小麦质核互作型雄性不育系及其保持系花药差异蛋白质组学分析  

Differential Proteomic Analysis of Anther Proteins Between Cytoplasmic-Nuclear Male Sterility Line and Its Maintainer in Wheat(Triticum aestivum L.)

作　　者：陈蕊红[1] 叶景秀[1] 张改生[1] 王俊生[1] 牛娜[1] 马守才[1] 赵继新[1] 朱建楚[1] 

机构地区：[1]西北农林科技大学,陕西省作物杂种优势研究与利用重点实验室/小麦育种教育部工程研究中心,杨凌712100

出　　处：《生物化学与生物物理进展》2009年第4期431-440,共10页Progress In Biochemistry and Biophysics

基　　金：国家自然科学基金(301705760);国家高技术研究发展计划(863)(2002AA207004);陕西省“13115”科技创新工程重大科技专项(2007ZDKG-020);国家杨凌农业生物技术育种中心专项基金(99-1A);西北农林科技大学拔尖人才支持计划项目资助项目~~

摘　　要：为了能从蛋白质水平揭示小麦细胞质雄性不育的分子遗传机制,采用IEF/SDS-PAGE双向凝胶电泳技术,对小麦质核互作型雄性不育系(S)-1376A及其保持系(A)-1376B在花药发育的单核期、二核期蛋白质进行了差异蛋白质组学研究,经考马斯亮蓝染色,得到了重复性较好的双向电泳图谱.PDQuest软件在分子质量9.0～100.0ku、等电点4～7线性范围内,可识别约610个蛋白质点,对28个差异表达的蛋白质点采用基质辅助激光解吸分离飞行时间质谱进行肽指纹图谱分析,并利用Mascot软件在NCBInr数据库搜索,鉴定出12个差异表达蛋白,其中5个差异表达蛋白可能与雄性不育有关,分别是泛素结合酶E2、甘氨酸富集蛋白、抗坏血酸过氧化物酶、假定半胱氨酸蛋白酶抑制剂及1,5-二磷酸核酮糖羧化酶/加氧酶小链克隆512,它们参与了物质能量代谢、细胞程序化死亡及花发育调控等过程,推测不育系(S)-1376A雄性不育性可能与这些生理生化代谢有关.研究结果为揭示雄性不育机理提供了理论依据.Cytoplasmic male sterility （CMS） is the maternally inherited inability to produce functional pollen in a range of crop plants and has been widely used for the production of hybrid seed in plant breeding .To better under-stand the molecular mechanism on protein level and find the crucial proteins which related to fertillity, total anther proteins extracted from uninucleate and binucleate stage of （S）-1376A and （A）-1376B were separated by two- dimensional electrophoresis, the coomassie brilliant blue stained protein spots were analyzed using PDQuest software, among 610 reproducibly protein spots could be visualized on each 2D-gel within M 9.0 - 100.0 ku and pI 4 -7. A total of 28 differentially expressed proteins were identified by matrix-assisted laser desorption ionization time of flight mass spectrometry （MALDI-TOF-MS）, 5 of them were identified following NCBInr database queries, which are ubiqutin-protein ligase E2, putative glycine-rich protein, ascorbate peroxidase, putative cysteine proteinase inhibitor, ribulose bisphosphate carboxylase small chain clone 512. These proteins were involved in the process of energy metabolism, programmed cell death （PCD）, regulating flower development.These results provide a clue for elucidating the mechanism of CMS.

关 键 词：小麦 质核互作型雄性不育 差异蛋白质组学 双向凝胶电泳 基质辅助激光解吸飞行时间质谱 

分 类 号：Q946.1[生物学—植物学] Q51