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解析根系蛋白质组变化揭示萝卜(Raphanus sativus L.)镉胁迫响应的新见解。

Dissecting Root Proteome Changes Reveals New Insight into Cadmium Stress Response in Radish (Raphanus sativus L.).

机构信息

National Key Laboratory of Crop Genetics and Germplasm Enhancement, Key Laboratory of Horticultural Crop Biology and Genetic Improvement (East China) of MOA, College of Horticulture, Nanjing Agricultural University, Nanjing 210095, P.R. China.

The UWA Institute of Agriculture, and School of Agriculture and Environment, The University of Western Australia, Perth, WA 6009, Australia.

出版信息

Plant Cell Physiol. 2017 Nov 1;58(11):1901-1913. doi: 10.1093/pcp/pcx131.

Abstract

Cadmium (Cd) is a widespread heavy metal of particular concern with respect to the environment and human health. Although intensive studies have been conducted on Cd-exposed transcriptome profiling, little systematic proteome information is available on the molecular mechanism of Cd stress response in radish. In this study, the radish root proteome under Cd stress was investigated using a quantitative multiplexed proteomics approach. Seedlings were grown in nutrient solution without Cd (control) or with 10 or 50 μM CdCl2 for 12 h (Cd10 and Cd50, respectively). In total, 91 up- and 66 down-regulated proteins were identified in the control vs Cd10 comparison, while 340 up- and 286 down-regulated proteins were identified in the control vs Cd50 comparison. Functional annotation indicated that these differentially expressed proteins (DEPs) were mainly involved in carbohydrate and energy metabolism, stress and defense and signal transduction processes. Correlation analysis showed that 33 DEPs matched with their transcripts, indicating a relatively low correlation between transcript and protein levels under Cd stress. Quantitative real-time PCR evidenced the expression patterns of 12 genes encoding their corresponding DEPs. In particular, several pivotal proteins associated with carbohydrate metabolism, ROS scavenging, cell transport and signal transduction were involved in the coordinated regulatory network of the Cd stress response in radish. Root exposure to Cd2+ activated several key signaling molecules and metal-containing transcription factors, and subsequently some Cd-responsive functional genes were mediated to reduce Cd toxicity and re-establish redox homeostasis in radish. This is a first report on comprehensive proteomic characterization of Cd-exposed root proteomes in radish. These findings could facilitate unraveling of the molecular mechanism underlying the Cd stress response in radish and provide fundamental insights into the development of genetically engineered low-Cd-content radish cultivars.

摘要

镉(Cd)是一种广泛存在的重金属,尤其与环境和人类健康有关。尽管已经对 Cd 暴露的转录组谱进行了深入研究,但关于萝卜 Cd 胁迫响应的分子机制,系统的蛋白质组信息却很少。在这项研究中,采用定量多重蛋白质组学方法研究了 Cd 胁迫下萝卜根的蛋白质组。将幼苗在不含 Cd 的营养液中生长(对照)或在 10 或 50 μM CdCl2 中生长 12 小时(分别为 Cd10 和 Cd50)。在对照与 Cd10 的比较中,共鉴定出 91 个上调和 66 个下调蛋白,而在对照与 Cd50 的比较中,鉴定出 340 个上调和 286 个下调蛋白。功能注释表明,这些差异表达蛋白(DEPs)主要参与碳水化合物和能量代谢、应激和防御以及信号转导过程。相关性分析表明,33 个 DEP 与它们的转录物相匹配,表明在 Cd 胁迫下,转录物和蛋白质水平之间的相关性较低。定量实时 PCR 证实了 12 个编码其对应 DEP 的基因的表达模式。特别是,与碳水化合物代谢、ROS 清除、细胞转运和信号转导相关的几个关键蛋白参与了萝卜 Cd 胁迫响应的协调调控网络。根暴露于 Cd2+ 激活了几个关键信号分子和含金属的转录因子,随后一些 Cd 响应的功能基因被介导以降低 Cd 毒性并在萝卜中重新建立氧化还原稳态。这是首次全面蛋白质组学描述萝卜根暴露于 Cd 后的蛋白质组。这些发现可以帮助揭示萝卜 Cd 胁迫响应的分子机制,并为培育低 Cd 含量的萝卜遗传工程品种提供基础见解。

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