The Role of Ascorbate Free Radical as an Electron Acceptor to Cytochrome b-Mediated Trans-Plasma Membrane Electron Transport in Higher Plants

The action of ascorbate free radical as an electron acceptor to cytochrome b-mediated trans-plasma membrane electron transport is demonstrated. Addition of ascorbate free radical to ascorbate-loaded plasma membrane vesicles caused a rapid oxidation of the cytochrome, followed by a slower re-reduction. The fully reduced dehydroascorbate was ineffective.     

Environmental Dissolved Organic Matter Governs Biofilm Formation and Subsequent Linuron Degradation Activity of a Linuron-Degrading Bacterial Consortium

It was examined whether biofilm growth on dissolved organic matter (DOM) of a three-species consortium whose members synergistically degrade the phenylurea herbicide linuron affected the consortium''s integrity and subsequent linuron-degrading functionality. Citrate as a model DOM and three environmental DOM (eDOM) formulations of different quality were used. Biofilms developed with all DOM formulations, and the three species were retained in the biofilm. However, biofilm biomass, species composition, architecture, and colocalization of member strains depended on DOM and its biodegradability. To assess the linuron-degrading functionality, biofilms were subsequently irrigated with linuron at 10 mg liter⁻¹ or 100 μg liter⁻¹. Instant linuron degradation, the time needed to attain maximal linuron degradation, and hence the total amount of linuron removed depended on both the DOM used for growth and the linuron concentration. At 10 mg liter⁻¹, the final linuron degradation efficiency was as high as previously observed without DOM except for biofilms fed with humic acids which did not degrade linuron. At 100 μg liter⁻¹ linuron, DOM-grown biofilms degraded linuron less efficiently than biofilms receiving 10 mg liter⁻¹ linuron. The amount of linuron removed was more correlated with biofilm species composition than with biomass or structure. Based on visual observations, colocalization of consortium members in biofilms after the DOM feed appears essential for instant linuron-degrading activity and might explain the differences in overall linuron degradation. The data show that DOM quality determines biofilm structure and composition of the pesticide-degrading consortium in periods with DOM as the main carbon source and can affect subsequent pesticide-degrading activity, especially at micropollutant concentrations.     

Functional aspects of Abscisic Acid Metabolism in Cotyledons of Phaseolus vulgaris L. Seedlings

During the early stages of germination and vegetative development,cotyledons of intact bean (Phaseolus vulgaris L.) seedlingsshowed active ABA catabolism causing a low endogenous ABA content.At the end of the substrate mobilizing phase, when the cotyledonsbecame senescent, a drastic increase of the endogenous ABA contentlinked with a decrease of the ABA catabolic activity was observed.This indicates that a close connection exists between senescenceand endogenous ABA content and metabolism in bean cotyledons. Removal of the apical growth region induced in the cotyledonsactivation of the ABA catabolism and the endogenous ABA concentrationdecreased below the detection limit (0.1 ng/g fr wt) at theonset of the outgrowth of the axillary buds. From then, apicaldominance was restored and the cotyledons returned to the senescentstate, which was correlated with a drastic increase of theirendogenous ABA content. ¹ Bevoegd verklaard navorser N. F. W. O. ² Wetenschappelijk medewerker F. K. F. O. (Received November 25, 1980; Accepted February 13, 1981)     

Facilitated diffusion drives transport of oxidised ascorbate molecules into purified plasma membrane vesicles of Phaseolus vulgaris

Recently, the presence of a carrier‐mediated transport system for ascorbate was demonstrated in the plant plasma membrane. To investigate the possible physiological importance of this system in apoplastic ascorbate metabolism we further characterized this carrier. Transport of Asc was measured by incubating freshly‐purified plasma membrane vesicles from hypocotylar hooks of Phaseolus vulgaris together with [¹⁴C]‐labelled Asc. In this paper we show that ascorbate transport is detectable over a relatively broad pH range (6 to 7.5) and is not affected by protonophore addition. [¹⁴C]‐Ascorbate is not taken up into vesicle fractions consisting of sealed inside‐out oriented vesicles, suggesting that it is transported only from the apoplast to the cytoplasm. Asc uptake into vesicles previously loaded with ascorbate was also tested. Surprisingly, uptake of radioactive molecules was up to 3‐fold higher in the ascorbate‐loaded vesicles compared to non‐loaded control vesicles ( P < 0.001). The uptake of [¹⁴C]‐ascorbate in both the ascorbate‐loaded as the non‐loaded membrane vesicles was inhibited by addition of DTT and not by glutathione or ferricyanide. Based on various observations such as cis ‐inhibition, trans ‐stimulation and insensitivity towards proton gradients, a facilitated uptake mechanism is suggested. Our results strongly indicate that dehydroascorbate is the preferred transported species from the apoplastic to the cytoplasmic side of the membrane. This transport system is possibly involved in the regeneration of apoplastic ascorbate.     

Copper-mediated oxidative burst in Nicotiana tabacum L. cv. Bright Yellow 2 cell suspension cultures

In cell suspension cultures of Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) a rapid and concentration-dependent accumulation of H(2)O(2) is induced by excess concentrations of copper (up to 100 microM). This specific and early response towards copper stress was shown to be extracellular. Addition of 300 U of catalase per ml decreased the level of H(2)O(2). Superoxide dismutase (5 U/ml) induced an increase in H(2)O(2) production by 22.2%. This indicates that at least part of the H(2)O(2) is produced by dismutation of superoxide. Pretreatment of the cell cultures with the NAD(P)H oxidase inhibitors diphenylene iodonium (2 and 10 microM) and quinacrine (1 and 5 mM) prevented the generation of H(2)O(2) under copper stress for 90%. The influence of the pH on the H(2)O(2) production revealed the possible involvement of cell-wall-dependent peroxidases in the generation of reactive oxygen species after copper stress.     

Transport of ascorbate into protoplasts ofNicotiana tabacum Bright Yellow-2 cell line

Summary The uptake of ascorbate into protoplasts isolated from aNicotiana tabacum Bright Yellow-2 (BY-2) cell suspension culture was investigated. Addition of¹⁴C-labelled ascorbate to freshly isolated protoplasts resulted in a time- and substrate-dependent association of radioactive molecules with the protoplasts. The kinetic characterisation of this presumptive uptake revealed kinetics of Michaelis-Menten type with an apparent maximal uptake activity of 24 pmol/min·10⁶ protoplasts and an apparent affinity constant of 139 M. The amount of ascorbate molecules transported into_{N. tabacum} protoplasts decreased when nonlabelled dehydroascorbate or iso-ascorbate were added but was not affected by addition of 5,6-o-cyclohexylidene ascorbate or ascorbate-2-sulfate. These data indicate a carrier-mediated uptake of ascorbate into the protoplasts that shows a high structural specificity. To investigate which redox status of ascorbate is preferentially taken up by theN. tabacum protoplasts, transport was tested in the presence of various compounds that can affect the redox status of ascorbate. Testing uptake in the presence of a reductant, dithiothreitol, resulted in a significant and concentration-dependent inhibition of the amount of ascorbate molecules transported into the protoplasts. On the other hand, ascorbate uptake was significantly stimulated in the presence of the enzyme ascorbate oxidase. Ferricyanide did not affect ascorbate transport. Inhibition studies revealed that ascorbate uptake in the protoplasts is sensitive to addition of sulfhydryl reagents N-ethyl maleimide andp-chloro-mercuribenzenesulfonic acid and to a disruption of the proton gradient by the protonophore carbonylcyanide-3-chlorophenylhydrazone. The uptake of ascorbate was also inhibited by addition of cytochalasin B but not sensitive to addition of phloretin or sulfinpyrazone. Taken together these data indicate the presence of an ascorbate transport system in the plasma membrane ofN. tabacum protoplasts and suggest dehydroascorbate as the preferentially transported redox species. The putative presence of different carriers for reduced and oxidised ascorbate in the plasma membrane is discussed.Abbreviations Asc ascorbate - BY-2 Bright Yellow 2 - CCCP carbonylcyanide-3-chlorophenylhydrazone - DHA dehydroascorbate - DTT dithiothreitol - MS medium Murashige and Skoog medium - NEM N-ethylmaleimide - pCMBS p-chloromercuribenzenesulfonic acid     

Dehydroascorbate and glucose are taken up into Arabidopsis thaliana cell cultures by two distinct mechanisms

The possible involvement of glucose (Glc) carriers in the uptake of vitamin C in plant cells is still a matter of debate. For the first time, it was shown here that plant cells exclusively take up the oxidised dehydroascorbate (DHA) form. DHA uptake is not affected by 6-bromo-6-deoxy-ascorbate, an ascorbate (ASC) analogue, specifically demonstrating ASC uptake in animal cells. There is no competition between Glc and DHA uptake. Moreover, DHA and Glc carriers respond in the opposite manner to different inhibitors (cytochalasin B, phloretin and genistein). In conclusion, the plant plasma membrane DHA carrier is distinct from the plant Glc transporters.     

Acute effects of whole-body vibration on jump force and jump rate of force development: a comparative study of different devices

The goal of this study was to compare the acute effects of whole-body vibration (WBV) delivered by 3 devices with different mechanical behavior on jump force (JF) and jump rate of force development (JRFD). Twelve healthy persons (4 women and 8 men; age 30.5 ± 8.8 years; height 178.6 ± 7.3 cm; body mass 74.8 ± 9.7 kg) were exposed to WBV for 15 and 40 seconds using 2 professional devices (power plate [PP; vertical vibration] and Galileo 2000 [GA; oscillatory motion around the horizontal axis in addition to vertical vibration]) and a home-use device [Power Maxx, PM; horizontal vibration]). The JF and JRFD were evaluated before, immediately after, and 5 minutes after WBV. The JF measured immediately after 40 seconds of vibration by the GA device was reduced (3%, p = 0.05), and JRFD measured after 5 minutes of rest after 40 seconds of vibration by the PM device was reduced (12%, p < 0.05) compared with the baseline value. The acute effects of WBV (15 or 40 seconds) on JF and JRFD were not significantly different among the 3 devices. In conclusion, our hypothesis that WBV devices with different mechanical behaviors would result in different acute effects on muscle performance was not confirmed.