in protein molecular functions, mainly with functions of catalytic activity and combining with the differences of proteins involved in biological functions mainly cell metabolism.5. Energy shortage caused microspore abortive.The COG comments found that differences in protein, the proportion of the highest general function, in the key-mononuclear stage abortive, associated with the generation and transformation of energy differences in protein proportion is higher also, and the difference in the two nuclear issue is relatively low, this suggests that the critical period in the abortive due to energy generation and transformation of energy supply is insufficient, cause in the process of the microspore development, caused by a lack of energy, make its are stunted, and then abortion.6、The PPP is the key reason to abortion, after pathway enrichment.It’s not significantly different G6 PD in oxidation stage of PPP, but there are four key enzyme in the non-oxidative phase were down regulation: FBA、TAL、RpiA、PGM. It also found that autophagy regulation and endocytosis were relative to fertility. Alkaloid metabolism may affect the CMS. There no different protein that fertility-related at binucleate stage. And also, not all different protein are fertility-related, some are related with plant growth.7. Autophagy,especially the autophagy pathway VAC8 proteins,associated with T –CMS,by cytology observation and iTRAQ analysis.