普通小麦具有偏凸山羊草(Ae.ventricosa)细胞质的不育系为 Ven 型胞质雄性不育系(Ven-cytoplasmic male sterility,Ven-CMS),是粘类小麦CMS的一种类型。该研究对小麦 Ven型雄性不育系冀5418A及其同型保持系冀5419B的单核期和二核期的花药进行差异蛋白质组学分析,探讨小麦质核互作雄性不育的分子机制。通过双向电泳分离花药蛋白,基质辅助激光解析飞行时间串联质谱(MALDI-TOF-TOF)对差异表达蛋白进行质谱鉴定,利用生物信息学进行差异表达蛋白鉴定和功能注释分析。结果表明,在分子量19.0~100.0 kD、等电点4~7线性范围内,共检测到约2000个蛋白点。2个时期共检测到差异蛋白98个,其中两个时期差异表达变化一致的蛋白点56个；数据库搜索获得鉴定的蛋白点41个,其中18个蛋白的表达量在冀5418A 中显著下调,23个在冀5418B 中明显下调。在不育系和可育系中均有参与能量代谢、活性氧代谢、核糖体合成、花粉物质合成的差异蛋白。GO分析预测差异蛋白生物学过程多涉及电子传递和能量代谢、核糖体代谢、活性氧代谢等,细胞组成主要是在膜区域和线粒体,分子功能主要是DNA和 RNA结合功能和水解酶等。KEGG分析表明,较多蛋白分布于碳水化合物代谢、活性氧代谢和蛋白组装和折叠途径。推测不育系冀5418A 的雄性不育性除了涉及能量代谢、活性氧清除过程,核糖体蛋白、伴侣蛋白等也有重要作用,雄性不育性可能还与蛋白质加工、物质合成过程的紊乱有关。
Comparative Proteomics Analysis of Anther Proteins between Ven Cytoplasmic Male Sterile and Fertile Plants on the Early Stages of Pollen Development in Wheat
The sterile line of common wheat (Triticum aestivum)with the cytoplasm of Ae.ventricosa is Ven cytoplastmic male sterile (Ven CMS),and it is one type of Sv CMS in wheat.To better understand the molecular mechanism on nucleus-cytoplasmic interaction male sterility in wheat,we analyzed differenti-al proteomics at uninucleate and binucleate stages in sterile line JI5418A and its maintainer line JI5418B. Another proteins were separated by two dimensional electrophoresis.Differential protein spots were identi-fied by matrix-assisted laser desorption/ionization tandem time-of-flight mass spectrometry (MALDI-TOF-TOF).Identification and function annotation and analysis of these proteins were analyzed by bioin-formatics methods.About 2000 reproducibly protein spots could be visualized on these 2D-gels within M 19.0-100.0 kD and pI 4-7.Ninety eight differentially expressed proteins were found between JI5418A and JI5418B,and among these proteins 56 showed same differences at both stages.Forty one proteins were successfully identified in NCBI database.Eighteen and 23 proteins were significantly down-regulated expressed in JI5418A,and JI5418B,respectively.The differentially expressed proteins involving in energy metabolism,active oxygen scavenge,ribosome synthesis and pollen wall formation existed in both sterile and fertile lines.GO analysis predicted many differential expressed proteins involved in electron transport and energy metabolism,ribosome metabolism,and active oxygen metabolism,etc.in biological process, distributed in membrane region and mitochondria in cellular component,DNA and RNA binding function and hydrolytic enzymes in molecular function.KEGG pathways analysis indicated that more proteins in-volved in carbohydrate metabolism,active oxygen metabolism and protein assembly and folding than other pass ways.It is speculated that male sterility in JI5418B might be related to energy metabolism,active ox-ygen scavenging process.Ribosomal protein and chaperone protein also have great roles in controlling its fertility.Male sterility in Ven CMS might be related to the regulation disorders in protein processing and material synthesis.
Triticumaestivumcytoplasmic nuclear male sterilitydifferential proteomicstwo-dimension-al gel electrophoresisMALDI-TOF-TOF
NETL
NSTL
万方数据
维普
