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对过量表达AGG3基因的荠蓝种子进行定量蛋白质组学分析,以确定产量增加和胁迫耐受性增强的蛋白质组学基础。

Quantitative Proteomics Analysis of Camelina sativa Seeds Overexpressing the AGG3 Gene to Identify the Proteomic Basis of Increased Yield and Stress Tolerance.

作者信息

Alvarez Sophie, Roy Choudhury Swarup, Sivagnanam Kumaran, Hicks Leslie M, Pandey Sona

机构信息

†Donald Danforth Plant Science Center, 975 North Warson Road, St. Louis, Missouri 63132, United States.

‡Department of Chemistry, University of North Carolina, Chapel Hill, North Carolina 27599, United States.

出版信息

J Proteome Res. 2015 Jun 5;14(6):2606-16. doi: 10.1021/acs.jproteome.5b00150. Epub 2015 May 19.

Abstract

Camelina sativa, a close relative of Arabidopsis, is an oilseed plant that is emerging as an important biofuel resource. The genome and transcriptome maps of Camelina have become available recently, but its proteome composition remained unexplored. A labeling LC-based quantitative proteomics approach was applied to decipher the Camelina seed proteome, which led to the identification of 1532 proteins. In addition, the effect of overexpression of the Arabidopsis G-protein γ subunit 3 (AGG3) on the Camelina seed proteome was elucidated to identify the proteomic basis of its increased seed size and improved stress tolerance. The comparative analysis showed a significantly higher expression of proteins involved in primary and secondary metabolism, nucleic acid and protein metabolism, and abscisic acid related responses, corroborating the physiological effects of AGG3 overexpression. More importantly, the proteomic data suggested involvement of the AGG3 protein in the regulation of oxidative stress and heavy metal stress tolerance. These observations were confirmed by the physiological and biochemical characterization of AGG3-overexpressing seeds, which exhibit a higher tolerance to exogenous cadmium in a glutathione-dependent manner. The activity of multiple redox-regulating enzymes is higher in seeds expressing enhanced levels of AGG3. Overall, these data provide critical evidence for the role of redox regulation by the AGG3 protein in mediating important seed-related traits.

摘要

亚麻荠是拟南芥的近亲,是一种正在成为重要生物燃料资源的油料作物。亚麻荠的基因组和转录组图谱最近已经公布,但其蛋白质组组成仍未得到探索。应用基于标记液相色谱的定量蛋白质组学方法来解析亚麻荠种子蛋白质组,从而鉴定出了1532种蛋白质。此外,还阐明了拟南芥G蛋白γ亚基3(AGG3)过表达对亚麻荠种子蛋白质组的影响,以确定其种子大小增加和胁迫耐受性提高的蛋白质组学基础。比较分析表明,参与初级和次级代谢、核酸和蛋白质代谢以及脱落酸相关反应的蛋白质表达显著更高,这证实了AGG3过表达的生理效应。更重要的是,蛋白质组学数据表明AGG3蛋白参与了氧化应激和重金属胁迫耐受性的调节。这些观察结果通过过表达AGG3的种子的生理和生化特性得到了证实,这些种子以谷胱甘肽依赖性方式对外源镉表现出更高的耐受性。在表达增强水平AGG3的种子中,多种氧化还原调节酶的活性更高。总体而言,这些数据为AGG3蛋白的氧化还原调节在介导重要种子相关性状中的作用提供了关键证据。

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