High temperature often leads to failure of grain filling in rice (Oryza sativa) causing yield loss, but the underlying mechanisms are still not elucidated. Here, we report that two genes encoding ...seed-specific NAM/ATAF/CUC (NAC) domain transcription factors, ONAC127 and ONAC129, are responsive to heat stress and involved in the grain filling process of rice. ONAC127 and ONAC129 are dominantly expressed in the pericarp and can form a heterodimer during rice grain filling. CRISPR/Cas9 induced mutants and overexpression lines were then generated to investigate the function of these two transcription factors. Interestingly, both knock-out and overexpression plants showed incomplete grain filling and shrunken grains, which became more severe under heat stress. Transcriptome analysis revealed that ONAC127 and ONAC129 mainly regulate stimulus response and nutrient transport. ChIP-seq analysis identified that the direct target genes of ONAC127 and ONAC129 in developing rice seeds include monosaccharide transporter gene OsMST6, sugar transporter gene OsSWEET4, calmodulin-like protein gene OsMSR2 and AP2/ERF factor gene OsEATB. These results suggest that ONAC127 and ONAC129 regulate grain filling by affecting sugar transportation and abiotic stress responses. Overall, this study demonstrates a transcriptional regulatory network with ONAC127 and ONAC129 coordinating multiple pathways to modulate seed development and heat stress responses at rice reproductive stages.
• Far-red (FR) light promotes fruit growth by increasing dry mass partitioning to fruits, but the mechanism behind this is unknown. We hypothesise that it is due to an increased fruit sink strength ...as FR radiation enhances sugar transportation and metabolism.
• Tomato plants were grown with or without 50–80 μmol m−2 s−1 of FR radiation added to a common background 150–170 μmol m−2 s−1 red + blue light-emitting diode lighting. Potential fruit growth, achieved by pruning each truss to one remaining fruit, was measured to quantify fruit sink strength. Model simulation was conducted to test whether the measured fruit sink strength quantitatively explained the FR effect on dry mass partitioning. Starch, sucrose, fructose and glucose content were measured. Expression levels of key genes involved in sugar transportation and metabolism were determined.
• FR radiation increased fruit sink strength by 38%, which, in model simulation, led to an increased dry mass partitioned to fruits that quantitatively agreed very well with measured partitioning. FR radiation increased fruit sugar concentration and upregulated the expression of genes associated with both sugar transportation and metabolism.
• This is the first study to demonstrate that FR radiation stimulates dry mass partitioning to fruits mainly by increasing fruit sink strength via simultaneous upregulation of sugar transportation and metabolism.
Chloroplasts have important roles in photosynthesis, stress sensing and retrograde signaling. However, the relationship between chloroplast peptide chain release factor and ROS-mediated plant growth ...is still unclear. In the present study, we obtained a loss-of-function mutant
by EMS mutation. The
mutant has few lateral roots and a pale green leaf phenotype. By map-based cloning, the
gene was located on AT3G62910, with a point mutation leading to amino acid substitution in functional release factor domain. Using yeast-two-hybrid and BiFC, we confirmed
protein was characterized locating in chloroplast by co-localization with plastid marker and interacting with ribosome-related proteins. Through observing by transmission electron microscopy, quantifying ROS content and measuring the transport efficiency of plasmodesmata in dig8 mutant, we found that abnormal thylakoid stack formation and chloroplast dysfunction in the
mutant caused increased ROS activity leading to callose deposition and lower PD permeability. A local sugar supplement partially alleviated the growth retardation phenotype of the mutant. These findings shed light on chloroplast peptide chain release factor-affected plant growth by ROS stress.
Sugar unloading in grape berries is a crucial step in the long-distance transport of carbohydrates from grapevine leaves to berries. Brassinosteroids (BRs) mediate many physiological processes in ...plants including carbohydrate metabolism. Here, ‘Cabernet Sauvignon’ (Vitis vinifera L.) grape berries cultivated in clay loam fields were treated with an exogenous BR (24-epibrassinolide; EBR), a BR synthesis inhibitor (brassinazole; Brz), Brz + EBR (sprayed with EBR 24 h after a Brz treatment), and deionized water (control) at the onset of véraison. The EBR treatment sharply increased the soluble sugars content in the berries, but decreased it in the skins. The EBR and Brz + EBR treatments significantly promoted the activities of both invertases (acidic and neutral) and sucrose synthase (sucrolytic) at various stages of ripening. The mRNA levels of genes encoding sucrose metabolic invertase (VvcwINV), and monosaccharide (VvHT3, 4, 5 and 6) and disaccharide (VvSUC12 and 27) transporters were increased by the EBR and/or Brz + EBR treatments. Generally, the effects of the Brz treatment on the measured targets contrasted with the effects of the EBR treatments. The EBR and Brz treatments inhibited the biosynthesis of the endogenous BRs 6-deoxocastastarone and castasterone. Both EBR and Brz + EBR treatments increased the brassinolide contents, down-regulated the expression of genes encoding BRs biosynthetic enzymes BRASSINOSTEROID-6-OXIDASE and DWARF1, (VvBR6OX1 and VvDWF1) and induced BR receptor gene BRASSINOSTEROID INSENSITIVE 1 (VvBRI1) expression in deseeded berries. Together, these results show that BRs are involved in controlling sugar unloading in grape berries during véraison.
The EBR and Brz + EBR treatments significantly increased the reducing sugars in ‘Cabernet Sauvignon’ (Vitis vinifera L.) grape berries. Display omitted
•24-Epibrassinolide treatment rapidly increased the sugars content in grape berries.•24-Epibrassinolide treatment increased sucrose synthase and invertase activities.•24-Epibrassinolide treatment altered the mRNA levels of sugar transporter genes.•24-Epibrassinolide treatment altered the biosynthesis of endogenous brassinosteroids.
The eastern foot of Helan Mountains in Ningxia belongs to the semiarid area and has been identified as the best wine grape producing area in China. In order to solve the problems of a high sugar ...content, organic acid falling too fast and severe shortage of acidity in the berries during ripening, which lead to lack of wine harmony, this study took Vitis. vinifera L. cv. 'Cabernet Sauvignon' as the experimental material and expanded nine treatments from setting to harvest. Nine rows were selected and divided into three groups, treated with mild (-0.20 MPa ≥ Ψb ≥ -0.40 MPa), moderate (-0.40 MPa ≥ Ψb ≥ -0.60 MPa), severe (Ψb ≥ -0.60 MPa) water stress from setting to veraison separately. From veraison to harvest, within each group, the mild, moderate, severe water stress were set, forming light-light (CK), light-medium (T1), light-heavy (T2), medium-light (T3), medium-medium (T4), medium-heavy (T5), heavy-light (T6), heavy-medium (T7), heavy-heavy (T8) treatments. The results showed that when the medium treatment was used from setting to veraison and the medium or severe treatment was used during post-veraison to harvest, it would facilitate the transportation and accumulation of sugar and improve the fruit quality.
The ability of Streptococcus mutans to survive and cause dental caries is dependent on its ability to metabolize various carbohydrates, accompanied by extracellular polysaccharide synthesis and ...biofilm formation. Here, the role of an
competence-related regulator (RcrR) in the regulation of multiple sugar transportation and biofilm formation is reported. The deletion of the
gene in S. mutans caused delayed growth, decreased biofilm formation ability, and affected the expression level of its multiple sugar transportation-related genes. Transcriptional profiling revealed 17 differentially expressed genes in the
mutant. Five were downregulated and clustered with the sugar phosphotransferase (PTS) systems (mannitol- and trehalose-specific PTS systems). The conserved sites bound by the
promoter were then determined by electrophoretic mobility shift assays (EMSAs) and DNase I footprinting assays. Furthermore, a potential binding motif in the promoters of the two PTS operons was predicted using MEME Suite 5.1.1. RcrR could bind to the promoter regions of the two operons
, and the sugar transporter-related genes of the two operons were upregulated in an
-overexpressing strain. In addition, when RcrR-binding sites were deleted, the growth rates and final yield of S. mutans were significantly decreased in tryptone-vitamin (TV) medium supplemented with different sugars, but not in absolute TV medium. These results revealed that RcrR acted as a transcription activator to regulate the two PTS systems, accompanied by multiple sugar transportation and biofilm formation. Collectively, these results indicate that RcrR is a critical transcription factor in S. mutans that regulates bacterial growth, biofilm formation, and multiple sugar transportation.
The human oral cavity is a constantly changing environment. Tooth decay is a commonly prevalent chronic disease mainly caused by the cariogenic bacterium Streptococcus mutans. S. mutans is an oral pathogen that metabolizes various carbohydrates into extracellular polysaccharides (EPSs), biofilm, and tooth-destroying lactic acid. The host diet strongly influences the availability of multiple carbohydrates. Here, we showed that the RcrR transcription regulator plays a significant role in the regulation of biofilm formation and multiple sugar transportation. Further systematic evaluation of how RcrR regulates the transportation of various sugars and biofilm formation was also conducted. Notably, this study decrypts the physiological functions of RcrR as a potential target for the better prevention of dental caries.
Dans son environnement la plante est confrontée à une variété de microorganismes bénéfiques, neutres et pathogènes, qui sont fortement dépendants des ressources carbonées qu’elle libère dans le sol. ...Le transport de sucres, processus clé de la physiologie de la plante, est essentiel pour les interactions plantes-microorganismes et leur devenir. Au cours de l'évolution, les plantes ont acquis des mécanismes leur permettant de percevoir les signaux microbiens du milieu extérieur, et aboutissant à la transduction d’un signal spécifique puis à des réponses biologiques adaptées (défense versus mutualisme) à la stratégie du microorganisme. Ces réponses assurent la survie et le développement des plantes. Mes travaux de thèse, menés avec un système « d’interaction » simplifié, contribuent à une meilleure compréhension des mécanismes sous-jacents au déterminisme des interactions plantes-microorganismes. Ce système a permis d’étudier, sur des suspensions cellulaires de N. tabacum, les réponses cellulaires précoces déclenchées suite à la perception de molécules microbiennes provenant de microorganismes à stratégie pathogène avirulent ou à stratégie mutualiste. Nous avons mesuré des évènements de signalisation et des flux de sucres induits en réponse à ces molécules microbiennes. Nos résultats ont mis en évidence que les chitotétrasaccharides (CO4), sécrétés par les champignons mycorhiziens à arbuscules dans les stades pré-symbiotiques de l’interaction, mobilisent les mêmes événements de signalisation précoce (H2O2 dépendant de la protéine rbohD, Ca2+ cytosolique, activation de MAPK) que la cryptogéine, un éliciteur des réactions de défense ; mais avec des réponses différentes en terme d’intensité et de cinétique. Les CO4 et la cryptogéine ont par ailleurs montré des impacts distincts sur les flux de sucres et l’expression de transporteurs impliqués. En complément nous avons montré un effet de la modification de la dynamique membranaire associée à la clathrine sur des évènements de signalisation déclenchés par la cryptogéine, ainsi que dans les flux entrants de sucres et l’expression de gènes de transporteurs de sucres. Enfin, l’analyse in silico de l’interactome de transporteurs de sucres chez la plante modèle A. thaliana, nous a permis d’apporter des connaissances supplémentaires quant aux évènements de régulations des transporteurs de sucres et l’identification de protéines régulatrices putatives en interaction avec ces derniers. L’ensemble de ces travaux ouvrent la voie à de nouvelles recherches visant à élucider les mécanismes cellulaires et moléculaires impliqués dans la mise en place des interactions entre plantes et microorganismes.
In their natural environment plants are in close interaction with beneficial, neutral, or pathogenic microbes, which are highly dependent on carbon resources exuded by plant roots. Sugar transport, which is a key process of plant physiology, is essential to support the fate of plant-microbe interactions. During evolution, plants have acquired the ability to perceive microbial molecules, initiating specific signal transduction cascades and leading to adapted response for microbe lifestyles (avirulent, virulent, or benefic). Plant survival will depend on the nature of the induced mechanisms. My PhD work, carried out on a simplified experimental system, contributes to the understanding of mechanisms underlying the determinism of plant-microbe interactions. We used Nicotiana tabacum cells in suspension exposed to microbial molecules derived from mutualistic or avirulent microbes. Using such a simplified system, we analyzed elements of the early signaling cascade and sugar fluxes. We have shown that CO4, which is originating from AMF, initiate early signaling components (rbohD-dependent H2O2, cytosolic Ca2+, MAPK activation) as cryptogein, a defense elicitor, but with distinct profile and amplitude. Those two molecules (CO4 and cryptogein) are responsible of different effects on sugar fluxes and the expression of the underlying sugar transporter genes. In addition, we presented an impact of the alteration of clathrin-mediated process on early signaling events triggered by cryptogein, as well as inward sugar fluxes and expression of sugar transporter genes. Finally, in silico analyses of sugar transporter interactome in Arabidopsis thaliana has provided some possible regulation mechanisms through the identification of new candidate proteins involved in sugar transporter regulation. These information open new perspectives towards a better understanding of the cellular and molecular mechanisms involved in plant-microbe interactions.
La distribution des sucres est un processus clé dans le développement de la plante et cours des interactions plantes/microorganismes.Une recherche des acteurs moléculaires impliqués dans la ...répartition des ressources carbonées au cours de l'interaction avec le champignon nécrotrophe B. cinerea a été réalisée. Plusieurs familles de transporteurs de sucres et d'invertases ont été ciblées, permettant d'établir une cartographie des gènes régulés transcriptionnellement lors de l'interaction. Le rôle de certains gènes candidats a été étudié par une approche de génomique fonctionnelle afin de mettre en évidence une fonction biologique de l'allocation du carbone dans la résistance de la plante aux champignons nécrotrophes. Un système d'interaction simplifié, basé sur un dialogue moléculaire sans contact physique entre une culture cellulaire d'A. thaliana et B. cinerea, a été développé. Il a permis de mesurer les flux de sucres ainsi que les activités enzymatiques et métaboliques pour chaque partenaire. Nos résultats montrent que B. cinerea entraine une forte augmentation de l'activité invertasique pariétale dans les tissus infectés, indiquant qu'une transition source/puits a lieu. Plusieurs transporteurs de sucres sont différentiellement exprimés, certains d'entre eux modulant le devenir de l'interaction. L'activité d'absorption d'hexoses et le métabolisme primaire des cellules hôtes sont fortement stimulés, démontrant l'importance de la compétition pour les sucres à l'interface plante/agent pathogène. En conclusion, l'absorption des sucres alimente le métabolisme énergétique des cellules hôtes et participe aux mécanismes de défense de la plante.
During plant development and upon pathogen infection, sugar allocation is a key process in plant physiology. Cell wall invertases and sugar transporters, involved in the sink strength, likely play a major role in the metabolic plant response. Molecular actors involved in carbohydrates allocation upon B. cinerea interaction have been identified using a transcriptional approach. Some gene families of sugar transporters and invertases have been targeted, allowing the establishment of a cartography of genes regulated during the interaction. To understand the biological role of carbon allocation during the interaction between plants and necrotrophic fungi, candidate genes have been studied using a functional genomics approach.A simplified interaction system has been developped, allowing a molecular dialogue between Arabidopsis and B. cinerea cells, without any physical contact. This system enables the monitoring of radiolabelled sugar uptake rates and some enzymatic and metabolomic activities for both the host cells and the pathogen, independently.Globally, our results demonstrate that B. cinerea infection leads to the transition from a source to a sink tissue, with a strong increase in cell wall invertase activity. The expression of some sugar transporter genes is also affected, while some of them (AtSTP1 and 13) are involved in the disease development. Besides the increase in hexose uptake activity, primary metabolism is deeply affected, highlighting the competition for apoplastic sugars that takes place at the plant/pathogen interface. Sugar retrieval appears to be a key process, fuelling host cells with energy and signal molecules, contributing to the plant defense mechanisms.
Le potassium (K) est un élément majeur connu pour contribuer à la résistance des plantes à la sècheresse. L'étudede son influence sur la réponse physiologique du maïs (Zea mays L.) sous contrainte ...hydrique est essentiellepour prédire la future productivité dans un contexte de changements climatiques, en particulier de la diminutiondes précipitations. Des modalités d'apports en K et en eau ont été croisées et soumises à des plants de maïs,élevés en condition contrôlées ou cultivés au champ. La croissance (biomasses aériennes et racinaires,rendements en grain) ainsi que les mécanismes écophysiologiques du métabolisme carboné (photosynthèse,transport des sucres) et du statut hydrique (transpiration, conductance stomatique, potentiels hydriques) ont étéétudiés. L'apport de K a contribué à l'augmentation de la croissance, le développement et le rendement grain quelque soit le régime hydrique imposé au maïs et les conditions d'expérimentation. Les résultats attendus sur lameilleure régulation stomatique en cas de déficit hydrique sont moins évidents. L'effet du stress hydrique ou dela déficience en K tendent à diminuer la photosynthèse. Cependant, ces effets ressortent plus sur les feuillesâgées que sur les feuilles jeunes. Dans ces mêmes conditions, le transport des sucres ne semble pas être unélément limitant de la croissance. Plusieurs résultats convergent pour attribuer au K un rôle dans la maîtrise despertes en eau (par unité de surface foliaire) et sur la meilleure efficience d'utilisation de l'eau. Néanmoins, cetteefficience est imputée à des meilleurs rendements, liés à une surface foliaire plus importante, et non pas à unemoindre consommation de l'eau.
Potassium (K) is a major nutrient known to help plants resist drought. In the context of climate change,quantifying the role of K on maize physiological acclimation to reduced precipitations is essential to betterpredict future productivity. Maize (Zea mays L.) plants grown under controlled or field conditions weresubmitted to different K and water levels. Plant growth (shoot and root biomass, grain yield) as well as plantwater status (transpiration, stomatal conductance, water potential) and ecophysiological mechanisms of Carbonmetabolism (photosynthesis, sugar transport) were studied. Regardless of the water regime and experimentalconditions, K nutrition increased growth and whole-plant development and improved grain yield. The effect ofwater stress on stomatal regulation was not straightforward and depended on the level of K fertilization. Theeffects of water or K deficit tend to decrease photosynthesis. Drought or K nutrition affected more leafphotosynthesis in old than in young leaves, and sugar transport did not seem to be a growth limiting factor. Ourresults demonstrated a strong effect of K on biomass production and a higher water use efficiency with less of animpact on leaf-level physiology. This better water use was mainly the consequence of the positive effect of leafarea on yield, and not due to a reduce water use.
Chez les plantes supérieures, la distribution des sucres entre organes sources et puits requiert l'activité de transporteurs membranaires de sucre. Les flux de sucre variant au cours du développement ...de la plante et en réponse à des stress, il est logique de penser que les transporteurs de sucres sont impliqués dans ces changements. Le but du travail était de suivre la répartition des sucres et l'expression des transporteurs correspondants dans les racines de la plante modèle Arabidopsis thaliana, en réponse à un stress osmotique. Afin de pouvoir récolter des racines, un système de culture en hydroponie a été mis en place. Après avoir vérifié l'homogénéité des plantes cultivées dans ce système, une étude de l'expression des gènes de transporteurs de sucre a été effectuée au cours du développement des plantes et de d'une journée (24h), en utilisant la technique de macroarray. Cette étude a révélé l'expression dans les racines de 3 transporteurs de saccharose (AtSUC1, AtSUC2 et AtSUC5), 2 transporteurs de polyols (AtPLT5 et AtPLT6) et 2 transporteurs d'hexoses (AtSTP7 et AtSTP13). Le suivi de la teneur en sucres solubles et en amidon ainsi que du transport à longue distance de U-14C-saccharose a permis d'émettre des hypothèses quant au rôle des transporteurs de sucres exprimés dans la racine. Afin de mimer un stress hydrique en hydroponie, un protocole d'application progressif d'un agent osmotique (polyéthylène glycol 6000) dans le milieu de culture a été élaboré. Ce système a permis de mettre en évidence que 5 des gènes de transporteurs de sucre identifiés dans la racine ont une expression qui varie dans ces conditions. Trois d'entre eux sont fortement réprimés : AtSUC1
In plants, sugar allocation between source and sink organs is based on the activity of membrane transporters for sugars. As sugar fluxes are changing during development and in response to stress, sugar transporters are supposed to be involved in those changes. The aim of this work was to study sugar allocation and the corresponding transporters gene expression in the roots of the model plant Arabidopsis thaliana during an osmotic stress. In order to have access to the roots, an hydroponic culture system was developped. The homogeneity of the plants obtained with this system was checked and the expression of sugar tranporter genes was followed during development and during a 24h period was studied by a macro-array technique. The expression in the roots of the following genes was found: 3 sucrose transporters (AtSUC1, AtSUC2 and AtSUC5), 2 polyol transporters (AtPLT5 and AtPLT6) and 2 hexose transporters (AtSTP7 and AtSTP13). The sugar and starch content and the long distance of 14C-sucrose were measured and used to build some hypothesis on the role of sugar transporters in the roots. To mimick a water stress, an osmotic stress due to the progressive addition of Polyethylene-glycol was applied. This system demonstrated that 5 of the identified transporter genes display a change in expression: 3 are repressed (AtSUC1, AtSUC5 and AtPLT6) and 2 are over expressed (AtSUC2 and AtSTP13). Moreover, soluble sugar and starch accumulate in the leaves and 14C-sucrose transport to the roots is decreased in plants subjected to an osmotic stress. The respective role of transporters is discussed. The gene expression data were also confirmed with plants grown in rhizoboxes.
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