水稻二核期花药蛋白质组差异与花粉败育的相关性

Differences in expression of proteins from the binucleate-staged anthers of rice and its relationship with pollen abortion

对红莲型细胞质雄性不育水稻小孢子发育二核期花药总蛋白质进行双向电泳分离,通过考马斯亮蓝染色,获得分辨率和重复性较好的双向电泳图谱。采用基质辅助激光解析电离飞行时间质谱鉴定12个蛋白质点,液相色谱质谱/质谱联用技术鉴定另外7个蛋白质点,这些蛋白质分别参与糖类物质的合成及代谢(5个点)、蛋白质生物合成及代谢(3个点)、转录(2个点)、次生代谢产物合成(2个点)、信号转导(1个点)、细胞疾病和死亡(1个点)及未知功能蛋白(5个点)。除1个蛋白酶亚基(点9)仅在不育系中表达外,其他18个蛋白质点在不育系中缺失或表达量降低,因而花粉发育过程中糖类及蛋白质的正常积累受到影响,与花粉不能正常发育密切相关。

Total proteins expressed in binucleate stage anthers of Honglian rice CMS line during the development of microspores were separated by two-dimensional electrophoresis, through coomassie blue staining, two dimensional gel electrophoresis profiles with high resolution and reproducibility were obtained. Twelve protein spots were identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF/MS） and 7 were identified by liquid chromatography coupled with tandem mass spectrometry （LC-MS/MS）. Among these protein spots, 5 spots were involved in biosynthesis or metabolism of carbonhydroxydate, 3 spots involved in biosynthesis, 5 spots involved in transcription, 2 spots involved in biosynthesis of plant secondary metabolites, 2 spots involved signal transduction, 2 spots involved cell death/disease, and 5 spots with unknown function. Among these 19 protein spots, the expression of 18 spots were down-regulated or absent in the male-sterile line, only one spot, standing for a proteasome subunit （spot 9） was regularly expressed, indicating the abnormal accumulation of carbonhydroxydates and proteins in male-sterile line anthers during the development of pollens might relate to pollen sterility.