Proteomic characterization of copper stress response in Elsholtzia splendens roots and leaves. Plant Mol Biol
Department of Environmental Engineering, Zhejiang University, 310029 Hangzhou, People's Republic of China. Plant Molecular Biology
(Impact Factor: 4.26).
08/2009; 71(3):251-63. DOI: 10.1007/s11103-009-9521-y
Elsholtzia splendens is generally considered as a Cu-tolerant and -accumulating plant species, and a candidate for phytoremediation of Cu-contaminated soils. To better understand the Cu tolerance/accumulation mechanisms in E. splendens, proteomic analysis was performed on E. splendens roots and leaves exposed to 100 muM CuSO(4) for 3 and 6 days. After 6 days of treatment, Cu accumulation in roots increased much more than that in leaves. SDS-PAGE analysis showed that the proteins changed more intensively in roots than did in leaves upon Cu stress. Two-dimensional gel electrophoresis (2-DE) and image analyses found that 45 protein spots were significantly changed in roots, but only six protein spots in leaves. The abundance of protein spots mostly showed temporal changes. MALDI-TOF MS and LTQ-ESI-MS/MS were used to identify the differently expressed protein spots. The identified root proteins were involved in various cellular processes such as signal transduction, regulation of transcription and translation, energy metabolism, regulation of redox homeostasis and cell defense. The leaf proteins were mainly degraded fragments of RuBisCo and antioxidative protein. The roles of these proteins in Cu tolerance/accumulation were discussed. The resulting differences in protein expression pattern suggested that redirection of root cellular metabolism and redox homeostasis might be important survival mechanisms of E. splendens upon Cu stress.
Available from: Shaoshan Li
- "The approach can therefore help to elucidate new aspects of plant heavy metal stress (Cvjetko et al. 2014; Hossain and Komatsu 2013). Using this method, the proteomic changes induced by Cu have been conducted in several species, including Cannabis sativa (Bona et al. 2007), Arabidopsis (Haensch and Mendel 2009), Elsholtzia splendens (Li et al. 2009), Triticum aestivum L. (Li et al. 2013), Oryza sativa L. (Song et al. 2013, 2014), cyanobacterium Cyanothece (Mota et al. 2015), Oryza sativa L. (Chen et al. 2015), and Sargassum fusiforme (Zou et al. 2015). Despite the literature available, knowledge of plant responses to Cu stress at the proteomic level is scarce, and little is known about the responses of sensitive plants to Cu stress at an early stage, especially, for non-model plants without genomic background information. "
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In the present study, the effects of Cu (2.0 and 8.0 μM) on root growth of Allium cepa var. agrogarum L. were addressed and protein abundance levels were analyzed using the technology of proteomics combined with transcriptomics, in order to go deeper into the understanding of the mechanism of Cu toxicity on plant root systems at the protein level and to provide valuable information for monitoring and forecasting the effects of exposure to Cu in real scenarios conditions.
Protein extraction; Two-dimensional electrophoresis (2-DE) analysis; Mass spectrometry analysis; Establishment of the in-house database; Restriction enzyme map of the in-house database and protein identification.
Root growth was dramatically inhibited after 12 h Cu treatment. By establishing an in-house database and using mass spectrometry analysis, 27 differentially abundant proteins were identified. These 27 proteins were involved in multiple biological processes including defensive response, transcription regulation and protein synthesis, cell wall synthesis, cell cycle and DNA replication, and other important functions.
Our results provide new insights at the proteomic level into the Cu-induced responses, defensive responses and toxic effects, and provide new molecular markers of the early events of plant responses to Cu toxicity. Moreover, the establishment of an in-house database provides a big improvement for proteomics research on non-model plants.
Available from: Frank Bedon
- "These processes may work cooperatively to re-establish the cellular and redox homeostasis upon Cu stress  . Most of these temporal studies, however, were carried out using short-term, high Cu exposures (e.g. 100 M Cu, 3–6 days ; 601 M Cu, 6 weeks  "
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ABSTRACT: Differential expression of soluble proteins was explored in roots of metallicolous (M) and non-metallicolous (NM) plants of Agrostis capillaris L. exposed to increasing Cu to partially identify molecular mechanisms underlying higher Cu tolerance in M plants.Plants were cultivated 2 months on perlite with a CuSO4 (1–30 μM) spiked-nutrient solution. Soluble proteins extracted by the TCA/acetone procedure were separated with 2-DE (linear 4–7 pH gradient). After CCB staining and image analysis, 19 proteins differentially expressed were identified using LC-MS/MS and ESTs databases.At supra-optimal Cu exposure (15–30 μM), glycolysis was likely altered in NM roots with increased production of glycerone-P and methylglyoxal based on over-expression of Triosephosphate Isomerase and Fructose bisphosphate aldolase. Changes in Tubulins and higher expressions of 5-methyltetrahydropteroyltriglutamatehomocysteine methyltransferase and S-Adenosylmethionine synthase respectively underpinned impacts on the cytoskeleton and stimulation of ethylene metabolism. Increased L-methionine and S-Adenosylmethionine amounts may also facilitate production of nicotianamine, which complexes Cu, and L-cysteine, needed for metallothioneins and GSH. In M roots, the increase of [Cu/Zn] Superoxide dismutase suggested a better detoxification of superoxide, when Cu exposure rose. Higher Cu-tolerance of metallicolous plants would rather results from simultaneous cooperation of various processes than from a specific mechanism.This article is protected by copyright. All rights reserved
Available from: Lin Weipeng
- "However, this kinase exhibited an increase at 24 h post infection of Colletotrichum fragariae in strawberry (Fang et al. 2012). The study of Vacuolar ATPase subunit B1 is a major focus using cells and tissues in vivo (Liu et al. 1996; Miller et al. 2005, 2009; Păunescu et al. 2012). Our results showed that expression of this protein in tomato roots was down-accumulated by pathogen infection, which is similar to the study by Dahal et al. (2010) in tomato stems, but up-accumulated after Si addition to R. solanacearum inoculation. "
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ABSTRACT: Background and aims Silicon has an important role in enhancing resistance of plants against multiple environmental stresses including pathogen infection, but the mechanism is still not completely understood. In the present study, the role of Si-mediated resistance to Ralstonia solanacearum in tomato (Solanum lycopersicum) root was explored by a proteomics approach. Methods Treatments consisted of pathogen inoculation with or without 2.0 mM Si amendments, and two controls with or without 2.0 mM Si amendments. Proteins from the tomato roots with different treatments were extracted and identified by two-dimensional gel electrophoresis (2-DE) and liquid chromatography-mass spectrometry (LC-MS/MS). Results Fifty-three protein spots were identified at least two-fold differences in abundance on 2-DE maps under R. solanacearum inoculation and/or Si application. Among these proteins, 40 were significantly altered (6 were up-accumulated and 34 were down-accumulated) by R. solanacearum inoculation only. And 26 were altered (16 were increased and 10 were decreased) when Si was added to R. solanacearum-inoculated tomato plants. More than half of the altered proteins (62 %) were associated with energy/metabolism including glycolytic pathway and TCA cycle. Five proteins were grouped into defense-response, of which four were membrane-associated proteins. Conclusions These findings provide insights into molecular mechanisms responsible for Si-mediated resistance of tomato against R. solanacearum.
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