48 research outputs found

    The RPN12a proteasome subunit is essential for the multiple hormonal homeostasis controlling the progression of leaf senescence

    Get PDF
    The 26S proteasome is a conserved multi-subunit machinery in eukaryotes. It selectively degrades ubiquitinated proteins, which in turn provides an efficient molecular mechanism to regulate numerous cellular functions and developmental processes. Here, we studied a new loss-of-function allele of RPN12a, a plant ortholog of the yeast and human structural component of the 19S proteasome RPN12. Combining a set of biochemical and molecular approaches, we confirmed that a rpn12a knock-out had exacerbated 20S and impaired 26S activities. The altered proteasomal activity led to a pleiotropic phenotype affecting both the vegetative growth and reproductive phase of the plant, including a striking repression of leaf senescence associate cell-death. Further investigation demonstrated that RPN12a is involved in the regulation of several conjugates associated with the auxin, cytokinin, ethylene and jasmonic acid homeostasis. Such enhanced aptitude of plant cells for survival in rpn12a contrasts with reports on animals, where 26S proteasome mutants generally show an accelerated cell death phenotype.A loss-of-function mutation in the RPN12a proteasome subunit in Arabidopsis thaliana shows that this protein is essential for hormonal homeostasis, thereby modulating plant's development and the progression of leaf senescence

    Manipulating photorespiration to increase plant productivity:recent advances and perspectives for crop improvement

    Get PDF
    Recycling of the 2-phosphoglycolate generated by the oxygenase reaction of Rubisco requires a complex and energy-consuming set of reactions collectively known as the photorespiratory cycle. Several approaches aimed at reducing the rates of photorespiratory energy or carbon loss have been proposed, based either on screening for natural variation or by means of genetic engineering. Recent work indicates that plant yield can be substantially improved by the alteration of photorespiratory fluxes or by engineering artificial bypasses to photorespiration. However, there is also evidence indicating that, under certain environmental and/or nutritional conditions, reduced photorespiratory capacity may be detrimental to plant performance. Here we summarize recent advances obtained in photorespiratory engineering and discuss prospects for these advances to be transferred to major crops to help address the globally increasing demand for food and biomass production

    Metabolic control of arginine and ornithine levels paces the progression of leaf senescence

    Get PDF
    Pools of arginine and ornithine generated during protein degradation can pace the progression of leaf senescence by affecting the TCA cycle, polyamine biosynthesis and the ethylene signaling pathway.Leaf senescence can be induced by stress or aging, sometimes in a synergistic manner. It is generally acknowledged that the ability to withstand senescence-inducing conditions can provide plants with stress resilience. Although the signaling and transcriptional networks responsible for a delayed senescence phenotype, often referred to as a functional stay-green trait, have been actively investigated, very little is known about the subsequent metabolic adjustments conferring this aptitude to survival. First, using the individually darkened leaf (IDL) experimental setup, we compared IDLs of wild-type (WT) Arabidopsis (Arabidopsis thaliana) to several stay-green contexts, that is IDLs of two functional stay-green mutant lines, oresara1-2 (ore1-2) and an allele of phytochrome-interacting factor 5 (pif5), as well as to leaves from a WT plant entirely darkened (DP). We provide compelling evidence that arginine and ornithine, which accumulate in all stay-green contexts-likely due to the lack of induction of amino acids (AAs) transport-can delay the progression of senescence by fueling the Krebs cycle or the production of polyamines (PAs). Secondly, we show that the conversion of putrescine to spermidine (SPD) is controlled in an age-dependent manner. Thirdly, we demonstrate that SPD represses senescence via interference with ethylene signaling by stabilizing the ETHYLENE BINDING FACTOR1 and 2 (EBF1/2) complex. Taken together, our results identify arginine and ornithine as central metabolites influencing the stress- and age-dependent progression of leaf senescence. We propose that the regulatory loop between the pace of the AA export and the progression of leaf senescence provides the plant with a mechanism to fine-tune the induction of cell death in leaves, which, if triggered unnecessarily, can impede nutrient remobilization and thus plant growth and survival

    Triglyceride-rich lipoproteins and high-density lipoprotein cholesterol in patients at high risk of cardiovascular disease: evidence and guidance for management

    Get PDF
    Even at low-density lipoprotein cholesterol (LDL-C) goal, patients with cardiometabolic abnormalities remain at high risk of cardiovascular events. This paper aims (i) to critically appraise evidence for elevated levels of triglyceride-rich lipoproteins (TRLs) and low levels of high-density lipoprotein cholesterol (HDL-C) as cardiovascular risk factors, and (ii) to advise on therapeutic strategies for management. Current evidence supports a causal association between elevated TRL and their remnants, low HDL-C, and cardiovascular risk. This interpretation is based on mechanistic and genetic studies for TRL and remnants, together with the epidemiological data suggestive of the association for circulating triglycerides and cardiovascular disease. For HDL, epidemiological, mechanistic, and clinical intervention data are consistent with the view that low HDL-C contributes to elevated cardiovascular risk; genetic evidence is unclear however, potentially reflecting the complexity of HDL metabolism. The Panel believes that therapeutic targeting of elevated triglycerides (≥1.7 mmol/L or 150 mg/dL), a marker of TRL and their remnants, and/or low HDL-C (<1.0 mmol/L or 40 mg/dL) may provide further benefit. The first step should be lifestyle interventions together with consideration of compliance with pharmacotherapy and secondary causes of dyslipidaemia. If inadequately corrected, adding niacin or a fibrate, or intensifying LDL-C lowering therapy may be considered. Treatment decisions regarding statin combination therapy should take into account relevant safety concerns, i.e. the risk of elevation of blood glucose, uric acid or liver enzymes with niacin, and myopathy, increased serum creatinine and cholelithiasis with fibrates. These recommendations will facilitate reduction in the substantial cardiovascular risk that persists in patients with cardiometabolic abnormalities at LDL-C goal

    Étude des mécanismes cellulaires lors de la sénescence foliaire

    No full text
    When switching from green to yellow, a leaf undergoes both morphological and metabolic changes. This process is known as senescence and improved understanding of its mechanisms is important both from a fundamental scientific perspective but also for biotechnological applications. The present thesis reports on several important aspects regarding the cellular and metabolic mechanisms occurring during leaf senescence with an emphasis on the mitochondrial contribution to this process. As a first step, we developed methods to isolate either highly functional crude mitochondria or highly purified mitochondria from leaves of Arabidopsis thaliana. These methods were further used to study mitochondrial contributions to cellular redox homeostasis and to estimate the mitochondrial capacities in leaves undergoing senescence. In particular, we compared the induction of senescence by different dark treatments in Arabidopsis. The comparison between individually darkened leaves and leaves from whole darkened plants revealed different metabolic strategies in response to darkness. Integrating data from measurements of photosynthesis, respiration and confocal laser microscopy with transcriptomic and metabolomic profiling, we suggested that metabolism in leaves of the whole darkened plants enter a ?stand-by mode? with low mitochondrial activity in order to maintain the photosynthetic machinery for as long as possible. In contrast, mitochondria from individually darkened leaves are more active and may provide energy and carbon skeletons for the degradation of cell constituents, facilitating the retrieval of nutrients. We also investigated the dynamics of the microtubular cytoskeleton during dark-induced senescence. Mitochondrial mobility was affected by an early disruption of the microtubules in individually darkened leaves but not in whole darkened plants. In addition, several microtubules associated proteins (MAPs) seemed to be involved in the bundling of the microtubules around the chloroplasts. Altogether, the work presented in this thesis highlights several important steps regarding the metabolic adjustments and the cellular mechanisms in Arabidopsis leaves submitted to prolonged darkness. In particular, we suggest the mitochondria to fulfill specific and important functions during leaf senescence since the role of mitochondria in leaves experiencing prolonged darkness appears very dependant on the whole metabolic status of the plant.Lors de son jaunissement, une feuille subit aussi bien des modifications morphologiques que métaboliques. Ce processus est appelé « sénescence ». Une meilleure compréhension des mécanismes de la sénescence représente un challenge très important non seulement pour la recherche fondamentale mais aussi pour de futures applications en biotechnologie. La thèse présentée ici porte sur d?importants aspects relatifs aux mécanismes cellulaires et métaboliques rencontrés lors de la sénescence foliaire et ce, en apportant une attention particulière à l'implication des mitochondries lors de ce processus. Dans un premier temps, nous avons développé deux méthodes pour isoler, à partir de feuilles d'Arabidopsis, soit des mitochondries conservant leurs fonctionnalités, soit des mitochondries hautement purifiées. Ces méthodes furent utilisées afin d'étudier le rôle des mitochondries dans l'équilibre redox des cellules mais aussi dans le but de déterminer les capacités mitochondriales lors de la sénescence foliaire. Plus précisément, nous avons comparé l'induction de la sénescence foliaire grâce à différents traitements à l'obscurité. Cette comparaison entre des feuilles individuellement placées à l'obscurité et des feuilles provenant d'une plante entièrement disposée à l'obscurité révéla des stratégies métaboliques très différentes. En intégrant des mesures de l'activité photosynthétique, de la respiration et de microscopie laser confocale avec des analyses de transcriptomique et de métabolomique, nous suggérons que le métabolisme d'une feuille provenant d'une plante placée longuement à l'obscurité entre dans un état de « veille » dans le but de maintenir la machinerie photosynthétique fonctionnelle le plus longtemps possible; dans ce cas, les capacités mitochondriales diminuent. A contrario, les mitochondries issues de feuilles individuellement soumises à l'obscurité sont beaucoup plus actives et peuvent par conséquent fournir l'énergie et les squelettes carbonés nécessaires à la dégradation des constituants cellulaires facilitant ainsi la remobilisation des nutriments. Par ailleurs, nous avons aussi mené des investigations sur la dynamique du cytosquelette lors d'une sénescence induite par l'obscurité. La mobilité mitochondriale fut affectée dans les feuilles individuellement soumises à l'obscurité par la dégradation précoce des microtubules ce qui ne fut pas le cas dans les feuilles issues d?une plante entièrement placée à l'obscurité. De plus, un certain nombre de MAPS (microtubules-associated proteins) semblent être impliquées dans l'agrégation des microtubules autour des chloroplastes. Dans son ensemble, cette thèse apporte d?importantes informations quant aux ajustements métaboliques ainsi qu'aux mécanismes cellulaires prenant place lorsque des feuilles d'Arabidopsis sont soumises à une obscurité prolongée. En particulier, nous pensons que les mitochondries ont un rôle prépondérant lors de la sénescence foliaire et que selon le statut métabolique de la plante, les régulations mitochondriales peuvent apparaître divergentes

    From green to yellow : a leaf story

    No full text
    När ett blad gulnar genomgår det både morfologiska och metaboliska förändringar. Denna process benämns senescence och en förbättrad förståelse av dess mekanismer är viktiga både ur ett grundvetenskapligt perspektiv och för potentiella bioteknologiska applikationer. Denna avhandling rapporterar om flera viktiga aspekter relaterade till de cellulära och metaboliska mekanismer som sker under senescencen med tonvikt på mitokondriernas bidrag till denna process. I ett första steg utvecklade vi metoder för att isolera antingen mycket funktionella mitokondrier eller mycket rena mitokondrier från blad av Arabidopsis thaliana. Dessa metoder användes sedan till för att studera mitokondriella bidrag till cellens redox balans och att uppskatta mitokondriernas kapacitet under senescence-processen. Framför allt jämfördes induktionen av senescencen berodende på olika mörkerbehandlingar av Arabidopsis. Jämförelse mellan individuellt mörklagda blad med hela mörklagda växter visade en betydande skillnad i metabolisk strategi mellan de två mörkerbehandlingarna. Genom att integrera data från mätningar av fotosyntes, respiration och konfokal mikroskopi med transcriptomics- och metabolomics-profiler föreslår vi att metabolismen hos blad från helt mörklagda växter antar ett ”stand-by läge” för att kunna bibehålla fotosynteskapaciteten så länge som möjligt. I kontrast till detta visar mitokondrier från individuellt mörklagda blad en hög aktivitet och kan därmed producera energi och kolskelett för degraderingen av cellkomponenter, vilket möjliggör återvinning av näringsämnen. Vi har även studerat dynamiken av det mikrotubulibaserade cytoskelettet under mörkerindicerad senescence. Mitokondriernas rörlighet påverkades av en tidig nedbrytning av mikrotubuli hos individuellt mörklagda blad men inte hos blad där hela växten mörkerbehandlats. Dessutom verkade ett flertal mikrotubuliassocierade proteiner (MAP’s) att vara involverade i buntningen av mikrotubuli runt kloroplasterna. Sammanfattningsvis belyser det arbete som presenteras i denna avhandling ett flertal viktiga steg med avseende på metabolisk anpassning och andra cellulär mekanismer i Arabidopsisblad som utsätts för långvarig mörkerbehendling. Specifikt föreslår vi att mitokondrierna bidrar med speciella och viktiga funktioner under bladens senescence eftersom mitokondriernas roll under långvarig mörkerbehandling av blad verkar bero på den totala statusen av metabolismen hos växten.When switching from green to yellow, a leaf undergoes both morphological and metabolic changes. This process is known as senescence and improved understanding of its mechanisms is important both from a fundamental scientific perspective but also for biotechnological applications. The present thesis reports on several important aspects regarding the cellular and metabolic mechanisms occurring during leaf senescence with an emphasis on the mitochondrial contribution to this process. As a first step, we developed methods to isolate either highly functional crude mitochondria or highly purified mitochondria from leaves of Arabidopsis thaliana. These methods were further used to study mitochondrial contributions to cellular redox homeostasis and to estimate the mitochondrial capacities in leaves undergoing senescence. In particular, we compared the induction of senescence by different dark treatments in Arabidopsis. The comparison between individually darkened leaves and leaves from whole darkened plants revealed different metabolic strategies in response to darkness. Integrating data from measurements of photosynthesis, respiration and confocal laser microscopy with transcriptomics and metabolomics profiling, we suggested that metabolism in leaves of the whole darkened plants enter a “stand-by mode” with low mitochondrial activity in order to maintain the photosynthetic machinery for as long as possible. In contrast, mitochondria from individually darkened leaves are more active and may provide energy and carbon skeletons for the degradation of cell constituents, facilitating the retrieval of nutrients. We also investigated the dynamic of the microtubular cytoskeleton during dark-induced senescence. Mitochondrial mobility was affected by an early disruption of the microtubules in individually darkened leaves but not in whole darkened plants. In addition, several microtubules associated proteins (MAPs) seemed to be involved in the bundling of the microtubules around the chloroplasts. Altogether, the work presented in this thesis highlights several important steps regarding the metabolic adjustments and the cellular mechanisms in Arabidopsis leaves submitted to prolonged darkness. In particular, we suggest the mitochondria to fulfill specific and important functions during leaf senescence since the role of mitochondria in leaves experiencing prolonged darkness appears very dependant on the whole metabolic status of the plant

    Tissue-specific isolation of tagged Arabidopsis plastids

    No full text
    Plastids are found in all plant cell types. However, most extraction methods to study these organelles are performed at the organ level (e.g., leaf, root, fruit) and do not allow for tissue-specific resolution, which hinders our understanding of their physiology. Therefore, IPTACT (Isolation of Plastids TAgged in specific Cell Types) was developed to isolate plastids in a tissue-specific manner in Arabidopsis thaliana (Arabidopsis). Plastids are biotinylated using one-shot transgenic lines, and tissue specificity is achieved with a suitable promoter as long as such a promoter exists. Cell-specific biotinylated plastids are then isolated with 2.8-µm streptavidin beads. Plastids extracted by IPTACT are suitable for RNA or protein isolation and subsequent tissue-specific OMICs analyses. This method provides the user with a powerful tool to investigate plastidial functions at cell-type resolution. Furthermore, it can easily be combined with studies using diverse genetic backgrounds and/or different developmental or stress conditions

    From green to yellow (a leaf story)

    No full text
    Lors de son jaunissement, une feuille subit aussi bien des modifications morphologiques que métaboliques. Ce processus est appelé sénescence . Une meilleure compréhension des mécanismes de la sénescence représente un challenge très important non seulement pour la recherche fondamentale mais aussi pour de futures applications en biotechnologie. La thèse présentée ici porte sur d importants aspects relatifs aux mécanismes cellulaires et métaboliques rencontrés lors de la sénescence foliaire et ce, en apportant une attention particulière à l implication des mitochondries lors de ce processus. Dans un premier temps, nous avons développé deux méthodes pour isoler, à partir de feuilles d Arabidopsis, soit des mitochondries conservant leurs fonctionnalités, soit des mitochondries hautement purifiées. Ces méthodes furent utilisées afin d étudier le rôle des mitochondries dans l équilibre redox des cellules mais aussi dans le but de déterminer les capacités mitochondriales lors de la sénescence foliaire. Plus précisément, nous avons comparé l induction de la sénescence foliaire grâce à différents traitements à l obscurité. Cette comparaison entre des feuilles individuellement placées à l obscurité et des feuilles provenant d une plante entièrement disposée à l obscurité révéla des stratégies métaboliques très différentes. En intégrant des mesures de l activité photosynthétique, de la respiration et de microscopie laser confocale avec des analyses de transcriptomique et de métabolomique, nous suggérons que le métabolisme d une feuille provenant d une plante placée longuement à l obscurité entre dans un état de veille dans le but de maintenir la machinerie photosynthétique fonctionnelle le plus longtemps possible; dans ce cas, les capacités mitochondriales diminuent. A contrario, les mitochondries issues de feuilles individuellement soumises à l obscurité sont beaucoup plus actives et peuvent par conséquent fournir l énergie et les squelettes carbonés nécessaires à la dégradation des constituants cellulaires facilitant ainsi la remobilisation des nutriments. Par ailleurs, nous avons aussi mené des investigations sur la dynamique du cytosquelette lors d une sénescence induite par l obscurité. La mobilité mitochondriale fut affectée dans les feuilles individuellement soumises à l obscurité par la dégradation précoce des microtubules ce qui ne fut pas le cas dans les feuilles issues d une plante entièrement placée à l obscurité. De plus, un certain nombre de MAPS (microtubules-associated proteins) semblent être impliquées dans l agrégation des microtubules autour des chloroplastes. Dans son ensemble, cette thèse apporte d importantes informations quant aux ajustements métaboliques ainsi qu aux mécanismes cellulaires prenant place lorsque des feuilles d Arabidopsis sont soumises à une obscurité prolongée. En particulier, nous pensons que les mitochondries ont un rôle prépondérant lors de la sénescence foliaire et que selon le statut métabolique de la plante, les régulations mitochondriales peuvent apparaître divergentes.When switching from green to yellow, a leaf undergoes both morphological and metabolic changes. This process is known as senescence and improved understanding of its mechanisms is important both from a fundamental scientific perspective but also for biotechnological applications. The present thesis reports on several important aspects regarding the cellular and metabolic mechanisms occurring during leaf senescence with an emphasis on the mitochondrial contribution to this process. As a first step, we developed methods to isolate either highly functional crude mitochondria or highly purified mitochondria from leaves of Arabidopsis thaliana. These methods were further used to study mitochondrial contributions to cellular redox homeostasis and to estimate the mitochondrial capacities in leaves undergoing senescence. In particular, we compared the induction of senescence by different dark treatments in Arabidopsis. The comparison between individually darkened leaves and leaves from whole darkened plants revealed different metabolic strategies in response to darkness. Integrating data from measurements of photosynthesis, respiration and confocal laser microscopy with transcriptomic and metabolomic profiling, we suggested that metabolism in leaves of the whole darkened plants enter a stand-by mode with low mitochondrial activity in order to maintain the photosynthetic machinery for as long as possible. In contrast, mitochondria from individually darkened leaves are more active and may provide energy and carbon skeletons for the degradation of cell constituents, facilitating the retrieval of nutrients. We also investigated the dynamics of the microtubular cytoskeleton during dark-induced senescence. Mitochondrial mobility was affected by an early disruption of the microtubules in individually darkened leaves but not in whole darkened plants. In addition, several microtubules associated proteins (MAPs) seemed to be involved in the bundling of the microtubules around the chloroplasts. Altogether, the work presented in this thesis highlights several important steps regarding the metabolic adjustments and the cellular mechanisms in Arabidopsis leaves submitted to prolonged darkness. In particular, we suggest the mitochondria to fulfill specific and important functions during leaf senescence since the role of mitochondria in leaves experiencing prolonged darkness appears very dependant on the whole metabolic status of the plant.NANCY1-Bib. numérique (543959902) / SudocSudocFranceF
    corecore