A Comparative Proteomics Survey of Proteins Responsive to Phosphorous Starvation in Roots of Hydroponically-grown Rice Seedlings
A Comparative Proteomics Survey of Proteins Responsive to Phosphorous Starvation in Roots of Hydroponically-grown Rice Seedlings
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DOI：
10.3839/jksabc.2011.102
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发表时间：
2011-10-01
影响因子：
--
通讯作者：
Kim, Sun Tae
中科院分区：
文献类型：
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作者：
Kim, Sang Gon;Wang, Yiming;Kim, Sun Tae
水稻生长发育需要磷素作为主要营养源。为了更好地了解磷饥饿的潜在机制并开发潜在的蛋白质生物标志物，研究了完全培养基中水培生长的水稻幼苗和磷缺乏（磷饥饿）的磷营养液的生理和蛋白质组变化。磷饥饿表现出显着减少根生长在三周大的幼苗相比，各自的完全媒体。此外，磷饥饿还显示了增加的酸性磷酸酶活性的根中的一个和三个周龄的幼苗，这表明实验设计是适合的蛋白质组学研究磷饥饿反应蛋白。双向凝胶电泳结合基质辅助激光解吸/电离飞行时间（MALDI-TOF）质谱分析的总根蛋白从3周龄的幼苗确定了10个P-饥饿响应蛋白点的140个高质量的蛋白点。共鉴定出10个蛋白质，这些蛋白质参与代谢和防御/应激反应。在10个蛋白点中，分别发现2个和8个蛋白点被上调和下调。半定量逆转录聚合酶链反应（RT-PCR）分析四种随机选择的蛋白质的相应基因，包括假定的甘油醛-3-磷酸脱氢酶（G3 PDH，斑点R1），S-腺苷-L-甲硫氨酸合成酶（SAMS，斑点R4）、ATP合酶亚基α（斑点R6）和根特异性病程相关蛋白10（PR-10，斑点R8），表明正如蛋白质丰度一样，这些蛋白质也在转录水平上受到调控。结果表明，鉴定的磷饥饿响应蛋白参与维持营养稳态和/或与缺磷条件下根系生理变化相关。
Rice takes up phosphorous (P) as major nutrient source for its growth and development when grown under anaerobic water-logged soil conditions. To better understand the underlying mechanisms and to develop potential protein biomarkers of P-starvation, hydroponically-grown rice seedlings in the complete media and phosphorus absence (P-starvation) of phosphorous nutrient solutions were investigated for physiological and proteome changes. The P-starvation manifested significant reduction in root growth in three-week-old seedlings compared to respective complete media. Furthermore, P-starvation also showed increased activity of acid phosphatase in roots of one- and three-week-old seedlings, suggesting that experimental design is suitable for proteomics survey of P-starvation responsive proteins. Two-dimensional gel electrophoresis coupled with matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry analysis of total root protein from three-week-old seedlings identified 10 P-starvation responsive protein spots out of 140 high-quality protein spots. Identified 10 proteins were involved in metabolism and defense/stress response. Out of 10, 2 and 8 protein spots were found to be up- and down-regulated, respectively. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis of corresponding genes of four randomly selected proteins, including putative glyceraldehydes-3-phophate dehydrogenase (G3PDH, spot R1), S-adenosyl-L-methionine synthetase (SAMS, spot R4), ATP synthase subunit alpha (spot R6), and root-specific pathogenesis-related protein 10 (PR-10, spot R8), showed that just as protein abundance, these proteins are also regulated at the transcript level. Results suggest identified P-starvation responsive proteins are involved in maintaining nutrient homeostasis and/or associated with changes in root physiology under the absence of P.