Comparative Proteomic Analysis of Early-Stage Soybean Seedlings Responses to Flooding by Using Gel and Gel-Free Techniques
Comparative Proteomic Analysis of Early-Stage Soybean Seedlings Responses to Flooding by Using Gel and Gel-Free Techniques
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DOI：
10.1021/pr100179f
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发表时间：
2010-08-01
影响因子：
4.4
通讯作者：
Komatsu, Setsuko
中科院分区：
文献类型：
--
作者：
Nanjo, Yohei;Skultety, Ludovit;Komatsu, Setsuko
利用凝胶和无凝胶蛋白质组学技术研究了大豆幼苗根和下胚轴对淹水胁迫的早期反应。从2日龄大豆幼苗淹水12小时的蛋白质提取和分析。两个质谱为基础的蛋白质组学分析，二维荧光差异凝胶电泳，和纳米液体色谱法确定了32个17个点和81个蛋白质，分别为响应洪水胁迫。根据所鉴定的蛋白质的数量和功能，糖酵解和发酵酶以及热激蛋白的诱导物是对淹水胁迫的早期反应的关键因素。对淹水幼苗酶活性和碳水化合物含量的分析表明，淹水期间糖酵解的激活和蔗糖降解酶的下调导致葡萄糖降解和蔗糖积累加速。此外，甲基乙二醛途径，这是一个与糖酵解有关的解毒系统，被上调。此外，基于二维聚丙烯酰胺凝胶电泳的磷酸蛋白质组学分析表明，参与蛋白质折叠和合成的蛋白质在洪水条件下被去磷酸化。这些结果表明，翻译和翻译后控制在淹水过程中可能会导致蛋白质的表达不平衡，参与几个代谢途径，包括碳水化合物代谢，可能会导致大豆幼苗淹水伤害。
Gel-based and gel-free proteomics techniques were used to investigate early responses to flooding stress in the roots and hypocotyls of soybean seedlings. Proteins from 2-day-old soybean seedlings flooded for 12 h were extracted and analyzed. Two mass-spectroscopy-based proteomics analyses, two-dimensional fluorescence difference gel electrophoresis, and nanoliquid chromatography identified 32 from 17 spots and 81 proteins, respectively, as responsive to flooding stress. On the basis of the number and function of proteins identified, glycolysis and fermentation enzymes and inducers of heat shock proteins were key elements in the early responses to flooding stress. Analysis of enzyme activities and carbohydrate contents in flooded seedlings showed that glucose degradation and sucrose accumulation accelerated during flooding due to activation of glycolysis and down-regulation of sucrose degrading enzymes. Additionally, the methylglyoxal pathway, which is detoxification system linked to glycolysis, was up-regulated. Furthermore, two-dimensional polyacrylamide gel electrophoresis-based phosphoproteomics analysis showed that proteins involved in protein folding and synthesis were dephosphorylated under flooding conditions. These results suggest that translational and post-translational control during flooding possibly induces an imbalance in the expression of proteins involved in several metabolic pathways including carbohydrate metabolism that might cause flooding injury of soybean seedlings.