College of Agronomy, Northwest A&F University, Yangling 712100, Shaanxi, China.
Int J Mol Sci. 2018 Jan 23;19(2):324. doi: 10.3390/ijms19020324.
Cytoplasmic male sterility (CMS) where no functional pollen is produced has important roles in wheat breeding. The anther is a unique organ for male gametogenesis and its abnormal development can cause male sterility. However, the mechanisms and regulatory networks related to plant male sterility are poorly understood. In this study, we conducted comparative analyses using isobaric tags for relative and absolute quantification (iTRAQ) of the pollen proteins in a CMS line and its wheat maintainer. Differentially abundant proteins (DAPs) were analyzed based on Gene Ontology classifications, metabolic pathways and transcriptional regulation networks using Blast2GO. We identified 5570 proteins based on 23,277 peptides, which matched with 73,688 spectra, including proteins in key pathways such as glyceraldehyde-3-phosphate dehydrogenase, pyruvate kinase and 6-phosphofructokinase 1 in the glycolysis pathway, isocitrate dehydrogenase and citrate synthase in the tricarboxylic acid cycle and nicotinamide adenine dinucleotide (NADH)-dehydrogenase and adenosine-triphosphate (ATP) synthases in the oxidative phosphorylation pathway. These proteins may comprise a network that regulates male sterility in wheat. Quantitative real time polymerase chain reaction (qRT-PCR) analysis, ATP assays and total sugar assays validated the iTRAQ results. These DAPs could be associated with abnormal pollen grain formation and male sterility. Our findings provide insights into the molecular mechanism related to male sterility in wheat.
细胞质雄性不育(CMS)中花粉不能正常产生,在小麦育种中具有重要作用。花药是雄性配子体发生的独特器官,其发育异常可导致雄性不育。然而,与植物雄性不育相关的机制和调控网络仍知之甚少。在这项研究中,我们使用相对和绝对定量同位素标记(iTRAQ)技术对 CMS 系及其小麦保持系的花粉蛋白进行了比较分析。根据基因本体论分类、代谢途径和转录调控网络,利用 Blast2GO 对差异丰度蛋白(DAP)进行了分析。我们基于 23277 个肽段鉴定了 5570 种蛋白质,与 73688 个光谱相匹配,其中包括糖酵解途径中的甘油醛-3-磷酸脱氢酶、丙酮酸激酶和 6-磷酸果糖激酶 1、三羧酸循环中的异柠檬酸脱氢酶和柠檬酸合酶以及氧化磷酸化途径中的烟酰胺腺嘌呤二核苷酸(NADH)脱氢酶和腺苷三磷酸(ATP)合酶等关键途径中的蛋白质。这些蛋白质可能构成了一个调节小麦雄性不育的网络。实时定量聚合酶链反应(qRT-PCR)分析、ATP 测定和总糖测定验证了 iTRAQ 结果。这些 DAPs 可能与花粉粒异常形成和雄性不育有关。我们的研究结果为小麦雄性不育相关的分子机制提供了新的见解。