水稻红莲型细胞质雄性不育系小孢子不同发育时期花药总蛋白质比较分析

A Comparative Analysis about Two-dimensional Gel Electrophoresis of Anther Total Proteins from Honglian Cytoplasmic Male Sterile Rice during the Different Developing Stages of the Microspores

采用双向凝胶电泳对水稻红莲型细胞质雄性不育的不育系小孢子发育单核期和二核期花药总蛋白进行了分离，通过银染显色，获得了分辨率和重复性较好的双向电泳图谱，且单核期和二核期花药总蛋白质在双向电泳胶上分布的图谱十分相似。PDQuest 2DE图像分析软件在等电点（pI）3．0—10．0、分子量（M．W．）9．0—98．0kD之间可识别约1800个蛋白质点。比较分析发现单核期和二核期花药中共有241个差异表达的蛋白质点，其中仅在单核期中表达的点数为125，仅在二核期中表达的为13点；表现为表达量差异的105点，其中在二核期表达下调的点数为70点，表达上调的为33点。还对蛋白质点集中的区域（p14．5—8．0，M．W．25．0—70．0kD）中的41个差异蛋白质点进行了分子量和等电点分析。

Cytoplasmic male sterility （CMS）, which eliminates the possibility of self-pollination, is a commonly observed phenomenon in the plant kingdom and has been exploited for hybrid seed production. Rice （ Oryza sativa L. ） is one of the most important food crops in which CMS has been used successfully for commercial hybrid seed production. Two dimensional gel electrophoresis technique was used to investigate the protein expression profile in uninucleate-stage and binucleate-stage anthers of the rice CMS line （YTA）. After silver staining and analysis by PDQuest 2DE software, about 1 800 protein spots, including 241 differentially accumulated spots, were detected reproducibly. Out of those differentially accumulated spots, 125 spots were specially accumulated in uninucleate-stage, 13 spots were specially accumulated in binucleate-stage, and 70 spots were down-regulated and 33 spots were up-regualted in binucleatestage. The results showed that proteins distribution of uninucleate-stage and binucleate-stage anthers were similar and the protein spots were mainly focusing on pI 4.5 - 8.0, and molecular weight ranging from 25.0 kD to 70.0 kD, and 41 protein spots in this area were analyzed.