63 research outputs found
The PIP peptide of INFLORESCENCE DEFICIENT IN ABSCISSION enhances Populus leaf and Elaeis guineensis fruit abscission
The programmed loss of a plant organ is called abscission, which is an important cell separation process that occurs with different organs throughout the life of a plant. The use of floral organ abscission in Arabidopsis thaliana as a model has allowed greater understanding of the complexities of organ abscission, but whether the regulatory pathways are conserved throughout the plant kingdom and for all organ abscission types is unknown. One important pathway that has attracted much attention involves a peptide ligand-receptor signalling system that consists of the secreted peptide IDA (INFLORESCENCE DEFICIENT IN ABSCISSION) and at least two leucine-rich repeat (LRR) receptor-like kinases (RLK), HAESA (HAE) and HAESA-LIKE2 (HSL2). In the current study we examine the bioactive potential of IDA peptides in two different abscission processes, leaf abscission in Populus and ripe fruit abscission in oil palm, and find in both cases treatment with IDA peptides enhances cell separation and abscission of both organ types. Our results provide evidence to suggest that the IDAâHAEâHSL2 pathway is conserved and functions in these phylogenetically divergent dicot and monocot species during both leaf and fruit abscission, respectively
ĐĐœĐŽŃĐșŃĐžĐŸĐœĐœŃĐč ĐșĐŸŃДл ĐŽĐ»Ń ĐœĐ°ĐłŃĐ”ĐČĐ° ĐČĐŸĐŽŃ
ĐĐ°ŃĐ”ŃĐžĐ°Đ»Ń XVIII ĐДжЎŃĐœĐ°Ń. ĐœĐ°ŃŃ.-ŃĐ”Ń
Đœ. ĐșĐŸĐœŃ. ŃŃŃĐŽĐ”ĐœŃĐŸĐČ, Đ°ŃпОŃĐ°ĐœŃĐŸĐČ Đž ĐŒĐŸĐ»ĐŸĐŽŃŃ
ŃŃĐ”ĐœŃŃ
, ĐĐŸĐŒĐ”Đ»Ń, 26â27 Đ°ĐżŃ. 2018 Đł
Redox regulation of PEP activity during seedling establishment in Arabidopsis thaliana
Activation of the plastid-encoded RNA polymerase is tightly controlled
and involves a network of phosphorylation and, as yet unidentified,
thiol-mediated events. Here, we characterize PLASTID REDOX INSENSITIVE2,
a redox-regulated protein required for full PEP-driven transcription.
PRIN2 dimers can be reduced into the active monomeric form by
thioredoxins through reduction of a disulfide bond. Exposure to light
increases the ratio between the monomeric and dimeric forms of PRIN2.
Complementation of prin2-2 with different PRIN2 protein variants
demonstrates that the monomer is required for light-activated
PEP-dependent transcription and that expression of the nuclear-encoded
photosynthesis genes is linked to the activity of PEP. Activation of PEP
during chloroplast development likely is the source of a retrograde
signal that promotes nuclear LHCB expression. Thus, regulation of
PRIN2 is the thiol-mediated mechanism required for full PEP activity,
with PRIN2 monomerization via reduction by TRXs providing a mechanistic
link between photosynthetic electron transport and activation of
photosynthetic gene expression.</p
Nectin-4 is a new histological and serological tumor associated marker for breast cancer
Abstract
Introduction
Breast cancer is a complex and heterogeneous disease at the molecular level. Evolution is difficult to predict according to classical histoclinical prognostic factors. Different studies highlight the importance of large-scale molecular expression analyses to improve taxonomy of breast cancer and prognostic classification. Identification of new molecular markers that refine this taxonomy and improve patient management is a priority in the field of breast cancer research.
Nectins are cell adhesion molecules involved in the regulation of epithelial physiology. We present here Nectin-4/PVRL4 as a new histological and serological tumor associated marker for breast carcinoma.
Methods
Expression of Nectin-4 protein was measured on a panel of 78 primary cells and cell lines from different origins and 57 breast tumors by FACS analysis and immunohistochemistry (IHC), respectively. mRNA expression was measured by quantitative PCR.
Serum Nectin-4 was detected by ELISA and compared with CEA and CA15.3 markers, on panels of 45 sera from healthy donors, 53 sera from patients with non-metastatic breast carcinoma (MBC) at diagnosis, and 182 sera from patients with MBC. Distribution of histological/serological molecular markers and histoclinical parameters were compared using the standard Chi-2 test.
Results
Nectin-4 was not detected in normal breast epithelium. By contrast, Nectin-4 was expressed in 61% of ductal breast carcinoma vs 6% in lobular type. Expression of Nectin-4 strongly correlated with the basal-like markers EGFR, P53, and P-cadherin, and negatively correlated with the luminal-like markers ER, PR and GATA3. All but one ER/PR-negative tumors expressed Nectin-4. The detection of Nectin-4 in serum improves the follow-up of patients with MBC: the association CEA/CA15.3/Nectin-4 allowed to monitor 74% of these patients compared to 67% with the association CEA/CA15.3. Serum Nectin-4 is a marker of disease progression, and levels correlate with the number of metastases (P = 0.038). Serum Nectin-4 is also a marker of therapeutic efficiency and correlates, in 90% of cases, with clinical evolution.
Conclusion
Nectin-4 is a new tumor-associated antigen for breast carcinoma. Nectin-4 is a new bio-marker whose use could help refine breast cancer taxonomy and improve patients' follow-up. Nectin-4 emerges as a potential target for breast cancer immunotherapy.http://deepblue.lib.umich.edu/bitstream/2027.42/112418/1/12885_2007_Article_723.pd
Effects of saffron extract supplementation on mood, well-being, and response to a psychosocial stressor in healthy adults: A randomized, double-blind, parallel group, clinical trial
Anxiety, stress, and low mood are closely related and may contribute to depressive symptoms. Among non-pharmacological solutions to improve subclinical mood symptoms and resilience to stress, natural products such as saffronâidentified as promising following preliminary beneficial effects in major depressive disorderârepresent a relevant strategy. This study aimed to assess the efficacy of 8 weeks' supplementation with 30 mg standardized saffron extract on emotional well-being in healthy adults with subclinical feelings of low mood and anxiety and/or stress and evaluate the acute effect of saffron in response to a lab-based psychosocial stressor. The study adopted a double-blind, randomized, parallel groups design in which 56 healthy male and female individuals (18â54 years) received either a saffron extract or a placebo for 8 weeks. Chronic effects of saffron on subjective anxiety, stress, and depressive feelings were assessed using a questionnaire battery [including Profile of Mood State-2, (POMS)] and acute effects in response to a lab-based psychosocial stressor were measured through psychological and physiological parameters. Urinary crocetin levels were quantified. Participants who received the saffron extract reported reduced depression scores and improved social relationships at the end of the study. Urinary crocetin levels increased significantly with saffron supplementation and were correlated with change in depression scores. The typical stress-induced decrease in heart rate variability (HRV) during exposure to the stressor was attenuated following acute saffron intake. Saffron extract appears to improve subclinical depressive symptoms in healthy individuals and may contribute to increased resilience against the development of stress-related psychiatric disorders. Clinical trials number: NCT03639831
CBP-HSF2 structural and functional interplay in Rubinstein-Taybi neurodevelopmental disorder
Rubinstein-Taybi syndrome (RSTS) is a neurodevelopmental disorder with unclear underlying mechanisms. Here, the authors unravel the contribution of a stress-responsive pathway to RSTS where impaired HSF2 acetylation, due to RSTS-associated CBP/EP300 mutations, alters the expression of neurodevelopmental players, in keeping with hallmarks of cell-cell adhesion defects.Patients carrying autosomal dominant mutations in the histone/lysine acetyl transferases CBP or EP300 develop a neurodevelopmental disorder: Rubinstein-Taybi syndrome (RSTS). The biological pathways underlying these neurodevelopmental defects remain elusive. Here, we unravel the contribution of a stress-responsive pathway to RSTS. We characterize the structural and functional interaction between CBP/EP300 and heat-shock factor 2 (HSF2), a tuner of brain cortical development and major player in prenatal stress responses in the neocortex: CBP/EP300 acetylates HSF2, leading to the stabilization of the HSF2 protein. Consequently, RSTS patient-derived primary cells show decreased levels of HSF2 and HSF2-dependent alteration in their repertoire of molecular chaperones and stress response. Moreover, we unravel a CBP/EP300-HSF2-N-cadherin cascade that is also active in neurodevelopmental contexts, and show that its deregulation disturbs neuroepithelial integrity in 2D and 3D organoid models of cerebral development, generated from RSTS patient-derived iPSC cells, providing a molecular reading key for this complex pathology.</p
Extracorporeal Membrane Oxygenation for Severe Acute Respiratory Distress Syndrome associated with COVID-19: An Emulated Target Trial Analysis.
RATIONALE: Whether COVID patients may benefit from extracorporeal membrane oxygenation (ECMO) compared with conventional invasive mechanical ventilation (IMV) remains unknown. OBJECTIVES: To estimate the effect of ECMO on 90-Day mortality vs IMV only Methods: Among 4,244 critically ill adult patients with COVID-19 included in a multicenter cohort study, we emulated a target trial comparing the treatment strategies of initiating ECMO vs. no ECMO within 7 days of IMV in patients with severe acute respiratory distress syndrome (PaO2/FiO2 <80 or PaCO2 â„60 mmHg). We controlled for confounding using a multivariable Cox model based on predefined variables. MAIN RESULTS: 1,235 patients met the full eligibility criteria for the emulated trial, among whom 164 patients initiated ECMO. The ECMO strategy had a higher survival probability at Day-7 from the onset of eligibility criteria (87% vs 83%, risk difference: 4%, 95% CI 0;9%) which decreased during follow-up (survival at Day-90: 63% vs 65%, risk difference: -2%, 95% CI -10;5%). However, ECMO was associated with higher survival when performed in high-volume ECMO centers or in regions where a specific ECMO network organization was set up to handle high demand, and when initiated within the first 4 days of MV and in profoundly hypoxemic patients. CONCLUSIONS: In an emulated trial based on a nationwide COVID-19 cohort, we found differential survival over time of an ECMO compared with a no-ECMO strategy. However, ECMO was consistently associated with better outcomes when performed in high-volume centers and in regions with ECMO capacities specifically organized to handle high demand. This article is open access and distributed under the terms of the Creative Commons Attribution Non-Commercial No Derivatives License 4.0 (http://creativecommons.org/licenses/by-nc-nd/4.0/)
Mode of action of ÎČ-aminobutyric acid in grapevine : an inducer of resistance to pathogens and Mechanisms involved in the susceptibility to pathogens of the Arabidopsis PAD2 mutant impaired in glutathione production
La comprĂ©hension des mĂ©canismes de dĂ©fense mis en place lors de la rĂ©sistance des plantes vis-Ă -vis d'agents pathogĂšnes a pour objectif de proposer des alternatives Ă l'utilisation de produits phytosanitaires utilisĂ©s en agriculture. Dans une premiĂšre partie, nous avons Ă©tudiĂ© les mĂ©canismes molĂ©culaires impliquĂ©s dans la rĂ©sistance induite aux pathogĂšnes par l'acide ÎČ-aminobutyrique (BABA) chez la vigne. En effet, cet acide aminĂ© non protĂ©ique favorise un Ă©tat physiologique particulier, appelĂ© potentialisation, dans lequel la plante est capable de mobiliser plus rapidement et/ou plus intensĂ©ment ses rĂ©actions de dĂ©fense en rĂ©ponse Ă un stress. Contrairement aux Ă©liciteurs comme les oligogalacturonates (OG), nous avons montrĂ© que le BABA seul nâinduisait pas les Ă©vĂ©nements prĂ©coces de signalisation sur suspensions cellulaires de vigne, tels que les variations de la concentration en calcium cytosolique libre ([Ca2+]cyt), la production de monoxyde dâazote (NO), la production dâH2O2, la phosphorylation de MAPkinases, ni lâexpression de gĂšnes de dĂ©fense. Seules la production dâH2O2 et lâexpression plus intense du gĂšne RbohD codant une NADPH oxydase sont potentialisĂ©es par le BABA dans les suspensions cellulaires Ă©licitĂ©es par les OG. In planta, le BABA potentialise Ă©galement une production dâH2O2 en rĂ©ponse Ă lâinfection par lâoomycĂšte Plasmopara viticola. Lâutilisation dâun inhibiteur de NADPH oxydase abolit complĂštement cette production dâH2O2 et bloque partiellement la rĂ©sistance induite par le BABA. Nous montrons donc que la potentialisation de la production dâH2O2 dĂ©pendante dâune NADPH oxydase contribue Ă lâĂ©tablissement de la rĂ©sistance induite par le BABA chez la vigne. Une deuxiĂšme partie a permis dâapprĂ©hender les Ă©vĂ©nements cellulaires impliquĂ©s dans la rĂ©sistance des plantes en se focalisant sur le mutant pad2 (phytoalexin deficient) dâArabidopsis thaliana. Ce mutant prĂ©sente une sensibilitĂ© accrue Ă diffĂ©rents pathogĂšnes et contient un taux de glutathion de lâordre de 20 % par rapport Ă lâĂ©cotype sauvage. Nous avons tout dâabord montrĂ© que le faible taux de glutathion dĂ©pendait dâune quantitĂ© rĂ©duite de la premiĂšre enzyme de sa biosynthĂšse, la glutamate-cystĂ©ine ligase. Le glutathion Ă©tant impliquĂ© dans la mise en place des rĂ©actions de dĂ©fense, nous avons tentĂ© de dĂ©finir le lien entre la dĂ©ficience en glutathion et la sensibilitĂ© de pad2 aux pathogĂšnes. Nous avons tout dâabord montrĂ© que pad2 possĂ©dait un Ă©tat redox du glutathion plus oxydĂ© que le sauvage. Une analyse transcriptomique Ă lâĂ©tat basal a rĂ©vĂ©lĂ© que la plupart des gĂšnes diffĂ©rentiellement exprimĂ©s Ă©taient rĂ©primĂ©s chez pad2. Parmi ces gĂšnes, certains codent des protĂ©ines impliquĂ©es dans les flux dâions qui pourraient dĂ©rĂ©guler la dĂ©polarisation membranaire. Nous avons ainsi confirmĂ© que la dĂ©polarisation de la membrane plasmique est amoindrie chez pad2 en rĂ©ponse aux OG. De plus, des Ă©vĂ©nements en aval tels que la production dâH2O2 et la production de NO sont Ă©galement plus faibles chez le mutant par rapport au sauvage. Cette absence de la production dâH2O2 a Ă©galement Ă©tĂ© spĂ©cifiquement observĂ©e sur plantes pad2 infectĂ©es par lâoomycĂšte Phytophthora brassicae. Il en rĂ©sulte un dĂ©veloppement accru du pathogĂšne corrĂ©lĂ© Ă une absence de rĂ©ponse hypersensible, une mort cellulaire localisĂ©e normalement observĂ©e dans le cas du sauvage rĂ©sistant. En rĂ©ponse aux OG ou Ă lâinfection par P. brassicae, les analyses transcriptomiques font ressortir un fort enrichissement de gĂšnes relatifs Ă la dĂ©gradation des protĂ©ines chez pad2. De maniĂšre globale, nos rĂ©sultats suggĂšrent que la dĂ©ficience en glutathion chez pad2 pourrait profondĂ©ment modifier le turn-over des protĂ©ines, perturbant ainsi la signalisation cellulaire et les rĂ©ponses biologiques associĂ©es.Alternative strategies are required to reduce pesticide input into the environment for effective and sustainable plant protection. One solution is the activation of plant basal resistance that relies on the application of resistance inducer molecules. In the first part of this study, we analyzed the mode of action of ÎČ-aminobutyric acid (BABA), a non-protein amino acid, in the grapevine induced resistance. BABA confers a physiological state, called priming, during which plants are able to mobilize better and/or more rapidly defense responses to biotic or abiotic stress. Unlike oligogalacturonides (OG), we showed that BABA did not induce early signaling events in grapevine cells such as variations of cytosolic free calcium concentration, H2O2 and nitric oxide production, MAPkinase phosphorylation, nor the expression of defense-related genes. Among them, only H2O2 production and the expression of RbohD gene, which encodes a NADPH oxidase, are primed by BABA in OG-treated cells. Moreover, BABA-treated plants display a stronger accumulation of H2O2 in response to the oomycete Plasmopara viticola. Application of an NADPH oxidase inhibitor completely abolishes this H2O2 production and leads to a reduction of BABA-induced resistance against P. viticola. These data suggest that the priming of an NADPH oxidase-dependent H2O2 production contributes to BABA-induced resistance in grapevine. The second part consisted to analyze molecular events involved in plant resistance by using the pad2 (phytoalexin deficient) mutant of Arabidopsis thaliana which is susceptible to a broad range of pathogens. We showed that the glutathione depletion depends on the low amount of glutamate-cysteine ligase protein, the first enzyme involved in its biosynthesis. We studied molecular events, which are involved in defense mechanisms, to understand the impact of the glutathione content on pad2 susceptibility. Our results show that the redox state of glutathione is more oxidized in pad2 than in wild type Col-0. Since cellular redox state change is known to regulate gene expression, a basal transcriptome analysis has been performed in pad2 and wild type plants. Interestingly, most of the identified genes in pad2 are down-regulated, some of them encoding proteins involved in ion fluxes. As expected, the plasma membrane depolarization and events downstream like H2O2 and NO production are impaired in pad2 in response to OG. During infection with Phytophthora brassicae, the lack of H2O2 production is concomitant with an absence of the hypersensitive response, a localize cell death observed in the resistant wild type. After OG treatment or P. brassicae infection, microarray analysis brings out genes related to protein machinery including degradation in pad2. Taken together, these data suggest that the depletion of glutathione has an impact on protein turn-over which disturbs cell signaling events and related biological responses
Mode d'action de l'acide Ă-aminobutirique chez la vigne : un inducteur de rĂ©sistance aux pathogĂšnes et Ă©tude des mĂ©canismes impliquĂ©s dans la sensibilitĂ© aux pathogĂšnes du mutant PAD2 d'arabidopsis dĂ©ficient en glutathion
Alternative strategies are required to reduce pesticide input into the environment for effective and sustainable plant protection. One solution is the activation of plant basal resistance that relies on the application of resistance inducer molecules. In the first part of this study, we analyzed the mode of action of ÎČ-aminobutyric acid (BABA), a non-protein amino acid, in the grapevine induced resistance. BABA confers a physiological state, called priming, during which plants are able to mobilize better and/or more rapidly defense responses to biotic or abiotic stress. Unlike oligogalacturonides (OG), we showed that BABA did not induce early signaling events in grapevine cells such as variations of cytosolic free calcium concentration, H2O2 and nitric oxide production, MAPkinase phosphorylation, nor the expression of defense-related genes. Among them, only H2O2 production and the expression of RbohD gene, which encodes a NADPH oxidase, are primed by BABA in OG-treated cells. Moreover, BABA-treated plants display a stronger accumulation of H2O2 in response to the oomycete Plasmopara viticola. Application of an NADPH oxidase inhibitor completely abolishes this H2O2 production and leads to a reduction of BABA-induced resistance against P. viticola. These data suggest that the priming of an NADPH oxidase-dependent H2O2 production contributes to BABA-induced resistance in grapevine. The second part consisted to analyze molecular events involved in plant resistance by using the pad2 (phytoalexin deficient) mutant of Arabidopsis thaliana which is susceptible to a broad range of pathogens. We showed that the glutathione depletion depends on the low amount of glutamate-cysteine ligase protein, the first enzyme involved in its biosynthesis. We studied molecular events, which are involved in defense mechanisms, to understand the impact of the glutathione content on pad2 susceptibility. Our results show that the redox state of glutathione is more oxidized in pad2 than in wild type Col-0. Since cellular redox state change is known to regulate gene expression, a basal transcriptome analysis has been performed in pad2 and wild type plants. Interestingly, most of the identified genes in pad2 are down-regulated, some of them encoding proteins involved in ion fluxes. As expected, the plasma membrane depolarization and events downstream like H2O2 and NO production are impaired in pad2 in response to OG. During infection with Phytophthora brassicae, the lack of H2O2 production is concomitant with an absence of the hypersensitive response, a localize cell death observed in the resistant wild type. After OG treatment or P. brassicae infection, microarray analysis brings out genes related to protein machinery including degradation in pad2. Taken together, these data suggest that the depletion of glutathione has an impact on protein turn-over which disturbs cell signaling events and related biological responses.La comprĂ©hension des mĂ©canismes de dĂ©fense mis en place lors de la rĂ©sistance des plantes vis-Ă -vis d'agents pathogĂšnes a pour objectif de proposer des alternatives Ă l'utilisation de produits phytosanitaires utilisĂ©s en agriculture. Dans une premiĂšre partie, nous avons Ă©tudiĂ© les mĂ©canismes molĂ©culaires impliquĂ©s dans la rĂ©sistance induite aux pathogĂšnes par l'acide ÎČ-aminobutyrique (BABA) chez la vigne. En effet, cet acide aminĂ© non protĂ©ique favorise un Ă©tat physiologique particulier, appelĂ© potentialisation, dans lequel la plante est capable de mobiliser plus rapidement et/ou plus intensĂ©ment ses rĂ©actions de dĂ©fense en rĂ©ponse Ă un stress. Contrairement aux Ă©liciteurs comme les oligogalacturonates (OG), nous avons montrĂ© que le BABA seul nâinduisait pas les Ă©vĂ©nements prĂ©coces de signalisation sur suspensions cellulaires de vigne, tels que les variations de la concentration en calcium cytosolique libre ([Ca2+]cyt), la production de monoxyde dâazote (NO), la production dâH2O2, la phosphorylation de MAPkinases, ni lâexpression de gĂšnes de dĂ©fense. Seules la production dâH2O2 et lâexpression plus intense du gĂšne RbohD codant une NADPH oxydase sont potentialisĂ©es par le BABA dans les suspensions cellulaires Ă©licitĂ©es par les OG. In planta, le BABA potentialise Ă©galement une production dâH2O2 en rĂ©ponse Ă lâinfection par lâoomycĂšte Plasmopara viticola. Lâutilisation dâun inhibiteur de NADPH oxydase abolit complĂštement cette production dâH2O2 et bloque partiellement la rĂ©sistance induite par le BABA. Nous montrons donc que la potentialisation de la production dâH2O2 dĂ©pendante dâune NADPH oxydase contribue Ă lâĂ©tablissement de la rĂ©sistance induite par le BABA chez la vigne. Une deuxiĂšme partie a permis dâapprĂ©hender les Ă©vĂ©nements cellulaires impliquĂ©s dans la rĂ©sistance des plantes en se focalisant sur le mutant pad2 (phytoalexin deficient) dâArabidopsis thaliana. Ce mutant prĂ©sente une sensibilitĂ© accrue Ă diffĂ©rents pathogĂšnes et contient un taux de glutathion de lâordre de 20 % par rapport Ă lâĂ©cotype sauvage. Nous avons tout dâabord montrĂ© que le faible taux de glutathion dĂ©pendait dâune quantitĂ© rĂ©duite de la premiĂšre enzyme de sa biosynthĂšse, la glutamate-cystĂ©ine ligase. Le glutathion Ă©tant impliquĂ© dans la mise en place des rĂ©actions de dĂ©fense, nous avons tentĂ© de dĂ©finir le lien entre la dĂ©ficience en glutathion et la sensibilitĂ© de pad2 aux pathogĂšnes. Nous avons tout dâabord montrĂ© que pad2 possĂ©dait un Ă©tat redox du glutathion plus oxydĂ© que le sauvage. Une analyse transcriptomique Ă lâĂ©tat basal a rĂ©vĂ©lĂ© que la plupart des gĂšnes diffĂ©rentiellement exprimĂ©s Ă©taient rĂ©primĂ©s chez pad2. Parmi ces gĂšnes, certains codent des protĂ©ines impliquĂ©es dans les flux dâions qui pourraient dĂ©rĂ©guler la dĂ©polarisation membranaire. Nous avons ainsi confirmĂ© que la dĂ©polarisation de la membrane plasmique est amoindrie chez pad2 en rĂ©ponse aux OG. De plus, des Ă©vĂ©nements en aval tels que la production dâH2O2 et la production de NO sont Ă©galement plus faibles chez le mutant par rapport au sauvage. Cette absence de la production dâH2O2 a Ă©galement Ă©tĂ© spĂ©cifiquement observĂ©e sur plantes pad2 infectĂ©es par lâoomycĂšte Phytophthora brassicae. Il en rĂ©sulte un dĂ©veloppement accru du pathogĂšne corrĂ©lĂ© Ă une absence de rĂ©ponse hypersensible, une mort cellulaire localisĂ©e normalement observĂ©e dans le cas du sauvage rĂ©sistant. En rĂ©ponse aux OG ou Ă lâinfection par P. brassicae, les analyses transcriptomiques font ressortir un fort enrichissement de gĂšnes relatifs Ă la dĂ©gradation des protĂ©ines chez pad2. De maniĂšre globale, nos rĂ©sultats suggĂšrent que la dĂ©ficience en glutathion chez pad2 pourrait profondĂ©ment modifier le turn-over des protĂ©ines, perturbant ainsi la signalisation cellulaire et les rĂ©ponses biologiques associĂ©es
Mode d'action de l'acide Ă-aminobutirique chez la vigne : un inducteur de rĂ©sistance aux pathogĂšnes et Ă©tude des mĂ©canismes impliquĂ©s dans la sensibilitĂ© aux pathogĂšnes du mutant PAD2 d'arabidopsis dĂ©ficient en glutathion
Alternative strategies are required to reduce pesticide input into the environment for effective and sustainable plant protection. One solution is the activation of plant basal resistance that relies on the application of resistance inducer molecules. In the first part of this study, we analyzed the mode of action of ÎČ-aminobutyric acid (BABA), a non-protein amino acid, in the grapevine induced resistance. BABA confers a physiological state, called priming, during which plants are able to mobilize better and/or more rapidly defense responses to biotic or abiotic stress. Unlike oligogalacturonides (OG), we showed that BABA did not induce early signaling events in grapevine cells such as variations of cytosolic free calcium concentration, H2O2 and nitric oxide production, MAPkinase phosphorylation, nor the expression of defense-related genes. Among them, only H2O2 production and the expression of RbohD gene, which encodes a NADPH oxidase, are primed by BABA in OG-treated cells. Moreover, BABA-treated plants display a stronger accumulation of H2O2 in response to the oomycete Plasmopara viticola. Application of an NADPH oxidase inhibitor completely abolishes this H2O2 production and leads to a reduction of BABA-induced resistance against P. viticola. These data suggest that the priming of an NADPH oxidase-dependent H2O2 production contributes to BABA-induced resistance in grapevine. The second part consisted to analyze molecular events involved in plant resistance by using the pad2 (phytoalexin deficient) mutant of Arabidopsis thaliana which is susceptible to a broad range of pathogens. We showed that the glutathione depletion depends on the low amount of glutamate-cysteine ligase protein, the first enzyme involved in its biosynthesis. We studied molecular events, which are involved in defense mechanisms, to understand the impact of the glutathione content on pad2 susceptibility. Our results show that the redox state of glutathione is more oxidized in pad2 than in wild type Col-0. Since cellular redox state change is known to regulate gene expression, a basal transcriptome analysis has been performed in pad2 and wild type plants. Interestingly, most of the identified genes in pad2 are down-regulated, some of them encoding proteins involved in ion fluxes. As expected, the plasma membrane depolarization and events downstream like H2O2 and NO production are impaired in pad2 in response to OG. During infection with Phytophthora brassicae, the lack of H2O2 production is concomitant with an absence of the hypersensitive response, a localize cell death observed in the resistant wild type. After OG treatment or P. brassicae infection, microarray analysis brings out genes related to protein machinery including degradation in pad2. Taken together, these data suggest that the depletion of glutathione has an impact on protein turn-over which disturbs cell signaling events and related biological responses.La comprĂ©hension des mĂ©canismes de dĂ©fense mis en place lors de la rĂ©sistance des plantes vis-Ă -vis d'agents pathogĂšnes a pour objectif de proposer des alternatives Ă l'utilisation de produits phytosanitaires utilisĂ©s en agriculture. Dans une premiĂšre partie, nous avons Ă©tudiĂ© les mĂ©canismes molĂ©culaires impliquĂ©s dans la rĂ©sistance induite aux pathogĂšnes par l'acide ÎČ-aminobutyrique (BABA) chez la vigne. En effet, cet acide aminĂ© non protĂ©ique favorise un Ă©tat physiologique particulier, appelĂ© potentialisation, dans lequel la plante est capable de mobiliser plus rapidement et/ou plus intensĂ©ment ses rĂ©actions de dĂ©fense en rĂ©ponse Ă un stress. Contrairement aux Ă©liciteurs comme les oligogalacturonates (OG), nous avons montrĂ© que le BABA seul nâinduisait pas les Ă©vĂ©nements prĂ©coces de signalisation sur suspensions cellulaires de vigne, tels que les variations de la concentration en calcium cytosolique libre ([Ca2+]cyt), la production de monoxyde dâazote (NO), la production dâH2O2, la phosphorylation de MAPkinases, ni lâexpression de gĂšnes de dĂ©fense. Seules la production dâH2O2 et lâexpression plus intense du gĂšne RbohD codant une NADPH oxydase sont potentialisĂ©es par le BABA dans les suspensions cellulaires Ă©licitĂ©es par les OG. In planta, le BABA potentialise Ă©galement une production dâH2O2 en rĂ©ponse Ă lâinfection par lâoomycĂšte Plasmopara viticola. Lâutilisation dâun inhibiteur de NADPH oxydase abolit complĂštement cette production dâH2O2 et bloque partiellement la rĂ©sistance induite par le BABA. Nous montrons donc que la potentialisation de la production dâH2O2 dĂ©pendante dâune NADPH oxydase contribue Ă lâĂ©tablissement de la rĂ©sistance induite par le BABA chez la vigne. Une deuxiĂšme partie a permis dâapprĂ©hender les Ă©vĂ©nements cellulaires impliquĂ©s dans la rĂ©sistance des plantes en se focalisant sur le mutant pad2 (phytoalexin deficient) dâArabidopsis thaliana. Ce mutant prĂ©sente une sensibilitĂ© accrue Ă diffĂ©rents pathogĂšnes et contient un taux de glutathion de lâordre de 20 % par rapport Ă lâĂ©cotype sauvage. Nous avons tout dâabord montrĂ© que le faible taux de glutathion dĂ©pendait dâune quantitĂ© rĂ©duite de la premiĂšre enzyme de sa biosynthĂšse, la glutamate-cystĂ©ine ligase. Le glutathion Ă©tant impliquĂ© dans la mise en place des rĂ©actions de dĂ©fense, nous avons tentĂ© de dĂ©finir le lien entre la dĂ©ficience en glutathion et la sensibilitĂ© de pad2 aux pathogĂšnes. Nous avons tout dâabord montrĂ© que pad2 possĂ©dait un Ă©tat redox du glutathion plus oxydĂ© que le sauvage. Une analyse transcriptomique Ă lâĂ©tat basal a rĂ©vĂ©lĂ© que la plupart des gĂšnes diffĂ©rentiellement exprimĂ©s Ă©taient rĂ©primĂ©s chez pad2. Parmi ces gĂšnes, certains codent des protĂ©ines impliquĂ©es dans les flux dâions qui pourraient dĂ©rĂ©guler la dĂ©polarisation membranaire. Nous avons ainsi confirmĂ© que la dĂ©polarisation de la membrane plasmique est amoindrie chez pad2 en rĂ©ponse aux OG. De plus, des Ă©vĂ©nements en aval tels que la production dâH2O2 et la production de NO sont Ă©galement plus faibles chez le mutant par rapport au sauvage. Cette absence de la production dâH2O2 a Ă©galement Ă©tĂ© spĂ©cifiquement observĂ©e sur plantes pad2 infectĂ©es par lâoomycĂšte Phytophthora brassicae. Il en rĂ©sulte un dĂ©veloppement accru du pathogĂšne corrĂ©lĂ© Ă une absence de rĂ©ponse hypersensible, une mort cellulaire localisĂ©e normalement observĂ©e dans le cas du sauvage rĂ©sistant. En rĂ©ponse aux OG ou Ă lâinfection par P. brassicae, les analyses transcriptomiques font ressortir un fort enrichissement de gĂšnes relatifs Ă la dĂ©gradation des protĂ©ines chez pad2. De maniĂšre globale, nos rĂ©sultats suggĂšrent que la dĂ©ficience en glutathion chez pad2 pourrait profondĂ©ment modifier le turn-over des protĂ©ines, perturbant ainsi la signalisation cellulaire et les rĂ©ponses biologiques associĂ©es
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