红莲型细胞质雄性不育水稻单核期花粉蛋白差异表达分析

Differential Protein Expression Analyses of Pollens of Honglian CMS Rice at Uninucleate Stage

为在孢子发育时期从蛋白质水平更好地理解细胞质雄性不育的分子机理,对红莲型细胞质雄性不育水稻的不育系粤泰A、保持系粤泰B及其F1(红莲优6号)单核期花粉总蛋白质采用固相pH梯度等电聚焦/SDS-PAGE(3～10非线性胶条)进行双向电泳分离,利用基质辅助激光解析电离飞行时间质谱(MALDI-TOF/MS)或者LC-MS/MS分析对其中15个差异点进行鉴定,并对已鉴定的蛋白质点进行亚细胞定位和功能分析。发现与可育系相比,不育系有部分参与物质和能量代谢、细胞周期、转录、物质转运等的蛋白质缺失或表达量降低。这些蛋白质包括K+/H+协同运输蛋白、锌指蛋白和WD重复蛋白等。这些蛋白质的表达下调或缺失可能与线粒体供能不足而导致的花粉不能正常发育有关。

To better understand the molecular mechanism of cytoplasmic male sterility （CMS） at the protein level during the development of microspores, the protein expression profile was analyzed in pollens of the CMS rice line Yuetai A, its fertile maintainer line Yuetai B and their hybrid F1 （Honglianyou 6）. The proteins were separated by two dimensional electrophesis with immobilized pH （3-10 non-linear, NL） gradients as the first dimension and SDS-PAGE as the second, and 15 differentially expressed proteins were identified by Matrix assisted laserdesorption/ionizaton time of flight mass spectrometry （MALDI-TOF/MS） or liquid chromatography tandem mass spectrometry （LC-MS/MS） analyses. Among these proteins, some involved in substance and energy metabolism, cell cycle, transcription and cellular transporters etc. were down-regulated or absent in the male-sterile line, including a putative potassium/proton antiporter-like protein, a zinc knuckle domain like, a WD-repeat protein etc. It seemed to suggest that the downregulation or absence of these proteins in pollen might relate to the pollen sterility caused by the insufficient mitoehondrial energy.