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Quantitative analysis of proteome extracted from barley crowns grown under different drought conditions

机译:不同干旱条件下从大麦冠上提取的蛋白质组学定量分析

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摘要

Barley cultivar Amulet was used to study the quantitative proteome changes through different drought conditions utilizing two-dimensional difference gel electrophoresis (2D-DIGE). Plants were cultivated for 10 days under different drought conditions. To obtain control and differentially drought-treated plants, the soil water content was kept at 65, 35, and 30% of soil water capacity (SWC), respectively. Osmotic potential, water saturation deficit, 13C discrimination, and dehydrin accumulation were monitored during sampling of the crowns for proteome analysis. Analysis of the 2D-DIGE gels revealed 105 differentially abundant spots; most were differentially abundant between the controls and drought-treated plants, and 25 spots displayed changes between both drought conditions. Seventy-six protein spots were successfully identified by tandem mass spectrometry. The most frequent functional categories of the identified proteins can be put into the groups of: stress-associated proteins, amino acid metabolism, carbohydrate metabolism, as well as DNA and RNA regulation and processing. Their possible role in the response of barley to drought stress is discussed. Our study has shown that under drought conditions barley cv. Amulet decreased its growth and developmental rates, displayed a shift from aerobic to anaerobic metabolism, and exhibited increased levels of several protective proteins. Comparison of the two drought treatments revealed plant acclimation to milder drought (35% SWC); but plant damage under more severe drought treatment (30% SWC). The results obtained revealed that cv. Amulet is sensitive to drought stress. Additionally, four spots revealing a continuous and significant increase with decreasing SWC (UDP-glucose 6-dehydrogenase, glutathione peroxidase, and two non-identified) could be good candidates for testing of their protein phenotyping capacity together with proteins that were significantly distinguished in both drought treatments.
机译:使用大麦品种Amulet,通过二维差异凝胶电泳(2D-DIGE)研究了在不同干旱条件下的蛋白质组定量变化。在不同的干旱条件下将植物培养10天。为了获得对照植物和经过不同干旱处理的植物,土壤含水量分别保持在土壤含水量(SWC)的65%,35%和30%。在冠的取样过程中监测渗透势,水饱和度不足, 13 C分辨力和脱水素积累,以进行蛋白质组分析。对2D-DIGE凝胶的分析显示105个差异丰富的斑点。对照和经过干旱处理的植物之间的差异最大,并且在两种干旱条件下都有25个斑点出现变化。通过串联质谱法成功鉴定出76个蛋白质斑点。鉴定出的蛋白质中最常见的功能类别可以分为以下组:应激相关蛋白质,氨基酸代谢,碳水化合物代谢以及DNA和RNA调节与加工。讨论了它们在大麦对干旱胁迫的响应中的可能作用。我们的研究表明,在干旱条件下大麦简历。护身符降低了它的生长和发育速度,显示了从有氧代谢到厌氧代谢的转变,并且几种保护性蛋白质的含量增加。两种干旱处理的比较表明,植物适应了较轻的干旱(SWC为35%)。但在更严格的干旱处理下(30%SWC),植物会受到损害。获得的结果表明,cv。护身符对干旱压力敏感。此外,四个斑点显示出随SWC的降低而连续且显着增加(UDP-葡萄糖6-脱氢酶,谷胱甘肽过氧化物酶和两个未鉴定的斑点)可能是测试其蛋白质表型能力的好候选物,同时在两个方面均得到显着区分干旱处理。

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