Isoenzymes Analysese Of Cytoplasm Male Sterile Line Sensitived To Temperature In Chinese Cabbage

The male sterile line of Chinese cabbage with Brassica napus cytoplasm CMS3411-7,CMS7311 and their maintainers were used in this experiment, and the isozymes related with sterility changeover were analyzed in order to provide the necessary clues of the mechanisms of temperature sensitive male sterility and changeover of sterility. The POD, ATP, AMY, EST isozymes of leaves and roots in seedling stage with six leaves and that of buds in flowering stage were analyzed in the male sterile lines and their maintainers by using vertical slab polyacrylamide gel electrophoresis. Research on the sterile line CMS7311 dealt with about 9℃and 3000lux light in the whole press (from the sterile appearance changes into fertile appearance and the fertile appearance changes into the sterile appearance), in the period, the stamen sterility was observed and β-EST was analyzed continuously, in order to find out the change of β-EST accompanied by the sterility changeover. Additionally, the aimed zymograms bands were electrically transferred to PVDF membrane and it's N-Amino acids was analyzed, which is favorable to the further research in future. The main results of the experiment were shown as follows: 1. The EST isozymes in buds are bound up with the expression of the stamen sterility by the research on lots isozymes of the different organ in sterile line CMS3411-7, CMS7311. 2. Find out preliminarily that disappearing of the β-EST zymograms bands(Rf 0.718, 0.742) in Chinese Cabbage small buds(≤1.5mm) is bound up with the stamen sterility in CMS3411-7, CMS7311 . 3. Initially discover that the appearing of the β-EST zymogram band(Rf 0.759)is bound up with sterility changeover of sterile line sensitized to temperature. 4. Analyze of the SDS zymogram fined out that protein molecular weight of β-EST isozymes(Rf 0.718,0.742,0.759)are about 14KD. Initially speculate that the central enzyme subunits of all the EST are the same. 5.Established a correspond easy way of recovering the aimed isozymes bands in Chinese cabbage. The isozyme band can't be dyed before electroelution. After electrophoresis, cut the bands of the both slab sides and dye them, then replace them on their original place. Based on the aimed zymogram bands on the each side cut out the aimed zymogram bands in middle,　and then reclaimed by　electroelution.