Comparative proteomic analysis of melon phloem exudates in response to viral infection

Serra-Soriano, Marta; Navarro, J. A.; Genovés, Ainhoa; Pallás, Vicente

doi:10.1016/j.jprot.2015.04.008

Cited by 30 publications

(24 citation statements)

References 91 publications

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“…These include comparative phloem proteomes derived from poplar (Populus spp.) and pumpkin upon wounding stress (Dafoe et al, 2009;Gaupels et al, 2012), rice (Oryza sativa) exposed to plant-hopper insects , salt-stressed cucumber (Fan et al, 2015), melon (Cucumis melo) responding to viral infection (Serra-Soriano et al, 2015), and iron-limited Brassica napus (Gutierrez-Carbonell et al, 2015). A common theme among these proteomes, including this study, is the accumulation of redox-related proteins during stress.…”

Section: Discussion Phloem Proteomicsmentioning

confidence: 92%

“…Of these proteins, JR1 and FLA8 are down-regulated in distant leaves of SAR-induced plants (Gruner et al, 2013;Bernsdorff et al, 2016), and analysis of cpn60B knockout mutants demonstrated a constitutive SAR-like response to P. syringae pv maculicola (Ishikawa et al, 2003). Interestingly, CPN60, a chloroplastic chaperon protein, also was suppressed in melon phloem during viral infection (Serra-Soriano et al, 2015), hinting that CPN60 may act as a negative regulator of disease resistance responses in the phloem.…”

Section: Proteins Suppressed In the Sar Phloem Proteomementioning

confidence: 99%

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Comparative Proteomics Analysis of Arabidopsis Phloem Exudates Collected During the Induction of Systemic Acquired Resistance

et al. 2016

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ORCID IDs: 0000-0002-5467-7290 (P.C.); 0000-0002-3422-4083 (J.M.-P.).Systemic acquired resistance (SAR) is a plant defense response that provides long-lasting, broad-spectrum pathogen resistance to uninfected systemic leaves following an initial localized infection. In Arabidopsis (Arabidopsis thaliana), local infection with virulent or avirulent strains of Pseudomonas syringae pv tomato generates long-distance SAR signals that travel from locally infected to distant leaves through the phloem to establish SAR. In this study, a proteomics approach was used to identify proteins that accumulate in phloem exudates in response to the induction of SAR. To accomplish this, phloem exudates collected from mock-inoculated or SAR-induced leaves of wild-type Columbia-0 plants were subjected to label-free quantitative liquid chromatography-tandem mass spectrometry proteomics. Comparing mock-and SAR-induced phloem exudate proteomes, 16 proteins were enriched in phloem exudates collected from SAR-induced plants, while 46 proteins were suppressed. SAR-related proteins THIOREDOXIN h3, ACYL-COENZYME A-BINDING PROTEIN6, and PATHOGENESIS-RELATED1 were enriched in phloem exudates of SAR-induced plants, demonstrating the strength of this approach and suggesting a role for these proteins in the phloem during SAR. To identify novel components of SAR, transfer DNA mutants of differentially abundant phloem proteins were assayed for SAR competence. This analysis identified a number of new proteins (m-type thioredoxins, major latex protein-like protein, ULTRAVIOLET-B RESISTANCE8 photoreceptor) that contribute to the SAR response. The Arabidopsis SAR phloem proteome is a valuable resource for understanding SAR long-distance signaling and the dynamic nature of the phloem during plant-pathogen interactions.

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Section: Discussion Phloem Proteomicsmentioning

confidence: 92%

Section: Proteins Suppressed In the Sar Phloem Proteomementioning

confidence: 99%

Comparative Proteomics Analysis of Arabidopsis Phloem Exudates Collected During the Induction of Systemic Acquired Resistance

et al. 2016

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“…Various methods have been employed to study the composition of the angiosperm phloem translocation stream, including insect stylectomy (Fisher and Frame, ; Gaupels et al ., ), EGTA‐mediated release of phloem sap (PS) (King and Zeevaart, ; Tetyuk et al ., ; Du et al ., ) and spontaneous exudation following an incision into the phloem (Hall and Baker, ; Beyenbac et al ., ; Richardson et al ., ; Fan et al ., ; Serra‐Soriano et al ., ). The latter method has been widely applied to analysis of the phloem translocation stream of such plants as lupin and various cucurbits.…”

Section: Introductionmentioning

confidence: 97%

Proteomics and metabolomics analyses reveal the cucurbit sieve tube system as a complex metabolic space

Ham

El‐shabrawi

et al. 2016

The Plant Journal

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These two authors made equal contributions. SUMMARYThe plant vascular system, and specifically the phloem, plays a pivotal role in allocation of fixed carbon to developing sink organs. Although the processes involved in loading and unloading of sugars and amino acids are well characterized, little information is available regarding the nature of other metabolites in the sieve tube system (STS) at specific sites along the pathway. Here, we elucidate spatial features of metabolite composition mapped with phloem enzymes along the cucurbit STS. Phloem sap (PS) was collected from the loading (source), unloading (apical sink region) and shoot-root junction regions of cucumber, watermelon and pumpkin. Our PS analyses revealed significant differences in the metabolic and proteomic profiles both along the source-sink pathway and between the STSs of these three cucurbits. In addition, metabolite profiles established for PS and vascular tissue indicated the presence of distinct compositions, consistent with the operation of the STS as a unique symplasmic domain. In this regard, at various locations along the STS we could map metabolites and their related enzymes to specific metabolic pathways. These findings are discussed with regard to the function of the STS as a unique and highly complex metabolic space within the plant vascular system.

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“…Based on this rationale, 200 μm‐pixel laser/PMT imaging using similar Typhoon imagers has commonly been used in 2D‐DIGE analyses, presumably in an attempt to maximise sensitivity as recommended in the Typhoon User's Guide v.3.0 (Amersham Biosciences / GE Healthcare, Buckinghamshire, UK), though seemingly without consideration for the impacts of low image resolution on software‐dependent analysis outcomes. For instance, studies have used 200 μm imaging followed by analyses with DeCyder™ (GE Healthcare, Buckinghamshire, UK) , the user guide(s) of which explicitly stipulate that images with no smaller than 100 μm pixels should be subject to analysis (https://www.gelifesciences.com).…”

Section: Resultsmentioning

confidence: 99%

Coomassie staining provides routine (sub)femtomole in‐gel detection of intact proteoforms: Expanding opportunities for genuine Top‐down Proteomics

et al. 2017

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Modified colloidal Coomassie Brilliant Blue (cCBB) staining utilising a novel destain protocol and near-infrared fluorescence detection (nIRFD) rivals the in-gel protein detection sensitivity (DS) of SYPRO Ruby. However, established DS estimates are likely inaccurate in terms of 2DE-resolved proteoform 'spots' since DS is routinely measured from comparatively diffuse protein 'bands' following wide-well 1DE. Here, cCBB DS for 2DE-based proteomics was more accurately determined using narrow-well 1DE. As precise estimates of protein standard monomer concentrations are essential for accurate quantitation, coupling UV absorbance with gel-based purity assessments is described. Further, as cCBB is compatible with both nIRFD and densitometry, the impacts of imaging method (and image resolution) on DS were assessed. Narrow-well 1DE enabled more accurate quantitation of cCBB DS for 2DE, achieving (sub)femtomole DS with either nIRFD or densitometry. While densitometry offers comparative simplicity and affordability, nIRFD has the unique potential for enhanced DS with Deep Imaging. Higher-resolution nIRFD also improved analysis of a 2DE-resolved proteome, surpassing the DS of standard nIRFD and densitometry, with nIRFD Deep Imaging further maximising proteome coverage. cCBB DS for intact proteins rivals that of mass spectrometry (MS) for peptides in complex mixtures, reaffirming that 2DE-MS currently provides the most routine, broadly applicable, robust, and information-rich Top-down approach to Discovery Proteomics.

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Comparative proteomic analysis of melon phloem exudates in response to viral infection

Cited by 30 publications

References 91 publications

Comparative Proteomics Analysis of Arabidopsis Phloem Exudates Collected During the Induction of Systemic Acquired Resistance

Comparative Proteomics Analysis of Arabidopsis Phloem Exudates Collected During the Induction of Systemic Acquired Resistance

Proteomics and metabolomics analyses reveal the cucurbit sieve tube system as a complex metabolic space

Coomassie staining provides routine (sub)femtomole in‐gel detection of intact proteoforms: Expanding opportunities for genuine Top‐down Proteomics

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