A Research Into The Expression Of Proteins Related To Wheat T-Type Cytoplasmic Male Sterility(T-Cms) And The Determination Of Molecular Markers Of Fertility Restorer Gene Rf3

Male sterility became one of the hottest topics in genetics because the significance in both theoretical and applied researches. The relationship between sterile genes and their interactions with restorer had been paid more interest. It was in our investigation, the system of nearly isogenic lines ( NILs) was employed, which composed of 2 pure wheat strains with different fertility properties. The experiment was one secondary project of the comparative phonemics system analysis in our lab taking the nearly isogenic lines as experimental materials, combining the theory of developmental genetics with the practice of the two dimensional polyacrylamide gel electrophoresis (PAGE), the different proteins were analyzed at different developmental stages in young spikes of common wheat, to research on the molecular mechanism of T type cytoplasmic male sterility and its fertility restoration in common wheat. In order to compare the difference of their genomes and find the molecular markers, which were closely linked to Rf3 gene, AFLP was used in this experiment.After the research on the single-nucleur stage and the meiosis stage, the soluble proteins were concerned at the sporule stage and the microsporocyte stage. The result of SDS-PAGE indicated: six polypeptides disappeared specially in the male sterile line and the polypeptide of 40.6KD expressed specifically in B type cytoplasm; according to the result of IEF-2D, three polypeptides expressed specially at the sporule stage and four polypeptides disappeared specifically at the microsporocyte stage in the male sterile line; the polypeptide of 20.8KD disappeared only in male sterile line; the polypeptides of 51.6KD and 56KD were detected only in 1031; the result of NEPHGE-2D displayed one different protein.The point of separate gene system about the mechanism of male sterility and restoration in developmental process was putted according to the result. It convinced that the proteins had been expressed seperatly by male sterile gene and fertile restorer. the obvious characteristic of expressing genes was timely and spacial, in timliness, the differences between the sporule stage and the microsporocyte stage in the same line were not many, but those at the same stage in different lines were more; in spacialiness, synthesizing the results from each stage, the differences of the nutritional organism(flag leaf) were not many ,those of reproductive organism(young spide) were more.Totally, 32 different bands were isolated by using 6 out of 16 primer combinations. Eight were labeled as probes for Southern blot to confirm the results and 3 of them were proved positive. By DNA squencing and Blast analysis, two DNA sequences have been identified. One of them was a new DNA sequence which had no homologous sequence and known conserved putative domain in GenBank. The other fragment of DNA codes a peptide which shares the same conserved domain with Hip1/Sla2 protein, indicating that male sterility had something to do with cell membrane remolding, apoptosis and energy matabolism. According to the sequence information, gene cloning and functional analysis would be performed. Therefore, this study provided a basic clue for Rf3 gene cloning and investigation into the mechanism of male sterility and restoration.NEPHGE gel electrophoresis was applied and improved firstly and the result of it was good enough in addition, the article posed the mind of the IPG gel electrophoresis and the three- dimensional gel electrophoresis.