The Molecular Mechanism Of Fertility Conversion On Thermo-sensitive Cytoplasmic Male-sterile Wheat KTM3315A By ITRAQ-based Proteomics Analysis

Wheat is one of the major crops in the world,the distribution range and the planting area of the most extensive,most of the total yield,high nutritional value of food crops,human food security depends largely on the production of wheat.Among them,the most of the research is cytoplasmic male sterility.The K-type cytoplasmic male sterile line is Aegilops Kotschyi cytoplasm-based on a hybrid system developed,a K-type cytoplasmic male sterile materials,KTM3315 A,exhibits completely male sterility and it can be used to produce hybrid wheat seeds during the normal wheat-growing season,whereas it propagates via self-pollination at high temperatures.Used of the thermo sensitive male sterile line breeding,seed production,two-line hybrid system,it has important application value in cross breeding,but its fertility transformation mechanism needs further study.This test used the K-type thermo-sensitive male sterile wheat KTM3315 A as experimental materials,the morphological and cytological observation of the different fertility condition of anther;iTRAQ-based proteomics analyses KTM3315 A in different fertility condition of anther,using bioinformatics comparison of different express proteins of fertility and sterility condition,find out the some proteins and some metabolic pathways relate to fertility conversion;in addition,selected some genes in different developmental stages of the qRT-PCR test,the main results obtained are as follows:1.Cytological observation and morphological characteristics of anther and microspore,K-type thermo sensitive cytoplasmic male sterile wheat KTM3315 A in anther development of abnormal process.Infertile conditions,anthers can not be pollinated normally,the fertile conditions,anthers can be pollinated normally.The anthers and microspore of cytological observation showed;during the TNP stage,the fertile anther skin cells were significantly larger than the sterile anther cells,while the fertile anthers were significantly plumper with a more rounded phenotype,whereas the sterile grains appeared to be deformed and shrunken2.By sequencing of KTM3315 A of different conditions for proteins,a total 186,000 spectra,and 11,292 unique peptides were obtained.The protein abundance of different samples were compared and analyzed.The results showed that the protein abundance level is basically the same.According to the measured expression of protein samples,4639 different abundant proteins(DAPs)were identified.By classification and functional annotation of differences between the measured protein obtained in different periods,many increased or decreased proteins by differential protein functional classification of enrichment analysis showed that the protein mainly concentrated in energy metabolism,synthesis and degradation and formation of exine protein of the.The similar enrichment analysis showed that these differential proteins and metabolic pathways played an important role in fertility transformation in three periods,which provided a basis for further research on the fertility transformation mechanism of thermo sensitive cytoplasmic male sterility.3.In order to evaluate the correlations between the mRNA and protein levels,we employed quantitative real-time PCR(qRT-PCR)to confirm the transcript levels of the DAPs,where DAPs were analyzed by qRT-PCR.It was found that the quantitative results of some DAPs have similar expression trends with their sequencing results,and many DAPs that the results of quantitative PCR showed that their transcript levels were poor indicators of the corresponding proteins levels.These different express proteins change in different periods,may play an important role in the fertility conversion process of the wheat KTM3315 A.