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

文李1, 3,*,刘盖2,王坤2 ,谭碧君1 ,陶钧1

doi:10.3969/j.issn.10017216.2012.05.003

中国水稻科学 >

2012 , Vol. 26 >Issue 5: 529 - 536

DOI: https://doi.org/10.3969/j.issn.10017216.2012.05.003

研究报告

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

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¹长沙理工大学化学与生物工程学院，湖南长沙 410114；² 武汉大学生命科学学院植物发育学教育部重点实验室，湖北武汉 430072；³ 湖南农业大学国家油料作物研究所，湖南长沙 410128；

收稿日期: 2012-01-09

修回日期: 2012-02-04

网络出版日期: 2012-09-10

基金资助

中国博士后基金面上重点项目（20110490147）；湖南省科技厅项目（2011RS4007）；湖南省教育厅项目（07C069）。

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

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¹ Chemical and Biological Engineering College, Changsha University of Science and Technology, Changsha 410114, China; ²Key Laboratory of MOE for Plant Developmental Biology, College of Life Science, Wuhan University, Wuhan 430072, China; ³Institute of Oilseed Crops, Hunan Agricultural University, Changsha 410128, China;

Received date: 2012-01-09

Revised date: 2012-02-04

Online published: 2012-09-10

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摘要

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。

关键词： 水稻; 双向凝胶电泳; 蛋白质组学分析

本文引用格式

文李1, 3,*,刘盖2,王坤2 ,谭碧君1 ,陶钧1 . 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[J]. 中国水稻科学, 2012 , 26(5) : 529 -536 . DOI: 10.3969/j.issn.10017216.2012.05.003

Abstract

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 downregulation or absence of these proteins in pollen might relate to the pollen sterility caused by the insufficient mitochondrial energy.

Key words： Oryza sativa; two dimensional gel electrophoresis; proteomic analysis

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