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See detailDifferential cadmium and zinc distribution in relation to their physiological impact in the leaves of the accumulating Zygophyllum fabago L
LEFÈVRE, Isabelle; VOGEL‐MIKUŠ, Katarina; JEROMEL, Luka et al

in Plant, cell & environment (in press)

Cadmium and zinc share many similar physiochemical properties, but their compartmentation, complexation and impact on other mineral element distribution in plant tissues may drastically differ. In this ... [more ▼]

Cadmium and zinc share many similar physiochemical properties, but their compartmentation, complexation and impact on other mineral element distribution in plant tissues may drastically differ. In this study, we address the impact of 10-μM Cd or 50-μM Zn treatment on ion distribution in leaves of a metallicolous population of the non-hyperaccumulating species Zygophyllum fabago at tissue and cell level, and the consequences on the plant response through a combined physiological, proteomic and metabolite approach. Micro-proton induced X-ray emission and laser ablation inductively coupled mass spectrometry analyses indicated hot spots of Cd concentrations in the vicinity of vascular bundles in response to Cd treatment, essentially bound to S-containing compounds as revealed by extended X-Ray absorption fine structure and non-protein thiol compounds analyses. A preferential accumulation of Zn occurred in vascular bundle and spongy mesophyll in response to Zn treatment, and was mainly bound to O/N-ligands. Leaf proteomics and physiological status evidenced a protection of photosynthetically active tissues and the maintenance of cell turgor through specific distribution and complexation of toxic ions, reallocation of some essential elements, synthesis of proteins involved in photosynthetic apparatus or C-metabolism, and metabolite synthesis, with some specificities regarding the considered heavy metal treatment. [less ▲]

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See detailA multiple-level study of metal tolerance in Salix fragilis and Salix aurita clones
Evlard, Aricia ULg; Sergeant, Kjell; Printz, Bruno et al

in Journal of Proteomics (2014), 101C

The response of two willow clones (Salix fragilis (Sf) and S. aurita (Sa)) to the presence of metals (Zn, Cu, Cd, Ni) was studied. Rooted cuttings were planted in control and contaminated soil. After 100 ... [more ▼]

The response of two willow clones (Salix fragilis (Sf) and S. aurita (Sa)) to the presence of metals (Zn, Cu, Cd, Ni) was studied. Rooted cuttings were planted in control and contaminated soil. After 100 days, different parameters (biomass, chlorophyll fluorescence (Fv/Fm), pigment and sugar concentrations, electrolyte leakage and proteome-level changes) were analyzed. The growth of Sa was not influenced by metals whereas Sf produced significantly less biomass when exposed to the pollutants. Furthermore, although Sa did not show a growth reduction in the presence of metals, the overall view of the physiological results among others the changes in the accumulation of sugars and pigments indicated that metals had a more severe impact on this clone. The response at the proteome level confirmed these observations. The growth reduction and the proteomic changes in Sf indicate that this clone adjusts its metabolism to maintain cellular homeostasis. Sa on the contrary maintains growth but the physiological and proteomics data suggests that this can only be done at the cost of cellular deregulation. Therefore high biomass is not linked with a good tolerance strategy. In a long-term study the survival of Sa might be compromised making it a poorer candidate for phytoremediation efforts. [less ▲]

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See detailPhysiological and proteomic responses of different willow clones (Salix fragilis x alba) exposed to dredged sediment contaminated by heavy metals
Evlard, Aricia ULg; Sergeant, Kjell; Ferrandis, Salvador et al

in International Journal of Phytoremediation (2014), 16(11), 1148-1169

High biomass producing species are considered as tools for remediation of contaminated soils. Willows (Salix spp.) are prominent study subjects in this regard. In this study, different willow clones ... [more ▼]

High biomass producing species are considered as tools for remediation of contaminated soils. Willows (Salix spp.) are prominent study subjects in this regard. In this study, different willow clones (Salix fragilis x alba) were planted on heavy-metal polluted dredging sludge. A first objective was assessment of the biomass production for these clones. Using a Gupta statistic, four clones were identified as high biomass producers (HBP). For comparison, a group of four clones with lowest biomass production were selected (LBP). A second objective was to compare metal uptake as well as the physiological and proteomic responses of these two groups. All these complementary data's allow us to have a better picture of the health of the clones that would be used in phytoremediation programs. Cd, Zn, and Ni total uptake was higher in the HBPs but Pb total uptake was higher in LBPs. Our proteomic and physiological results showed that the LBPs were able to maintain cellular activity as much as the HBPs although the oxidative stress response was more pronounced in the LBPs. This could be due to the high Pb content found in this group although a combined effect of the other metals cannot be excluded. [less ▲]

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See detailIMPACT OF HEAVY METALS ON PHYSIOLOGICAL AND PROTEOMIC RESPONSES OF WILLOWS (SALIX SP.)
Evlard, Aricia ULg; Sergeant, Kjell; Ferrandis, Salvador et al

Poster (2012)

In 2010, around 3,800 sites in Wallonia (Belgium) were identified as potentially contaminated by heavy metals, a result of their past industrial and agricultural activities [1]. The technique of using ... [more ▼]

In 2010, around 3,800 sites in Wallonia (Belgium) were identified as potentially contaminated by heavy metals, a result of their past industrial and agricultural activities [1]. The technique of using plants to remediate contaminated sites (phytoremediation) has been studied for over twenty years. In particular, the use of trees (alder, willow, poplar) has been considered because of their large biomass production [2] [3] [4] [5]. The aim of this study was to identify the potential of local willow ecotypes to grow in the presence of heavy metals using lysimeters filled with dredging sludge. Several willow ecotypes were compared realizing morphological and physiological measurements (chlorophyll fluorescence, electrolyte leakage, carbohydrate content). The plant responses to metal stress were also investigated using a proteomic approach. Heavy metals contents in leaves and stems were analysed. With this study, the potential of these trees to valorise contaminated, abandoned sites in Belgium was evaluated. [1] Cellule Etat de l'environnement wallon (2010). Tableau de bord de l'environnement wallon. SPW-DGARNE-DEMNA-DEE. [2] Dickinson N. M. (2000). Strategies for sustainable woodland on contaminated soils. Chemosphere 41(1-2): 259-263. [3] Meers E., Lamsal S., Vervaeke P., Hopgood M., Lust N. and Tack F. M. G. (2005). Availability of heavy metals for uptake by Salix viminalis on a moderately contaminated dredged sediment disposal site. Environ Pollut 137(2): 354-364. [4] Meers E., Vandecasteele B., Ruttens A., Vangronsveld J. and Tack F. M. G. (2007). Potential of five willow species (Salix spp.) for phytoextraction of heavy metals. Environ Exp Bot 60(1): 57-68. [5] Rosselli W., Keller C. and Boschi K. (2003). Phytoextraction capacity of trees growing on a metal contaminated soil. Plant Soil 256(2): 265-272. [less ▲]

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See detailBiocontrol proteomics:Implication of the pentoses phosphates pathway in the antagonist effect of Pichia anomala against Botrytis cinerea on apple
Kwasiborski, Anthony ULg; Renaut, Jenny; Lepoivre, Philippe ULg et al

Poster (2011, September 21)

The growing interest of the consumers for the wholesome food and the protection of the environment as well as the development of resistant pathogens to pesticides, stimulate the interest of growers to ... [more ▼]

The growing interest of the consumers for the wholesome food and the protection of the environment as well as the development of resistant pathogens to pesticides, stimulate the interest of growers to apply biological control methods. Pichia anomala strain K was previously identified as an efficient biocontrol agent of the main apple pathogens, Botrytis cinerea and Penicillum expansum. Further study demonstrated the complexicity of the mode of action of P. anomala against B. cinerea. A cDNA-AFLP and gene disruption study revealed implication of exo-β-1,3-glucanases in the mode of action of P. anomala strain Kh6 (a haploid form of P. anomala strain K displaying the same biocontrol properties). However, these studies suggested also implication of other factors. The present study aims to increase our knowledge of the mode of action of P. anomala strain Kh6 against B. cinerea using an in situ approach allowing the triple interaction, host/pathogen/antagonist and the proteomic tool allowing to study the ultime expression of the genome without a priori. One 50mm wound per apple were covered by a membrane and inoculated by a P. anomala suspension then by B. cinerea or not. Samples were collected during the exponential and stationary phase to identify the early and later responses to the presence of B. cinerea. After extraction, proteins were separated on 2-D gels. Spots influenced by the presence of B. cinerea in exponential and stationary phases were identified by MALDI-ToF. One hundred five and sixty spots of proteins were influenced by the presence of B. cinerea in exponential and stationary phases respectively. In exponential phase, P. anomala Kh6 in absence of B. cinerea uses mainly the glycolysis pathway, whereas in presence of pathogen, it orientates its energetic metabolism to the oxidative phosphorylation and sets up the pentose phosphate pathway. Thanks to this new orientation, P. anomala Kh6 probably obtains energy and nucleic acids allowing to colonize the wound as fast as in absence of B. cinerea and prevents the use of nutrients by the pathogen. In stationary phase, no differences in the P. anomala Kh6 energetic metabolism, in absence and in presence of B. cinerea were observed. During that phase, P. anomala Kh6 seems to use the alcoholic fermentation in order to face the nutrients impoverishment of the substrate. [less ▲]

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See detailBiocontrol proteomics:Implication of the pentoses phosphates pathway in the antagonist effect of Pichia anomala against Botrytis cinerea on apple.
Kwasiborski, Anthony ULg; renaut, Jenny; Delaplace, Pierre ULg et al

Conference (2011, August 31)

Pichia anomala strain K was previously identified as an efficient biocontrol agent of the apple pathogen, Botrytis cinerea. Further study demonstrated the complexicity of the mode of action of P. anomala ... [more ▼]

Pichia anomala strain K was previously identified as an efficient biocontrol agent of the apple pathogen, Botrytis cinerea. Further study demonstrated the complexicity of the mode of action of P. anomala against B. cinerea. A molecular study revealed implication of exo-β-1,3-glucanases in the mode of action of P. anomala but suggested implication of other factors. The present study aims to increase our knowledge of the mode of action of P. anomala strain Kh6 against B. cinerea using an in situ approach allowing interactions between organisms, and the proteomic tool allowing to study the ultime expression of the genome. One wound per apple were covered by a membrane and inoculated by P. anomala then by B. cinerea or not. Samples were collected at different incubation times to identify the responses of P. anomala to the presence of B. cinerea. After extraction, proteins were separated on 2-D gels. Spots influenced were identified by MALDI-ToF. In exponential phase, P. anomala Kh6 orientates its energetic metabolism from the glycolysis to the oxidative phosphorylation and sets up the pentose phosphate pathway. Thanks to this new pathway, P. anomala probably obtains energy and nucleic acids allowing to colonize the wound as fast as in absence of B. cinerea. In stationary phase, no differences in the P. anomala energetic metabolism, in absence and in presence of B. cinerea were observed. During that phase, P. anomala seems to use the alcoholic fermentation in order to face the nutrients impoverishment of the substrate. [less ▲]

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See detailIn situ proteome study of Pichia anomala strain K, an antagonist of the apple pathogen Botrytis cinerea
Kwasiborski, Anthony ULg; Renaut, Jenny; Delaplace, Pierre ULg et al

Conference (2011, May 24)

Postharvest fungal pathogens, mainly Botrytis cinerea, Penicillum expansum and Gloeosporium spp., annually cause 5-20% losses of fruit. Control measures against these diseases include chemical fungicide ... [more ▼]

Postharvest fungal pathogens, mainly Botrytis cinerea, Penicillum expansum and Gloeosporium spp., annually cause 5-20% losses of fruit. Control measures against these diseases include chemical fungicide applications, but the development of resistant fungal explains the growing interest for biological control. Pichia anomala strain K was previously identified as an efficient antagonist of pathogens on apples. Indeed, the percentage of protection of P.anomala against B.cinerea reached from 90 to 100% on apple wounds according to the experimental conditions. Microbiological, biochemical and molecular approaches demonstrated the implication of exo-β-1,3-glucanases in the mechanism of action of P.anomala. However, study of these mechanisms could be improved by observations under natural infection conditions in order to take into account the tripartite interactions host/antagonist/pathogen. The proteomic tool allowed an overview of process implicated in the antagonism against B.cinerea in such conditions. One 50mm wound per apple were covered by a membrane and inoculated by a P.anomala suspension then by B.cinerea or not. Samples were collected during the exponential and stationary phase to identify the early and later responses to the presence of B.cinerea. After extraction, proteins were separated on 2DE gels. Spots influenced by the presence of B.cinerea in exponential and stationary phases were identified by MALDI-ToF. One hundred five and sixty spots of proteins were influenced by the presence of B.cinerea in exponential and stationary phase respectively. In exponential phase, influenced proteins were implicated in the different steps of the proteins biosynthesis: amino acid synthesis, translation or mRNA maturation and in energy synthesis. On the other hand, in stationary phase, influenced proteins were mainly implicated in energy metabolic pathway: glycolysis, alcoholic fermentation or gluconeogenesis. The presence of B.cinerea seemed to induce a slowdown in the metabolism of P.anomala without influenced its growth. Further studies have to be realized to understand the complexity of these modes of action. [less ▲]

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See detailPotential Therapeutic Target Discovery by 2D-DIGE Proteomic Analysis in Mouse Models of Asthma
QUESADA CALVO, Florence ULg; Fillet, Marianne ULg; Renaut, Jenny et al

in Journal of Proteome Research (2011), 10(9), 4291-4301

As asthma physiopathology is complex and not fully understood to date; it is expected that new key mediators are still to be unveiled in this disease. The main objective of this study was to discover ... [more ▼]

As asthma physiopathology is complex and not fully understood to date; it is expected that new key mediators are still to be unveiled in this disease. The main objective of this study was to discover potential new target proteins with a molecular weight >20 kDa by using two-dimensional differential in-gel electrophoresis (2D-DIGE) on lung parenchyma extracts from control or allergen-exposed mice (ovalbumin). Two different mouse models leading to the development of acute airway inflammation (5 days allergen exposure) and airway remodeling (10 weeks allergen exposure) were used. This experimental setting allowed the discrimination of 33 protein spots in the acute inflammation model and 31 spots in the remodeling model displaying a differential expression. Several proteins were then identified by MALDI-TOF/TOF MS. Among those differentially expressed proteins, PDIA6, GRP78, Annexin A6, hnRPA3, and Enolase display an increased expression in lung parenchyma from mice exposed to allergen for 5 days. Conversely, Apolipoprotein A1 was shown to be decreased after allergen exposure in the same model. Analysis on lung parenchyma of mice exposed to allergens for 10 weeks showed decreased calreticulin levels. Changes in the levels of those different mediators were confirmed by Western blot and immunohistochemical analysis. Interestingly, alveolar macrophages isolated from lungs in the acute inflammation model displayed enhanced levels of GRP78. Moreover, intratracheal instillation of anti-GRP78 siRNA in allergen-exposed animals led to a decrease in eosinophilic inflammation and bronchial hyperresponsiveness. This study unveils new mediators of potential importance that are up- and down-regulated in asthma. Among up-regulated mediators, GRP-78 appears as a potential new therapeutic target worthy of further investigations. [less ▲]

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See detailThe proapoptotic C16-ceramide-dependent pathway requires the death-promoting factor Btf in colon adenocarcinoma cells.
Renert, Anne-Francoise; Leprince, Pierre ULg; Dieu, Marc et al

in Journal of Proteome Research (2009), 8(10), 4810-22

Ceramides are central molecules in sphingolipid metabolism. They are involved in the regulation of cancer-cell growth, differentiation, senescence and apoptosis. To better understand how these secondary ... [more ▼]

Ceramides are central molecules in sphingolipid metabolism. They are involved in the regulation of cancer-cell growth, differentiation, senescence and apoptosis. To better understand how these secondary messengers induce their biological effects, adenocarcinoma cells (HCT116) were treated with exogenous long-chain ceramides (C16-ceramide) in order to mimic endogenous sphingolipids. This treatment induced a decrease of cell viability partly due to apoptosis as shown by PARP cleavage and a decrease of pro-caspase 3. Two-dimensional differential in-gel electrophoresis (2D-DIGE) revealed the differential expression of 51 proteins in response to C16-ceramide. These proteins are notably involved in cell proliferation, apoptosis, protein transport and transcriptional regulation. Among them, the cell death-promoting factor Btf was found to be implicated in the apoptotic signal triggered by ceramide. In adenocarcinoma cells, Btf regulates apoptosis related proteins such as Mdm2, p53, BAX and pBcl-2 and thus plays an important role in the ceramide mediated cell death. These findings bring new insight into the proapoptotic ceramide-dependent signaling pathway. [less ▲]

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See detailCombining proteomics and metabolite analyses to unravel cadmium stress-response in poplar leaves
Kieffer, Pol ULg; Planchon, Sébastien; Oufir, Mouhssin et al

in Journal of Proteome Research (2009), 8

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See detailProteomic and enzymatic response of poplar to cadmium stress
Kieffer, Pol; Schröder, Peter; Dommes, Jacques ULg et al

in Journal of Proteomics (2009), 72

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See detailProteomic changes in rat hippocampus and adrenals following short-term sleep deprivation.
Poirrier, Jean-Etienne; Guillonneau, Francois; Renaut, Jenny et al

in Proteome Science (2008), 6

ABSTRACT: BACKGROUND: To identify the biochemical changes induced by sleep deprivation at a proteomic level, we compared the hippocampal proteome of rats either after 4 hours of sleep or sleep deprivation ... [more ▼]

ABSTRACT: BACKGROUND: To identify the biochemical changes induced by sleep deprivation at a proteomic level, we compared the hippocampal proteome of rats either after 4 hours of sleep or sleep deprivation obtained by gentle handling. Because sleep deprivation might induce some stress, we also analyzed proteomic changes in rat adrenals in the same conditions. After sleep deprivation, proteins from both tissues were extracted and subjected to 2D-DIGE analysis followed by protein identification through mass spectrometry and database search. RESULTS: In the hippocampus, 87 spots showed significant variation between sleep and sleep deprivation, with more proteins showing higher abundance in the latter case. Of these, 16 proteins were present in sufficient amount for a sequencing attempt and among the 12 identified proteins, inferred affected cellular functions include cell metabolism, energy pathways, transport and vesicle trafficking, cytoskeleton and protein processing. Although we did not observe classical, macroscopic effect of stress in sleep-deprived rats, 47 protein spots showed significant variation in adrenal tissue between sleep and sleep deprivation, with more proteins showing higher abundance following sleep. Of these, 16 proteins were also present in sufficient amount for a sequencing attempt and among the 13 identified proteins, the most relevant cellular function that was affected was cell metabolism. CONCLUSION: At a proteomic level, short term sleep deprivation is characterized by a higher expression of some proteins in the hippocampus and a lower abundance of other proteins in the adrenals (compared to normal sleep control). Altogether, this could indicate a general activation of a number of cellular mechanisms involved in the maintenance of wakefulness and in increased energy expenditure during sleep deprivation. These findings are relevant to suggested functions of sleep like energy repletion and the restoration of molecular stocks or a more global homeostasis of synaptic processes. [less ▲]

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See detailQuantitative changes in protein expression of cadmium-exposed poplar plants
Kieffer, Pol; Dommes, Jacques ULg; Hoffmann, Lucien et al

in Proteomics (2008), 8

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