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生理和定量蛋白质组学分析揭示了空心莲子草(Alternanthera philoxeroides L.)根系低钾胁迫响应。

Physiological and quantitative proteomic analyses unraveling potassium deficiency stress response in alligator weed (Alternanthera philoxeroides L.) root.

机构信息

College of Agronomy, Sichuan Agricultural University, Chengdu, 611130, China.

出版信息

Plant Mol Biol. 2018 Jun;97(3):265-278. doi: 10.1007/s11103-018-0738-5. Epub 2018 May 18.

Abstract

Physiological and iTRAQ based proteomic analysis provided new insights into potassium deficiency stress response in alligator weed root. Alligator weed (Alternanthera philoxeroides) has a strong ability to adapt to potassium deficiency (LK) stress. Proteomic changes in response to this stress are largely unknown in alligator weed. In this study, we investigated physiological and molecular mechanisms under LK using isobaric tags for relative and absolute quantitation to characterize proteome-level changes in this plant. First, root physiology, 2, 3, 5-Triphenyl-trazolium chloride (TTC) assay and peroxidase activity were significantly altered after 10 and 15 days of LK treatment. The comparative proteomic analysis suggested a total of 375 proteins were differential abundance proteins. The proteomic results were verified by western blot assays and quantitative real-time PCR. Correlation analysis of transcription and proteomics suggested protein processing in the endoplasmic reticulum, endocytosis, and spliceosome pathways were significantly enriched. The protein responsible for energy metabolism, signal sensing and transduction and protein degradation played crucial roles in this stress. Twelve ubiquitin pathway related proteins were identified in our study, among them 11 proteins were up-regulated. All protein ubiquitination of lysine using pan antibodies were also increased after LK treatment. Our study provide a valuable insights of molecular mechanism underlying LK stress response in alligator weed roots and afford a vital basis to further study potassium nutrition molecular breeding of other plant species.

摘要

生理和 iTRAQ 基于蛋白质组学分析为水花生根中钾缺乏胁迫反应提供了新的见解。水花生(Alternanthera philoxeroides)具有很强的适应低钾(LK)胁迫的能力。水花生对这种胁迫的蛋白质组变化在很大程度上是未知的。在这项研究中,我们使用相对和绝对定量的同位素标记(iTRAQ)技术来研究 LK 下的生理和分子机制,以表征该植物的蛋白质组水平变化。首先,根生理、2,3,5-三苯基氯化四氮唑(TTC)测定和过氧化物酶活性在 LK 处理 10 和 15 天后明显改变。比较蛋白质组分析表明,共有 375 种蛋白质是差异丰度蛋白。蛋白质组学结果通过 Western blot 分析和定量实时 PCR 进行了验证。转录组和蛋白质组学的相关性分析表明,内质网、内吞作用和剪接体途径的蛋白质加工显著富集。负责能量代谢、信号感应和转导以及蛋白质降解的蛋白质在这种胁迫中起着至关重要的作用。在我们的研究中鉴定了 12 种泛素途径相关蛋白,其中 11 种蛋白上调。LK 处理后,泛抗体用于赖氨酸的所有蛋白质泛素化也增加。我们的研究为水花生根中 LK 胁迫反应的分子机制提供了有价值的见解,并为进一步研究其他植物物种的钾营养分子育种提供了重要基础。

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