Dark complexes of the Calvin-Benson cycle in a physiological perspective

: Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and phosphoribulokinase (PRK) are two enzymes of the Calvin Benson cycle that stand out for some peculiar properties they have in common: (i) they both use the products of light reactions for catalysis (NADPH for GAPDH, ATP for PRK), (ii) they are both light-regulated through thioredoxins and (iii) they are both involved in the formation of regulatory supramolecular complexes in the dark or low photosynthetic conditions, with or without the regulatory protein CP12. In the complexes, enzymes are transiently inactivated but ready to recover full activity after complex dissociation. Fully active GAPDH and PRK are in large excess for the functioning of the Calvin-Benson cycle, but they can limit the cycle upon complex formation. Complex dissociation contributes to photosynthetic induction. CP12 also controls PRK concentration in model photosynthetic organisms like Arabidopsis thaliana and Chlamydomonas reinhardtii. The review combines in vivo and in vitro data into an integrated physiological view of the role of GAPDH and PRK dark complexes in the regulation of photosynthesis

Vino e Ambiente: sostenibilità e qualità primaria nel sottobacino Iudeo-Bucari (TP).

In questa raccolta di scritti, vengono riportati i risultati dell’attività di ricerca realizzata con la collaborazione della cantina UVAM e dell’Istituto Regionale Vino e Olio di Sicili

Conformational Disorder Analysis of the Conditionally Disordered Protein CP12 from Arabidopsis thaliana in Its Different Redox States

CP12 is a redox-dependent conditionally disordered protein universally distributed in oxygenic photosynthetic organisms. It is primarily known as a light-dependent redox switch regulating the reductive step of the metabolic phase of photosynthesis. In the present study, a small angle X-ray scattering (SAXS) analysis of recombinant Arabidopsis CP12 (AtCP12) in a reduced and oxidized form confirmed the highly disordered nature of this regulatory protein. However, it clearly pointed out a decrease in the average size and a lower level of conformational disorder upon oxidation. We compared the experimental data with the theoretical profiles of pools of conformers generated with different assumptions and show that the reduced form is fully disordered, whereas the oxidized form is better described by conformers comprising both the circular motif around the C-terminal disulfide bond detected in previous structural analysis and the N-terminal disulfide bond. Despite the fact that disulfide bridges are usually thought to confer rigidity to protein structures, in the oxidized AtCP12, their presence coexists with a disordered nature. Our results rule out the existence of significant amounts of structured and compact conformations of free AtCP12 in a solution, even in its oxidized form, thereby highlighting the importance of recruiting partner proteins to complete its structured final folding

A Plant Bioreactor for the Synthesis of Carbon Nanotube Bionic Nanocomposites

Bionic composites are an emerging class of materials produced exploiting living organisms as reactors to include synthetic functional materials in their native and highly performing structures. In this work, single wall carboxylated carbon nanotubes (SWCNT-COOH) were incorporated within the roots of living plants of Arabidopsis thaliana. This biogenic synthetic route produced a bionic composite material made of root components and SWCNT-COOH. The synthesis was possible exploiting the transport processes existing in the plant roots. Scanning electrochemical microscopy (SECM) measurements showed that SWCNT-COOH entered the vascular bundles of A. thaliana roots localizing within xylem vessels. SWCNT-COOH preserved their electrical properties when embedded inside the root matrix, both at a microscopic level and a macroscopic level, and did not significantly affect the mechanical properties of A. thaliana roots

Moscato Cerletti, a rediscovered aromatic cultivar with oenological potential in warm and dry areas

Baron Antonio Mendola was devoted to the study of grapevine, applying ampelography and dabbling in crosses between cultivars in order to select new ones, of which Moscato Cerletti, obtained in 1869, was the most interesting. Grillo, one of the most important white cultivars in Sicily, was ascertained to be an offspring of Catarratto Comune and Zibibbo, the same parents which Mendola claimed he used to obtain Moscato Cerletti. Thus the hypothesis of synonymy between Moscato Cerletti and Grillo or the same parentage for both sets of parents needs to be verified. In the present study, historical documents were consulted and genetic analyses and ampelographic, agronomic and qualitative characterisation carried out to determine the distinctiveness of each cultivars. These were also compared with Catarratto Comune and Zibibbo in order to establish the Moscato Cerletti pedigree. Due to their different SSR profiles, Grillo and Moscato Cerletti were confirmed as two distinct cultivars; they also differed in ripening times and sugar storage ability, as well as in the aromatic grape produced by Moscato Cerletti only. The trio genotype genetic analysis confirmed that Zibibbo is a parent of Moscato Cerletti (justifying the aromatic grape), whilst the SSR profiles did not show Catarratto Comune to be a second parent. Moscato Cerletti was found to have oenological potential in the production of sparkling muscat wines due to its ability to adapt to a changing climate in warm and dry environments and in different winegrowing regions

The Thioredoxin-Regulated α-Amylase 3 of Arabidopsis thaliana Is a Target of S-Glutathionylation

Reactive oxygen species (ROS) are produced in cells as normal cellular metabolic by-products. ROS concentration is normally low, but it increases under stress conditions. To stand ROS exposure, organisms evolved series of responsive mechanisms. One such mechanism is protein S-glutathionylation. S-glutathionylation is a post-translational modification typically occurring in response to oxidative stress, in which a glutathione reacts with cysteinyl residues, protecting them from overoxidation. α-Amylases are glucan hydrolases that cleave α-1,4-glucosidic bonds in starch. The Arabidopsis genome contains three genes encoding α-amylases. The sole chloroplastic member, AtAMY3, is involved in osmotic stress response and stomatal opening and is redox-regulated by thioredoxins. Here we show that AtAMY3 activity was sensitive to ROS, such as H2O2. Treatments with H2O2 inhibited enzyme activity and part of the inhibition was irreversible. However, in the presence of glutathione this irreversible inhibition was prevented through S-glutathionylation. The activity of oxidized AtAMY3 was completely restored by simultaneous reduction by both glutaredoxin (specific for the removal of glutathione-mixed disulfide) and thioredoxin (specific for the reduction of protein disulfide), supporting a possible liaison between both redox modifications. By comparing free cysteine residues between reduced and GSSG-treated AtAMY3 and performing oxidation experiments of Cys-to-Ser variants of AtAMY3 using biotin-conjugated GSSG, we could demonstrate that at least three distinct cysteinyl residues can be oxidized/glutathionylated, among those the two previously identified catalytic cysteines, Cys499 and Cys587. Measuring the pKa values of the catalytic cysteines by alkylation at different pHs and enzyme activity measurement (pKa1 = 5.70 ± 0.28; pKa2 = 7.83 ± 0.12) showed the tendency of one of the two catalytic cysteines to deprotonation, even at physiological pHs, supporting its propensity to undergo redox post-translational modifications. Taking into account previous and present findings, a functional model for redox regulation of AtAMY3 is proposed

Le prove di vinificazione della cultivar grillo su tre vigneti sperimentali site nel sottobacino Iudeo - Bucari (TP).

Gli Autori, dopo aver descrittole caratteristiche ambientali del sottobacino Iudeo Bucari e l’attività agricola prevalentemente legataalla vitivinicoltura, descrivono alcune prove di vinificazione di uve Grillo provenienti da tre vignesite nel sottobacino Iudeo – Bucari gestite in modo ordinario, delle quali si è monitorata la maturazione delle uve. L'obiettivo è quello di caratterizzare il prodotto vino ottenuto attraverso una tecnica di vinificazione uniforme ed ordinaria nella zona, ma che mira a mettere in evidenza la diversa qualità tecnologica delle uve utilizzate espressione dicontesti eco-pedologici diversi

Redox homeostasis in photosynthetic organisms: Novel and established thiol-based molecular mechanisms

Redox homeostasis consists of an intricate network in which reactive molecular species (RMS), redox modifications and redox proteins act in concert to allow both physiological responses and adaptation to stress conditions. This review highlights established and novel thiol-based regulatory pathways underlying the functional facets and significance of redox biology in photosynthetic organisms. This cannot be all-encompassing, but is intended to provide a comprehensive overview on the structural/molecular mechanisms governing the most relevant thiol switching modifications with emphasis on the large genetic and functional diversity of redox controllers (i.e. redoxins). We also summarize the different proteomic-based approaches aimed at investigating the dynamics of redox modifications and the recent evidence that extends the possibility to monitor the cellular redox state in vivo. Lastly, the physiological relevance of redox transitions is discussed based on reverse genetic studies confirming the importance of redox homeostasis in plant growth, development, and stress responses