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URI permanente para esta coleçãohttps://locus.ufv.br/handle/123456789/11846
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Item Enhanced photosynthesis and growth in atquac1 knockout mutants are due to altered organic acid accumulation and an increase in both stomatal and mesophyll conductance(Plant Physiology, 2015-11-05) Medeiros, David B.; Martins, Samuel C.V.; Cavalcanti, João Henrique F.; Daloso, Danilo M.; Martinoia, Enrico; Nunes-Nesi, Adriano; DaMatta, Fábio M.; Fernie, Alisdair R.; Araújo, Wagner L.Stomata control the exchange of CO2 and water vapor in land plants. Thus, whereas a constant supply of CO2 is required to maintain adequate rates of photosynthesis, the accompanying water losses must be tightly regulated to prevent dehydration and undesired metabolic changes. Accordingly, the uptake or release of ions and metabolites from guard cells is necessary to achieve normal stomatal function. The AtQUAC1, an R-type anion channel responsible for the release of malate from guard cells, is essential for efficient stomatal closure. Here, we demonstrate that mutant plants lacking AtQUAC1 accumulated higher levels of malate and fumarate. These mutant plants not only display slower stomatal closure in response to increased CO2 concentration and dark but are also characterized by improved mesophyll conductance. These responses were accompanied by increases in both photosynthesis and respiration rates, without affecting the activity of photosynthetic and respiratory enzymes and the expression of other transporter genes in guard cells, which ultimately led to improved growth. Collectively, our results highlight that the transport of organic acids plays a key role in plant cell metabolism and demonstrate that AtQUAC1 reduce diffusive limitations to photosynthesis, which, at least partially, explain the observed increments in growth under well-watered conditions.Item Impaired malate and fumarate accumulation due to the mutation of the tonoplast dicarboxylate transporter has little effects on stomatal behavior(Plant Physiology, 2017-09-12) Medeiros, David B.; Barros, Kallyne A.; Barros, Jessica Aline S.; Omena-Garcia, Rebeca P.; Arrivault, Stéphanie; Sanglard, Lílian M.V.P.; Detmann, Kelly C.; Silva, Willian Batista; Daloso, Danilo M.; DaMatta, Fábio M.; Nunes-Nesi, Adriano; Fernie, Alisdair R.; Araújo, Wagner L.Malate is a central metabolite involved in a multiplicity of plant metabolic pathways, being associated with mitochondrial metabolism and playing significant roles in stomatal movements. Vacuolar malate transport has been characterized at the molecular level and is performed by at least one carrier protein and two channels in Arabidopsis (Arabidopsis thaliana) vacuoles. The absence of the Arabidopsis tonoplast Dicarboxylate Transporter (tDT) in the tdt knockout mutant was associated previously with an impaired accumulation of malate and fumarate in leaves. Here, we investigated the consequences of this lower accumulation on stomatal behavior and photosynthetic capacity as well as its putative metabolic impacts. Neither the stomatal conductance nor the kinetic responses to dark, light, or high CO2 were highly affected in tdt plants. In addition, we did not observe any impact on stomatal aperture following incubation with abscisic acid, malate, or citrate. Furthermore, an effect on photosynthetic capacity was not observed in the mutant lines. However, leaf mitochondrial metabolism was affected in the tdt plants. Levels of the intermediates of the tricarboxylic acid cycle were altered, and increases in both light and dark respiration were observed. We conclude that manipulation of the tonoplastic organic acid transporter impacted mitochondrial metabolism, while the overall stomatal and photosynthetic capacity were unaffected.Item Autophagy deficiency compromises alternative pathways of respiration following energy deprivation in Arabidopsis thaliana(Plant Physiology, 2017-09-22) Barros, Jessica A.S.; Cavalcanti, João Henrique F.; Medeiros, David B.; Nunes-Nesi, Adriano; Avin-Wittenberg, Tamar; Fernie, Alisdair R.; Araújo, Wagner L.Under heterotrophic conditions, carbohydrate oxidation inside the mitochondrion is the primary energy source for cellular metabolism. However, during energy-limited conditions, alternative substrates are required to support respiration. Amino acid oxidation in plant cells plays a key role in this by generating electrons that can be transferred to the mitochondrial electron transport chain via the electron transfer flavoprotein/ubiquinone oxidoreductase system. Autophagy, a catabolic mechanism for macromolecule and protein recycling, allows the maintenance of amino acid pools and nutrient remobilization. Although the association between autophagy and alternative respiratory substrates has been suggested, the extent to which autophagy and primary metabolism interact to support plant respiration remains unclear. To investigate the metabolic importance of autophagy during development and under extended darkness, Arabidopsis (Arabidopsis thaliana) mutants with disruption of autophagy (atg mutants) were used. Under normal growth conditions, atg mutants showed lower growth and seed production with no impact on photosynthesis. Following extended darkness, atg mutants were characterized by signatures of early senescence, including decreased chlorophyll content and maximum photochemical efficiency of photosystem II coupled with increases in dark respiration. Transcript levels of genes involved in alternative pathways of respiration and amino acid catabolism were up-regulated in atg mutants. The metabolite profiles of dark-treated leaves revealed an extensive metabolic reprogramming in which increases in amino acid levels were partially compromised in atg mutants. Although an enhanced respiration in atg mutants was observed during extended darkness, autophagy deficiency compromises protein degradation and the generation of amino acids used as alternative substrates to the respiration.Item Impaired malate and fumarate accumulation due the mutation of tonoplast dicarboxylate transporter(Plant Physiology, 2017-09-20) Medeiros, David B.; Barros, Kallyne; Barros, Jessica AS; Omena-Garcia, Rebeca P; Detmann, Kelly C; Silva, Willian Batista; DaMatta, Fabio; Nunes-Nesi, Adriano; Araújo, Wagner L.; Arrivault, Stéphanie; Sanglard, Lilian Vincis Pereira; Daloso, Danilo M.; Fernie, Alisdair R.Malate is a central metabolite involved in a multiplicity of plant metabolic pathways, being associated with mitochondrial metabolism and playing significant roles n stomatal movements. Vacuolar malate transport has been characterized at the molecular level and is performed by at least one carrier protein and two channels in Arabidopsis vacuoles. The absence of the Arabidopsis thaliana tonoplast Dicarboxylate Transporter (tDT) in tdt knockout mutant was previously associated with an impaired accumulation of malate and fumarate in leaves. Here, we investigated the consequences of this lower accumulation on stomatal behaviour and photosynthetic capacity as well as ts putative metabolic impacts. Neither the stomatal conductance (g s ) nor the kinetic responses to dark, light or high CO 2 were highly affected in tdt plants. In addition, we did not observe any impact on stomatal aperture following incubation with either bscisic acid (ABA), malate or citrate. Further, an effect on photosynthetic capacity was not observed in the mutant lines. However, leaf mitochondrial metabolism was affected in the tdt plants. Levels of the intermediates of the tricarboxylic acid cycle were altered and increases in both light and dark respiration were observed. We conclude that manipulation of the tonoplastic organic acid transporter impacted mitochondrial metabolism, while the overall stomatal and photosynthetic capacity were unaffected.Item Knockout mutants are due to altered organic acid accumulation and an increase in both stomatal and mesophyll conductance(Plant Physiology, 2003-11-30) Medeiros, David B.; Martins, Samuel C.V.; Cavalcanti, João Henrique F.; Daloso, Danilo M.; Martinoia, Enrico; Nunes-Nesi, Adriano; DaMatta, Fábio M.; Fernie, Alisdair R.; Araújo, Wagner L.Stomata control the exchange of CO 2 and water vapor in land plants. Thus, whereas a constant supply of CO 2 is required to maintain adequate rates of photosynthesis, the accompanying water losses must be tightly regulated to prevent dehydration and undesired metabolic changes. Accordingly, the uptake or release of ions and metabolites from guard cells is necessary to achieve normal stomatal function. The AtQUAC1, an R-type anion channel responsible for the release of malate from guard cells, is essential for efficient stomatal closure. Here, we demonstrate that mutant plants lacking AtQUAC1 accumulated higher levels of malate and fumarate. These mutant plants not only display slower stomatal closure in response to increased CO 2 concentration and dark but are also characterized by improved mesophyll conductance. These responses were accompanied by increases in both photosynthesis and respiration rates, without affecting the activity of photosynthetic and respiratory enzymes and the expression of other transporter genes in guard cells, which ultimately led to improved growth. Collectively, our results highlight that the transport of organic acids plays a key role in plant cell metabolism and demonstrate that AtQUAC1 reduce diffusive limitations to photosynthesis, which, at least partially, explain the observed increments in growth under well-watered conditions.