Capillary Electrophoresis Separation Of Proteins Of Seeds And Needles Of Chinese Pine

Chinese pine is the main tree species in northern China, the wood is resin-rich, anticorrosive and suitable for construction, furniture, sleepers, pillar, pole, man-made fibers. Resin can also be adopted for industrial use. The research of its development mechanism has important significance.Capillary electrophoresis separation as a relatively new technology for its high throughput, high sensitivity, fast low-power, high auto-control of the advantages maked up for the deficiencies of traditional two-dimensional gel electrophoresis. Following the existing research results, in this article the six standard proteins were used to further optimize the conditions for protein separation by capillary electrophoresis. Results showed that the following conditions can be better in six standard protein separation:50μm diameter silica capillary column, length 50cm/58cm (effective length/column length), 30kV separation voltage,25℃electrophoresis temperature,50mbar inject sample 5seconds at 50mbar, electrophoresis buffer 50mmol, pH 2.5 phosphate buffer containing 0.05% hydroxyethyl cellulose, detection wavelength 191nm.While explored the conditions for capillary electrophoresis separation of pine seeds during germination protein, we found that protein extraction methods played great role in the separation. Compared of three kinds of method after repeated experiments a more suitable method for protein extraction of pine seeds was determined:TCA-acetone. This method overcomed the problem of low extraction mass of plant tissue protein, retained many low abundance proteins, was more suitable for sample preparation for capillary electrophoresis.Proteins were separated from dry seeds, seeds 3d after germinating and seeds 6d after germinating using capillary electrophoresis. The results showed that the technology could detect changes in the protein with the seed germination. From the capillary electrophoresis profiles big difference between the protein components could be seen between dry seeds, seeds 3d after germinating, seeds 6d after germinating.1# protein with the retention time 6min in the dry seeds down-regulated when 3d after germinating, and restored when 6d after germinating.7#,8#,9# proteins in the dry seeds down-regulated during the germination, until disappeared when 6d after germinating.14#,18#,19# proteins in the dry seeds d down-regulated during the germination, but still expressed when 6d after germinating.26# protein did not down-regulated when 3d after germinating, but disappeared when 6d after germinating.21#,22#,23# proteins which were low abundance did not appeared in the dry seeds, but were produced while germinating.The method was used to separate proteins of the female fertile and female sterile pine needles. The results showed that this technique can separate proteins of pine needles, but the results of the stability and resolution of the separation needed to be improved. Because the protein extraction conditions were optimized for prior to seeds, to continue to study this problem, the re-optimize of the protein extraction conditions for the needles is needed. Or two-dimensional electrophoresis analysis can be used, as a more detailed analysis to identify differences in protein. Further optimized the conditions, if there was still no improvement, then infer that the different nature of female fertility did not perform in the pine needles, the female reproductive organs should be specific variation in sterile plants.