陕农138小孢子发育过程中蛋白表达差异的比较分析

Analysis of Protein Expression at Different Microspore Development Stages in Wheat Cultivar Shaannong 138

为了寻找小麦单核中晚期愈伤组织诱导率较高的原因,探讨小麦花药发育的蛋白质代谢机制,本试验利用双向电泳技术对陕农138小麦小孢子单核中晚期和双核期的全蛋白分析表明,在等电点4-7之间可识别约450个以上清晰的蛋白质点,检测到差异点26个,对其中14个质量较好、重复性较高的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（MALDI-TOF-MS）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11个蛋白质点,另3个蛋白质点未得到有意义的鉴定,11个蛋白质点分别被鉴定为UDP-葡萄糖焦磷酸化酶（1个蛋白点）、叶绿素抗体结合蛋白（1个蛋白点）、pentatricopeptide重复蛋白PPR566-6（1个蛋白点）、半胱氨酸蛋白酶抑制剂（1个蛋白点）、泛素（1个蛋白点）、S-腺苷-L-半胱氨酸水解酶（2个蛋白点）、放氧增强蛋白（2个蛋白点）和假定蛋白（2个蛋白点）。这些蛋白质点的功能涉及到糖代谢、蛋白质降解、细胞防卫等代谢过程。

To understand the metabolic mechanism of wheat anther development, we analyzed the holoprotein of Shaannong 138 at mid-late mononuclear pollen and binuclear pollen stages using two-dimensional electrophoresis technique. More than 450 clear protein spots were identified, including 26 differentially expressed protein spots. We selected 14 protein spots with expression difference at least two times for further separation by means of matrix-assisted laser desorption time of flight mass spectrometry peptide mass fingerprint analysis （MALDI-TOF-MS） in isoelectric point （pI） of 4–7. After indexing database NCBInr using soft-ware Mascot, we identified 11 significant protein spots. These protein spots were classified into several groups based on function, i.e., UDP-glucose pyrophosphorylase （one spot）, chlorophyll antibody-binding protein （one spot）, pentatricopeptide repeat protein PPR566-6 （one spot）, cysteine protease inhibitor （one spot）, ubiquitin （one spot）, S-adenosyl-L-cysteine hydrolase （two spots）, oxygen-enhanced proteins （two spots）, and putative proteins （two spots）. These proteins are involved in glycometabolism, protein degradation, cell defense, and some other metabolic processes.