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Differential proteomics analysis to identify proteins and pathways associated with male sterility of soybean using iTRAQ-based strategy

机译:使用基于iTRAQ的策略进行差异蛋白质组学分析,以鉴定与大豆雄性不育相关的蛋白质和途径

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摘要

To further elucidate the molecular mechanism of cytoplasmic male sterility (CMS) in soybean, a differential proteomic analysis was completed between the CMS line NJCMS1A and its maintainer NJCMS1B using iTRAQ-based strategy. As a result, 180 differential abundance proteins (DAPs) were identified, of which, 60 were down-regulated and 120 were up-regulated in NJCMS1A compared with NJCMS1B. Bioinformatic analysis showed that 167 DAPs were annotated in 41 Gene Ontology functional groups, 106 DAPs were classified into 20 clusters of orthologous groups of protein categories, and 128 DAPs were enrichment in 53 KEGG pathways. Fifteen differential level proteins/genes with the same expression pattern were identified in the further conjoint analysis of DAPs and the previously reported differential expression genes. Moreover, multiple reaction monitoring test, qRT-PCR analysis and enzyme activity assay validated that the iTRAQ results were reliable. Based on functional analysis of DAPs, we concluded that male sterility in NJCMS1A might be related to insufficiencies in energy supply, unbalance of protein synthesis and degradation, disruption of flavonoid synthesis, programmed cell death, abnormalities of substance metabolism, etc. These results might facilitate our understanding of the molecular mechanisms behind CMS in soybean.
机译:为了进一步阐明大豆细胞质雄性不育(CMS)的分子机制,使用基于iTRAQ的策略完成了CMS品系NJCMS1A及其维持者NJCMS1B之间的差异蛋白质组分析。结果,与NJCMS1B相比,在NJCMS1A中鉴定出180种差异丰度蛋白(DAP),其中60种被下调,而120种被上调。生物信息学分析表明,在41个基因本体论功能组中注释了167个DAP,将106个DAP分为20个直系同源蛋白类别的簇,并且在53条KEGG途径中富集了128个DAP。在DAP和先前报道的差异表达基因的进一步联合分析中,鉴定出十五种具有相同表达模式的差异水平蛋白质/基因。此外,多反应监测测试,qRT-PCR分析和酶活性测定验证了iTRAQ结果的可靠性。根据DAP的功能分析,我们得出结论,NJCMS1A中的雄性不育可能与能量供应不足,蛋白质合成和降解失衡,类黄酮合成破坏,程序性细胞死亡,物质代谢异常等有关。这些结果可能有助于我们对大豆CMS背后的分子机制的了解。

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