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Quantitative Proteomic Analysis of Wheat Seeds during Artificial Ageing and Priming Using the Isobaric Tandem Mass Tag Labeling

机译:利用等压串联质量标签标记对小麦种子进行人工老化和启动过程中的蛋白质组学定量分析。

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摘要

Wheat (Triticum aestivum L.) is an important crop worldwide. The physiological deterioration of seeds during storage and seed priming is closely associated with germination, and thus contributes to plant growth and subsequent grain yields. In this study, wheat seeds during different stages of artificial ageing (45°C; 50% relative humidity; 98%, 50%, 20%, and 1% Germination rates) and priming (hydro-priming treatment) were subjected to proteomics analysis through a proteomic approach based on the isobaric tandem mass tag labeling. A total of 162 differentially expressed proteins (DEPs) mainly involved in metabolism, energy supply, and defense/stress responses, were identified during artificial ageing and thus validated previous physiological and biochemical studies. These DEPs indicated that the inability to protect against ageing leads to the incremental decomposition of the stored substance, impairment of metabolism and energy supply, and ultimately resulted in seed deterioration. Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that the up-regulated proteins involved in seed ageing were mainly enriched in ribosome, whereas the down-regulated proteins were mainly accumulated in energy supply (starch and sucrose metabolism) and stress defense (ascorbate and aldarate metabolism). Proteins, including hemoglobin 1, oleosin, agglutinin, and non-specific lipid-transfer proteins, were first identified in aged seeds and might be regarded as new markers of seed deterioration. Of the identified proteins, 531 DEPs were recognized during seed priming compared with unprimed seeds. In contrast to the up-regulated DEPs in seed ageing, several up-regulated DEPs in priming were involved in energy supply (tricarboxylic acid cycle, glycolysis, and fatty acid oxidation), anabolism (amino acids, and fatty acid synthesis), and cell growth/division. KEGG and protein-protein interaction analysis indicated that the up-regulated proteins in seed priming were mainly enriched in amino acid synthesis, stress defense (plant-pathogen interactions, and ascorbate and aldarate metabolism), and energy supply (oxidative phosphorylation and carbon metabolism). Therefore, DEPs associated with seed ageing and priming can be used to characterize seed vigor and optimize germination enhancement treatments. This work reveals new proteomic insights into protein changes that occur during seed deterioration and priming.
机译:小麦(Triticum aestivum L.)是全世界重要的农作物。种子在储存和启动过程中的生理恶化与发芽密切相关,因此有助于植物生长和随后的谷物产量。在这项研究中,对小麦种子在人工老化的不同阶段(45°C; 50%相对湿度; 98%,50%,20%和1%的发芽率)和启动处理(加氢启动处理)进行了蛋白质组学分析通过基于等压串联质量标签标记的蛋白质组学方法。在人工衰老过程中共鉴定出主要涉及代谢,能量供应和防御/应激反应的总共162种差异表达蛋白(DEP),从而验证了先前的生理和生化研究。这些DEPs表明不能抗衰老导致所储存物质的增量分解,代谢和能量供应的损害,并最终导致种子变质。京都基因与基因组百科全书(KEGG)分析显示,参与种子衰老的上调蛋白质主要富含核糖体,而下调的蛋白质主要蓄积在能量供应(淀粉和蔗糖代谢)和胁迫防御(抗坏血酸)中和藻酸盐代谢)。包括血红蛋白1,油质蛋白,凝集素和非特异性脂质转移蛋白在内的蛋白质首先在衰老的种子中鉴定出来,并可能被视为种子变质的新标志。在鉴定的蛋白质中,与未引种的种子相比,在引种种子期间识别出531个DEP。与种子老化中上调的DEP相比,引发中几个上调的DEP涉及能量供应(三羧酸循环,糖酵解和脂肪酸氧化),合成代谢(氨基酸和脂肪酸合成)和细胞增长/部门。 KEGG和蛋白质-蛋白质相互作用分析表明,种子启动过程中上调的蛋白质主要富含氨基酸合成,胁迫防御(植物-病原体相互作用,抗坏血酸和藻酸盐代谢)和能量供应(氧化磷酸化和碳代谢)。 。因此,与种子老化和引发相关的DEP可用于表征种子活力并优化发芽增强处理。这项工作揭示了对蛋白质变质的新见解,这些蛋白质变化发生在种子变质和启动过程中。

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