Fruit cell culture as a model system to study cell wall changes during strawberry fruit ripening

Strawberry (Fragaria x ananassa, Duch.) fruit is characterized by its fast ripening and soft texture at the ripen stage, resulting in a short postharvest shelf life and high economic losses. It is generally believed that the disassembly of cell walls, the dissolution of the middle lamella and the reduction of cell turgor are the main factors determining the softening of fleshy fruits. In strawberry, several studies indicate that the solubilisation and depolymerisation of pectins, as well as the depolymerisation of xyloglucans, are the main processes occurring during ripening. Functional analyses of genes encoding pectinases such as polygalacturonase and pectate lyase also point out to the pectin fraction as a key factor involved in textural changes. All these studies have been performed with whole fruits, a complex organ containing different tissues that differ in their cell wall composition and undergo ripening at different rates. Cell cultures derived from fruits have been proposed as model systems for the study of several processes occurring during fruit ripening, such as the production of anthocyanin and its regulation by plant hormones. The main objective of this research was to obtain and characterize strawberry cell cultures to evaluate their potential use as a model for the study of the cell wall disassembly process associate with fruit ripening. Cell cultures were obtained from cortical tissue of strawberry fruits, cv. Chandler, at the stages of unripe-green, white and mature-red. Additionally, a cell culture line derived from strawberry leaves was obtained. All cultures were maintained in solid medium supplemented with 2.5 mg.l-1 2,4-D and incubated in the dark. Cell walls from the different callus lines were extracted and fractionated to obtain CDTA and sodium carbonate soluble pectin fractions, which represent polyuronides located in the middle lamella or the primary cell wall, respectively. The amounts of homogalacturonan in both fractions were estimated by ELISA using LM19 and LM20 antibodies, specific against demethylated and methyl-esterified homogalacturonan, respectively. In the CDTA fraction, the cell line from ripe fruit showed a significant lower amount of demethylated pectins than the rest of lines. By contrast, the content of methylated pectins was similar in green- and red-fruit lines, and lower than in white-fruit and leaf lines. In the sodium carbonate pectin fraction, the line from red fruit also showed the lowest amount of pectins. These preliminary results indicate that cell cultures obtained from fruits at different developmental stages differ in their cell wall composition and these differences resemble to some extent the changes that occur during strawberry softening. Experiments are in progress to further characterize cell wall extracts with monoclonal antibodies against other cell wall epitopes.Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

Nonequilibrium evolution of volatility in origination and extinction explains fat-tailed fluctuations in Phanerozoic biodiversity

Fluctuations in biodiversity, large and small, pervade the fossil record, yet we do not understand the processes generating them. Here, we extend theory from nonequilibrium statistical physics to describe the fat-tailed form of fluctuations in Phanerozoic marine invertebrate richness. Using this theory, known as superstatistics, we show that heterogeneous rates of origination and extinction between clades and conserved rates within clades account for this fat-tailed form. We identify orders and families as the taxonomic levels at which clades experience interclade heterogeneity and within-clade homogeneity of rates, indicating that families are subsystems in local statistical equilibrium, while the entire system is not. The separation of timescales between within-clade background rates and the origin of major innovations producing new orders and families allows within-clade dynamics to reach equilibrium, while between-clade dynamics do not. The distribution of different dynamics across clades is consistent with niche conservatism and pulsed exploration of adaptive landscapes.Fil: Rominger, Andrew J.. No especifíca;Fil: Fuentes, Miguel Angel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Filosóficas. - Sociedad Argentina de Análisis Filosófico. Instituto de Investigaciones Filosóficas; Argentina. Universidad San Sebastián; ChileFil: Marquet, Pablo A.. Pontificia Universidad Católica de Chile; Chile. Instituto de Ecología y Biodiversidad; Chile. Universidad Nacional Autónoma de México; Méxic

Preyssler Heteropolyacids in the Self-Etherification of 5-Hydroxymethylfurfural to 5,5′-[Oxybis(methylene)]bis-2-furfural Under Mild Reaction Conditions

The synthesis of 5,5′-[oxybis(methylene)]bis-2-furfural (OBMF) from 5-hydroxymethylfurfural (5-HMF) was studied using bulk and alumina-supported Preyssler heteropolyacids (HPAs). The formation of OBMF was related to the amount of Brønsted acid sites, and the lowest yield of OBMF was obtained with supported HPAs. However, the Lewis acidity of the HPA supported on Al2O3 favored the formation of 2,5-dimethylfurane. The effects of solvent, catalyst loading, temperature, and reaction time on the selectivity to OBMF from 5-HMF were studied to optimize OBMF production using bulk Preyssler HPAs; a yield of 84 % to OBMF was obtained at 5 h and 343 K. These results demonstrate that bulk Preyssler HPA is a good candidate for OBMF synthesis under mild reaction conditions.Fil: Páez, Alexander. Universidad Pedagógica y Tecnológica de Colombia; ColombiaFil: Rojas, Hugo A.. Universidad Pedagógica y Tecnológica de Colombia;Fil: Portilla Zúñiga, Omar Miguel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Ciencias Aplicadas ; ArgentinaFil: Sathicq, Angel Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Ciencias Aplicadas ; ArgentinaFil: Afonso, Carlos A. M.. Universidade de Lisboa; PortugalFil: Romanelli, Gustavo Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Ciencias Aplicadas ; ArgentinaFil: Martínez, José J.. Universidad Pedagógica y Tecnológica de Colombia; Colombi

Rhamnogalacturonase lyase gene downregulation in strawberry and its potential on mechanical fruit properties

Strawberry softening is one of the main factors that reduces fruit quality and leads to economically important losses. Textural changes during fruit ripening are mainly due to the dissolution of middle lamellae, a reduction in cell-to-cell adhesion and the weakening of parenchyma cell walls as a result of the action of cell wall modifying enzymes. Functional studies of genes encoding pectinase enzymes (polygalacturonase, pectate lyase and -galactosidase) support a key role of pectin disassembly in strawberry softening. Evidence that RG-I may play an important role in strawberry texture has been obtained from the transient silencing of a RG-lyase gene. Pectins are major components of fruit cell walls and highly dynamic polysaccharides, but due to their heterogeneity the precise relation between the structures and functions is incomplete. In this work, stable transgenic strawberry lines with a rhamnogalacturonate lyase gene (FaRGLyase1) down-regulated have been analyzed. Several transgenic lines showing more than 95% silencing of FaRGLyase1 displayed fruit firmness values higher than control. Cell walls from these lines were extracted and analyzed by ELISA and Epitope Detection Chromatography (EDC). This last technique is based on the detection of specific cell wall oligosaccharide epitopes and provides information on sub-populations of pectins containing homogalacturonan and RG-I domains, but also reveals potential links with other cell wall polysaccharides such as xyloglucan. The results obtained indicate that the silencing of FaRGLyase1 reduces degradation of RG-I backbones, but also homogalacturonan, in cell walls, especially in pectin fractions covalently bound to the cell wall. These changes contribute to the increased firmness of transgenic fruits.This research was supported by FEDER EU Funds and the Ministerio de Economía y Competitividad of Spain (grant reference AGL2014-55784-C2), a Marie Curie IEF within the 7th European Community Framework Programme (reference: PIEF-2013-625270) for SP and a FPI fellowship (BES-2015-073616) to support PR-V. Universidad de Málaga. Campus de Excelencia Internacional Andalucía Tech

The Glycerol-Dependent Metabolic Persistence of Pseudomonas putida KT2440 Reflects the Regulatory Logic of the GlpR Repressor

The growth of the soil bacterium Pseudomonas putida KT2440 on glycerol as the sole carbon source is characterized by a prolonged lag phase, not observed with other carbon substrates. We examined the bacterial growth in glycerol cultures while monitoring the metabolic activity of individual cells. Fluorescence microscopy and flow cytometry, as well as the analysis of the temporal start of growth in single-cell cultures, revealed that adoption of a glycerol-metabolizing regime was not the result of a gradual change in the whole population but rather reflected a time-dependent bimodal switch between metabolically inactive (i.e., nongrowing) and fully active (i.e., growing) bacteria. A transcriptional Φ(glpD-gfp) fusion (a proxy of the glycerol-3-phosphate [G3P] dehydrogenase activity) linked the macroscopic phenotype to the expression of the glp genes. Either deleting glpR (encoding the G3P-responsive transcriptional repressor that controls the expression of the glpFKRD gene cluster) or altering G3P formation (by overexpressing glpK, encoding glycerol kinase) abolished the bimodal glpD expression. These manipulations eliminated the stochastic growth start by shortening the otherwise long lag phase. Provision of glpR in trans restored the phenotypes lost in the ΔglpR mutant. The prolonged nongrowth regime of P. putida on glycerol could thus be traced to the regulatory device controlling the transcription of the glp genes. Since the physiological agonist of GlpR is G3P, the arrangement of metabolic and regulatory components at this checkpoint merges a positive feedback loop with a nonlinear transcriptional response, a layout fostering the observed time-dependent shift between two alternative physiological states

Hydride precipitation and stresses in zircaloy-4 observed by synchrotron X-ray diffraction

The grain stresses within hydrides precipitated in rolled zircaloy-4 plates were determined by synchrotron X-ray diffraction experiments using an 80 keV photon beam and a high-speed area detector placed in transmission geometry. Results showed large compressive stresses (360 ± 20 MPa) in the hydrides along the plate rolling direction. The origin of these stresses was investigated by performing hydride dissolution/precipitation in situ for thermal cycles between room temperature and 400 C. A large stress hysteresis was observed, with a steady decrease on heating and an abrupt change on cooling. The observed stresses are explained by the constraint imposed by grain boundaries on the growth of hydride platelets on the rolling–transverse plane of the rolled plates.Fil: Santisteban, Javier Roberto. Comisión Nacional de Energía Atómica. Centro Atómico Bariloche; Argentina. Comisión Nacional de Energía Atómica. Gerencia del Área de Energía Nuclear. Instituto Balseiro; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; ArgentinaFil: Vicente Alvarez, Miguel Angel. Comisión Nacional de Energía Atómica. Gerencia del Área de Energía Nuclear. Instituto Balseiro; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Patagonia Norte; ArgentinaFil: Vizcaino, Pablo. Comisión Nacional de Energía Atómica. Centro Atómico Ezeiza; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Banchik, A. D.. Comisión Nacional de Energía Atómica. Centro Atómico Ezeiza; ArgentinaFil: Almer, J. D.. No especifíca

Effect of Sr, Mg and Fe substitution on thephysico-chemical and biological properties ofSi Ca P multilayer scaffolds

tIn this work, a new combination of ceramic materials is proposed for bone tissue engi-neering applications. Multilayer scaffolds consisting of a core composed mainly of calciumpyrophosphate and external coatings of silica and calcium doped with Fe3+, Sr2+and Mg2+were prepared. To study the influence of the arrangement of dopant ions in the externalcoatings, two different scaffolds were developed: scaffolds 3J consisting of a single exter-nal coating with 9 mol% of Fe3+, Sr2+and Mg2+ions; and scaffolds 3S comprising threeexternal coatings, each containing 3 mol% of Fe3+, Sr2+and Mg2+ions. Scaffolds were physico-chemically characterized and evaluated for in vitro bioactivity and cellular response in thepresence of MG-63 cells. The results showed that the core scaffold displayed no in vitro bioac-tivity or good cellular response, but served as a support for the external coatings given itsmechanical resistance. The cell viability of scaffolds 3J and 3S increased more than 100%in relation to the core, and also improved cell proliferation and adhesion resulting in adense layer of cells that covered the scaffolds’ entire surface. The arrangement of ions inthe external coatings did not influence the cellular response, but determined the bioactivityrate

Ternary hydrotalcites in the multicomponent synthesis of 4H-pyrans

Lamellar double hydroxides (LDH) with double divalent cations were synthesized by the co-precipitation method and studied in the multicomponent synthesis of 4H-pyrans. The solids obtained were characterized by X-ray diffraction (XRD), infrared spectroscopy (FTIR), N2 adsorption isotherms, and thermogravimetric analysis (TGA). The XRD patterns confirmed the formation of LDHs in which the incorporation of Ni2+ or Co2+ improves their crystalline and textural properties. The results of catalytic activity showed that the synthesis of 4H-pyrans is favored in solvent-free conditions with the LDH–Ni catalyst, avoiding calcination processes. In addition, it was found that hydrotalcite with double divalent cations can conduct this reaction through multicomponent synthesis or by the Michael addition reaction, which can be performed by different types of basicity that depend on the composition of another divalent cation in the brucite layer or a calcination process.Fil: Nope Vargas, Eliana Rocio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Ciencias Aplicadas "Dr. Jorge J. Ronco". Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Ciencias Aplicadas; ArgentinaFil: Sathicq, Angel Gabriel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Ciencias Aplicadas "Dr. Jorge J. Ronco". Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Ciencias Aplicadas; ArgentinaFil: Martínez, José J.. Universidad Pedagógica y Tecnológica de Colombia; ColombiaFil: Rojas, Hugo A.. Universidad Pedagógica y Tecnológica de Colombia; ColombiaFil: Luque, Rafael. Universidad de Córdoba; EspañaFil: Romanelli, Gustavo Pablo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Centro de Investigación y Desarrollo en Ciencias Aplicadas "Dr. Jorge J. Ronco". Universidad Nacional de la Plata. Facultad de Ciencias Exactas. Centro de Investigación y Desarrollo en Ciencias Aplicadas; Argentin

Rendering Bio-inert Low-Density Polyethylene Amenable for Biodegradation via Fast High Throughput Reactive Extrusion Assisted Oxidation

An energy-efficient high throughput pre-treatment of low-density polyethylene (LDPE) using a fast, reactive extrusion (REX) assisted oxidation technique followed by bacterial attachment as an indicator for bio-amenability was studied. Silicon dioxide (SiO2) was selected as a model oxidizing and catalytic reagent with the REX process demonstrated to be effective both in the presence and absence of the catalyst. Optimized 5-min duration pre-treatment conditions were determined using Box-Behnken design (BBD) with respect to screws speed, operating temperature, and concentration of SiO2. The crystallinity index, carbonyl index and weight loss (%) of LDPE were used as the studied responses for BDD. FTIR and DSC spectra of the residual LDPE obtained after pre-treatment with the REX assisted oxidation technique showed a significant increase in residual LDPE carbonyl index from 0 to 1.04 and a decrease of LDPE crystallinity index from 29 to 18%. Up to fivefold molecular weight reductions were also demonstrated using gel permeation chromatography. Optimum LDPE pre-treatment with a duration of 5 min was obtained at low screw speed (50 rpm), operating temperature of 380-390 degrees C and variable concentration of SiO2 (0 and 2% (w/w)) indicating that effective pre-treatment can occur under noncatalytic and catalysed conditions. Biofilms were successfully formed on pre-treated LDPE samples after 14 days of incubation. Furthermore, the technique proposed in this study is expected to provide a high throughput approach for pre-treatment of pervasive recalcitrant PE-based plastics to reduce their bio inertness.Related to published version:[https://imagine.imgge.bg.ac.rs/handle/123456789/1575]Peer-reviewed manuscript: Ferrero, P., Attallah, O. A., Valera, M. Á., Aleksic, I., Azeem, M., Nikodinovic-Runic, J., & Fournet, M. B. (2022). Rendering Bio-inert Low-Density Polyethylene Amenable for Biodegradation via Fast High Throughput Reactive Extrusion Assisted Oxidation. Journal of Polymers and the Environment, 30(7), 2837–2846.[https://doi.org/10.1007/s10924-022-02400-w