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为了在蛋白质水平揭示小麦雄性不育的分子遗传机制,以小麦经杀雄剂SQ-1诱导的生理型雄性不育系,以及对应正常可育系所构建的等生理差异系为试材,应用差速离心和蔗糖密度梯度离心,首先分离出供试材料小花的完整叶绿体,制备蛋白样品后采用固相IEF/SDS-PAGE双向凝胶电泳技术对完整叶绿体蛋白质进行了分离、银染,得到了重复性较好的双向电泳图谱.PDQuest2DE软件分析识别出约150个较为清晰的蛋白质点,采用MALDI-TOF鉴定出的6个差异表达蛋白质分别是:PAP-fibrillin、ATRABB1A、底物同源结构域蛋白/RhoGAP结构域蛋白、铜锌超氧化物歧化酶(Cu/Zn-SOD)、R2R3-MYB转录因子及1个假定蛋白质.生物信息学功能分析暗示,这些蛋白直接参与了花药内激素调节、蛋白质转运、蛋白质互作、活性氧积累及花药的发育,表明SQ-1诱导的小麦生理型雄性不育其败育机理可能就与这些生理代谢过程的变异直接相关.
In order to reveal the molecular genetic mechanism of wheat male sterility at the protein level, the physiological male sterile lines induced by the antiserum SQ-1 and the physiological isolates constructed from the corresponding normal fertile lines were used as materials Differential centrifugation and sucrose density gradient centrifugation, the complete chloroplast was isolated from the floret of the test material and the whole chloroplast protein was separated and silver stained by solid-phase IEF / SDS-PAGE two-dimensional gel electrophoresis after silver protein preparation Repeatability of the two-dimensional electrophoresis.PDQuest2DE software analysis identified about 150 clear protein spots identified using MALDI-TOF six differentially expressed proteins are: PAP-fibrillin, ATRABB1A, the substrate homology domain Protein / RhoGAP domain protein, copper / zinc superoxide dismutase (Cu / Zn-SOD), R2R3-MYB transcription factor and a hypothetical protein.Analysis of bioinformatics indicated that these proteins were directly involved in the regulation of anther hormones, Protein transport, protein interaction, reactive oxygen species accumulation and anther development indicated that the abortion mechanism of SQ-1-induced physiological male sterile in wheat may be related to these physiological metabolism The process of mutation is directly related.