The effects of ascorbate (ASC) and dehydroascorbate (DHA) on cell proliferation were examined in the tobacco Bright Yellow 2 (TBY-2) cell line to test the hypothesis that the ASC-DHA pair is a specific regulator of cell division. The hypothesis was tested by measuring the levels of ASC and DHA or another general redox pair, glutathione (GSH) and glutathione disulfide (GSSG), during the exponential-growth phase of TBY-2 cells. A peak in ASC, but not GSH, levels coincided with a peak in the mitotic index. Moreover, when the cells were enriched with ascorbate, a stimulation of cell division occurred whereas, when the cells were enriched with DHA, the mitotic index was reduced. In contrast, glutathione did not affect the mitotic-index peak during this exponential-growth phase. The data are consistent in showing that the ASC-DHA pair acts as a specific redox sensor which is part of the mechanism that regulates cell cycle progression in this cell line.

Programmed cell death (PCD) is a genetically controlled process described both in eukaryotic and prokaryotic organisms. Even if it is clear that PCD occurs in plants, in response to various developmental and environmental stimuli, the signalling pathways involved in the triggering of this cell suicide remain to be characterized. In this review, the main similarities and differences in the players involved in plant and animal PCD are outlined. Particular attention is paid to the role of reactive oxygen species (ROS) as key inducers of PCD in plants. The involvement of different kinds of ROS, different sites of ROS production, as well as their interaction with other molecules, is crucial in activating PCD in response to specific stimuli. Moreover, the importance is stressed on the balance between ROS production and scavenging, in various cell compartments, for the activation of specific steps in the signalling pathways triggering this cell suicide process. The review focuses on the complexity of the interplay between ROS and antioxidant molecules and enzymes in determining the most suitable redox environment required for the occurrence of different forms of PCD.

O. 1997, Ascorbate synthesis and ascorbate peroxidase activity during the early stage of wheat germination. -Physiol. Plant. 100: 894-900.Embryos from dry caryopses of wheat {Triticum durum L. cv. Norba) are completely devoid of ascorbate (ASC) but contain a low amount of dehydroascorbate (DHA). The de novo biosynthesis of ASC starts in the wheat embryos after 8-10 h of germination but before the ASC biosynthetic pathway is completely restored the embryos can provide themselves with ASC by the reduction of the stored DHA. Three different proteins having DHA-reducing capability are present in the embryos during the early stages of germination. However, when the de novo ASC biosynthesis from sugar is completely restored, the DHA reduction capability largely drops and only one DHA-reducing protein remains active. The presence of three proteins having DHA-reducing capability and their behaviour during germination is discussed. Dry embryos are also devoid of ASC peroxidase (EC 1.11.1.11); this hydrogen peroxide scavenger enzyme appears after the same lag as ASC and increases during germination in parallel with the rise in ASC. When ASC biosynthesis is experimentally induced, the ASC peroxidase also appears earlier; moreover the affinities for ASC of the three ASC peroxidase isoenzymes that progressively appear during germination depend on the ASC available in the embryos: highest in the first isoenzyme, that appears when the ASC content is very low, lowest in the isoenzyme that is expressed last, when the ASC content is 10-11 times higher.

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Summary• The combined effects of the two pollutants, cadmium and ozone, on sunflower ( Helianthus annuus ) metabolism are analysed here.• Photosysnthetic processes and ascorbate metabolism were studied in sunflower plants grown for 15 d in the presence of cadmium and exposed to acute O 3 treatments.• CO 2 assimilation rate was reduced in plants subjected to Cd(II) and/or O 3 treatments, but no alterations in stomatal conductance and F v : F m ratio were observed. Rubisco activity was significantly reduced only in plants grown in the presence of cadmium indicating that the photosynthetic process is mainly altered by this factor. Photochemical quenching and the quantum efficiency of PSII in steady-state conditions were significantly depressed and nonphotochemical quenching increased in stressed plants. Cd(II) and O 3 also strongly affected ascorbate metabolism.• The changes in ascorbate redox state and the increase in ascorbate-redox enzymes strongly supported an ascorbate over-utilization in Cd(II) and/or O 3 -treated plants. However, the increase in ascorbate-based detoxification mechanisms did not provide complete protection against the oxidative stress imposed by the two pollutants, since an increase in lipid peroxidation and protein oxidation accompanied a decrease in photosynthesis under pollutant exposure.

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Kernel development and maturation involve several well-characterised events, such as changes in ascorbate (ASC) metabolism, protein synthesis and storage, programmed cell death (PCD) of starchy endosperm and tissue dehydration. Despite many studies focusing on these events, whether and how they are metabolically related to each other, remains to be elucidated. In the present investigation, the changes in ASC-related metabolism, PCD occurrence, kernel filling and dehydration have been analysed during kernel maturation, over a 3-year period in plants grown under normal conditions and in plants displaying modified ASC synthesis. The obtained results suggest that ASC plays a pivotal role in the network of events characterising kernel maturation. During this process, a decrease in ASC content occurs. When ASC biosynthesis is improved in the kernel, by feeding the plants with its immediate precursor, L-galactone-γ-lactone (GL), the decrease in ASC, observed during kernel maturation, is delayed. As a consequence, ascorbate peroxidase (APX) activity is also enhanced. Moreover, a delay in the ASC decrease permits a delay in PCD occurring in kernel storage tissues and in kernel dehydration. Interestingly, the data emerging from the present investigation suggest that the delay in the decrease in ASC content and APX activity also improves kernel filling. The relevance of the ascorbate-dependent redox regulation for kernel productivity is discussed.

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