| Chinese jujube is the No. 1 dry fruit tree in our country. Its blossom is very small which makes emasculation very difficult and hybridization less fruitful. Using male-sterility gcrmplasms can save the artificial emasculation and help cross pollination. To study the sterility-related proteins of those male sterility germplasms not only can reveal the mechanism of male sterility in Chinese jujube, but also comprehend the pollen development, cytoplasmic inheritance and nucleo-cytoplasmic interaction, and provide important references to artificial induction of male sterility.Based on the method of O'Farrcll, an improved method of two-dimensional gel electrophoresis applied in Chinese jujube was established through improving the preparation of sample, isoclcctric focusing, balance and other important steps. We adopted the improved two-dimensional gel electrophoresis to analyze the protein expressions of different development phases of bud from three sterility germplasms JMSI and JMS3, as well as two kinds of fertile materials LS and JS, and found the sterility-related proteins. The main results arc as follows:1. Wc constructed two kinds of protein-extracting methods which suitable to Chinese jujube. The protein amounts obtained by the two methods were not significant different. The protein amounts of leaf and phloem obtained by soluble protein extracting method (1.89mg/g and 1.11mg/g) were higher than that of total protein extracting method (1.80mg/g and 1.11mg/g), but thc protein amount of bud obtained by total protein extracting method (8.75mg/g) was higher than that of soluble protein extracting method (7.68mg/g).2. We optimized the important steps of two-dimensional gel electrophoresis, and set up a kind of platform which suitable to Chinese jujube: the best spotting amount was 50ul (about 220ug); the 2.5% of ampholyte and pH3.5～9: pH4～6:pH6～9=1:1:3 were more suitable to woody plant; the best balance time was 30min～1h; to change the "vertical pouting method" to "horizontal pouring method" could greatly reduce the operating difficulty; to cancelled the translational prefocusing and extended the focusing time appropriately (reached 10 000V-h) could obtained better result.3. After silver staining, a high distinct and steady electrophoretograms of different tissues (leaf, bud and phloem) were obtained.4. We adopted the improved two-dimensional gel electrophoresis to analyze the protein expressions of different development phases of bud from three sterility germplasms JMS1 and JMS3, as well as two kinds of fertile materials LS and JS. The results showed that: from small bud to middle bud to yellow bud, the protein expressing tendencies of LS and JS were high-low-high; the protein expressing tendency of LMS1 was high-low-low; the protein expressing tendency of LS was low-high-low.5. In JMS1 of sterility stage, there were 6 obvious spots of specific expression and 28 spots of hindered expression in JMS1 during the sterility stage, in which the 6 specific spots all distributed alkaline area of pH8.0～pH8.2 (100%), 6 sports in the 28 hindered spots distributed acid area of pH4.0～pH5.0 (21.4%), 18 spots distributed subacid area of pH5.0～pH7.0 (64.3%), 4 spots distributed alkaline area ofpH7.75～pH8.2(14.3%); in JMS3 of sterility stage, there were 6 specific alkaline spots which were same to JMS1 and 15 hindered spots which were also same to JMS1, in which 3 sports in the 15 hindered spots distributed acid area of pH4.0～pH5.0 (20.0%), 10 spots distributed subaeid area of pH5.0～pH7.0 (66.7%), 2 spots distributed alkaline area ofpH7.75～pH8.2(13.3%).
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