张志兴, 李忠, 李洲, 林文雄. 水稻生育后期根系响应氮肥调控的蛋白作用机制分析[J]. 中国生态农业学报(中英文), 2015, 23(11): 1413-1422. DOI: 10.13930/j.cnki.cjea.150760
引用本文: 张志兴, 李忠, 李洲, 林文雄. 水稻生育后期根系响应氮肥调控的蛋白作用机制分析[J]. 中国生态农业学报(中英文), 2015, 23(11): 1413-1422. DOI: 10.13930/j.cnki.cjea.150760
ZHANG Zhixing, LI Zhong, LI Zhou, LIN Wenxiong. Effects of nitrogen management on protein expression of rice (Oryza sativa L.) root in the late growth stage[J]. Chinese Journal of Eco-Agriculture, 2015, 23(11): 1413-1422. DOI: 10.13930/j.cnki.cjea.150760
Citation: ZHANG Zhixing, LI Zhong, LI Zhou, LIN Wenxiong. Effects of nitrogen management on protein expression of rice (Oryza sativa L.) root in the late growth stage[J]. Chinese Journal of Eco-Agriculture, 2015, 23(11): 1413-1422. DOI: 10.13930/j.cnki.cjea.150760

水稻生育后期根系响应氮肥调控的蛋白作用机制分析

Effects of nitrogen management on protein expression of rice (Oryza sativa L.) root in the late growth stage

  • 摘要: 以大穗型水稻‘金恢809’为试验材料, 设置前氮后移(施氮量的30%作基肥, 30%作蘖肥, 30%作穗肥, 10%作粒肥)及传统氮肥(施氮量的40%作基肥, 30%作蘖肥, 30%作穗肥)施用两种处理, 并采用双向电泳技术分析了水稻生育后期根系对前氮后移调控的蛋白响应机制。通过构建的不同氮肥处理下水稻开花后15 d根系蛋白2-DE电泳图谱, 并结合MALDI-TOF/MS分析, 共成功鉴定到57个出现差异表达的蛋白, 其中40个在前氮后移处理下上调表达, 17个下调表达。依据其蛋白功能可以归为12类, 主要涉及信号转导, 氨同化, 胁迫与防御, 糖酵解, 三羧酸循环, 氨基酸代谢、蛋白合成及折叠等代谢途径。研究发现水稻生育后期根系GTP结合的核蛋白、GDP解离抑制因子、液泡型H+-ATP酶亚基C会响应氮肥的调控而出现表达变化, 并将信号传递到电压依赖性离子通道蛋白、液泡型ATP酶亚基C以及膜联蛋白p35, 进而调节根系细胞间物质运输与信息的传递。本研究结果还表明, 前氮后移的施肥方式, 通过适当增加水稻生育后期氮肥的施用, 调节根系糖类转换、糖酵解及三羧酸循环相关蛋白的表达, 增加了根系ATP的供应; 通过增强根系中清除过氧化氢系统及防御相关蛋白的表达, 延缓了根系的衰老; 通过上调谷氨酸脱氢酶、天冬氨酸转氨酶和谷氨酞胺合成酶的表达丰度, 促进了根系对土壤中氮元素的吸收与转运。研究结果对于进一步明确水稻根系响应氮素调控的分子生态特性具有重要的理论与实际意义。

     

    Abstract: Application of nitrogen fertilizer is the most important agricultural measures in increasing paddy rice production. Recently, the postponed nitrogen application has developed as a modern cultivation measure, which not only is beneficial to the high yield formation, but also increases utilization efficiency of nitrogen fertilizer. However, the molecular basis of such adaptation remains unclear. In present study, the molecular mechanisms of rice ‘jinhui No.809’ (large-panicle type) root in response to the postponed nitrogen application were investigated. By keeping the total nitrogen supply constant and varying the early and late growth stage fertilizer application ratios, changes in the protein expressions of the rice root in late growth stage were determined. The two nitrogen fertilization treatments were traditional (NT) and postponed nitrogen application (NP). Using 2-DE and MALDI-TOF/MS, 57 proteins with 40 up-regulated and 17 down-regulated in response to NP treatment were successfully identified. According to the protein function, the identified proteins were classified into 12 categories, mainly including signal transduction, ammonia assimilation, stress and defense, glycolysis, tricarboxylic acid cycle, protein synthesis and folding, and amino acid metabolism. The results indicated that the signal proteins of GDP dissociation inhibitor protein and GTP-binding nuclear protein Ran-2 in rice root were responsive to the NP treatment, then transfer signal to the membrane protein, including voltage dependent anion selective channel protein, annexin p35 and vacuolar ATP synthase subunit C, leading to the changes of intercellular material transport and message transference. The protein abundance of glycolysis and tricarboxylic acid cycle related protein, including phosphoglycerate kinase, phosphoglucomutase, 6-phosphofructokinase, UDP-glucose pyrophosphorylase, NADP-isocitrate dehydrogenase and aconitate hydratase were increased by the NP treatment, in turn, induced more ATP were generated for the root growth. The up-regulated antioxidative enzymes and defence related protein delayed the root senescence in the late growth stage. Moreover, glutamate dehydrogenase, glutamines synthetase root isozyme and aspartate minotransferase, three of which were involved in the plant nitrogen metabolism, increased in abundance in the NP treatment, which were favorable for the nitrogen uptake and translocation. This study added new significant insights to our current understanding of the molecular ecology characteristic of rice root in response to nitrogen management.

     

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