| The seed’s oil content as a biological trait indicator, is the main economics value of the rape.lipid often store in the oil body in the form of TAG.Enhancing the oil content in response to the situation of our country’s oil crops in short supply is of great significance. The molecular mechanism of the forming of oil content is still insufficient understanding. Proteomics is an effective technology for the research of seed oil content character. This paper first established a suitable system for the observation of oil bodies, and analyzed the morphological characteristics of oil bodies in the Brassica napus seed with higher and lower oil content. At the same time the research set up a two-dimensional electrophoresis method for Brassica napus seed proteins. Then utilizing the established method,we researched the two-dimensional electrophoresis for oil body proteins in the Brassica napus seed with higher and lower oil content. And further making use of the software, analyzed the differences between the proteins of Brassica napus seed with higher and lower oil content and the differences between the oil body proteins of Brassica napus seed with higher and lower oil content,which provides a theoretical basis for revealing the molecular mechanism of forming the high oil content,so is of certain value. The main results are as follows:(1) Establishied a suitable system for observation of oil bodies. The system contained Paraffin section technique,the fluorescent dye Nile red staining and fluorescence microscopy observation, which formed a feasible method to observe and analyze the oil bodies’morphology of Brassica napus seed. Paraffin sections of the seed produced after the use of K2Cr2O7solution fixative processing can be more complete slice of the cell morphology.(2) Using fluorescence microscopic technology for the rape seed oil body cells, histologic observation.The result of the observation under ordinary light:the cell inclusion of the Brassica napus seed section with higher oil content tightly packed with no gap,which may indicating that the Brassica napus seed with higher oil content has ample storaged material. The cell inclusion of the Brassica napus seed section with lower oil content was less,scattered distribution in the cell. The result of the observation under mercury lamp:the cells of the section of the Brassica napus seed with higher oil content had more oil bodies,which emit red fluorescence after Nile red staining and green light excitation,bright and clear.And the cells of the section of the Brassica napus seed with lower oil content had less oil bodies which emitted darker red fluorescence,and the whole section’s fluorescence was weaker. After microscopic form measurement analysis, there are statistically significant differences in the mean optical density value of unit area between the the Brassica napus seed with higher and the lower oil content. So this experimental method can be used as a method of identifying the level of rapeseed’s oil content.(3) Established and optimized two-dimensional electrophoresis system based on the homemade tubular gel.We separated the total the proteins in the higher and lower oil content seed of the Brassica napus,and got their protein expression map. After the anlysis by the software PDQuest8.01, there were12differentially expressed proteins between the higher and the lower oil content seed.(4) Established and optimized the suitable oil body extraction method, and we got the oil body proteins in the higher and lower oil content seed of the Brassica napus utilizing the method. Using the above-mentioned established system for two-dimensional electrophoresis,we separated oil body proteins in the higher and lower oil content seed of the Brassica napus and got their protein expression map. After the anlysis by the software PDQuest8.01, there were16differentially expressed oil body proteins between the higher and the lower oil content seed. |
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