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基于iTRAQ技术的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通路

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

作者信息

Li Jiajia, Ding Xianlong, Han Shaohuai, He Tingting, Zhang Hao, Yang Longshu, Yang Shouping, Gai Junyi

机构信息

Soybean Research Institute, National Center for Soybean Improvement, MOA Key Laboratory of Biology and Genetic Improvement of Soybean (General), State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing, Jiangsu 210095, China.

出版信息

J Proteomics. 2016 Apr 14;138:72-82. doi: 10.1016/j.jprot.2016.02.017. Epub 2016 Feb 26.

Abstract

UNLABELLED

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.

BIOLOGICAL SIGNIFICANCE

Soybean is an important global crop that provides protein and oil. Heterosis is a significantly potential approach to increase the yield of soybean. Cytoplasmic male sterility (CMS) plays a vital role in the production of hybrid seeds. However, the genetic and molecular mechanisms of male sterility in soybean still need to be further elucidated. In the present paper, a differential proteomic analysis was carried out and the results showed that several key proteins involved in key pathways were associated with male sterility in soybean. This work provides a new insight to understand the genetic and molecular mechanisms underlying CMS in soybean.

摘要

未标记

为进一步阐明大豆细胞质雄性不育(CMS)的分子机制,采用基于iTRAQ的策略,对CMS系NJCMS1A及其保持系NJCMS1B进行了差异蛋白质组学分析。结果,共鉴定出180个差异丰度蛋白(DAP),其中与NJCMS1B相比,NJCMS1A中有60个下调,120个上调。生物信息学分析表明,167个DAP注释于41个基因本体功能组中,106个DAP分类到20个蛋白质直系同源簇类别中,128个DAP富集于53条KEGG通路中。在DAP与先前报道的差异表达基因的进一步联合分析中,鉴定出15个具有相同表达模式的差异水平蛋白/基因。此外,多反应监测试验、qRT-PCR分析和酶活性测定验证了iTRAQ结果的可靠性。基于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、蛋白质合成与降解失衡、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理解大豆CMS背后的分子机制。

生物学意义

大豆是一种重要的全球性作物,可提供蛋白质和油脂。杂种优势是提高大豆产量的一种极具潜力的方法。细胞质雄性不育(CMS)在杂交种子生产中起着至关重要的作用。然而,大豆雄性不育的遗传和分子机制仍需进一步阐明。在本文中,进行了差异蛋白质组学分析,结果表明参与关键途径的几个关键蛋白与大豆雄性不育有关。这项工作为理解大豆CMS的遗传和分子机制提供了新的见解。

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