Functional characterization of Sl-ERF.B3, a member of the large multi-gene family of Ethylene Response Factor in tomato (Solanum lycopersicum)

Les derniers acteurs de la voie de signalisation à l’éthylène sont des facteurs de transcription appelés ERF (Ethylene Response Factors). La connaissance de leur rôle spécifique dans la régulation des processus développementaux dépendant de l’éthylène reste limitée. Les travaux présentés dans la thèse concernent la caractérisation fonctionnelle du gène Sl-ERF.B3, un membre de cette grande famille de régulateurs transcriptonnels dans la tomate (Solanum lycopersicum). Utilisant une stratégie répresseur dominant ; il est montré en particulier que ce gène intervient dans la mise en place de la réponse à l’éthylène et dans le contrôle de la maturation du fruit. L’expression d’une construction ERF.B3-SRDX, une version chimérique de Sl-ERF.B3 fusionné à un domaine répresseur de type EAR, entraine des phénotypes pléotropiques aussi bien dans la signalisation de l’éthylène que dans le développement des parties végétatives et des organes reproducteurs. Ainsi, une altération de la triple réponse à l’éthylène est constatée chez les lignées transgéniques et au stade adulte, les plantes présentent des phénotypes d’épinastie des feuilles, de sénescence prématurée des fleurs et d’abscission accélérée des fruits. L’ensemble de ces observations est corrélée avec une modification de l’expression de gènes impliqués dans la biosynthèse et la réponse à l’éthylène. Ces données suggèrent que ERF.B3 intervient dans un mécanisme de rétro-control de la réponse à l’éthylène en agissant à la fois sur les gènes de biosynthèse et de signalisation de l’hormone. Au niveau du fruit, la sur-expression d’ERF.B3-SRDX entraine une modification du processus de maturation avec un retard notable de l’avènement de l’acquisition de la compétence à murir. Cependant, une fois la maturation initiée, elle s’accompagne d’une forte production d’éthylène et d’une accélération du ramollissement du fruit. A l’inverse, l’accumulation de pigment est inhibée par altération de la voie de biosynthèse des caroténoïdes. Ces données phénotypiques sont corrélées avec le niveau d’expression des gènes clés impliqués dans ces processus. Les résultats indiquent que dans les lignées transgéniques, il y a découplage de certaines caractéristiques de la maturation du fruit et permettent de mettre en lumière le rôle d’ERF.B3 dans la régulation des processus de développement dépendant de l’éthylène chez la tomate. ABSTRACT : Ethylene Response Factors (ERFs) are known to be the last transcription factors of the ethylene transduction pathway. Their specific role in ethylene-dependent developmental processes remains poorly understood. This work demonstrated a specific role of Sl- ERF.B3, a member of the ERF gene family in tomato (Solanum lycopersicum), in mediating ethylene response and fruit ripening through a dominant repressor strategy. ERF.B3-SRDX dominant repressor etiolated seedlings displayed partial constitutive ethylene-response in the absence of ethylene and adult plants exhibited typical ethylenerelated alterations such as leaf epinasty, premature flower senescence and accelerated fruit abscission. The multiple symptoms related to enhanced ethylene sensitivity correlate with the altered expression of ethylene biosynthesis and signaling genes, suggesting the involvement of Sl-ERF.B3 in a feedback mechanism regulating components of ethylene production and response. In addition, over-expression of ERF.B3-SRDX in tomato results in alterations in both fruit morphology and ripening process. The attainment of competence to ripen is dramatically delayed in ERF.B3-SRDX fruits but once ripening proceeds it is associated with high climacteric ethylene production and enhanced fruit softening while pigment accumulation is strongly reduced. Moreover, a number of genes involved in the fruit ripening process showed expression pattern deviating from that of wild type. These data suggest a putative role of Sl-ERF.B3 in the transcriptional network underlying the ripening process and uncover a mean for uncoupling some of the main features of fruit ripening such as fruit softening and pigment accumulation. Overall, the study highlighted the importance of an ERF gene in ethylene-mediated developmental processes such as plant growth and fruit ripening

Investigation of Blocking Characteristics by Particles in Heterogeneous Reservoir

A mathematical model of suspension filtration in porous media has been established based on mass conservation principle and characteristics of particles depositing and blocking. On this basis, percolation rules and blocking characteristics of suspension in heterogeneous reservoir were investigated. It is showed that suspension injection could remarkably reduce the permeable ratio and improve the heterogeneity significantly. Low-speed and low-viscosity injection could achieve shallow profile control, and high-speed and high-viscosity injection could achieve deep profile control. Adjusting the injection rate or viscosity of carrying fluid slug at the right time to make the particle retention concentration profile in thief zones and the water-flood front keep consistent could achieve dynamic profile control. For the reservoirs without a good interlayer, the optimum injection rate and viscosity of carrying fluid were chosen based on the connectivity of layers, and in the reservoirs with good interlayers the injection rate and viscosity should be lowered appropriately under the field permitting conditions. When the suspension concentration was constant, the instantaneous fractional flow of high permeable layer first decreased sharply and then ramped up with the increasing injection volume. Initial percolation coefficient is the basis of a high utilization of suspension and a good result in profile control. Key words: Heterogeneous reservoir; Suspension; Percolation; Profile control; Retention concentration; Blocking characteristic

Ethylene Control of Fruit Ripening: Revisiting the Complex Network of Transcriptional Regulation

The plant hormone ethylene plays a key role in climacteric fruit ripening. Studies on components of ethylene signaling have revealed a linear transduction pathway leading to the activation of ethylene response factors. However, the means by which ethylene selects the ripening-related genes and interacts with other signaling pathways to regulate the ripening process are still to be elucidated. Using tomato (Solanum lycopersicum) as a reference species, the present review aims to revisit the mechanisms by which ethylene regulates fruit ripening by taking advantage of new tools available to perform in silico studies at the genomewide scale, leading to a global view on the expression pattern of ethylene biosynthesis and response genes throughout ripening. Overall, it provides new insights on the transcriptional network by which this hormone coordinates the ripening process and emphasizes the interplay between ethylene and ripening-associated developmental factors and the link between epigenetic regulation and ethylene during fruit ripening

Correlation of Cytokine Levels and Microglial Cell Infiltration during Retinal Degeneration in RCS Rats

Microglial cells, which are immunocompetent cells, are involved in all diseases of the central nervous system. During their activation in various diseases, a variety of soluble factors are released. In the present study, the correlation between cytokine levels and microglial cell migration in the course of retinal degeneration of Royal College of Surgeons (RCS) rats was evaluated. MFG-E8 and CD11b were used to confirm the microglial cells. In the retina of RCS rats, the mRNA expression of seven genes (MFG-E8 and its integrins αυ and ß5, CD11b and the cytokines TNF-α, IL-1ß, and MCP-1) formed almost similar bimodal peak distributions, which were centred at P7 and P45 to P60. In contrast, in rdy rats, which comprised the control group, a unimodal peak distribution centred at P14 was observed. The gene expression accompanied the activation and migration of microglial cells from the inner to the outer layer of the retina during the process of degeneration. Principal component analysis and discriminant function analysis revealed that the expression of these seven genes, especially TNF-α and CD11b, positively correlated with retinal degeneration and microglial activity during retinal degeneration in RCS rats, but not in the control rats. Furthermore, linear regression analysis demonstrated a significant correlation between the expression of these genes and the activation of microglial cells in the dystrophic retina. Our findings suggest that the suppression of microglial cells and the blockade of their cytotoxic effects may constitute a novel therapeutic strategy for treating photoreceptor death in various retinal disorders

Effects of Dietary Protein Level on Growth Performance, Muscle Composition, Blood Composition, and Digestive Enzyme Activity of Wuchang Bream (Megalobrama amblycephala) Fry

The purpose of this study was to determine the dietary protein requirement and effects of dietary protein level on growth performance, muscle composition, blood composition, and digestive enzyme activity in Wuchang bream fry. Five isoenergetic and isolipidic semi-purified diets were formulated to contain 28%, 30%, 32%, 34%, or 36% (dry matter) dietary protein. Diets were fed to triplicate groups of 25 fishes (16.08±0.03 g) to near satiation three times a day in a closed recirculation system for 10 weeks. Weight gain, specific growth rate, and feed conversion ratio significantly improved as the dietary protein content increased up to 34%. The protein efficiency ratio, hepatosomatic index, and viscerosomatic index significantly dropped as the dietary protein rose while the Fulton condition factor was positively correlated to the dietary protein level. Increased dietary protein resulted in increased muscle protein content and decreased lipid content. Red blood cell, hemoglobin, and hematocrit counts increased significantly with the increase in dietary protein. Serum triiodothyronine and thyroxine significantly rose as the dietary protein rose but serum aspartate aminotransferase significantly dropped. Intestinal protease and amylase activity rose significantly with the increase in dietary protein while lipase tended to drop. On the basis of broken-line regression analysis of weight gain and FCR, the dietary protein requirement of Wuchang bream fry is 32-33%

Advances in the study of ELABELA in renal physiological functions and related diseases

ELABELA (ELA), also known as Toddler or Apela, is a novel endogenous ligand of the angiotensin receptor AT1-related receptor protein (APJ). ELA is highly expressed in human embryonic, cardiac, and renal tissues and involves various biological functions, such as embryonic development, blood circulation regulation, and maintaining body fluid homeostasis. ELA is also closely related to the occurrence and development of acute kidney injury, hypertensive kidney damage, diabetic nephropathy, renal tumors, and other diseases. Understanding the physiological role of ELA and its mechanism of action in kidney-related diseases would provide new targets and directions for the clinical treatment of kidney diseases

Methodological quality of radiomic-based prognostic studies in gastric cancer: a cross-sectional study

BackgroundMachine learning radiomics models are increasingly being used to predict gastric cancer prognoses. However, the methodological quality of these models has not been evaluated. Therefore, this study aimed to evaluate the methodological quality of radiomics studies in predicting the prognosis of gastric cancer, summarize their methodological characteristics and performance.MethodsThe PubMed and Embase databases were searched for radiomics studies used to predict the prognosis of gastric cancer published in last 5 years. The characteristics of the studies and the performance of the models were extracted from the eligible full texts. The methodological quality, reporting completeness and risk of bias of the included studies were evaluated using the RQS, TRIPOD and PROBAST. The discrimination ability scores of the models were also compared.ResultsOut of 283 identified records, 22 studies met the inclusion criteria. The study endpoints included survival time, treatment response, and recurrence, with reported discriminations ranging between 0.610 and 0.878 in the validation dataset. The mean overall RQS value was 15.32 ± 3.20 (range: 9 to 21). The mean adhered items of the 35 item of TRIPOD checklist was 20.45 ± 1.83. The PROBAST showed all included studies were at high risk of bias.ConclusionThe current methodological quality of gastric cancer radiomics studies is insufficient. Large and reasonable sample, prospective, multicenter and rigorously designed studies are required to improve the quality of radiomics models for gastric cancer prediction.Study registrationThis protocol was prospectively registered in the Open Science Framework Registry (https://osf.io/ja52b)

Comprehensive Profiling of Ethylene Response Factor Expression Identifies Ripening-Associated ERF Genes and Their Link to Key Regulators of Fruit Ripening in Tomato

Our knowledge of the factors mediating ethylene-dependent ripening of climacteric fruit remains limited. The transcription of ethylene-regulated genes is mediated by ethylene response factors (ERFs), but mutants providing information on the specific role of the ERFs in fruit ripening are still lacking, likely due to functional redundancy among this large multigene family of transcription factors. We present here a comprehensive expression profiling of tomato (Solanum lycopersicum) ERFs in wild-type and tomato ripening-impaired tomato mutants (Never-ripe [Nr], ripening-inhibitor [rin], and non-ripening [nor]), indicating that out of the 77 ERFs present in the tomato genome, 27 show enhanced expression at the onset of ripening while 28 display a ripeningassociated decrease in expression, suggesting that different ERFs may have contrasting roles in fruit ripening. Among the 19 ERFs exhibiting the most consistent up-regulation during ripening, the expression of 11 ERFs is strongly down-regulated in rin, nor, and Nr tomato ripening mutants, while only three are consistently up-regulated. Members of subclass E, SlERF.E1, SlERF.E2, and SlERF.E4, show dramatic down-regulation in the ripening mutants, suggesting that their expression might be instrumental in fruit ripening. This study illustrates the high complexity of the regulatory network connecting RIN and ERFs and identifies subclass E members as the most active ERFs in ethylene- and RIN/NOR-dependent ripening