| Plants suffer from various of biotic and abiotic stress during the grow process. Cold is one of the severe phenomena. Almost all plants undergo chilly stress.It restricts severely the distribution of the plants and affects yield. Every year,crop losing attains to hundreds of billion because of low temperature in the world. In China, the area that endures low temperature reaches to two million km2 each year. So it is crucial and valuable to improve the quality of cold resistance of the plants.1.In this paper, a series of biochemical and physiological indexes, such as electrolyte leakage, superoxidedimutase (SOD) activity, catalase (CAT) activity, soluble protein contents and malondialdehyde (MDA) contents,in the roots of 2 winter turnip rape (Brassica campetris) were examined in order to see the relationship between osmoticumand cold resistance of Brassica. The results showed that cold resistance of DQW-1 was stronger than thatof Tianyou 4, The SOD and CAT activities, soluble protein content and MDA content presented up-down trend withthe decrease of temperature. DQW-1 with stronger cold resistance under low temperature showed higher SOD andCAT activities, higher solutble protein content, and lower MDA content. Therefore, the activities of SOD and CAT,soluble protein content andMDA content maybe reflect the cold resistant of winter turnip rape in some degree.2. For establishing a technique method of 2-DE which is applied in oilseeding .The winter turnip has been used as tested materials to study on the discrepancy expression of proteome by the method of two-dimensional gel electrophoresis.Some important steps, such as sample preparation, the definition of protein quantity, isoelectric focusing and staining, were discussed and improved. The conclusion is as follow:Comparing the two methods of proteins extraction, the results demonstrate that the purified total protein by using acetone precipitation method has less impurities, two-dimensional electrophoresis images are more clear and it has high concentrations of protein solution. For 1D SDS-PAGE, coomassie brilliant blue staining on the protein quantity is about 35μg -45μg. For 17cm strip with a linear gradient of pH3-10, the protein quantity for coomassie brilliant blue staining method is about 900μg. Adopting this protein quantity, we can get 2-DE gels with fine focusing effect and limpid background. The method of Coomassie Brilliant Blue G250 staining. Before staining, fixed the profile of the gel with 12% TCA. the profile of gel didn't change the area , it will help software analysis .3. The discrepancy expression of oilseeding proteins that under cold-stress condition in 12h, with the improved method of two-dimensional gel electrophoresis, profiling These protein spots were analyzed, the result indicated 56 spots change under cold stress condition. 34 proteins were up-regulated, and 22 proteins were down-regulated. The study initially establish a technique method of 2-DE which is applied in oilseeding. This study gives us new insights into cold stress response in oilseeding and provide a reference for the proteomic approaches in oilseeding studies. |
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