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线粒体蛋白组解析烟草雄性不育分子机制
引用本文:柯渔洲,王平松,段丽丽,莫泽君,何轶,喻奇伟,熊晶,刘仁祥.线粒体蛋白组解析烟草雄性不育分子机制[J].核农学报,2022,36(8):1559-1568.
作者姓名:柯渔洲  王平松  段丽丽  莫泽君  何轶  喻奇伟  熊晶  刘仁祥
作者单位:1贵州大学烟草学院/贵州省烟草品质研究重点实验室,贵州 贵阳 5500252贵州省烟草公司毕节市公司,贵州 毕节 551700
基金项目:国家自然科学基金资助项目(32060510);贵州省科技厅资助项目([2016]5663);贵州省科技厅重点基金项目([2019]1405);贵州省烟草公司项目(201904)
摘    要:目前我国烟草杂交种种植越发广泛,不育系作为制备烟草杂交种的有利材料,在我国烟草种植中具有重要地位。为了探究烟草胞质雄性不育形成的分子机制,选用烟草不育系及其保持系,在花芽分化时期利用石蜡切片和线粒体蛋白组学技术结合生物信息学分析方法进行研究。结果表明,烟草雄性不育系的败育过程发生在发芽分化的雌雄蕊原基分化时期;蛋白组学分析共筛选出线粒体差异表达蛋白113个,呼吸代谢过程中的焦磷酸酶、异柠檬酸脱氢酶、苹果酸脱氢酶和磷酸己糖异构酶等关键调控蛋白酶的表达显著下调,ATP合酶的δ和α亚基表达上调;在线粒体蛋白的合成、修饰和运输过程中,核糖体RNA大亚基中L4e、L7和小亚基中SAe的表达下调,烯醇酶、内质网蛋白加工酶、蛋白二硫键异构酶等功能蛋白结构修饰酶表达下调,蛋白酶复合体的Rpt3和α5亚基表达下调。由上述结果推测,烟草胞质雄性不育由于线粒体蛋白的合成、修饰及导入过程受阻使线粒体功能紊乱,具体表现在线粒体呼吸代谢途径的蛋白酶表达下调及合成ATP受阻,不能为其在花粉形成时期小孢子的快速分裂分化提供充足的能量,抑制了小孢子的形成和发育,从而表现为雄性不育。本研究结果为进一步开展烟草雄性不育机理研究奠定了重要基础。

关 键 词:烟草  胞质雄性不育  线粒体蛋白组学  液质联用(LC-MS)  生物信息学  
收稿时间:2021-11-10

Mitochondrial Proteome Elucidates the Molecular Mechanism of Male Sterility in Tobacco
KE Yuzhou,WANG Pingsong,DUAN Lili,MO Zejun,HE Yi,YU Qiwei,XIONG Jing,LIU Renxiang.Mitochondrial Proteome Elucidates the Molecular Mechanism of Male Sterility in Tobacco[J].Acta Agriculturae Nucleatae Sinica,2022,36(8):1559-1568.
Authors:KE Yuzhou  WANG Pingsong  DUAN Lili  MO Zejun  HE Yi  YU Qiwei  XIONG Jing  LIU Renxiang
Institution:1College of Tobacco Science, Guizhou University/Guizhou Provincial Key Laboratory of Tobacco Quality Research, Guiyang, Guizhou 5500252Guizhou Province tobacco Company Bijie City Company, Bijie, Guizhou 551700
Abstract:At present, the planting area of tobacco hybrid and sterile line is more and more extensive in China, male sterile line is a favorable material for tobacco hybrid. However, the molecular mechanism of male sterility in tobacco is unclear. To investigate the molecular mechanism of cytoplasmic male sterility in tobacco,tobacco sterile lines and their maintainers were used in this study, paraffin sectioning and mitochondrial proteomics combined with bioinformatics analysis were conducted during flower bud differentiation. The results show that the abortive process of tobacco male sterile line occurs at the stage of stamen primordia germination and differentiation;A total of 113 differentially expressed mitochondrial proteins were screened out by proteomic analysis. During respiratory metabolism, the expression of key regulatory proteases such as pyrophosphatase, isocitrate dehydrogenase, malate dehydrogenase and phosphate hexose isomerase was down-regulated, and the expression of δ and α subunits of ATP synthase was abnormal;During the synthesis, modification, and transportation of mitochondrial proteins, abnormal expression of L4e and L7 in large subunit and SAe in small subunit of ribosomal RNA, and enzymes such as enolase, endoplasmic reticulum protein processing enzyme, protein disulfide isomerase and other functional protein structural modification enzymes were down-regulated.It is inferred from the above results that cytoplasmic male sterility of tobacco is caused by mitochondrial dysfunction due to the blocked synthesis, modification and import of mitochondrial proteins.Specifically, protease expression in mitochondrial respiratory metabolic pathway is down-regulated and ATP synthesis is blocked, which cannot provide sufficient energy for the rapid division and differentiation of microspores during pollen formation, and inhibited the formation and development of microspore, then displayzed as male sterility. The results of this study lay an important foundation for further research on male sterility mechanism.
Keywords:tobacco  cytoplasmic male sterility  mitochondrial proteomics  liquid chromatography-mass spectrometry(LC-MS)  bioinformatics  
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