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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。
In order to find out the reason of high induction rate of middle and late callus of wheat single nucleus, and to explore the protein metabolism mechanism of wheat anther development, we analyzed the whole protein analysis of middle and late stage and double nuclei of Shannong 138 wheat microspore by two-dimensional electrophoresis The results showed that more than 450 clear protein spots were identified between isoelectric points 4 and 7, and 26 differential spots were detected. Among 14 differentially expressed protein spots with good quality and high reproducibility, matrix-assisted Peptide fingerprinting was performed by laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF-MS). In the NCBInr database search using Mascot software, 11 protein spots were identified and the other three protein spots were not identified. 11 proteins The spots were identified as UDP-glucose pyrophosphorylase (1 protein spot), chlorophyll antibody binding protein (1 protein spot), pentatricopeptide repeat protein PPR566-6 (1 protein spot), cysteine protease inhibitor (1 protein spot), ubiquitin (1 protein spot), S-adenosyl-L-cysteine hydrolase (2 protein spots), oxygen boosting protein (2 protein spots) and putative protein 2 protein spots). The function of these protein spots involves the metabolism of sugar, protein degradation, cell defense and other metabolic processes.