The Exploration Of Proteins Related To Nuclear Male Sterility In Chinese Cabbage

In this research, the anthers from multiple-allele inherited male-sterility line in Chinese cabbage were used as the experiment material. The differentially expressed proteins were separated using the two-dimensional electrophoresis (2-DE) technology. The sequence and function of these proteins were identified using mass spectrometry techniques and biotechnology methods. The research provides some clues to reveal the male-sterility mechanism of Chinese cabbage. Results as following:1. The anthers from fertile and sterile Chinese cabbage were used to make the frozen sections. We can clearly find that the fertile pollen grains were normal growth and development, but the sterile pollen grains of the sterile buds were becoming abnormal gradually, and then becoming more and more dry and scarce from the secondary stage, so the anthers of the secondary stage were selected as the experimental materials.2. To optimize 2-DE of the anther proteins on sample quantity, we set up three grading, respectively 800ug、1000μg and 1200ug. The results show that the 10000μg protein loading quantity is appropriate to the study of high-resolution 2-DE maps.3. Anthers’2-DE mapping was analyzed using PDQuest8.0.1 software. Results showed that almost 350 protein spots were identified at pI 4.0-7.0 on average.38 differentially expressed proteins with 95% protein score were found,11 proteins were specially expressed in fertile anther, but not in sterile ones,14 differentially expressed proteins were up-regulated in fertile anther; 17 differentially expressed proteins were specially expressed in sterile anther, but not in fertile ones; 14 differentially expressed proteins were up-regulated in sterile ones.4.8 differentially expressed proteins from 2-DE mapping were selected to be analyzed by MS and proteins information was acquired. GO analysis shows that within 8 differentially expressed proteins,30% for cell component accounted,17% involved in molecular function, 53% for biological process account. In the biological processes, there were two pollen related protein, one is involved in pollen wall formation and another one is related to pollen tube growth.5. KEGG results showed that there was a protein participated in several pathways. It may be a critical protein related to nuclear male sterility.